Emulsions including a PEG-derivative of tocopherol

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DESCRIPTION

Emulsions comprising a PEG derivative of tocopherol

Field of the invention

Compositions and methods are provided for preparing foods and beverages that contain additives, such as nutraceuticals, pharmaceuticals, and supplements.

Background

Nonpolar compounds do not dissolve easily in aqueous solutions, such as water or any other polar solvent. A number of nonpolar compounds are used in compositions for human ingestion, eg, pharmaceuticals, nutraceuticals, and / or dietary supplements. Exemplary of the nonpolar compounds used in such compositions are vitamins and minerals, fatty acids and other nonpolar compounds, nonpolar active agents and nonpolar active ingredients.

Document US2006 / 0088558 describes a composition of Vitamin E concentrate and liquid TPGS, the concentrate having a significantly higher percentage of TPGS per unit volume than traditional liquid TPGS formulations while providing the TPGS in a liquid form that can be used in gels. soft or mixed with water to produce desired concentrations for commercial use in supplements, beverages, pharmaceutical preparations etc.

WO96 / 36316 describes self-emulsifying pharmaceutical pre-concentrate compositions comprising: (a) a lipophilic medicinal compound, (b) d-alpha-tocopherol polyethylene glycol 1000 succinate (TPGS) and (c) a lipophilic phase as well as pharmaceutical compositions which They comprise such pre-concentrates in combination with a sufficient quantity of water to produce a stable emulsion.

US2005 / 208082 describes an aqueous emulsion in which the liquid phase of the emulsion includes a combination of a therapeutically effective concentration of a lipophilic such as a concentration of Vitamin E and TPGS and a concentration of linoleic acid. The presence of linoleic acid increases the solubilizing effect of Vitamin E and TPGS in the lipophilic and thus reduces the amount of Vitamin E and TPGS that would otherwise be required in the aqueous emulsion.

Due to poor water solubility, the inclusion of nonpolar compounds in products for human consumption, eg supplements, food and beverages, is often a challenge. Available compositions containing nonpolar compounds, particularly aqueous compositions containing nonpolar compounds, and methods for formulating such compositions are limited. Thus, there remains a need to develop compositions containing nonpolar compounds and methods for preparing the compositions. Accordingly, it is among the objects of this document to provide compositions, including aqueous compositions, containing nonpolar compounds and methods for preparing the compositions.

Summary

The present invention provides a liquid nanoemulsion concentrate comprising:

a PEG derivative of Vitamin E in an amount between 16% and 30% by weight of the concentrate;

a polar solvent in an amount between 60% and 79% by weight of the concentrate;

a nonpolar active ingredient containing a compound or compounds selected from any one or more of polyunsaturated fatty acids, omega-3 fatty acids, omega-6 fatty acids, conjugated fatty acids, Coenzyme Q10 compounds, and phytosterols; and the non-polar active ingredient is present in an amount between 5% and 10% by weight of the concentrate.

First compositions (concentrates) containing non-polar compounds are provided, including liquid nanoemulsion concentrates. Also provided are methods that use such first compositions to prepare other compositions, such as beverages and other aqueous liquids, in which the first compositions are diluted to form liquid dilution compositions. Liquid dilution compositions containing the beverage or other aqueous liquid and the dilute concentrate are also provided. The concentrates contain dispersions and / or can be used to prepare dispersions, of effective amounts of additives, such as non-polar compounds, including non-polar active ingredients such as nutraceuticals, pharmaceuticals and supplements, such as essential fatty acids, including polyunsaturated fatty acids, such as omega-3 fatty acids, omega-6 fatty acids, conjugated fatty acids, and other fatty acids; phytochemicals, including phytosterols; other oils and coenzymes, including Coenzyme Q10 and other oil-based additives. The amounts in the resulting diluted compositions are effective in supplementing the diet. The compositions provided herein are stable dispersions without phase separation and / or other changes.

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For example, the compositions provided include concentrates containing non-polar active ingredients, surfactants, and polar solvents in amounts at which dilution of the concentrate in an aqueous medium such as a beverage, in a particular amount (eg, any of the specified amounts , concentrations and dilutions of the concentrates and any of the amounts of the non-polar active ingredients described below in this document), produces a liquid dilution composition that contains effective amounts of the nonpolar active ingredient and that has one or more desirable properties. The compositions provided further include liquid dilution compositions, including liquid dilution compositions made from the concentrates, which contain aqueous media, non-polar active ingredients in effective amounts, and polar solvents having the desired properties. The amount of the concentrate and / or the amount of the non-polar active ingredient can be specified. Desired properties include transparency of liquid dilution compositions such as compositions that are clear or nearly as clear as the aqueous medium in the absence of the concentrate and / or in the absence of the nonpolar active ingredient; particle size, such as a particle size less than 200 nm or less than about 200 nm, less than 100 nm or less than about 100 nm, less than 50 nm or less than about 50 nm, or less than 25 nm or less approximately 25 nm, maximum or average; turbidity, such as a value in Nephelometric Turbidity Units (NTU) less than 200 or about 200; less than 100 or about 100, less than 50 or about 50, less than 30 or about 30, less than 25 or about 25, or less than 10 or about 10; and the absence of visible particles, visible crystals, phase separation and / or ring formation.

The concentrates provided are liquid nanoemulsion concentrates, containing surfactants, nonpolar compound or compounds (which are typically a nonpolar active ingredient that differs from surfactant) and a polar solvent (for example, water or other edible aqueous liquid, such as a polar protic solvent, such as a dihydric or trihydric alcohol, for example propylene glycol and glycerin (glycerol)).

The amount of nonpolar compound in the concentrate is between 5% or about 5% and 10% or about 10% by weight (w / w) of the concentrate, for example, the or about 5, 5.2 , 5.25, 6, 7, 8, 9 or 10% by weight of the concentrate.

The surfactants in the concentrates provided have a Hydrophilic Lipophilic Balance (HLB) value of between 14 or about 14 and 20 or about 20, such as between 15 or about 15 and 18 or about 18, for example, at or about 15, 16, 17 or 18.

Surfactants include polyethylene glycol (PEG) derivatives of Vitamin E, as defined in the claims.

The amount of surfactant or surfactants in the concentrate is between 16% or about 16% and 30% or about 30% by weight of the concentrate, for example, about 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30% by weight (w / w) of the concentrate.

In some examples, the amount of surfactant (s) in the concentrate is between 17% or about 17% and 26% or about 26% by weight (w / w) of the concentrate, for example, between 18% or about 18% and 26% or about 26% or between 16% or about 16% and 18% or about 18%. In some examples, the amount of surfactant or surfactants in the concentrate is at or about 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, or 26% by weight (w / w) of the concentrate, such as for example 17.75%, 20.25%, 20.5%, 22.7% or 25.2% (w / w) of the concentrate.

The amount of polar solvent in the concentrate is between 60% or about 60% and 79% or about 79% by weight (w / w) of the concentrate, for example, at or about 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78 or 79% by weight (w / w) of the concentrate.

In one example, the amount of polar solvent in the concentrate is between 65% or about 65% and 79% or about 79%, between 65% or about 65% and 75% or about 75%. , between 65% or about 65% and 76% or about 76%, between 68% or about 68% and 76% or about 76% by weight (w / w) of the concentrate. For example, the amount of polar solvent in the concentrate can be or about 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, or 80% by weight. (w / w) of the concentrate, such as of or about 74.25%, of or about 75.8%, of or about 68.9%, of or about 71.74%, of or about 63.94%, of or about 68.79%, of or about 68.29%, of or about 69.02%, or of or about 71.49% by weight (w / w) of the concentrate.

Among the PEG-derived surfactants are the PEG derivatives of Vitamin E, such as surfactants of tocopherol and tocotrienol derivatives, in which the Vitamin E moiety represents the hydrophobic region of the surfactant, and is attached, through a linker, to a moiety such as a polyethylene glycol (PEG) moiety. Exemplary Vitamin E-derived surfactants include, but are not limited to, tocopherol-derived surfactants, including polyalkylene glycol derivatives of tocopherol, typically polyethylene glycol (PEG) derivatives of tocopherol, such as polyethylene glycol diesters of tocopherol (TPGd), eg, succinate. polyethylene glycol tocopherol (TPGS), analogs of

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TPGS, TPGS homologs, and TPGS derivatives. Exemplary surfactants also include other PEG derivatives that have similar properties, for example, PEG derivatives of sterols, for example, a cholesterol or sitosterol (including, for example, any of the PEG derivatives disclosed in US Pat.

States No. 6,632,443) and PEG derivatives of other fat-soluble vitamins, for example, some forms of

Vitamin A (for example, Retinol) or Vitamin D (for example, Vitamin D1-D5).

Surfactants include polyethylene glycol (PEG) derivatives of Vitamin E, for example, a polyethylene glycol tocopherol diester (TPGD). In one example, the TPGD is selected from tocopherol sebacate polyethylene glycol, tocopherol dodecanedioate polyethylene glycol, tocopherol suberate polyethylene glycol, tocopherol azelaate polyethylene glycol, citraconyethyl tocopherol glycol polyethylene glycol, tocopherol tocopherol toconyethylene glycopherol toconyethylene glycol methyl ester , Tocopherol Maleate Polyethylene Glycol, Tocopherol Glutarate Polyethylene Glycol, tocopherol glutaconate polyethylene glycol and tocopherol phthalate polyethylene glycol. In another example, the TPGD surfactant is a polyethylene glycol tocopherol succinate (TPGS), such as TPGS-1000 and / or a TPGS day. In another example, the surfactant is a TPGS analog. In one aspect, the surfactant is a TPGS homolog, such as, for example, a TPGS homolog that differs from a TPGS parent compound by the addition or removal of one or more methylene units, for example, - (CH2) n- -.

PEG moieties in PEG-derived surfactants, including PEG moieties in PEG derivatives of Vitamin E, include PEG moieties selected from any one or more of PEG-OH, PEG-NHS, PEG-aldehyde, PEG-SH, PEG- NH2, PEG-CO2H, methylated PEGs (m-PEG), and branched PEGs, and include PEG moieties having a molecular weight between 200 kDa or about 200 kDa and 20,000 kDa or about 20,000 kDa, typically between 200 kDa or about 200 kDa and 6000 kDa or about 6000 kDa, for example, between 600 kDa or about 600 kDa and 6000 kDa or about 6000 kDa, typically between 200 kDa or about 200 kDa and 2000 kDa or about 2000 kDa, between 600 kDa or about 600 kDa and 1500 kDa or about 1500 kDa, or between 600 kDa or about 600 kDa and 1000 kDa or about 1000 kDa.

Exemplary nonpolar compounds in the provided compositions (including liquid nanoemulsion concentrates) are nonpolar active ingredients including, but not limited to, omega-3 fatty acids, omega-6 fatty acids, conjugated fatty acids, Coenzyme Q10 (e.g. , ubidecarenone), phytosterols, and saw palmetto extracts. Non-polar active ingredients include, for example, non-polar compounds containing Docosahexaenoic Acid (DHA) and / or Eicosapentaenoic Acid (EPA), Alpha-Linolenic Acid (aLinolenic Acid; ALA), Conjugated Linolenic Acid (CLA), and Gamma-Acid. linolenic (GLA), including, but not limited to, fish oil, seaweed oil, linseed oil, borage oil, and saw palmetto extract.

Non-polar active ingredients include, but are not limited to, compounds containing any nutraceutical or pharmaceutical and / or fat soluble oil, such as, for example, drugs, hormones, vitamins, nutrients, including any other lipophilic compound containing essential fatty acids. , for example, polyunsaturated fatty acids (PUFA), including, for example, omega-3 fatty acids, for example, natural and synthetic omega3 fatty acids, for example, compounds containing omega-3 polyunsaturated long chain fatty acids, including Eicosapentaenoic Acid (EPA) (20: 5w3), Docosahexaenoic Acid (DHA) (22: 6w3), Eicosatetraenoic Acid (24: 4w3); Docosapentaenoic acid (DPA, Clupanodic acid) (22: 5w3), 16: 3w3 acid; 24: 5w3, and / or nisinic acid (24: 6w3), which may include, for example, fish oil, seaweed oil, krill oil, canola oil, linseed oil, soybean oil, and soybean oil. Walnut; compounds containing short chain omega-3 fatty acids, for example Alpha-Linolenic Acid (α-Linolenic acid; ALA) (18: 3w3) (for example, linseed oil) and Stearidonic acid (18: 4w3); esters of an omega-3 fatty acid and glycerol, for example monoglycerides, diglycerides and triglycerides; omega-3 fatty acid esters and a primary alcohol, for example, fatty acid methyl esters and fatty acid esters; omega-3 fatty acid oil precursors, eg, EPA precursor, DHA precursor; derivatives such as polyglycolized derivatives or polyoxyethylene derivatives; oils containing omega-3 fatty acids, for example fish oil (for example marine oil), including for example highly purified fish oil concentrates, perilla oil, krill oil and seaweed oil (for example , microalgae oil); compounds containing omega-6 fatty acids, for example, compounds containing linoleic acid (18: 2w6) (a short-chain fatty acid), gammalinolenic acid (GLA) (18: 3w6), dihomo gamma-linolenic acid (DGLA) ( 20: 3w6), Eicosadienoic Acid (20: 2w6), Arachidonic Acid (AA) (20: 4w6), Docosadienoic Acid (22: 2w6), Adrenic Acid (22: 4w6) and / or Docosapentaenoic Acid (22: 5w6), for example, borage oil, corn oil, cottonseed oil, grapeseed oil, peanut oil, evening primrose oil for example, evening primrose oil (Oenothera biennis), blackcurrant seed oil, hemp seed oil, spirulina extract, safflower oil, sesame oil and soybean oil;

compounds containing other fatty acids, eg triglycerides, including medium chain triglycerides, polar lipids, eg ether lipids, phosphoric acid, choline, fatty acids, glycerol, glycolipids, triglycerides, and phospholipids (eg, phosphatidylcholine (lecithin ), phosphatidylethanolamine and phosphatidylinositol); saw palmetto extract; and ethyl linoleate; and herbal oils, for example garlic and scoridine oils; Short chain saturated fatty acids (4: 0-10: 0), Lauric acid (12: 0), Myristic acid (14: 0), Pentadecanoic acid (15: 0), Palmitic acid (16: 0), Palmitoleic acid (16: 1w7), Heptadecanoic Acid (17: 0), Stearic Acid (18: 0), Oleic Acid (18: 1w9), Acid

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Arachidic (20: 0);

compounds containing micronutrients, eg vitamins, minerals, cofactors eg coenzymes such as coenzyme Q, eg coenzyme Q10 (CoQ10, also called ubiquinone eg ubidecarenone or a reduced form of CoQ10 eg ubiquinol) , turmeric extract (for example cucuminoids), saw palmetto liquid extract (for example, saw palmetto oil) echinaceae extract, hawthorn berry extract, ginseng extract, lipoic acid (such as thioctic acid), ascorbyl palmitate, kava extract, St. John's wort (such as St. John's wort, klamath herb, horny goat weed), quercetin extract, dehydroepiandrosterone, indole-3-carbinol ;

Carotenoid-containing compounds, including hydrocarbons and oxygenated alcoholic derivatives of hydrocarbons, eg beta carotene, mixed carotenoid complexes, lutein, lycopene, Zeaxanthin, Cryptoxanthin, eg beta-cryitoxanthin, beta carotene, mixed carotenoid complexes, astaxanthin, bixin, canthaxanthin, capsanthin, capsorubin, apo-carotene, beta-12'-apo-carotene, beta-carotene, Carotene (mixture of alpha and beta-carotene), gamma carotene, beta crytoxanthin, cyolerritrine, zeaxanthin, hydroxyl- or carboxyl-containing member esters thereof;

compounds containing fat-soluble vitamins, for example, Vitamins A, D, E and K and corresponding provitamins and derivatives of vitamins such as esters with an action similar to that of vitamin A, D, E or K, for example; retinol (vitamin A) and pharmaceutically acceptable derivatives thereof, for example, retinol palmitate ester and other retinol esters, and calciferol (vitamin D) and its pharmaceutically acceptable derivatives thereof and vitamin D precursors, d-alpha tocopherol (vitamin E) and derivatives thereof, including pharmaceutical derivatives thereof, for example, Tocotrienols, d-alpha tocopherol acetate and other esters of dalpha tocopherol and ascorbyl palmitate, a fat-soluble version of vitamin C;

compounds containing phytochemicals, including phytoestrogens for example genistein and daidzein, for example isoflavones, for example soy isoflavones, flavonoids, phytoalexins, for example Resveratrol (3,5,4'-trihydroxystilbene), red clove extract and phytosterols ;

lipid-soluble drug-containing compounds, including natural and synthetic forms of immunosuppressive drugs such as Cyclosporine, protease inhibitor such as Ritonavir, oil-soluble anesthetics and macrolide antibiotics such as Propofol, natural and synthetic forms of steroid hormones, for example estrogens , estradiols, progesterone, testosterone, cortisone, phytoestrogens, dehydroepiandrosterone (DHEA), growth hormones and other hormones;

compounds containing oil-soluble acids and alcohols, for example, tartaric acid, lactyl acid butylated hydroxyanisole, butylated hydroxytoluene, lignin, sterols, polyphenolic compounds, oryzanol, cholesterol, phytosterols, flavonoids such as, but not limited to, quercetin and reservatol and diallyl disulfides.

Non-polar active ingredients include ingredients that contain polyunsaturated fatty acids, such as compounds that contain one or more of omega-3 fatty acids, including docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA), and alpha-linolenic acid (ALA) ( eg fish oils, krill oils, algae oils and / or linseed oils), omega-6 fatty acids such as gamma-linolenic acid (GLA) (eg borage oils); conjugated fatty oils (eg, conjugated linoleic acid (CLA)) and saw palmetto extracts; and ingredients containing coenzymes such as coenzyme Q, eg, Coenzyme Q10 (eg, ubidecarenone); and ingredients containing phytosterols and combinations thereof.

In one example, the nonpolar active ingredient contains eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), or a combination thereof. In one example, the nonpolar active ingredient contains DHA in an amount between 20% or about 20% and 90% or about 90%; between 25% or about 25% and 85% or about 85%; between 35% or about 35% and 70% or about 70%; or between 25% or about 25% and 40% or about 40%, by weight (w / w), of the non-polar active ingredient. In another aspect, the non-polar active ingredient contains EPA in an amount between 5% or about 5% and 15% or about 15%; between 5% or about 5% and 13% or about 13%; or between 5% or about 5% and 10% or about 10% by weight (w / w) or of the non-polar active ingredient. In one aspect, the amount of EPA is not more than 10% or about 10% or not more than 13% or about 13% by weight (w / w) of the non-polar active ingredient. For example, non-polar active ingredients include fish oil and seaweed oil that contain any of these percentages of EPA and / or DHA.

In another example, the nonpolar active ingredient contains alpha-linolenic acid (ALA). In one example, the nonpolar active ingredient contains ALA in an amount of at least 50% or about 50% by weight (w / w) of the nonpolar active ingredient, such as between 50% or about 50% and 80% or about 80%, or between 65% or about 65% and 75% or about 75% by weight (w / w) of the non-polar active ingredient. For example, non-polar active ingredients include linseed oils that contain any of these percentages of ALA.

In another example, the nonpolar active ingredient contains gamma-linolenic acid (GLA). In one example, the nonpolar active ingredient contains GLA in an amount of at least 22% or about 22% by weight (w / w) of the nonpolar active ingredient. For example, non-polar active ingredients include a borage oil containing GLA in an amount of at least 22% or about 22% by weight (w / w) of the borage oil.

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In some examples, the concentrate contains more than one non-polar active ingredient, for example, two or more non-polar active ingredients, such as any non-polar compound described herein. In one example, the total amount of the non-polar active ingredient (s) is between about 5% and 10% by weight of the concentrate, for example, when the combined weight of the non-polar active ingredient and the additional non-polar active ingredient (s) it is not more than about 10% by weight (w / w) of the concentrate.

Polar solvents contained in concentrates include polar protic and polar aprotic solvents and typically are polar protic solvents such as polar solvents having a dielectric constant greater than 15 or about 15 or equal to 15 or about 15, or a dielectric constant between 20 or about 20 and 90 or about 90, such as between 20 or about 20 and 80 or about 80 (for example, year about or at least year about 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41 , 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 5, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66 , 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, or 81); and polar solvents having a polarity index of between at or about 3 and at or about 9 or a bipolar moment between at or about 1.8 and at or about 2.8. Polar solvents include water and alcohols, such as monohydric, dihydric, trihydric, and other alcohols, and typically alcohols other than monohydric alcohols, alcohols that have two or more hydroxyl groups such as dihydric (two hydroxy groups) and trihydric alcohols (three hydroxy groups). hydroxyl). Polar solvents include, but are not limited to, glycerin, ethylene glycols such as propylene glycol, ethylene glycol, tetraethylene glycol, triethylene glycol, and trimethylene glycol. Polar solvents can additionally include low molecular weight polyethylene glycols (PEG) such as PEG with molecular weights of or about or less than or about 600, 400 or 200 kDa. In some examples, the polar solvent is water, glycerin, or propylene glycol.

The concentrates provided may contain one or more additional ingredients. In one example, the concentrate additionally contains a cosurfactant in an amount sufficient to stabilize the concentrate, compared to the absence of the cosurfactant. In one aspect, the cosurfactant is a phospholipid, such as, but not limited to, a phosphatidylcholine. In one example, the amount of co-surfactant, eg, phospholipid, is between 0.1% or about 0.1% and 1% or about 1% by weight (w / w) of the concentrate.

In another example, the concentrate additionally contains a preservative, in an amount sufficient to preserve the concentrate, compared to the absence of the preservative. They are exemplary of natural preservatives, such as benzyl alcohol and preservatives containing benzyl alcohol. In one example, the amount of preservative is between 0.1% or about 0.1% and 1% or about 1% by weight (w / w) of the concentrate, for example, at or about 0.1 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9 or 1% by weight of the concentrate. In one example, the amount of benzyl alcohol is between 0.1% or about 0.1% and 1% or about 1% by weight (w / w) of the concentrate, for example, at or about 0.1 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9 or 1% by weight of the concentrate.

In another example, the concentrate contains a non-polar solvent, for example, a non-polar solvent that dissolves and differs from the non-polar active ingredient. Typically, the amount of nonpolar solvent is sufficient to dissolve the nonpolar active ingredient, for example, between 1% or about 1% and 6% or about 6%, for example, at or about 1.2 , 3, 4, 5, or 6% by weight (w / w) of the concentrate. The nonpolar solvent is typically an oil, such as any suitable oil for dissolving the nonpolar ingredient. Exemplary of the nonpolar solvents are Vitamin E oil, linseed oil, sunflower oil, and vegetable oils and other oils.

In another example, the concentrate contains an emulsion stabilizer. Typically, the emulsion stabilizer is included in the concentrate in an amount sufficient to stabilize the concentrate. Emulsion stabilizers include, but are not limited to, compositions containing a combination of gums, such as Saladizer® brand emulsion stabilizer. In one example, the emulsion stabilizer contains one or more guar gums, xanthan gum, and sodium alginate. In one example, the emulsion stabilizer contains guar gum, xanthan gum, and sodium alginate.

In another example, concentrates contain flavorings. Typically, the flavor (s) are included in an amount sufficient to enhance the flavor of the concentrate, the odor of the concentrate, or a combination thereof, as compared to the absence of the flavor. Flavors include, but are not limited to, flavors containing lemon oil and D-limonene or a combination thereof, or any other known flavors, such as the flavors described herein.

In another example, when the concentrates contain water as the polar solvent, for example, the concentrates contain pH adjusters. Typically, the pH adjuster contains an acid or base in an amount sufficient to affect the pH of the concentrate compared to the absence of the pH adjuster. PH adjusters include, but are not limited to, citric acid and phosphoric acid.

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The concentrates provided include, but are not limited to, concentrates containing non-polar PUFA-containing active ingredients, such as omega-3 fatty acids, omega-6 fatty acid, conjugated fatty acid, and non-polar active ingredients containing saw palmetto acid.

In one example, the concentrate provided contains a non-polar active ingredient which is a fish oil containing eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) (for example, a fish oil containing 10% EPA and 70% EPA). % DHA, a fish oil that contains approximately 13% EPA and approximately 13% DHA; or a fish oil containing 40% EPA and 20% DHA), a surfactant which is polyethylene glycol tocopherol succinate (TPGS) or a TPGS analog (for example, a TPGS homologue), and a solvent polar which is water. In one aspect of this example, the amount of non-polar active ingredient in fish oil is at or about 5% by weight (w / w) of the concentrate; the amount of TPGS or analogous surfactant is at or about 18% by weight (w / w) of the concentrate, and the amount of water is at or about 74.25% by weight (w / w) of the concentrate. In another aspect of this example, the amount of the non-polar active ingredient in fish oil is at or about 5% by weight (w / w) of the concentrate; the amount of surfactant TPGS or TPGS analog is or about 25.2% by weight (w / w) of the concentrate, and the amount of water is or about 68.79% by weight (w / w) of the concentrated. In another aspect of this example, the amount of the non-polar active ingredient in fish oil is at or about 10% by weight (w / w) of the concentrate; the amount of TPGS or TPGS analog surfactant is or about 20.2% by weight (w / w) of the concentrate and the amount of water is or about 68.79% by weight (w / w) of the concentrate . In one aspect, the concentrate additionally contains an emulsion stabilizer which is a combination of xanthan gum, guar gum, and / or sodium alginate; a benzyl alcohol preservative; and a pH adjuster which is citric acid. In another aspect, the concentrate additionally contains D-limonene and / or lemon oil flavors. In another aspect, the concentrate additionally contains a phosphatidylcholine cosurfactant.

In another example, the concentrate provided contains a non-polar active ingredient that is an algae oil, such as an algae oil that contains 35% DHA, a surfactant that is TPGS, or an analog of TPGS (eg, a homologue of TPGS) and a polar solvent which is water. In one aspect of this example, the amount of non-polar active ingredient in the algae oil is at or about 5% by weight (w / w) of the concentrate; the amount of TPGS or a TPGS analog surfactant is at or about 18% by weight (w / w) of the concentrate and the amount of water is at or about 75.81% by weight (w / w) of the concentrate. In another aspect of this example, the amount of algae oil non-polar active ingredient is at or about 5% by weight (w / w) of the concentrate; the amount of TPGS or TPGS analog surfactant is or about 25.2% by weight (w / w) of the concentrate and the amount of water is or about 68.79% by weight (w / w) of the concentrate . In another aspect of this example, the amount of the non-polar active ingredient in the algae oil is at or about 10% by weight (w / w) of the concentrate; the amount of TPGS or TPGS analog surfactant is or about 25.2% by weight (w / w) of the concentrate and the amount of water is or about 63.94% by weight (w / w) of the concentrate . In one aspect of this example, the concentrate additionally contains an emulsion stabilizer which is a combination of xanthan gum, guar gum, and / or sodium alginate; a benzyl alcohol preservative; a pH adjuster that is citric acid. In another aspect, the concentrate additionally contains D-limonene and lemon oil flavors.

In another example, the concentrate contains a nonpolar active ingredient that is flaxseed oil (for example, one that contains 50% or 55% omega-3 fatty acids, for example 50% or 55% acid alpha-linolenic (ALA)), a surfactant that is TPGS or an analog of TPGS (eg, a homologue of TPGS), and a polar solvent that is water. In one aspect of this example, the amount of linseed oil nonpolar active ingredient is at or about 5% by weight (w / w) of the concentrate; the amount of the TPGS or TPGS analog surfactant is or about 17.75% by weight (w / w) of the concentrate, and the amount of the water is or about 71.74% by weight (w / w) of the concentrated. In another aspect of this example, the amount of nonpolar active ingredient in linseed oil is at or about 5% by weight (w / w) of the concentrate; the amount of TPGs or TPGS analog surfactant is about 25.2% by weight (w / w) of the concentrate and the amount of water is or about 68.79% by weight (w / w) of the concentrate. In another aspect of this example, the amount of linseed oil nonpolar active ingredient is at or about 10% by weight (w / w) of the concentrate; the amount of TPGS or TPGS analog surfactant is or about 20.2% by weight (w / w) of the concentrate and the amount of water is or about 68.79% by weight (w / w) of the concentrate . In one aspect, the concentrate additionally contains an emulsion stabilizer which is a combination of xanthan gum, guar gum, and / or sodium alginate; a benzyl alcohol preservative and a pH adjuster that is citric acid. In another aspect, it additionally contains D-limonene and lemon oil flavors. In another aspect, the concentrate additionally contains a phosphatidylcholine cosurfactant. In another aspect, the concentrate additionally contains a non-polar solvent which is Vitamin E oil or another oil.

In another example, the concentrate contains a non-polar active ingredient that is linseed oil (for example, one that contains 50% or 55% omega-3 fatty acids, for example 50% or 55% ALA), a surfactant which is TPGS or an analog of TPGS (eg, a homologue of TPGS), and a polar solvent which is glycerin. In one aspect of this example, the amount of nonpolar active ingredient in linseed oil is at or about 5% by weight (w / w) of the concentrate; the amount of TPGS or TPGS analog surfactant is at or about

ES 2 396 946 T3 is 25.2% by weight (w / w) of the concentrate, and the amount of glycerin is at or about 69.02% by weight (w / w) of the concentrate. In one aspect, the concentrate additionally contains a benzyl alcohol preservative.

In another example, the concentrate contains a non-polar active ingredient that is linseed oil (for example, one that contains 50% or 55% omega 3 acids, for example 50% or 55% ALA), a surfactant which is TPGS or an analog of TPGS (eg, a homologue of TPGS), and a polar solvent which is propylene glycol. In one aspect of this example, the amount of the linseed oil nonpolar active ingredient is at or about 5% by weight (w / w) of the concentrate; the amount of TPGS or TPGS analog surfactant is or about 25.2% by weight (w / w) of the concentrate and the amount of propylene glycol is or about 69.02% by weight (w / w) of the concentrate . In one aspect, the concentrate additionally contains a benzyl alcohol preservative.

In another example, the concentrate contains a nonpolar active ingredient that contains gamma linolenic acid (GLA) (for example, a borage oil that contains GLA, for example 22% GLA), a surfactant that is TPGS, or an analog of TPGS (eg, a homologue of TPGS) and a polar solvent which is water. In one aspect of this example, the amount of GLA-containing non-polar active ingredient is at or about 5% by weight (w / w) of the concentrate; the amount of TPGS or the TPGS analog surfactant is about 17.75% by weight (w / w) of the concentrate, and the amount of water is or about 71.74% by weight (w / w) of the concentrated. In another aspect of this example, the amount of GLA-containing non-polar active ingredient is at or about 5% by weight (w / w) of the concentrate; the amount of TPGS or TPGS analog surfactant is or about 25.2% by weight (w / w) of the concentrate, and the amount of water is or about 68.79% by weight (w / w) of the concentrated. In another aspect of this example, the amount of GLA-containing non-polar active ingredient is at or about 10% by weight (w / w) of the concentrate; the amount of TPGs or TPGS analog surfactant is or about 20.2% by weight (w / w) of the concentrate, and the amount of water is or about 68.79% by weight (w / w) of the concentrated. In one aspect, the concentrate additionally contains an emulsion stabilizer which is a combination of xanthan gum, guar gum, and / or sodium alginate. A benzyl alcohol preservative and a pH adjuster that is citric acid. In another aspect, the concentrate additionally contains D-limonene and lemon oil flavors. In another aspect, the concentrate additionally contains a phosphatidylcholine cosurfactant. In another aspect, the concentrate additionally contains a non-polar solvent which is a Vitamin E oil or other oil.

In another example, the concentrate contains a nonpolar active ingredient that contains gamma linolenic acid (GLA) (for example, a borage oil that contains, for example, 22% GLA), a surfactant that is TPGS, or an analog of TPGS. (eg a TPGS homolog) and a polar solvent which is glycerin. In one aspect of this example, the amount of GLA-containing non-polar active ingredient is at or about 5% by weight (w / w) of the concentrate; the amount of TPGS or TPGS analog surfactant is or about 25.2% by weight (w / w) of the concentrate, and the amount of glycerin is or about 69.02% by weight (w / w) of the concentrate In one aspect, the concentrate additionally contains a benzyl alcohol preservative.

In another example, the concentrate contains a nonpolar active ingredient that contains gamma linolenic acid (GLA) (for example, a borage oil that contains, for example, 22% GLA), a surfactant that is TPGS, or an analog of TPGS. (eg a homolog of TPGS) and a polar solvent which is propylene glycol. In one aspect of this example, the amount of GLA-containing non-polar active ingredient is at or about 5% by weight (w / w) of the concentrate; the amount of TPGS or TPGS analog surfactant is at or about 25.2% by weight (w / w) of the concentrate, and the amount of propylene glycol is at or about 69.02% by weight (w / w) of the concentrate In one aspect, the concentrate additionally contains a benzyl alcohol preservative.

In another example, the concentrate contains a nonpolar active ingredient that contains conjugated linoleic acid (CLA) (for example, 80% CLA), a surfactant that is TPGS or an analog of TPGS (for example, a homologue of TPGS) and a polar solvent that is water. In one aspect of this example, the amount of the CLA-containing non-polar active ingredient is at or about 5% by weight (w / w) of the concentrate; the amount of TPGs or TPGS analog surfactant is or about 17.75% by weight (w / w) of the concentrate, and the amount of water is or about 71.74% by weight (w / w) of the concentrated. In another aspect of this example, the amount of the CLA-containing non-polar active ingredient is at or about 5% by weight (w / w) of the concentrate; the amount of TPGS or TPGS analog surfactant is or about 25.2% by weight (w / w) of the concentrate, and the amount of water is or about 68.79% by weight (w / w) of the concentrated. In another aspect of this example, the amount of the CLA-containing non-polar active ingredient is at or about 10% by weight (w / w) of the concentrate; the amount of TPGS or TPGS analog surfactant is or about 20.2% by weight (w / w) of the concentrate, and the amount of water is or about 68.79% by weight (w / w) of the concentrated. In one aspect, the concentrate further contains an emulsion stabilizer that is a combination of xanthan gum, guar gum, and / or sodium alginate, a benzyl alcohol preservative, and a pH adjuster that is citric acid. In another aspect, the concentrate additionally contains D-limonene and lemon oil flavors. In another aspect, the concentrate additionally contains a phosphatidylcholine cosurfactant. In another aspect, the concentrate additionally contains a non-polar solvent which is a Vitamin E oil or other oil.

In another example, the concentrate contains a nonpolar active ingredient that contains linoleic acid (CLA) (eg, 80% CLA), a surfactant that is TPGS, or a TPGS analog (eg, a TPGS homologue) and

ES 2 396 946 T3 a polar solvent which is glycerin. In one aspect of this example, the amount of the CLA-containing non-polar active ingredient is at or about 5% by weight (w / w) of the concentrate; the amount of TPGS or TPGS analog surfactant is or about 25.2% by weight (w / w) of the concentrate, and the amount of glycerin is or about 69.02% by weight (w / w) of the concentrate In one aspect, the concentrate additionally contains a benzyl alcohol preservative.

In another example, the concentrate contains a non-polar active ingredient such as conjugated linoleic acid (CLA) (eg, 80% CLA), a surfactant that is TPGS, or a TPGS analog (eg, a TPGS homologue), and a polar solvent which is propylene glycol. In one aspect of this example, the amount of the CLA-containing non-polar active ingredient is at or about 5% by weight (w / w) of the concentrate; the amount of TPGs or TPGS analog surfactant is or about 25.2% by weight (w / w) of the concentrate, and the amount of propylene glycol is or about 69.02% by weight (w / w) of the concentrated. In one aspect, the concentrate additionally contains a benzyl alcohol preservative.

In another example, the concentrate contains a nonpolar active ingredient that is a saw palmetto extract (for example, one that contains between or about 85% and or about 90% fatty acids), a surfactant that is TPGS. or a TPGS analog (eg, a TPGS homolog) and a polar solvent which is water. In one aspect of this example, the amount of saw palmetto extract of the non-polar active ingredient is at or about 5% by weight (w / w) of the concentrate; the amount of TPGS or analogous surfactant is at or about 17.75% by weight (w / w) of the concentrate and the amount of water is at or about 71.74% by weight (w / w) of the concentrate. In another aspect of this example, the amount of saw palmetto extract non-polar active ingredient is at or about 5% by weight (w / w) of the concentrate; the amount of TPGs or TPGS analog surfactant is or about 25.2% by weight (w / w) of the concentrate and the amount of water is or about 68.79% by weight (w / w) of the concentrate . In another aspect of this example, the amount of saw palmetto extract non-polar active ingredient is at or about 10% by weight (w / w) of the concentrate; the amount of TPGS or TPGS analog surfactant is or about 20.2% by weight (w / w) of the concentrate, and the amount of water is or about 68.79% by weight (w / w) of the concentrated. In one aspect, the concentrate further contains an emulsion stabilizer that is a combination of xanthan gum, guar gum, and / or sodium alginate, a benzyl alcohol preservative, and a pH adjuster that is citric acid. In another aspect, the concentrate additionally contains D-limonene and lemon oil flavors. In another aspect, the concentrate additionally contains a phosphatidylcholine cosurfactant. In another aspect, the concentrate additionally contains a non-polar solvent which is Vitamin E oil or another oil.

The concentrates provided additionally include concentrates having coenzyme Q nonpolar active ingredients.

In one example, the concentrate contains a non-polar active ingredient of Coenzyme Q10 (CoQ10) (for example, the compound marketed under the name Kaneka Q10 ™ (USP Ubidecarenone)), a TPGS or TPGS analog surfactant (for example, a homologue ), and a polar solvent which is water. In one aspect of this example, the amount of CoQ10 non-polar active ingredient is at or about 5% by weight (w / w) of the concentrate, the amount of TPGS or TPGS analog is at or about 17.75%. by weight (w / w) of the concentrate, and the amount of water is at or about 71.74% by weight (w / w) of the concentrate. In one aspect, the concentrate further contains an emulsion stabilizer which is a combination of xanthan gum, guar gum, and / or sodium alginate, a benzyl alcohol preservative, and a pH adjuster which is citric acid. In another aspect, the concentrate additionally contains D-limonene and lemon oil flavors. In another aspect, the concentrate additionally contains a phosphatidylcholine cosurfactant. In another aspect, the concentrate additionally contains a non-polar solvent which is a Vitamin E oil or other oil.

In another example, the concentrate contains a non-polar active ingredient of Coenzyme Q10 (CoQ10) (for example, the compound marketed under the name Kaneka Q10 ™ (USP Ubidecarenone)), a TPGS or a TPGS analog surfactant (for example, homolog ), and a polar solvent which is glycerin. In one aspect of this example, the amount of non-polar active ingredient CoQ10 is at or about 5.25% by weight (w / w) of the concentrate, the amount of TPGS or TPGS analog is at or about 17.75 % by weight (w / w) of the concentrate, and the amount of glycerin is at or about 71.89% by weight (w / w) of the concentrate. In one aspect, the concentrate additionally contains a benzyl alcohol preservative. In another aspect, the concentrate additionally contains D-limonene and lemon oil flavors. In another aspect, the concentrate additionally contains a phosphatidylcholine cosurfactant. In another aspect, the concentrate additionally contains a non-polar solvent which is a Vitamin E oil or other oil.

In another example, the concentrate contains a non-polar active ingredient of Coenzyme Q10 (CoQ10) (for example, the compound marketed under the name Kaneka Q10 ™ (USP Ubidecarenone)), a TPGS or a TPGS analog surfactant (for example, a homolog), and a polar solvent which is propylene glycol. In one aspect of this example, the amount of the non-polar active ingredient of CoQ10 is at or about 5.25% by weight (w / w) of the concentrate, the amount of TPGS or TPGS analog is at or about 17, 75% by weight (w / w) of the concentrate, and the amount of propylene glycol is at or about 71.89% by weight (w / w) of the concentrate. In one aspect, the concentrate additionally contains a benzyl alcohol preservative. In another respect, the

ES 2 396 946 T3 concentrate additionally contains D-limonene and lemon oil flavors. In another aspect, the concentrate additionally contains a phosphatidylcholine cosurfactant. In another aspect, the concentrate additionally contains a non-polar solvent which is a Vitamin E oil or other oil.

In one example, the concentrate contains a non-polar active ingredient of phytosterol or phytosterols, a TPGS or TPGS analog surfactant (eg, a homologue), and a polar solvent that is water. In one aspect of this example, the amount of phytosterol or phytosterols nonpolar active ingredient is at or about 5.25% by weight (w / w) of the concentrate, the amount of TPGS or TPGS analog is at or about the 20% by weight (w / w) of the concentrate, and the amount of water is at or about 68.29% by weight (w / w) of the concentrate. In one aspect, the concentrate additionally contains a benzyl alcohol preservative and an emulsion stabilizer which is a combination of xanthan gum, guar gum, and / or sodium alginate. In another aspect, the concentrate additionally contains a phosphatidylcholine cosurfactant. In another aspect, the concentrate additionally contains a nonpolar solvent which is linseed oil or other oil.

In some examples, the concentrate is formulated based on the desired properties of the resulting dilution compositions generated by diluting the concentrate in an aqueous liquid. Typically, the concentrate is formed so that it can be diluted in an aqueous medium to produce a liquid dilution composition that has one, more than one, all, or a combination of the following properties:

In one example, the concentrate is formulated so that dilution of a certain amount of the concentrate in a certain amount of aqueous medium produces a liquid dilution composition that has a desired particle size, typically a particle size that is no greater than a particular particle size or is smaller than a particular particle size. The specified particle size can be expressed as the mean particle size, or a larger particle size in the aqueous medium. For example, it may be desired that the liquid dilution composition contain less than a particular particle size on average or at most. For example, the concentrate can be formulated such that dilution of at least 0.5 grams (g) or about 0.5 g, at least 1 g or about 1 g, at least 2 g or about 2 g, at least 5 g or about 5 g, or at least 10 g or about 10 g of the concentrate in or about 8 fluid ounces (0.236588 liters) of an aqueous medium; or dilution of the concentrate in an aqueous medium, at a dilution no greater than 1:10 or approximately 1:10, no greater than 1:25 or approximately 1:25, no greater than 1:50 or approximately 1:50, no greater of 1: 100 or about 1: 100, not more than 1: 250 or about 1: 250 or not more than 1: 500 or about 1: 500; or diluting the concentrate in an aqueous medium to form a liquid dilution composition containing at least 25 mg or about 25 mg, at least 35 mg or about 35 mg, at least 50 mg or about 50 mg or at least 100 mg or about 100 mg, at least 250 mg or about 250 mg, or at least 500 mg or about 500 mg of the non-polar active ingredient per 8 fluid ounces (0.236588 liters) of the liquid dilution composition, produces a liquid dilution composition that has a particle size less than 500 nm or less than about 500 nm, less than 200 nm or less than about 200 nm, less than 100 nm or less than about 100 nm, less than 50 nm or less than about 50 nm or less than 25 nm or less than about 25 nm, at most or on average.

In another example, the concentrate is formulated such that dilution of a certain amount of the concentrate in an amount of aqueous medium produces a liquid dilution composition having a desired transparency, such as producing a dilution composition having an NTU value. desired, typically an NTU value that is not greater than or less than a given NTU value, or by producing a liquid dilution composition that is clear or about as clear as the aqueous medium prior to the addition of the concentrate (ie, in the absence of the concentrate). For example, the concentrate can be formulated such that dilution of at least 0.5 grams (g) or about 0.5 g, at least 1 g or about 1 g, at least 2 g or about 2 g, at least 5 g or about 5 g, or at least 10 g or about 10 g of the concentrate in or about 8 fluid ounces (0.236588 liters) of an aqueous medium; or the dilution of a concentrate in an aqueous medium, at a dilution no greater than 1:10 or approximately 1:10, no greater than 1:25 or approximately 1:25, no greater than 1:50 or approximately 1:50, not more than 1: 100 or about 1: 100, not more than 1: 250 or about 1: 250 or not more than 1: 500, or dilution of the concentrate in an aqueous medium to form a liquid dilution composition containing the minus 25 mg or about 25 mg, at least 35 mg or about 35 mg, at least 50 mg or about 50 mg or at least 100 mg or about 100 mg, at least 250 mg or about 250 mg, or at least 500 mg or about 500 mg of the non-polar active ingredient per 8 fluid ounces (0.236588 liters ) of the liquid dilution composition, produces a liquid dilution composition that has a value in Nephelometric Turbidity Units (NTU) of less than 200 or about 200, less than 100 or about 100, less than 50 or about 50, less than 30 or about 30, less than 25 or about 25 or less than 10 or about 10, or produces a liquid dilution composition that is at least as clear or at least about as clear as the aqueous medium in the absence of the concentrate (i.e. i.e. compared to the transparency of the aqueous medium prior to the addition of the concentrate).

In another example, the concentrate is formulated so that upon dilution it produces a stable dilution liquid composition, for example, a composition that does not contain visible particles, does not contain visible crystals, does not exhibit ring formation, or a combination of the two. themselves. Stability can be for a period of

ES 2 396 946 T3 specified time and / or when the concentrate or liquid dilution composition is kept at a particular temperature. For example, the concentrate can be formulated such that the dilution of at least 0.5 g or about 0.5 g, at least 1 g or about 1 g, at least 2 g or about 2 g, at least 5 g or about 5 g, or at least 10 g or about 10 g of the concentrate in 8 fluid ounces (0.236588 liters), or about 8 fluid ounces (0.236588 liters) of an aqueous medium; dilution of the concentrate in an aqueous medium, at a dilution no greater than 1:10 or approximately 1:10, no greater than 1:25 or approximately 1:25, no greater than 1:50 or approximately 1:50, no greater 1: 100 or about 1: 100, no more than 1: 250 or about 1: 250 or no more than 1: 500; and / or dilution of the concentrate in an aqueous medium to form a liquid dilution composition containing at least 25 mg or about 25 mg, at least 35 mg or about 35 mg, at least 50 mg or about 50 mg or at least 100 mg or about 100 mg, at least 250 mg or about 250 mg, or at least 500 mg or about 500 mg of the non-polar active ingredient per 8 fluid ounces (0.236588 liters) of the liquid dilution composition, produces a liquid dilution composition that does not contain visible particles, does not contain visible crystals, does not exhibit phase separation and / or does not exhibit ring formation and / or has a pleasant taste and / or odor. The concentrate can be formulated so that the liquid dilution composition remains free of visible particles, remains free of visible crystals, remains free from phase separation, remains free from ring formation, and / or has a pleasant taste and odor, when the concentrate and / or the liquid dilution composition is stored at room temperature, or at a refrigerated temperature, or at a frozen temperature. Storage can be for example, for at least one day, at least one week, at least thirty days or at least one year.

The aqueous medium can be a beverage such as, for example, water, juice, soda, tea, coffee, sports drinks, nutritional drinks, energy drinks, milk, and other beverages, including those described herein.

Liquid dilution compositions are also described, which contain the concentrates diluted in an aqueous medium, for example a beverage. Liquid dilution compositions can contain any of the concentrates provided. Thus, liquid dilution compositions contain the non-polar active ingredients in aqueous medium, such as beverages that are desirable for human consumption. Liquid dilution compositions include those made by diluting concentrates, such as those having the properties described above, such as a desired particle size, transparency, NTU value, and / or stability, for example, absence of formation of ring, visible crystals, phase separation and / or pleasant taste / odor, eg according to the specifications described above.

In one example, the aqueous medium contained in the liquid dilution composition is a beverage such as, for example, water, soda, milk, tea, coffee, juice, energy drink, or a sports or nutritional drink. In one aspect, the liquid dilution composition is as clear or about as clear as the aqueous medium, such as the beverage prior to the addition of the concentrate (for example, compared to the absence of the concentrate) and / or remains as clear or about as clear as beverage when stored at room temperature (for example, 25 ° C or about 25 ° C) or at a refrigerated temperature (for example, 0-10 ° C or about 0-10 ° C, e.g. at or about 4 ° C) or at a frozen temperature (e.g., -20 ° C or about -20 ° C) where storage is for at least one day, at least one week, at least thirty days or at least one year.

The amount of the concentrate in the liquid dilution composition can be specified. For example, liquid dilution compositions include those containing at least 0.5 grams (g) or about 0.5 g, at least 1 g or about 1 g, at least 2 g or about 2 g, at least 5 g or about 5 g. g, or at least 10 g or about 10 g of the concentrate per 8 fluid ounces (0.236588 liters) of the aqueous medium; or containing the concentrate in a dilution no greater than 1:10 or approximately 1:10, no greater than 1:25 or approximately 1:25, no greater than 1:50 or approximately 1:50, no greater than 1: 100 or about 1: 100, no more than 1: 250 or about 1: 250 or no more than 1: 500; or containing at least 25 mg or about 25 mg, at least 35 mg or about 35 mg, at least 50 mg or about 50 mg or at least 100 mg or about 100 mg, at least 250 mg or about 250 mg, or at least minus 500 mg or about 500 mg of the non-polar active ingredient per 8 fluid ounces (0.236588 liters) of the aqueous medium. Liquid dilution compositions typically have one or more desired properties, such as particle size, transparency, NTU value, stability, e.g., they are crystal-free, phase separation, ring formation, or unpleasant taste / odor, such as for at least a specified amount of time when stored under specified storage conditions.

For example, the compositions include liquid dilution compositions that have a particle size less than 500 or about 500, less than 300 or about 300, less than 200 or about 200 nm, less than 100 or about 100 nm, less than 50 or about about 50 nm or less than 25 or about 25 nm on average or at most; those that have an NTU value of less than 500 or about 500, less than 300 or about 300, less than 200 or about 200, less than 100 or about 100, less than 50 or about 50, less than 25 or about 25, or less than 10 or about 10; those that contain visible particles do not contain visible crystals, do not present ring formation and / or phase separation; and / or remain free of (or do not present) visible particles, crystals

ES 2 396 946 T3 visible, ring formation and / or phase separation and / or unpleasant taste / odor when stored at room temperature (e.g. 25 ° C or about 25 ° C) or at a refrigerated temperature (e.g.

0-10 ° C or about 0-10 ° C, e.g. at or about 4 ° C) or at freezing temperature (e.g. -20 ° C or about -20 ° C), when stored for at least one day, at least a week, at least thirty days, or at least a year.

Methods for preparing the concentrates and methods for preparing the liquid dilution compositions are also described. Generally, the methods for preparing the concentrates are performed by separately generating an oil phase and an aqueous phase, and mixing the two phases, typically by emulsion, to form the concentrate, which is a liquid nanoemulsion concentrate. The oil phase ingredients are added to form the oil phase and the aqueous phase ingredients are added to form the water phase. The ingredients are selected from the ingredients of the concentrates, as described herein, which typically include a nonpolar compound, a surfactant, and a polar solvent, as described herein. Typically, the oil phase ingredients include the nonpolar compound (s), typically a nonpolar active ingredient (s) of the concentrate, and the aqueous phase ingredients include the polar solvent. Ingredients are added in amounts within the appropriate concentration range to provide concentrates as described herein. In one example, the ingredients of the aqueous phase include the surfactant. In another example, the oil phase ingredients contain the surfactant. In one example, the aqueous phase ingredients and the oil phase ingredients contain the surfactant.

The amounts of the surfactant (s), the non-polar active ingredient (s), and the polar solvent are selected based on the appropriate concentration ranges of these ingredients in the resulting concentrate. For example, the non-polar active ingredient is included in an amount between 5% or about 5% and 10% or about 10% by weight (w / w) of the final concentrate; the surfactant is included in an amount that is between 16% or about 16% and 30% or about 30% by weight (w / w) of the final concentrate; and the polar solvent is included in an amount that is between 60% or about 60% and 79% or about 79% by weight (w / w) of the final concentrate as described above.

In one example, the oil phase ingredients additionally include the non-polar solvent (s). In one example, the concentrate is prepared with first and second oil phase ingredients and the first oil phase ingredient includes the nonpolar active ingredient and the solvent. In one example, the solvent contains an oil, other than the non-polar active ingredient, such as, for example, Vitamin E, linseed oil, and / or safflower oil.

In one example, the oil phase ingredients and / or the aqueous phase ingredients contain the co-surfactant in an amount sufficient to stabilize the concentrate, such as a phospholipid, eg, phosphatidylcholine. In one example, the amount of phospholipids is between 0.1% or about 0.1% and 1% or about 1% by weight (w / w) of the concentrate. In another example, the oil phase ingredients and / or the water phase ingredients additionally contain at least one preservative in an amount sufficient to preserve the concentrate, such as, for example, a preservative containing benzyl alcohol. In one example, the amount of the preservative and / or benzyl alcohol is between 0.1% or about 0.1% and 1% or about 1% by weight (w / w) of the concentrate.

In another example, the oil phase ingredients and / or the water phase ingredients additionally contain an emulsion stabilizer, in an amount sufficient to stabilize the concentrate, such as an emulsion stabilizer containing a combination of gums, such as any one or more of guar gum, xanthan gum, and sodium alginate.

In an exemplary method of making the concentrate, an oil phase is generated by mixing the oil phase ingredients in a first container and heating the oil phase ingredients; an aqueous phase is generated by mixing one or more aqueous phase ingredients in a second container and heating the aqueous phase ingredients; and the oil and water phases are emulsified to generate the concentrate.

In another exemplary method provided, an oil phase is generated by mixing one or more first oil phase ingredients in the first container and heating the first oil phase ingredients at least until the first oil phase ingredients dissolve; then one or more additional ingredients of the oil phase are added to the first container; and mixing and heating the first ingredient and additional ingredients of the oil phase; an aqueous phase is generated by mixing one or more aqueous phase ingredients in a second container and heating the aqueous phase ingredient (s); and the aqueous and oily phases are emulsified, to generate the concentrate.

Heating and mixing of the water and oil phases can be done simultaneously or sequentially, in any order.

In any of the methods for preparing the concentrates, the mixing steps (for example, mixing the oil and / or aqueous phases) can be carried out with a conventional mixer, such as any of the conventional mixers indicated in this document, or with any another mixer described in this

ES 2 396 946 T3 document, such as with a homogenizer. In any of the methods provided, heating can be accomplished using one or more heating devices such as, for example, a hot plate, a water jacket, or any of the heating devices indicated herein. In one example, the oil phase ingredients are heated with a first heating apparatus and the aqueous phase ingredients are heated with a second heating apparatus. In one example, heating involves heating the ingredients to 60 ° C or about 60 ° C, or to about 70 ° C, or to or about 71 ° C. In one example, the oil phase and / or aqueous phase ingredients are heated between about 45 ° C or about 45 ° C and between about 85 ° C or about 85 ° C, for example; year approximately 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68 , 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84 or 85 ° C.

In either method, the emulsifier can be made using a homogenizer, such as any homogenizer described herein. In one example, the emulsification is performed between 850 rpm or about 850 rpm and 1200 rpm or about 1200 rpm. In another example, it is done at or about 30 rpm.

In some examples, the methods further include rapidly cooling the emulsion being formed during the emulsifying step. In some examples, quenching results in the cooling of the emulsion being formed between 25 ° C or about 25 ° C and 43 ° C or about 43 ° C, such as 25, 26, 27, 28. , 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42 or at 44 ° C, for example, between at or about 25 ° C and at or at about 35 ° C, or between at or about 35 ° C and at or at about 43 ° C. In one example, cooling the emulsion results in less than or about 60 minutes, less than or about 30 minutes, or between or about 30 and in or about 60 minutes. Exemplary means for performing rapid cooling include repeatedly passing the phases through a cooling apparatus attached to a container.

The methods may further include adding one or more flavorings (eg lemon oil, D-limonene) and / or one or more pH adjusters (eg citric acid, phosphoric acid) to the concentrate, eg after emulsifying. the oil and water phases. The pH is typically measured simultaneously with, before and / or after the addition of the pH adjuster, and the amount of the pH adjuster is determined by the pH of the concentrate. Typically, the pH adjuster comprises an acid or a base in an amount sufficient to affect the pH of the concentrate.

In one example of the methods, the ingredients are added to the container (s) simultaneously or sequentially, in any order. In another example, the ingredients (for example, the oil phase and / or water phase ingredients) are added to the containers in a particular order, such as the specific order provided herein, for example, in the examples. individual provided. In one example, where the aqueous phase ingredients contain a polar solvent (for example, water, propylene glycol, glycerin, or another diol such as another sugar alcohol) and an emulsion stabilizer, the aqueous phase ingredients are added sequentially in The following order: 1) polar solvent (eg, water, propylene glycol, or glycerin); 2) emulsion stabilizer. In another example, where the oil phase ingredients contain the surfactant, the nonpolar compound, and a preservative, the oil phase ingredients are added sequentially, in the following order: 1) surfactant, 2) preservative; 3) non-polar compound (ie, non-polar active ingredient). In another example, where the oil phase ingredients contain the surfactant, the nonpolar compound, a preservative, and an emulsion stabilizer, the oil phase ingredients are added to the oil phase container sequentially, in the following order : 1) surfactant, 2) preservative; 3) nonpolar compound; and 4) emulsion stabilizer. In another example, where the oil phase ingredients contain the surfactant, the nonpolar compound, a preservative, a solvent, and an emulsion stabilizer, the oil phase ingredients are added to the oil phase container sequentially, in the following order : 1) surfactant, 2) preservative; 3) non-polar solvent; 4) nonpolar compound; and 5) emulsion stabilizer. In another example, where the oil phase ingredients contain the surfactant, the nonpolar compound, a preservative, a nonpolar solvent, a cosurfactant, and an emulsion stabilizer, the oil phase ingredients are added to the container of the oil phase sequentially, in the following order: 1) surfactant, 2) preservative; 3) non-polar solvent; 4) co-surfactant; 5) non-polar compound and 6) emulsion stabilizer. In another example, where the aqueous phase contains a polar solvent and a surfactant, the polar solvent and the surfactant are added sequentially in this order. In another example, where the oil phase contains a nonpolar solvent, a preservative, a cosurfactant, a surfactant, and a nonpolar compound, the nonpolar solvent, the preservative, and the cosurfactant are added and mixed until dissolved. the cosurfactant; the surfactant is added until it dissolves; and the non-polar active ingredient is added, in that order.

In one example, the methods for producing the concentrates are performed using experimental scale processes, as provided herein. In another example, particularly with large batches of the concentrates, the methods are performed with a large-scale process, as described later in this document, such as the exemplary large-scale processes illustrated in Figure 1.

In one example, where the polar solvent is water, the water is purified first by passing it through purifiers. In one example, water purification is done by passing it through purifiers,

ES 2 396 946 T3 sequentially, in the following order: a carbon filter, an ion exchange kit, a reverse osmosis kit, a 100 micron end-point filter, and a 50-micron point-of-use filter. In this example, after purification, the water is added with the other ingredients of the aqueous phase to an aqueous phase tank. The ingredient (s) of the aqueous phase are then mixed using a conventional mixer attached to the tank, for example, mounted on top of the tank. A heating apparatus (typically the water jacket in the aqueous phase tank) is used to heat the aqueous phase ingredients during the generation of the aqueous phase, typically at low heat (eg 60 ° C). To generate the oil phase, the oil phase ingredient (s) are weighed / measured and added to an oil phase tank. The oil phase ingredients are mixed using a conventional mixer attached to the oil phase tank, eg tank mounted. A heating apparatus (typically the water jacket in the oil phase tank) is used to heat the oil phase ingredients during generation of the water phase, typically at low heat (eg 60 ° C). Once the oil and water phases reach 60 ° C, and after the oil phase components have dissolved, the oil and water phases are combined by transferring the oil phase to the water phase container, through a medium of transfer. For this process, a homogenizer mounted on the aqueous phase tank is turned on, for example, at 850-1200 rpm. The ball valves are then opened and the transfer pump is switched on, thereby effecting the transfer of the liquid oil phase to the water phase tank through the transfer tube or tubes. As the phases combine, the mixture is homogenized by continuous mixing with the homogenizer. The homogenizer can be adjusted, for example, by adjusting the baffle plate in the homogenizer to achieve and maintain an emulsion, for example, by moving the baffle plate further during emulsion formation and / or further out of the forming emulsion. During the emulsion stage, the forming emulsion is cooled, typically rapidly cooled, by repeatedly passing the forming emulsion through a recirculating cooler, which is attached to the aqueous phase tank. The emulsion is transferred, via transfer medium to a containment / packaging tank, where additional ingredients can be added and / or the mixture can be evaluated. The additional ingredients are mixed into the concentrate using a conventional mixer. A final product filter is used to filter the concentrate before use.

Any of the methods of producing the concentrates can be used to prepare any of the provided concentrates, as described herein.

Methods for producing the liquid dilution compositions containing the concentrates, such as beverages containing the concentrates, are also described. These methods include methods of providing oil-based additives, for example, in a food or beverage. These methods include adding any of the provided concentrates, eg, liquid nanoemulsion concentrates, to an aqueous medium, such as a beverage. Typically, the concentrate is added to the medium eg, beverage such that the medium contains an effective amount of the additive (eg, the non-polar active ingredient).

The effective amount of the additive, such as the non-polar active ingredient, is the amount and / or concentration of the additive necessary to prevent, cure, ameliorate, arrest, or partially arrest a symptom of a disease or disorder or the amount and / or concentration desired to the intake of an individual such as daily intake and / or nutritional supplement, for example, an amount sufficient to enhance the nutritional, pharmaceutical, nutraceutical, health or energy properties of a food, beverage or other consumable product. In some examples, the concentrate is added to the aqueous medium such that the resulting dilution liquid composition contains an effective amount of a particular nonpolar compound, for example, a particular amount by volume or weight of the composition such as, for example, less 25 mg or about 25 mg, at least 35 mg or about 35 mg, at least 50 mg or about 50 mg or at least 100 mg or about 100 mg, at least 250 mg or about 250 mg, or at least 500 mg or about 500 mg of the non-polar active ingredient per 8 fluid ounces (0.236588 liters) of the liquid dilution composition.

In one example, an effective amount is a concentration or amount of the liquid nanoemulsion where at least 25 mg or about 25 mg, typically at least 35 mg, for example, 35, 40, 45, 50, 55, 60, 65, 70 , 75, 80, 85, 90, 95, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290 , 300, 325, 350, 375, 400, 425, 450, 475, 500, 550, 600, 700, 800, 900 1000, 1500, 2000 mg, or more of the nonpolar active ingredient is contained in at least 8 fluid ounces (0.236588 liters) of the aqueous medium.

The United States Provisional Application with Serial Number: 61 / 070,381, filed on March 20, 2008 entitled COMPOSITIONS CONTAINING NON-POLAR COMPOUNDS, and the Provisional Application of the United States with Serial No. 61 / 132,424, filed on June 16, 2008 entitled COMPOSITIONS CONTAINING NON-POLAR COMPOUNDS, each by Philip Bromley, disclose, for example, compositions containing non-polar compounds and surfactants such as PEG derivatives of Vitamin E, such as Polyethylene glycol tocopherol succinate (TPGS) and TPGS analogs, including TPGS homologs and methods for preparing the compositions.

Brief description of the drawings

Figure 1 sets forth an exemplary large-scale process 100 of the provided methods for preparing the liquid nanoemulsion concentrates. This process is exemplary and variations may be used. In this

ES 2 396 946 T3 example, water 101 is used as the polar solvent and the water is first purified by passing it through the following purifiers, sequentially, in the following order: a carbon filter 105, an exchange kit 106 ion, a reverse osmosis kit 107, a 100 micron end point filter 108, and a 50 micron point of use filter 109. After purification, the water is added, with the other ingredients of the aqueous phase, to the aqueous phase tank 103. In other examples, the polar solvent is another polar solvent, such as glycerin or propylene glycol. In the illustrated example the ingredient (s) of the aqueous phase are then mixed using a conventional mixer 111 attached to the tank, eg mounted on top of the tank. A heating apparatus (typically the water jacket in the aqueous phase tank) is used to heat the aqueous phase ingredients during aqueous phase generation, typically at low heat (eg 60 ° C). To generate the oil phase, the oil phase ingredient (s) are weighed / measured and added to the oil phase tank 102. The oil phase ingredients are mixed using a conventional mixer 111 attached to the oil phase tank, eg tank mounted. A heating apparatus (typically the water jacket in the oil phase tank) is used to heat the oil phase ingredients during generation of the water phase, typically at low heat (eg 60 ° C). Once the oil and water phases reach 60 ° C, and after the components of the oil phase have dissolved, the oil and water phases are combined by transferring the oil phase to the water phase container, through a medium. 112 transfer. For this process, a homogenizer 110 mounted in the aqueous phase tank is connected, for example, at 850-1200 rpm. The ball valves are then opened and the transfer pump is turned on, thereby effecting the transfer of the liquid aqueous phase to the aqueous phase tank through the transfer tube or tubes. As the phases are combined, the mixture is homogenized by continuous mixing with homogenizer 110. The homogenizer can be adjusted, for example, by adjusting the baffle plate in the homogenizer to achieve and maintain an emulsion, for example, by moving the baffle plate further into the forming emulsion and / or further out of the forming emulsion. During the emulsion stage, the emulsion being formed is cooled, typically rapidly cooled, by repeatedly passing the emulsion being formed through a recirculating cooler 115 (for example, Model No. OC-1000 RO, commercially available). by Turmoil, West Swanzey, NH), which is attached to the aqueous phase tank. The emulsion is transferred, via transfer means 112 to a containment / packaging tank 104, where additional ingredients can be added and / or the mixture can be evaluated. The additional ingredients are mixed into the concentrate using a conventional mixer 111. A final product filter 113 is used to filter the concentrate before use.

Detailed description

Outline

A. DEFINITIONS32

B. COMPOSITIONS CONTAINING NON-POLAR COMPOUNDS66

1. Liquid nanoemulsion concentrates containing nonpolar compounds 68

to. Formulation of liquid concentrates70

i. Typical Ingredients and Concentration Ranges 72 ii. Initial concentrate evaluation75 (1) Transparency75 (2) Empirical evaluation78 (3) Particle size78 (4) Turbidity measurement79 iii. Selection of a formulation and modification of 80 formulations

b. Nonpolar compounds82

i. Active ingredients containing Fatty Acid85

Polyunsaturated (PUFA) (1) Omega-387 fatty acid compounds (a) DHA / EPA87 (i) Fish oils 88 (ii) Seaweed oil90 (b) Flaxseed oil-omega 391 (ALA) (2) Omega compounds -692 (a) Borage Oil (Acid 92

Gamma-Linolenic (GLA)) (3) Extract of Saw Palmetto93

ES 2 396 946 T3 (4) Conjugated Linoleic Acid (CLA) 93 ii. Active Ingredients of Coenzyme Q94 (1) Coenzyme Q1094 iii. Active Ingredients Containing Phytosterol95

c. Surfactants96

i. Surfactants derived from Vitamin E98 (1) PEG derivatives of Vitamin E99 (a) Tocopherols and Tocotrienols100 (b) PEG moieties100 (c) Linkers101 (d) Polyethylene glycol of Tocopherol and 101 polyethylene glycol diesters of Tocotrienol Ethoxylic acid esters of Vitamin Dicarboxylic acid esters a PEG) _ (e) Other PEG Esters of 103

Vitamin E (f) Surfactants from TPGS104 ii. Surfactant concentration105 iii. HLB106

d. Co-surfactants (emulsifiers) 107

i. Phospholipids107

and. Polar solvents108

F. Preservatives and Sterilizers111

g. Emulsion stabilizers (co-emulsifier) ​​112

h. Non-polar solvents113

i. Scents114

j. PH adjusters115

2. Powder forms of compositions115

3. Liquid dilution compositions containing the dilute concentrates119

to. Transparency121

i. Transparency determined by empirical evaluation122 ii. Transparency determined by particle size123 or number of particles iii. Turbidity123

b. Stability124

c. Desirable characteristics for human consumption125

d. Security126

and. Oral bioavailability126

C. METHODS FOR PREPARING LIQUID NANOEMULSION CONCENTRATES THAT126

CONTAIN NON-POLAR COMPOUNDS

1. Equipment for preparing concentrates127

2. Scales127

to. Purifiers including filters128

b. Containers for mixing ingredients129

c. Mixers130

d. Heating devices132

and. Cooling devices132

F. Transfer media133

g. Assessment Team134

3. General methods for preparing liquid nanoemulsion concentrates134

to. Generate the Aqueous Phase 136

ES 2 396 946 T3

i. Ingredients of the aqueous phase 137

b. Generating the oil phase138

i. Oil Phase Ingredients139

c. Combine and emulsify the oil phase and the aqueous phase140

i. Combine the oil and aqueous phases 140 ii. Emulsify the oil and water phases141 iii. Cooling142

d. Additional stages142

i. Additional ingredients143 ii. Concentrate evaluation143 iii. Filter the concentrate143

Four. Experimental scale process144

5. Large-scale manufacturing process145

to. Water purification146

b. Generation of the aqueous phase and the oil phase146

c. Combination and emulsification of the phases147

d. Cooldown148

and. Additional stages149

D. METHODS FOR PREPARING THE LIQUID DILUTION COMPOSITIONS THAT149

CONTAIN THE DILUTED CONCENTRATES

1. Dilutions150

2. Analyze Aqueous Liquid Dilution Compositions Containing the 151 Liquid Concentrates

to. Transparency / turbidity152

i. Empirical evaluation152 ii. Particle size 153 iii. Turbidity measurement154

E. EXAMPLES

154

A. DEFINITIONS

Unless defined otherwise, all technical and scientific terms used in this document have the same meaning as commonly understood by one of ordinary skill in the art to which the invention or inventions pertain.

In the event that there are a plurality of definitions for the terms in this document, those in this section prevail. When referring to a URL or other identifier or address, it is understood that such identifiers may change and particular information on the Internet may come and go, although the equivalent information is known and can be easily accessed, such as searching the Internet and / or appropriate databases. The reference to these evidences the possibility and public dissemination of such information.

As used herein, colloid refers to a mixture containing two phases, a dispersed phase and a continuous phase, the dispersed phase containing particles (droplets) distributed throughout the continuous phase. Colloidal mixtures include aerosols, foams and dispersions, eg emulsions eg nanoemulsions. A liquid colloid, for example a nanoemulsion may have a similar appearance, for example transparency, to a solution, in which there is no dispersed phase.

As used herein, "emulsion" refers to a colloidal dispersion of two immiscible liquids, for example, an oil and water (or another aqueous liquid for example, a polar solvent), one of which is part of a continuous phase and the other is part of a dispersed phase. The compositions provided include emulsions, typically oil-in-water nanoemulsions (including any oil-soluble phase dispersed in any aqueous phase, also referred to as the aqueous phase), wherein the oil phase is the dispersed phase and the aqueous phase is the aqueous phase. keep going. Emulsions are typically stabilized by one or more surfactants and / or cosurfactants and / or emulsion stabilizers. Surfactants form an interfacial film between the oil and water phase of the emulsion, providing stability. Typically, the nanoemulsions of the provided compositions contain micelles, which contain one or more surfactants that surround a non-active ingredient.

ES 2 396 946 T3 polar, which are dispersed in the aqueous phase. Exemplary of the emulsions provided are the provided liquid nanoemulsion concentrates and liquid dilution compositions prepared by diluting the concentrates, typically in an aqueous medium.

As used herein, a nanoemulsion is an emulsion in which dispersed droplets, for example micelles, have a diameter (particle size) of less than 1000 nm or less than about 1000 nm, typically less than 500 nm or less. of about 500 nm, typically less than 300 nm or about 300 nm, for example, less than 250 nm or about 250 nm, for example, less than 200 nm or less than about 200 nm, for example, less than or less than about 5, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, or 200 nm. Exemplary of nanoemulsions are the provided liquid nanoemulsion concentrates and liquid dilution compositions, eg, liquid dilution compositions containing the dilute concentrates.

As used herein, "surfactant and surface active agent" refer synonymously to amphiphilic molecules of synthetic and natural origin having hydrophobic portion (s) and hydrophilic portion (s). Due to their amphiphilic (amphipathic) nature, surfactants and co-surfactants can typically reduce the surface tension between two immiscible liquids, eg, the oil and water phases in an emulsion, stabilizing the emulsion. Different surfactants can be characterized on the basis of their relative hydrophobicity and / or hydrophilicity. For example, relatively lipophilic surfactants are more soluble in fats, oils, and waxes, typically having HLB values ​​less than 10 or about 10, while relatively hydrophilic surfactants are more soluble in aqueous compositions, for example, water and typically have HLB values. HLBs older than 10 or about 10. Relatively amphiphilic surfactants are soluble in oil- and water-based liquids and typically have HLB values ​​near 10 or about 10.

Surfactants include, for example, soaps, detergents, lipids, emulsifiers, dispersing agents and wetting agents, molecules that emulsify liquids, for example, by forming an emulsion in an aqueous medium, or a diluting aqueous liquid composition, for example, by forming a dispersion colloidal of two immiscible liquids in the form of drops, for example, an emulsion such as a microemulsion; and compounds that form various macromolecular structures, eg, aggregates, in liquids, eg, micelles, liquid bilayer structures, including liposomes and reverse micelles.

Typically, surfactants used in compositions provided at an HLB value between 14 or about 14 and 20 or about 20, for example, at or about 14,15,16,17,18,19 or 20, and typically between at or about 15 and at approximately 18. Exemplary surfactants include, but are not limited to, nonionic surfactants, such as surfactants derived from polyethylene glycol (PEG), such as PEG derivatives of Vitamin E, for example, tocotrienol diesters or PEG diesters, such as TPGS ( eg, TPGS 1000) and TPGS analogs; and other known surfactants having HLB values ​​between 14 or about 14 and 20 or about 20, typically between at or about 15 and at or about 18, such as other surfactants described herein. Typically the surfactant is a natural surfactant, for example a surfactant that is GRAS (generally recognized as safe) by the FDA and / or certified by Kosher.

As used herein, a PEG derivative of Vitamin E is a compound containing one or more Vitamin E moieties (eg, tocopherol or tocotrienol) linked, eg, via an ester, ether, amide, or thioester bond with one or more polyethylene glycol (PEG) moieties through a linker, for example, a dicarboxylic or tricarboxylic acid. Exemplary of the PEG derivatives of Vitamin E are polyethylene glycol tocopherol succinate (TPGS), TPGS analogs, TPGS homologues, and TPGS derivatives.

As used herein, a tocopherol polyethylene glycol diester (TPGD) is a PEG derivative of tocopherol in which the linker is a dicarboxylic acid (a carboxylic acid having two carboxy groups, for example succinic acid) such as succinic acid. Exemplary of dicarboxylic acids that can be used as linkers in these surfactants are PEG diester of tocopherol and tocotrienol, succinic acid, sebacic acid, dodecanedioic acid, suberic acid or azelaic acid, citraconic acid, methyl citraconic acid, itaconic acid, maleic acid, acid glutaric, glutaconic acid, fumaric acids and phthalic acids. Exemplary of TPGD are tocopherol succinate polyethylene glycol (TPGS), tocopherol sebacate polyethylene glycol, tocopherol dodecanedioate polyethylene glycol, tocopherol suberate polyethylene glycol, tocopherol polyethylene glycol azelaate, tocopherol polyethylene glycol acetylene glycol, tocopherol tocopherol citol acetylene glycol ester tocopherol itaconate, tocopherol maleate polyethylene glycol, tocopherol glutarate polyethylene glycol, tocopherol glutaconate polyethylene glycol and tocopherol phthalate polyethylene glycol, among others.

As used herein, TPGS polyethylene glycol succinate, tocopheryl polyethylene glycol succinate surfactant, and TPGS surfactant refer to polyethylene glycol tocopherol (PEG) diesters, which are formed by bonding, by esterification, tocopherol succinate that by itself is an ester prepared by the esterification of tocopherol and succinic acid. The term tocopherol refers to any naturally occurring or synthetic form of vitamin E and can refer to a single compound or a mixture. Examples of tocopherols include,

ES 2 396 946 T3 for example, α-tocopherol, Da-tocopherol, β-tocopherol, g-tocopherol and δ-tocopherol. The PEG residue of the TPGS surfactant can be any PEG residue, for example, PEG residues between 200 kDa or about 200 kDa and 20,000 kDa or about 20,000 kDa, typically between 200 kDa or about 200 kDa and 6000 kDa or about 6000 kDa, for example , between 600 kDa or about 600 kDa and 6000 kDa or about 6000 kDa, typically between 200 kDa or about 200 kDa and 2000 kDa or about 2000 kDa, between 600 kDa or about 600 kDa and 1500 kDa or about 1500 kDa, or between 600 kDa or about 600 kDa and 1000 kDa or about 1000 kDa, for example, 200 kDa or about 200 kDa, 300 kDa or about 300 kDa, 400 kDa or about 400 kDa, 500 kDa or about 500 kDa, 600 kDa or about 600 kDa, 800 kDa or about 800 kDa, and 1000 kDa or about 1000 kDa; and PEG residues that are modified eg, methylated PEG residues (ιτιPEG) and / or PEG including other PEG analogs, eg, PEG-nHs, PEG-aldehyde, PEG-SH, PEG-NH2, PEG-CO2H, and PEG branched.

They are exemplary of the TPGS surfactants TPGS-1000, which is a 1000 kDa PEG residue. The TPGS can be any water soluble natural tocopherol polyethylene glycol succinate, eg, food grade TPGS marketed under the name TPGS® Vitamin E Eastman, food grade from Eastman Chemical Company, Kingsport, TN. This TPGS is a water soluble form of vitamin E from a natural source, which is prepared by esterifying the carboxyl group of crystalline Da-tocopheryl acid succinate with polyethylene glycol 1000 (PEG 1000), and contains between 260 and 300 mg / g of total tocopherol. A similar compound can be prepared by esterifying the carboxyl group of synthetic Vitamin E form d, 1 with PEG 1000. It forms a clear liquid when dissolved 20% in water. This tocopheryl polyethylene glycol is a water-soluble preparation of a fat-soluble vitamin (vitamin E), for example, as disclosed in US Patents No. 3,102,078, 2,680,749 and US Published Application No. 2007/0184117 and 2007/0141203. Also exemplary of the TPGS surfactant that can be used in the compositions provided is Natural Water Soluble Vitamin E (TPGS), available from ZMC-USA, The Woodlands, Texas. Any known source of TPGS can be used. Typically, TPGS surfactant is GRAS and Kosher certified. Typically the TPGS has an HLB value between 16 or about 16 and 18 or about 18.

As used herein, "analog" refers to a chemical compound that is structurally similar to another compound (called a parent compound), but differs slightly in composition, for example, by the variation, addition, or removal of an atom, a or more units (eg, methylene - (CH2) n- units) or one or more functional groups. The analog may have different chemical or physical properties compared to the parent compound and / or may have improved biological and / or chemical activity. Alternatively, the analog may have similar or identical chemical or physical properties compared to the parent compound and / or may have similar or identical biological and / or chemical activity. For example, the analog may be more hydrophilic or may have altered reactivity compared to the parent compound. The analog may be mimetic with the chemical and / or biological activity of the parent compound (ie, it may have similar or identical activity) or, in some cases, it may have greater or lesser activity. The analog can be of natural or non-natural origin (eg synthetic) which is a variant of the parent compound. Other types of analogs include isomers (eg, enantiomers, diastereomers) and other types of chiral variants of a compound as well as structural isomers. The analog can be a branched or cyclic variant of a linear compound. For example, a linear compound may have an analog that is branched or otherwise substituted to confer certain desirable properties (eg, improve hydrophobicity or bioavailability). Exemplary of these analogs used in the provided compositions and methods are the TPGS analogs, which can be used as surfactants in place of the TPGS in the provided compositions.

As used herein, "homologue" refers to an analog that differs from the parent compound only by the presence or absence of a single unit, such as a methylene unit or some multiples of such units, for example, - (CH2) n- . Typically, a homologue has similar chemical and physical properties as the parent compound. They are exemplary of the homologs used in the compositions provided and methods are homologs of TPGs.

As used herein, polyethylene glycol tocopherol succinate analog, TPGS analog, and TPGS analog surfactant refer to compounds, other than TPGS, that are similar to a parent TPGS compound, but differ slightly in composition, for for example, by varying, adding, or removing an atom, one or more units (eg, a methylene (CH2) n unit or units), or one or more functional groups. TPGS analogs include surfactants derived from Vitamin E, including PEG derivatives of Vitamin E, including PEG diesters of vitamin E, such as but not limited to, polyethylene glycol tocopherol sebacate (PTS), polyethylene glycol tocopherol dodecanedioate (PTD) , polyethylene glycol tocopherol suberate (PTSr), polyethylene glycol tocopherol azelaate (PTAz) and polyoxethanyl tocotrienol sebacate (PTrienS) as well as other PEG derivatives of Vitamin E. In one example, the surfactant in the provided compositions is a TPGS analog. Exemplary TPGS analogs compounds, other than TPGS compounds, having the formula shown in Scheme I are:

ES 2 396 946 T3

Scheme I

<img file="ES2396946T3_D0001.tif" />

in which each of R¹, R², R³ and R⁴ is, independently, hydrogen (H) or methyl (CH3); each dashed line is, independently, a single or double bond; n is an integer from 1 to 5000; each of myq is, independently, 0 or 1; and p is an integer from 1 to 20.

For example, TPGS analogs include, but are not limited to, compounds having the formula of Scheme I, where, when the bonds represented by the dashed lines marked A and B are single bonds, and m and q are both equal to 0, p is an integer from 2 to 20. TPGS analogs also include compounds in which the broken line in B or the broken line in A, or both broken lines, represents at least one double bond. For example, TPGS analogs include a compound as in Scheme I, where when the dashed line in A represents only single bonds, the dashed line in B represents one or more double bonds, for example, PEG diesters of tocotrienol . The TPGS analogs also include compounds as in Scheme I, where when the dashed line marked B represents only single bonds, the dashed line marked A represents one or more double bonds; or when the dashed line marked A does not represent double bonds, and m and q are both 0, p is greater than 1. For example, TPGS analogs include compounds in which one or more of the dashed lines represent a double bond, eg, PEG derivatives of tocotrienol esters (eg, PTrienS).

Also exemplary are compound TPGS analogs, other than TPGS compounds, having the formula shown in Scheme II:

Scheme II

<img file="ES2396946T3_D0002.tif" />

in which each of R¹, R², R³ and R⁴ is, independently, hydrogen (H) or methyl (CH3); the bond represented by the dashed line is a single or double bond; ym is an integer from 1 to 20, and n is an integer from 1 to 5000.

Also exemplary of TPGS analogs are those that include compounds other than TPGS, which have PEG residues that vary in chain length according to the formula shown in Scheme III:

Scheme III

OO

<td rowspan="2">J— ch₂ch₂--</td><td>k₀</td><td>Laugh</td><td></td><td></td>

<td>OR\</td><td></td><td></td><td></td>

<td></td><td>R³^</td><td></td><td></td><td>L lh</td>

<td></td><td></td><td>R⁴</td><td></td><td> 3</td>

ES 2 396 946 T3 in which each of R¹, R², R³ and R⁴ is independently hydrogen (H) or methyl (CH3) and n is an integer from 1 to 5000.

As used herein, TPGS-1000 analogs are compounds other than TPGS-1000 that are similar to a parent TPGS-1000 compound, but differ slightly in composition, for example, by variation, addition, or removal of an atom , one or more units (eg, a methylene (CH2) n unit or units), or one or more functional groups. In one example, the surfactant in the compositions provided herein is an analog of TPGS-1000. Suitable TPGS-1000 analogs include, but are not limited to, other TPGS compounds that have a PEG residue or residues that vary in chain length and molecular weight compared to TPGS-1000, including for example, TPGS compounds that have PEG residues between 200 or about 200 kDa and 20,000 kDa or about 20,000 kDa, typically between 200 kDa or about 200 kDa and 6000 kDa or about 6000 kDa, for example, between 600 kDa or about 600 kDa and 6000 kDa or about 6000 kDa, typically between 200 kDa or about 200 kDa and 2000 kDa or about 2000 kDa, between 600 kDa or about 600 kDa and 1500 kDa or about 1500 kDa, such as, although without limitation 200, 300,400, 500, 600, 800 and 1000 kDa. Also exemplary of TPGS-1000 analogs are TPGS compounds having PEG residues that are modified, eg, methylated PEG (m-PEG) and / or PEG residues including other PEG analogs, eg, PEG-NHS, PEG -aldehyde, PEG-SH, PEG-NH2, PEG-CO2H, and branched PEG. Also exemplary of TPGS-1000 analogs are any TPGS analogues, for example, surfactants derived from Vitamin E, including PEG derivatives of Vitamin E, including PEG diesters of vitamin E, such as, but not limited to, polyethylene glycol sebacate tocopherol (PTS), polyethylene glycol tocopherol dodecanedioate (PTD), polyethylene glycol tocopherol suberate (PTSr), Tocopherol polyethylene glycol azelaate (PTAz) and polyoxethanyl tocotrienyl sebacate (PTrienS) as well as other PEG derivatives of Vitamin E.

As used herein, TPGS homologs are TPGS analogs that differ from a TPGS precursor compound only by the presence or absence of a single unit, such as a methylene unit or some multiples of such units, for example, - (CH2) n-. In one aspect, the TPGS homologs are used as surfactants in the provided compositions. Typically, suitable TPGS homologs have similar surfactant properties compared to the parent compound (TPGS), eg, similar HLB values, eg, HLB values ​​between 14 or about 14 and 20 or about 20. TPGS homologs Exemplary are polyethylene glycol tocopherol sebacate (PTS), polyethylene glycol tocopherol dodecanedioate (PTD), polyethylene glycol tocopherol suberate (PTSr), and polyethylene glycol tocopherol azelaate (PTAz). Exemplary TPGS homologs are compounds having the formula of Scheme I (above), in which neither dotted line A or B represents a double bond and in which, when my and q are both 0, p is greater than 1.

As used herein, TPGS-1000 homologs are analogs of TPGS-1000 that differ from the parent compound TPGS-1000 only by the presence or absence of a single unit such as a methylene unit, or some multiple of such units. , for example, - (CH2) n-. Suitable TPGS-1000 homologues have similar surfactant properties compared to the parent compound (TPGS-1000), eg, similar HLB values, eg, HLB values ​​between 14 or about 14 and 20 or about 20. Homologs Suitable TPGS-1000s include homologs of TPGS-1000 with slight variations in the chain length of the PEG moiety, and me-TPGS-1000, which is a TPGS-1000 that has a methyl end on the PEG moiety.

As used herein, HLB refers to a value that is used to index and describe a surfactant according to its relative hydrophobicity / hydrophilicity, relative to other surfactants. An HLB value of the surfactant is an indication of the molecular balance of the hydrophobic and lipophilic portions of the surfactant, which is an amphipathic molecule. Each surfactant and surfactant mixture (and / or co-surfactants) have an HLB value which is a numerical representation of the relative weight percent of the hydrophobic and hydrophilic portions of the surfactant molecule (s). The HLB values ​​are derived from a semi-empirical formula. The relative weight percentages of the hydrophobic and hydrophilic groups are indicative of surfactant properties, including molecular structure, eg, the types of aggregates that the surfactant will form, and the solubility of the surfactant. See, for example, Griffin, WCJ Soc. Cos. Chem. 1: 311 (1949).

HLB values ​​for surfactant range from 1-45, while the range for nonionic surfactants is typically 1-20. A surfactant is more lipophilic the lower its HLB value. Conversely, the more hydrophilic a surfactant is, the higher its HLB value. Lipophilic surfactants have higher solubility in oil and lipophilic substances, while hydrophilic surfactants dissolve more easily in aqueous media. In general, surfactants with HLB values ​​greater than 10 or greater than about 10 are called hydrophilic surfactants, while surfactants having HLB values ​​less than 10 or less than about 10 are called hydrophobic surfactants. HLB values ​​have been determined and are available for a plurality of surfactants (see, for example, US Patent No. 6,267,985). It should be appreciated that the HLB values ​​for a given surfactant or co-surfactant can vary, depending on the empirical method used to determine the value. Thus, the HLB values ​​of surfactants and cosurfactants provide a rough guide for formulating compositions based on relative hydrophobicity / hydrophilicity. For example, a surfactant is typically selected from surfactants that have HLB values ​​within

ES 2 396 946 T3 of a particular range of surfactant or co-surfactant that can be used for guide formulations. The

Table 1A shows HLB values ​​of exemplary surfactants and co-surfactants.

Table 1A: Exemplary Surfactants and Co-Surfactants HLB Values

<td>Surfactant / co-surfactant</td><td>HLB</td><td>Surfactant / co-surfactant</td><td>HLB</td>

<td>PEG-2 Hydrogenated Castor Oil</td><td> 1,7</td><td>PEG-10 Oleyl Ether</td><td> 12,4</td>

<td>Sorbitan trioleate</td><td> 1,8</td><td>PEG-8 isooctylphenyl ether</td><td> 12,4</td>

<td>Sorbitan Tristearate</td><td> 2,1</td><td>PEG-10 stearyl ether</td><td> 12,4</td>

<td>Glyceryl Stearate</td><td> 3,5</td><td>PEG-35 Castor Oil</td><td> 12,5</td>

<td>Sorbitan Sesquioleate</td><td> 3,7</td><td>PEG-10 cetyl ether</td><td> 12,9</td>

<td>Labrafil</td><td> 4</td><td>Nonoxynol-9</td><td> 12,9</td>

<td>Sorbitan Oleate</td><td> 4,3</td><td>PEG-40 Castor Oil</td><td> 13</td>

<td>Sorbitan monostearate</td><td> 4,7</td><td>PEG-10 isooctylphenyl ether</td><td> 13,5</td>

<td>PEG-2 oleyl ether</td><td> 4,9</td><td>PEG-40 Hydrogenated Castor Oil</td><td> 14</td>

<td>PEG-2 stearyl ether</td><td> 4,9</td><td>Labrasol</td><td> 14</td>

<td>PEG-7 Hydrogenated Castor Oil</td><td> 5</td><td>Nonoxynol-15</td><td> 14,2</td>

<td>PEG-2 cetyl ether</td><td> 5,3</td><td>PEG-12 tridecyl ether</td><td> 14,5</td>

<td>PEG-4 Sorbitan Stearate</td><td> 5,5</td><td>PEG-18 tridecyl ether</td><td> 14,5</td>

<td>PEG-2 sorbitan isostearate</td><td> 6</td><td>Polysorbate 60</td><td> 14,9</td>

<td>Sorbitan Palmitate</td><td> 6,7</td><td>Polysorbate 80</td><td> 15</td>

<td>Triton SP-135</td><td> 8</td><td>PEG- Glyceryl Stearate twenty</td><td> 15</td>

<td>Sorbitan monolaurate</td><td> 8,6</td><td>PEG-20 stearate</td><td> 15</td>

<td>PEG-40 Sorbitan Peroleate</td><td> 9,5</td><td>PEG-20 stearyl ether</td><td> 15,3</td>

<td>PEG-4 Lauryl Ether</td><td> 9,7</td><td>PEG-20 oleyl ether</td><td> 15,3</td>

<td>Polysorbate 81</td><td> 10</td><td>Polysorbate 40</td><td> 15,6</td>

<td>PEG-40 Sorbitan Hexaoleate</td><td> 10</td><td>PEG20 Cetyl Ether</td><td> 15,7</td>

<td>PEG-40 Sorbitan Perisostarate</td><td> 10</td><td>PEG Hexadecyl Ether (20)</td><td> 15,7</td>

<td>Olive Glycerides of PEG-10</td><td> 10</td><td>PEG-60 Hydrogenated Castor Oil</td><td> 16</td>

<td>PEG Sorbitol Hexaoleate</td><td> 10,2</td><td>PEG-30 stearate</td><td> 16,5</td>

<td>Polysorbate 65</td><td> 10,5</td><td>Polysorbate 20</td><td> 16,7</td>

<td>PEG-25 Hydrogenated Castor Oil</td><td> 10,8</td><td>PEG-75 Lanolin</td><td> 16,7</td>

<td>Polysorbate 85</td><td> 11</td><td>PEG23 Lauryl Ether</td><td> 16,9</td>

<td>PEG-7 Glyceryl Cocoate</td><td> 11</td><td>PEG-40 stearate</td><td> 17,3</td>

<td>PEG-8 stearate</td><td> 11,1</td><td>PEG-50 stearate</td><td> 17,7</td>

<td>PEG Sorbitan Tetraoleate</td><td> 11,4</td><td>PEG40 Isooctylphenyl Ether</td><td> 17,9</td>

<td>PEG-15 Glyceryl Isostearate</td><td> 12</td><td>PEG-100 stearate</td><td> 18,8</td>

<td>PEG-35 Almond Glycerides</td><td> 12</td><td>Pluronic F68</td><td> 29</td>

<td>Tocopherol Polyethylene Glycol Succinate (TPGS)</td><td> 16-18</td><td>Phosphatidylcholine</td><td> 7,6</td>

ES 2 396 946 T3

The surfactants and HLB values ​​set forth in Table 1A are exemplary. Any known surfactant or cosurfactant can be used with the provided compositions (see, for example, US Patent No. 6,267,985). The surfactant (s) contained in the compositions provided typically have an HLB value between 14 or about 14 and 20 or about 20, eg, 14 or about 14, 15 or about 15, 16 or about 16, 17 or about 17, 18 or about 18, 19 or about 19 and 20 or about 20.

As used herein, the term "micelle" refers to aggregates formed by surfactants that typically form when the surfactant is present in an aqueous composition, typically when the surfactant is used at a concentration above the critical micelle concentration (CMC). . In micelles, the hydrophilic portions of the surfactant molecules come into contact with the aqueous phase or water, while the hydrophobic portions form the core of the micelle, which can encapsulate the non-polar ingredient (s), for example, the compounds nonpolar in the compositions provided. Typically, the surfactants in the provided compositions form micelles containing the nonpolar ingredient at their center in aqueous liquid dilution compositions. Typically, the micelles in the compositions provided have a particle size of about 1000 nm, typically less than 500 nm or less than about 500 nm, typically less than 300 or about 300 nm, for example, less than 250 nm or about 250 nm. , for example, less than 200 nm or less than about 200 nm, for example, less than or less than about 5, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190 or 200 nm.

As used herein, reverse micelles are surfactant aggregates that are typically formed in lipophilic solution, with the hydrophilic portions forming the core. When the cross-sectional area of ​​the hydrophobic region of the surfactant molecule is greater than that of the hydrophilic part of the molecule, the formation of micelles, which can be hexagonal phase structures, is favored.

As used herein, liposomes are surfactant aggregates composed of lipid bilayers, typically having an aqueous core. Liposomes are typically formed by lipid surfactants, typically phospholipids, which are phosphate-containing amphipathic lipids, eg, molecules containing a phosphate, a glycerol, and one or more fatty acids and similar surfactants. Alternatively, phospholipid surfactants can be used as co-surfactants, which can be incorporated into aggregates of other surfactant or surfactants, eg, micelles. Lipid bilayers are two-dimensional sheets in which all hydrophobic portions, eg, acyl side chains, are protected from interaction with an aqueous liquid, except those at the ends of the sheet. An energetically unfavorable interaction of the acyl chains with water results in the folding of the bilayers to form liposomes, three-dimensional lipid bilayer vesicles. In the example, the liposome is formed as a single bilayer that encloses a single aqueous space (small unilamellar vesicles; SUVS). In another example, the liposome is composed of concentric bilayers with many aqueous spaces that alternate with the bilayers (multilamellar vesicles; MLVS). Liposomes that can be used to encapsulate hydrophobic and hydrophilic active ingredients. In liposomes, the non-polar active ingredients are typically divided between the bilayers while the hydrophilic active ingredients are typically trapped within the aqueous compartments. In the example, liposomes can be advantageous as transport / encapsulation systems because they are stable and can protect the active ingredients from degradation, for example, by oxygen and digestive enzymes.

As used herein, co-surfactant is used to refer to a surfactant, typically a phospholipid, which is used in the compositions provided, in combination with a surfactant (eg, a major surfactant) for example, to improve the emulsifying the provided compositions and / or compounds, for example to emulsify the ingredients. In one example, the compositions provided contain at least one surfactant and at least one co-surfactant. Typically, the cosurfactant is a lipid, eg, a phospholipid, eg, phosphatidylcholine. In one example, the cosurfactant has an HLB value between 7 or about 7 and 8 or about 8. Typically the cosurfactant represents a lower percentage by weight (w / w) of the compositions provided, compared to the surfactant. . Thus, the compositions provided typically have a lower concentration of the cosurfactant (s) than the surfactant.

As used herein, a phospholipid is a phosphate-containing amphipathic lipid, eg, a molecule containing a phosphate, a glycerol, and one or more fatty acids. In one example, one or more phospholipids are used as a co-surfactant in the compositions provided. Exemplary phospholipids used in the compositions provided are lecithin, including phosphatidylcholine (PC), phosphatidylethanolamine (PE), distearoylphosphatidylcholine (DSPC), phosphatidylserine (PS), phosphatidylglycerol (PG), phosphatidic acid (PA), phosphatidyl acid (PA), phosphatidyl choline (PI). SPM) or a combination thereof. Typically, the phospholipid is phosphatidylcholine (PC), which is sometimes referred to by the general name lecithin. Exemplary of the phospholipids that can be used as co-surfactants in the provided compositions are the phospholipids marketed by Lipoid, LLC, Newark, NJ, for example, Purified Egg Lecithins, Purified Soybean Lecithins, Egg Lecithins, and Soybean Seed. Hydrogenated, Egg Phospholipids, Soy Seed Phospholipids, Hydrogenated Egg

ES 2 396 946 T3 and Soya Seed Phospholipids. Synthetic Phospholipids, PEGylated Phospholipids, and phospholipid combinations sold by Lipoid, LLC. Exemplary of the phosphatidylcholine that is used as a co-surfactant in the compositions provided is the phosphatidylcholine composition sold by Lipoid, LLC, under the name

Lipoid S100, which is obtained from soybean extract and contains more than 95% or more of approximately the

95% phosphatidylcholine.

Typically, for micelle formation, a surfactant or surfactants are used in which the cross-sectional area of ​​the hydrophilic portion of the surfactant molecule is greater than that of the hydrophobic portion of the molecule. For example, TPGS surfactants having an HLB in the range of or about 14 to or about 20, typically between or about 15 and or about 18, are surfactants for stabilizing oil-in-water emulsions. containing the non-polar active ingredients, for example, in nano-sized droplets suspended or dispersed in an aqueous phase or aqueous liquid, for example, an aqueous medium, in the form of spherical micelles containing the hydrophilic portions of the molecule (s) facing the aqueous phase and the hydrophobic portions in the center of the spherical micelles, for example, surrounding the non-polar active ingredient. Typically, the surfactants and / or co-surfactants in the provided compositions aggregate into the nanoemulsions and aqueous liquids to form micelles, which contain the nonpolar compound (s). The hydrophilic portion or portions of the surfactant molecules are oriented toward the outside of the micelle, in contact with the aqueous medium, while the hydrophobic portion or portions of the surfactant molecules are oriented toward the center of the micelle, in contact with the nonpolar compound or compounds, which is contained in the center of the micelle. Micelles can contain more than one surfactant.

As used herein, "particle size" and "average particle size" refer synonymously to the average size of the particles in a given liquid, for example, the droplet diameter or the micelle diameter in an emulsion. Typically, the nanoemulsion concentrates provided, and the liquids made from the concentrates, have a particle size less than about 1000 nm, typically less than 500 nm, or less than about 500 nm, typically less than 300 nm or about 300 nm. , for example, less than 250 nm or about 250 nm, for example, less than 200 nm or less than about 200 nm, for example, less than or less than about 5, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, or 200 nm. In one example, dilution compositions produced by diluting the liquid nanoemulsion concentrates have a particle size between 10 nm or about 10 nm and 1000 nm or about 1000 nm, for example, between 15 nm or about 15 nm and 500 nm or about 500 nm, for example, between 15 nm or about 15 nm and 300 nm or about 300 nm, for example, 15, 16, 17, 18, 19, 20, 21,22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200 nm or greater . Typically, the liquid nanoemulsion concentrates provided are formulated such that dilution of the liquid nanoemulsion concentrates in an aqueous medium produces the liquid dilution composition having an appropriate particle size, for example, between 15 nm or about 15 nm and 500 nm or about 500 nm. Information on particles in liquid dilution compositions may alternatively be expressed in terms of particle density, eg, ppm (parts per million) or percent solids, in liquids.

As used herein, visible particles are particles, eg, in a liquid, eg, an emulsion, that are visible when the liquid is viewed with the naked eye (eg, without magnification). In one example, visible particles are particles that are observed by the skilled person formulating the compositions, eg, concentrates or aqueous liquid dilution compositions containing dilute concentrates. In one example, the compositions provided do not contain visible particles. In another example, the compositions contain few visible particles, eg, no more visible particles than another liquid, eg, a beverage. The presence of visible particles and the number of visible particles is determined by empirical observation.

As used herein, visible crystals are crystals that are visible when a liquid is viewed with the naked eye (eg, without magnification). The presence or absence of visible crystals is typically determined by empirical observation and can be observed by the skilled person formulating the compositions, eg, concentrates or aqueous liquid dilution compositions containing dilute concentrates. In one example, the compositions provided do not contain visible crystals. In another example, the compositions contain few visible crystals, eg, no more visible crystals than are contained in another liquid, eg, a beverage.

As used herein, haze is a measure of the haze or haze of a liquid, caused by particles suspended in the liquid. Turbidity can be measured optically, for example, using a nephelometer, an instrument with a light, and a detector. The nephelometer measures turbidity by detecting scattered light resulting from exposure of the liquid to incident light. The amount of scattered light correlates with the amount of particulate matter in the liquid. For example, a ray of light will pass through a sample with low turbidity with little alteration. Other methods for measuring turbidity are well known and can be used with

ES 2 396 946 T3 methods and compositions provided. The units of a turbidity value measured with a nephelometer are Nephelometric Turbidity Units (NTU). In one example, the compositions provided, eg, the aqueous liquid dilution compositions containing the dilute nanoemulsion liquid concentrates, have a low haze, eg, a haze value (NTU) of 30 or about 30; or an NTU value less than 30 or about 30, for example, less than 29 or about 29, less than 28 or about 28, less than 27 or about 27, less than 26 or about 26, less than 25 or about 25, less than 24 or about 24, less than 23 or about 23, less than 22 or about 22, less than 21 or about 21, less than 20 or about 20, less than 19 or about 19, less than 18 or about 18, less than 17 or about 17, less than 16o about 16, less than 15 or about 15, less than 14 or about 14, less than 13o about 13, less than 12 or about 12, less than 11 or about 11, less than 10o about 10, less than 9 or about 9, less than 8 or about 8, less than 7 or about 7, less than 6 or about 6, less than 5 or about 5, less than 4 or about 4, less than 3 or about 3 , less than 2 or about 2, less than 1 or about 1; or 29 or about 29, 28 or about 28, 27 or about 27, 26 or about 26, 25 or about 25, 24 or about 24, 23 or about 23, 22 or about 22, 21 or about 21, 20 or about 20, 19 or about 19, 18 or about 18, 17 or about 17, 16 or about 16, 15 or about 15, 14 or about 14, 13 or about 13, 12 or about 12, 11 or about 11, 10 or about 10, 9 or about 9, 8 or about 8, 7 or about 7, 6 or about 6, 5 or about 5, 4 or about 4, 3 or about 3, 2 or about 2, 1 or about 1, or 0 or about 0. In another example, the haze value of the aqueous liquid dilution composition is less than 1000 or less than about 1000, less than 500 or less than about 500, less than 300 or less than about 300, less than 250 or less than about 250, 200 or less than about 200, for example 200, 175, 150, 100, 50, 25 or less.

As used herein, a cloudy liquid is one that is thick or opaque with visible suspended particles, for example, a liquid that is fuzzy or muddy in appearance.

As used herein, "transparent" can be used to describe a composition as provided herein, for example, aqueous dilution liquid compositions containing the dilute nanoemulsion concentrates and / or the nanoemulsion concentrates themselves. In one example, a clear liquid is one that does not appear cloudy by empirical observation (eg, to the naked eye) and / or does not contain particles or crystals that are visible to the naked eye, or does not exhibit ring formation. In another example, a clear liquid is one that has a low or relatively low haze value, eg, an NTU value that is less than or equal to a desired NTU value. In one example, a clear liquid has an NTU value of less than 300 or less than about 300, typically less than 250 or less than about 250, typically less than 200 or less than about 200, for example, 200, 175, 150, 100, 50, 25 or less. In another example, a liquid is clear if it has a turbidity value (NTU) of 30 or about 30; or an NTU value of less than 30 or about 30, for example, less than 29 or about 29, less than 28 or about 28, less than 27 or about 27, less than 26 or about 26, less than 25 or about 25, less than 24 or about 24, less than 23 or about 23, less than 22 or about 22, less than 21 or about 21, less than 20 or about 20, less than 19 or about 19, less than 18 or about 18, less than 17 or about 17, less than 16 or about 16, less than 15 or about 15, less than 14 or about 14, less than 13 or about 13, less than 12 or about 12, less than 11 or about 11, less than 10 or about 10, less than 9 or about 9, less than 8 or about 8, less than 7 or about 7, less than 6 or about 6, less than 5 or about 5, less than 4 or about 4, less than about 3 3, less than 2 or about 2, less than 1 or about 1; or 29 or about 29, 28 or about 28, 27 or about 27, 26 or about 26, 25 or about 25, 24 or about 24, 23 or about 23, 22 or about 22, 21 or about 21, 20 or about 20 , 19 or about 19, 18 or about 18, 17 or about 17, 16 or about 16, 15 or about 15, 14 or about 14, 13 or about 13, 12 or about 12, 11 or about 11,10 or about 10, 9 or about 9, 8 or about 8, 7 or about 7, 6 or about 6, 5 or about 5, 4 or about 4, 3 or about 3, 2 or about 2, 1 or about 1, or 0 or about 0. In another example, a clear liquid is one that has a small or relatively small average particle size (eg, less than 1000 nm or about 1000 nm, typically less than 500 nm or less than about 500 nm, typically less than 300 nm. or about 300 nm, typically less than 250 nm or about 250 nm, typically less than 200 nm or about 200 nm, for example, less than 150 or about 150 nm, less than 100 nm or about 100 nm, less than 75 nm or about 75 nm, less than 50 nm or about 50 nm, less than 25 nm or about 25 nm or less than 10 nm or about 10 nm), for example, less than or less than about 5, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 60, 70, 80, 90, 100, 110, 120,130, 140, 150, 160, 170, 180, 190 or 200 nm.

ES 2 396 946 T3

In another example, transparency is expressed relatively. For example, it may be desired that a particular composition is just as clear, about as clear, or more clear than another liquid (measured empirically, or by measuring haze value or particle size). For example, transparency can be evaluated relative to another aqueous dilution liquid composition, eg, a beverage. For example, in one example, a liquid is clear if it looks similar to another clear liquid, for example a beverage, for example water. For example, it may be desired that a composition have a particle size that is less than or equal to another liquid, eg, a beverage. In another example, it may be desired that a composition have a haze value that is less than or equal to another liquid, eg, a beverage. In another example, it may be desired for a composition to appear more transparent or as transparent as another liquid, eg, a beverage, for example, by having no more visible particles, no more crystal formation, and / or no more opacity than another liquid. In one example, the compositions provided are transparent. In another example, they are relatively clear or as clear as or about as clear as another liquid, for example, a beverage that does not contain the nonpolar compound or the liquid nanoemulsion composition.

As used herein, hydrophilic and polar refer synonymously to ingredients and / or compounds that have greater solubility in aqueous liquids, eg, water, than in fats, oils, and / or organic solvents (eg, methanol, ethanol, ethyl ether, acetone and benzene).

Exemplary of the polar ingredients in the provided compositions are polar solvents which are solvents more readily miscible with water and other polar ingredients. In this way, polar ingredients dissolve more easily in polar solvents than in non-polar solvents. Polar solvents are well known. The polarity of a solvent can be evaluated by measuring a number of different parameters according to well known methods as described herein (see, for example, Prizbytek, High Purity Solvent Guide, Burdick and Jackson Laboratories, Inc., 1980). Polar solvents generally have high dielectric constants, typically dielectric constants greater than or about 15, such as from or about 15 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, or greater than 90 and generally have high polarity indices, typically greater of or about 3, such as of or about 3, 4, 5, 6, 7, 8 or 9 or greater than 9. Polar solvents generally have large dipole moments, typically greater than or about 1.4. Debye, such as or about 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4 , 2.5, 2.6, 3.0, 3.5, 4 or greater than 4 Debye. Polar solvents include polar protic solvents and polar aprotic solvents.

As used herein, a polar protic solvent is a polar solvent that contains a hydrogen atom attached to an electronegative atom, such that the hydrogen has a proton-like character and / or the bond between the hydrogen and the atom. electronegative is polarized. Exemplary polar protic solvents include, but are not limited to, water, alcohols including monohydric, dihydric, and trihydric alcohols including, but not limited to, methanol, ethanol, glycerin, and propylene glycol.

Dihydric alcohols are alcohols that contain two hydroxyl groups. Exemplary dihydric alcohols include, but are not limited to, glycols for example, propylene glycol, ethylene glycol, tetraethylene glycol, triethylene glycol, trimethylene glycol.

Trihydric alcohols are alcohols that contain three hydroxyl groups. Exemplary trihydric alcohols include, but are not limited to, glycerin, butane-1,2,3-triol, pentane-1,3,5-triol, 2-amino-2-hydroxymethylpropane-1,3diol.

Monohydric alcohols are alcohols that contain a single hydroxyl group including but not limited to methanol, ethanol, propanol, isopropanol, n-butanol, and t-butanol. In one example, the polar solvent in the compositions provided is not a monohydric alcohol.

As used herein, nonpolar, lipophilic and lipid soluble refer synonymously to compounds (eg, nonpolar compounds) and / or ingredients, eg nonpolar active ingredients that have increased solubility in organic solvents ( for example, ethanol, methanol, ethyl ether, acetone and benzene) and in fats and oils than in aqueous liquids, for example water. Nonpolar compounds include drugs, hormones, vitamins, nutrients, and other lipophilic compounds. Typically, the non-polar compounds used in the provided compositions are poorly soluble in water, for example, water insoluble compounds or compounds that have low water solubility. Exemplary non-polar compounds include non-polar active ingredients, eg, lipid soluble drugs, hormones, essential fatty acids, eg, polyunsaturated fatty acids (PUFAs) eg, omega-3 and omega-6 fatty acids, vitamins, nutrients , mineral nutraceuticals and other compounds. Additional exemplary nonpolar compounds are described herein. The compositions provided can be formulated with any non-polar compound, for example, a non-polar active ingredient.

As used herein, "non-polar active ingredient" refers to a non-polar compound that, when administered to a subject, eg, a human, induces or is proposed to induce a desired response, such as altering function. body at the cellular, tissue, organ or other level, and / or altering the cosmetic or other aspect

ES 2 396 946 T3 property, or a nonpolar compound that is ingested to achieve a desired effect. The non-polar active ingredients can be synthetic or natural non-polar ingredients or compounds, including a pharmaceutical, drug, therapeutic, nutritional supplement, herbal preparation, hormone, or other ingredient. Nonpolar active ingredients can include the nonpolar active ingredients shown herein, as well as other pharmaceutically acceptable or food grade active derivatives of the active ingredients, for example salts, esters, amides, prodrugs, active metabolites, isomers, fragments and the like. Active ingredients can include compounds that have been shown to have a desired effect and also compounds that are thought to produce such effects, for example, compounds typically ingested for nutritional supplementation purposes.

As used herein, a subject includes an animal, typically a mammal, typically a human.

As used herein, an additive includes any that can be added to a food, drink or other product that can be consumed by humans, to enhance one or more of its nutritional, pharmaceutical, dietary, health, and nutraceutical properties beneficial to health. , which provide energy, treatment, holistic or other properties. For example, provided herein are compositions and methods for preparing human-consumable aqueous foods, beverages, and other products that include one or more additives, typically oil-based additives (eg, nonpolar compounds), such as nutraceuticals. , pharmaceuticals, vitamins typically oil-soluble vitamins, for example, Vitamin D, Vitamin E and Vitamin A, minerals, fatty acids such as fatty acids such as essential fatty acids for example polyunsaturated fatty acids for example omega-3 fatty acids and omega-6 fatty acids for example alpha-linolenic acid (ALA), docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA ), gamma-linolenic acid GLA, CLA, saw palmetto extract, flaxseed oil, fish oil, seaweed oil, phytosterols and Coenzymes, eg Coenzyme Q10 and other additives.

As used herein, an effective amount of an additive, such as a nonpolar compound, such as a nonpolar active ingredient, refers to the amount and / or concentration of the additive necessary to prevent, cure, ameliorate, stop or partially stop a symptom of a disease or disorder, or the amount and / or concentration desired by an individual who wants to ingest, such as a daily intake and / or nutritional supplement, for example, an amount sufficient to enhance the nutritional, health, or energy, pharmaceutical, nutraceutical property of a food, beverage, or other consumable product. In some examples, it is desired that the compositions provided, for example liquid nanoemulsion concentrates and / or liquid dilution compositions, contain an effective amount of a particular nonpolar compound, for example a particular amount by volume or weight of the composition.

In one example, an effective amount is a concentration or amount of a liquid nanoemulsion composition where at least 25 mg or about 25 mg, typically at least 35 mg, for example, 35, 40, 45, 50, 55, 60, 65 , 70, 75, 80, 85, 90, 95, 100,110,120,130,140,150,160,170, 180,190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 325, 350, 375, 400, 425, 450 , 475, 500, 550, 600, 700, 800, 900, 1000, 1500, 2000 mg, or greater of the non-polar active ingredient is contained in at least 8 fluid ounces (0.236588 liters) of an aqueous medium, eg, a beverage.

As used herein, a unit dose form refers to physically discrete units suitable for human and animal subjects and individually packaged as is known in the art.

As used herein, "water soluble" refers to a property of a compound, none of which dissolves when the compound is mixed with water, for example, when it is mixed with water at room temperature, for example, between 25 and 50 ° C or between 25 and 50 ° C. In one example, nonpolar compounds are insoluble in water. In another example, the nonpolar compounds in the provided compositions are slightly soluble in water, for example, having low water solubility.

As used herein, low water solubility refers to water solubility less than 30 or about 30 mg / ml, typically less than 20 mg / ml or about 20 mg / ml, typically less than 10 mg / ml or about 10 mg / ml, typically less than 1 mg / ml or about 1 mg / ml, for example, water solubility of 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19 , 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1 mg / ml or less, for example, when mixed with water at room temperature, for example, between 25 and 50 ° C or between about 25 and 50 ° C. As used herein, poorly soluble in water can be used to refer to compounds, for example, nonpolar compounds that are insoluble in water or have low solubility in water.

As used herein, concentrate, liquid concentrate and liquid nanoemulsion concentrate are used synonymously to refer to the compositions provided that contain the non-polar compounds, they are liquid at room temperature, for example 25 ° C or about 25 ° C. ° C, or at a temperature between 25 ° C or about 25 ° C and 50 ° C or about 50 ° C, and can be diluted in aqueous media to form the aqueous liquid dilution compositions. Typically, the liquid nanoemulsion concentrate is an emulsion concentrate that has a particle size (droplet) (or that can be diluted to form a composition

ES 2 396 946 T3 aqueous dilution liquid having a particle size) that is less than 1000 or about 1000, typically less than 500 or about 500, typically less than 300 or about 300 nm, typically less than 250 or about 250 nm , for example, less than 200 or about 200, for example, less than 150 or about 150 nm, for example, a particle size equal to or less than or less than about 5, 10, 11, 12, 13, 14, 15 , 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190 or 200 nm.

As used herein, "liquid composition" is used to refer to any liquid, for example, a composition that is a liquid at room temperature, for example, at 25 ° C or about 25 ° C, or at a temperature between 25 ° C. C or about 25 ° C and 50 ° C or about 50 ° C. Exemplary provided liquid dilution compositions are aqueous liquid dilution compositions in which one or more liquid nanoemulsion concentrates have been diluted, eg, aqueous liquid dilution compositions containing the dilute concentrates. In this example, the nonpolar compound and other lipophilic compounds in the concentrate form a dispersion phase within the aqueous liquid that is an emulsion (eg, nanoemulsion).

As used herein, liquid dilution composition dilution composition and liquid dilution composition are used synonymously to refer to a composition containing one or more of the provided nanoemulsion liquid concentrates (e.g., the liquid nanoemulsion concentrates that contain the nonpolar compound (s)), diluted in a liquid, for example an aqueous medium. Exemplary of the liquid dilution compositions provided are aqueous liquid dilution compositions, eg, beverages or other liquids containing the liquid nanoemulsion concentrates, eg, water, sauces, soups, syrups, soda, juice, eg, juice. fruit, milk, coffee, tea, nutritional drinks, sports drinks, energy drinks, vitamin fortified drinks, flavored water and other drinks that contain the diluted concentrates.

As used herein, aqueous liquid dilution compositions are liquid dilution compositions that are primarily aqueous, eg, a composition comprising a liquid nanoemulsion concentrate diluted in an aqueous medium, eg, water or other beverage. The aqueous liquid dilution composition need not be fully aqueous. For example, aqueous liquid dilution compositions may contain an aqueous portion, eg, a continuous phase, as well as an additional portion, eg, a dispersion phase, eg, a lipophilic dispersion phase. Typically, the lipophilic dispersion phase contains one or more lipophilic substances, eg, one or more non-polar compounds, eg, non-polar active ingredients. Exemplary of the aqueous liquid dilution compositions provided are beverages containing the active ingredients, for example water, soda, juice, for example fruit juice, milk, coffee, tea, nutritional drinks, sports drinks, energy drinks, fortified drinks with vitamins, flavored water and other drinks. Typically, the aqueous liquid dilution compositions are beverages including the nonpolar compound, eg, beverages containing the dilute concentrates.

As used herein "oil phase" is used to refer to the portion (or phase) of a composition such as those provided herein that contain one or more lipophilic ingredients and / or amphiphilic ingredients (ingredients in the oil phase) and that in general it is the soluble phase in the lipid. In the provided emulsion compositions (eg, nanoemulsion concentrates and dilution compositions), the oil phase typically represents the dispersion phase. The oil phase can also be used to refer to the liquid containing the oil phase ingredients that are generated, typically in an oil phase container while performing the methods for preparing the liquid nanoemulsion concentrates. For example, the oil phase can refer to the mixture of the components (oil phase ingredients) that are combined, mixed and heated, for example, in the oil phase container (for example, a tank), before mixing it with the aqueous phase. The oil phase can refer to the oil phase mixture that is formed after all the ingredients dissolve; alternatively, it may refer to the mixture being formed, eg while mixing / heating.

As used herein, the oil phase ingredient (s) refers to the components of the provided compositions that are included in the oil phase in the provided methods for preparing the compositions. Typical oil phase ingredients include nonpolar compounds, eg, nonpolar active ingredients; surfactants; co-surfactants, oils such as non-polar solvents, preservatives, and emulsion stabilizers. Other lipophilic and / or amphiphilic ingredients may be included in the oil phase.

As used herein "aqueous phase" is used to refer to the portion (phase) of a composition such as those provided herein that contains one or more hydrophilic ingredients and / or amphiphilic ingredients (aqueous phase ingredients) and, in Generally, it is the water soluble phase. Typically, in the emulsion compositions provided, eg, nanoemulsion concentrates and dilution compositions, the aqueous phase is the continuous phase. The aqueous phase is also used to refer to the liquid containing the aqueous phase ingredients that are generated while performing the methods to form the liquid nanoemulsion concentrates. For example, the aqueous phase may refer to the mixture of the

ES 2 396 946 T3 components (ingredients of the aqueous phase) that are combined, mixed and heated, for example, in the aqueous phase tank before mixing with the oil phase. The aqueous phase can refer to the aqueous phase mixture that is formed after all the ingredients are dissolved; alternatively, the aqueous phase may refer to the mixture being formed, eg, as it is being mixed / heated.

As used herein, the aqueous phase ingredient (s) refer to the components of the compositions provided that are included in the aqueous phase (e.g., added to the aqueous phase container) in the methods provided for prepare the compositions. Typical aqueous phase ingredients include, but are not limited to, polar solvents, typically protic polar solvents, such as water and alcohols, typically alcohols having more than one hydroxy group such as dihydroxy and trihydroxy alcohols, eg, glycerol and propylene glycol; surfactants; co-surfactants; preservatives and emulsion stabilizers. Other hydrophilic and / or amphiphilic ingredients can be included in the aqueous phase.

As used herein, an initial concentrate is a concentrate (eg, a nanoemulsion liquid concentrate) that is prepared in the provided methods of formulating the provided concentrates, typically by selecting the ingredients eg, a surfactant or surfactants, a nonpolar compound (s), a polar solvent and optionally other ingredients, and selecting starting concentrations of the ingredients from an appropriate concentration range as described herein.

As used herein, stability refers to a desirable property of the compositions provided, for example the ability of the compositions provided to remain free from one or more changes over a period of time for example, at least or for 1 , 2, 3, 4, 5, 6 or more days, at least or for 1, 2, 3, 4, or more weeks, at least or for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 or more months, or at least for 1, 2, 3, 4 or more years. In one example, the composition is stable if it is formulated so that it remains free from oxidation or substantial oxidation over time. In another example, stable compositions remain transparent over time. In another example, stable compositions remain safe and / or desirable for human consumption over time. In one example, stability refers to the absence of precipitates that form in the compositions over the period of time. In a related example, stability refers to the absence of ring formation over the period of time. In another example, the composition is stable if it does not exhibit any visible phase separation for a period of time, eg, after 24 hours, after a week, or after a month. In one example, the compositions are stable and exhibit one or more of these described characteristics, over time, when held at a particular temperature. In one example, the compositions remain stable at room temperature, eg, 25 ° C or about 25 ° C. In another example, the compositions remain stable between 19 ° C and 25 ° C. In another example the compositions remain stable at refrigerated temperatures eg 4 ° C or about 4 ° C or freezing temperature eg at -20 ° C or about -20 ° C.

As used herein, "stabilizing" means increasing the stability of one of the provided compositions.

As used herein, room temperature and room temperature are used to describe a temperature that is common in one or more enclosed spaces in which humans typically are or reside. Ambient temperature can vary, although it generally refers to temperatures between 19 ° C or about 19 ° C and 25 ° C or about 25 ° C. When a composition is stored at room temperature, it should be understood that it is generally maintained at a temperature within or approximately within this range.

As used herein, "chilled temperature" refers to a temperature that is common in a refrigerator, for example, a home or restaurant refrigerator, for example, a temperature that is colder than room temperature, but is typically about degrees above the freezing point of water (0 ° F (-17.77 ° C) or about 0 ° F (-17.77 ° C), or -19 ° C or -20 ° C). Typically, refrigerated temperatures are between about 10 ° C or about 10 ° C and 0 ° C or about 0 ° C, for example, 4 ° C or about 4 ° C. When a composition is stored at a refrigerated temperature, it should be understood that it is maintained at a temperature common to domestic or industrial refrigerators.

As used herein, frozen temperature refers to a temperature of about or below the freezing point of water, for example, a temperature commonly used in a home freezer, for example, 0 ° F (-17.77 ° C) or about 0 ° F (-17.77 ° C), for example, -19 ° C or about -19 ° C or -20 ° C or about -20 ° C or colder.

As used in this document, the singular forms un, an el, and la include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to a compound comprising an extracellular domain includes compounds with one or a plurality of extracellular domains.

As used herein, ranges and amounts can be expressed as approximately, a particular value or range. Approximately also includes the exact amount. In this way about 5

ES 2 396 946 T 3 grams means about 5 grams and also 5 grams. The ranges expressed herein are also understood to include integers within the ranges and fractions thereof. For example, a range between 5 grams and 20 grams includes whole number values ​​such as 5, 6, 7, 8, 9, 10,

11, 12, 13, 14, 15, 16, 17, 18, 19 and 20 grams, and fractions within the range, for example, but not limited to,

5.25, 6.72, 8.5 and 11.95 grams.

As used herein, optionally or optionally means that the event or circumstance described below does or does not occur and that the description includes instances in which such event or circumstance occurs and instances in which it does not. For example, an optionally variant portion means that the portion is variant or non-variant. In another example, an optional ligation step means that the process includes a ligation step or does not include a ligation step.

As used herein, "ring formation" refers to the formation of an off-white or opaque ring around a container containing a liquid, for example, an aqueous liquid, for example, a beverage, for example, a liquid composition of dilution containing an emulsion or nanoemulsion. Typically, the ring is formed around the perimeter of the container, typically at the surface level of the liquid in the container, eg, at the mouth of the container. Ring formation can occur over time and, if it occurs in a short period of time, it can be a sign of instability. Ring formation is typically undesirable, particularly in the case of a liquid for human consumption, for example a beverage. Typically the compositions provided are either non-ring or stable, without ring formation for a long period of time, eg, days, weeks, months or years. In one example, the compositions are free of ring formation over time, when kept, for example, at room temperature, refrigerated and / or frozen.

As used herein, "fatty acid" refers to straight chain hydrocarbon molecules with a carboxyl group (COOH) at one end of the chain.

As used herein, polyunsaturated fatty acid and PUFA are used synonymously to refer to fatty acids that contain more than one carbon-carbon double bond in the carbon chain of the fatty acid. PUFAs, particularly essential fatty acids, are useful as supplements to the diet.

As used herein, essential fatty acids are PUFAs that mammals, including humans, cannot synthesize using any known chemical route. In this way, essential fatty acids must be obtained from the diet or supplementation. Exemplary essential PUFA fatty acids are omega-3 fatty acids (w3; n-3) and omega-6 fatty acids (w-6; n-6).

As used herein, omega-3 (w3; n-3) fatty acids are methylene interrupted polyethylenes, having two or more cis double bonds separated by a single methylene group, and in which the first double bond appears on the third carbon from the last carbon (w). Omega-3 fatty acids are used as supplements to the diet, for example, for the treatment and prevention of diseases. In one example, the compositions provided contain non-polar active ingredients that contain at least one omega-3 fatty acid. Exemplary omega-3 fatty acids are Alpha-Linolenic Acid (α-Linolenic Acid; ALA) (18: 3w3) (a short-chain fatty acid); Stearidonic acid (18: 4w3) (a short chain fatty acid); Eicosapentaenoic acid (EPA) (20: 5w3); Docosahexaenoic acid (DHA) (22: 6w3); Eicosatetraenoic acid (24: 4w3); Docosapentaenoic acid (DPA, Clupanodic acid) (22: 5w3); 16: 3 w3; 24: 5w3 and nisinic acid (24: 6w3). Longer chain Omega-3 fatty acids can be synthesized from ALA (the short chain omega-3 fatty acid). Exemplary non-polar active ingredients containing omega-3 fatty acids are non-polar active ingredients containing DHA and / or EPA, for example, containing fish oil, krill oil and / or seaweed oil, for example oil of microalgae, non-polar active ingredients containing alpha-linolenic acid (ALA), for example containing flaxseed oil.

As used herein, omega-6 (w-6; n-6) fatty acids are methylene interrupted polyenes, having two or more cis double bonds, separated by a single methylene group, and in which the first double bond appears on the sixth carbon from the last carbon (w). In one example, the compositions provided contain non-polar active ingredients that contain at least one omega-3 fatty acid. Exemplary Omega-6 fatty acids are Linoleic acid (18: 2w6) (a short chain fatty acid); Gamma-linolenic acid (GLA) (18: 3w6); Dihomo gamma linolenic acid (DGLA) (20: 3w6); Eicosadienoic acid (20: 2w6); Arachidonic acid (AA) (20: 4w6); Docosadienoic acid (22: 2w6); Adenic acid (22: 4w6); and Docosapentaenoic acid (22: 5w6). Exemplary non-polar active ingredients containing omega-6 fatty acids are ingredients that contain GLA, eg, borage oil. Also exemplary non-polar active ingredients containing PUFA are compounds containing conjugated fatty acids, for example, Conjugated linoleic acid (CLA) and compounds containing saw palmetto extract.

As used herein, "algal oil" refers to any oil derived from marine dinoflagellates eg microalgae, eg Crypthecodinium sp, particularly Crypthecodinium cohnii. In one example, algae oil is used as a nonpolar compound, for example, as an active ingredient in

ES 2 396 946 T3 compositions provided. Seaweed oil typically contains DHA. In one example, algae oil is also a source of EPA.

As used herein, "fish oil" refers to any oil derived from any fish, typically a cold water fish, for example, from fish tissue, for example, from frozen fish tissue, for example, cod liver. . In one example, fish oil is used as a nonpolar compound, eg, an active ingredient in the compositions provided. Fish oil typically contains DHA. In one example, fish oil also contains EPA.

As used herein, preservative and preservative are used synonymously to refer to ingredients that can improve the stability of the compositions provided. Preservatives, particularly food and beverage preservatives, are well known. Any known preservative can be used in the compositions provided. Exemplary of preservatives that can be used in the provided compositions are all oil-soluble preservatives, for example Benzyl Alcohol, Benzyl Benzoate, Methyl Paraben, Propyl Paraben, antioxidants, for example Vitamin E, Vitamin A Palmitate and Beta Carotene . Typically, a preservative is selected that is safe for human consumption, for example in food and beverages, for example a GRAS certified and / or Kosher certified preservative, for example benzyl alcohol.

As used herein, a solvent is an ingredient that can be used to dissolve another ingredient. For example, solvents include polar and nonpolar solvents. Nonpolar solvents include oils and other nonpolar ingredients that dissolve nonpolar compounds. In one example, the nonpolar active ingredient is dissolved in a nonpolar solvent in practicing the methods of producing the provided compositions. In this example, the compositions provided contain polar solvents in amounts sufficient to dissolve the non-polar active ingredient. More than one nonpolar solvent can be used. Typically, the nonpolar solvent is an oil that is included in the composition in addition to the nonpolar compound. For example, the non-polar solvent is typically not the non-polar compound itself, eg, it is other than the non-polar solvent. Certain compounds, for example linseed oil or safflower oil can be non-polar solvents and non-polar active ingredients. Typically, the nonpolar solvent contains one or more oils, typically oils other than the nonpolar active ingredient or an oil or oils that are not contained in the active ingredient. Exemplary non-polar solvents include, but are not limited to, oils (in addition to the non-polar active ingredient), for example, Vitamin E oil, linseed oil, CLA, Borage Oil, D-limonene, Canola oil, corn oil. , MCT oil and oat oil. Other oils can also be used. Exemplary of the Vitamin E oil are the oil marketed by ADM Natural Health and Nutrition, Decatur, IL, under the name Novatol ™ 5-67 Vitamin E (D-alpha-Tocopherol; product code ADM 410217). This Vitamin E oil contains at least 67.2% Tocopherol and approximately 32.8% soybean oil. In one example, the nonpolar solvent is synonymously referred to as the nonpolar solubilizer.

As used herein, w / w, weight for weight, wt% wt, and wt% are used synonymously to express the ratio of the mass of one component of a composition compared to the mass of the entire composition. For example, when the amount of a particular ingredient represents 1% by weight (w / w) of a concentrate, the mass of that ingredient is 1% of the mass of the entire concentrate. Similarly, when the amount of an ingredient is 50% (w / w) of the concentrate, the mass of that ingredient is 50% of the entire mass of the concentrate. Similarly, when a composition and / or a compound contains 10% by weight of an ingredient, the mass of the ingredient is 10% of the total mass of the composition or compound. When only a concentration, quantity or percentage is indicated (without units), the concentration or percentage should be understood to be a concentration or percentage by weight.

Similarly, as used herein v / v, volume by volume, percent by volume, and percent by volume are used synonymously to express the ratio of the volume of one component of a composition and the volume of the entire composition.

As used herein, "emulsion stabilizer" refers to compounds that can be used to stabilize and / or emulsify and / or change the viscosity of the compositions provided, for example, liquid nanoemulsion concentrate and / or aqueous compositions. containing diluted concentrates. In one example, the emulsion stabilizer increases the viscosity of the liquid concentrate. In one example, one or more emulsion stabilizers are added during formulation, after evaluation of an initial concentrate, particularly if the oil and aqueous phases of the initial concentrate (or the aqueous liquid dilution composition resulting from dilution of the initial concentrate) it seems that they are separating. The addition of the emulsion stabilizer can prevent the separation of the oil and water phases.

Exemplary of emulsion stabilizers that can be used in the provided compositions is a composition containing a combination of gums, for example gums used as emulsifying agents, for example a combination containing one or more of xanthan gum, guar gum and sodium alginate, eg, the emulsion stabilizer marketed under the trade name SALADIZER®, available from TIC Gums, Inc. (Belcamp, MD). Other gums can be included in the emulsion stabilizer, for example gum

ES 2 396 946 T3 arabic, sugar beet pectin. Other similar gum combinations can be used as emulsion stabilizers.

As used herein, a pH adjuster is any compound, typically an acid or a base that is capable of changing the pH of the compositions provided, for example, to lower the pH of the composition or increase the pH of the composition. , typically without altering other properties of the composition, or without substantially altering other properties. PH adjusters are well known. Exemplary of acidic pH adjusters are, for example, citric acid and phosphoric acid and bases.

As used herein, a flavor is any ingredient that changes, typically enhances, the taste and / or odor of the provided composition, eg, aqueous liquid dilution compositions, eg, beverages.

As used herein, not greater than and NMT refer to an amount that is less than or equal to the listed amount. Similarly not less than and NLT refer to an amount that is greater than or equal to the listed amount.

As used herein, "natural" is used to refer to a composition and / or the ingredients in the composition that can be found in nature and that are not exclusively man-made. For example, benzyl alcohol is a natural preservative. Similarly, tocopheryl polyethylene glycol is a natural surfactant. In one example, the natural composition / ingredient is GRAS and / or Kosher certified. Typically, the compositions provided are natural, semi-natural, and / or contain one or more natural ingredients.

As used herein, GRAS and GRAS are used synonymously to refer to compounds, compositions, and ingredients that are Generally Considered Safe by the USDA, FDA for use as additives, for example, in foods, beverages, and / or other substances for human consumption, for example, any substance that meets the criteria of sections 201 (s) and 409 of the United States Federal Food, Drug, and Cosmetic Act. Typically, the compositions provided in this document are GRAS certified.

As used in this document, kosher is used to refer to substances that conform to the laws of the Jewish Kosher diet, for example, substances that do not contain ingredients derived from animals that are not kosher or ingredients that were not prepared following the procedures kosher. Typically, the compositions provided in this document are certified Kosher.

As used herein, "container" refers to any container, eg, tank, jar, vial, flask, cylinder, and beaker, that can be used to contain the ingredients and / or phases of the compositions provided, during the methods. to repair the compositions. In one example (for example, the large-scale methods provided), the container is a tank that is used to mix and / or heat one or more ingredients and / or phases of the compositions, for example, aqueous phase tanks and tanks. oil phase. Typically, the oil and water phases are mixed and heated in separate tanks, before combining the phases to form an emulsion. In another example, the tank is a container or containment tank, which contains the compositions provided after forming the compositions, for example, the emulsions. A number of tanks are available for mixing ingredients. Tanks are typically cleaned, eg, rinsed, soaped and / or sanitized according to known procedures, before use and between uses. Typically, the tanks are equipped with one or more mixers, for example a conventional mixer and / or homogenizer, which are used to mix the ingredients added to the tank. In one example, the tank is additionally equipped with a heating and / or cooling device. For example, the tank can be a water jacketed tank. The temperature of the water jacketed tank is controlled through the water jacket, eg to heat the contents eg while mixing.

As used herein, an aqueous phase container refers to the container used to mix and / or heat the aqueous phase ingredients to generate the aqueous phase of the compositions provided. In one example (eg, for large-scale methods), the aqueous phase vessel is an aqueous phase tank. In one example, the aqueous phase tank is a water jacketed tank, which is equipped with a water jacket that can be used to heat the contents of the tank.

As used herein, an oil phase container refers to the container used to mix and / or heat the oil phase ingredients to generate the oil phase of the provided compositions. Typically, the oil phase vessel is an oil phase tank. In one example, the oil phase tank is a water jacketed tank.

As used herein, "transfer medium" refers to any equipment, combination of equipment and / or system that can be used to transfer a liquid, for example, from one tank to another tank, in the methods provided for preparing the compositions. Exemplary of the transfer means are a pump of

ES 2 396 946 T3 transfer and appropriate accessories, for example sanitary fittings, ball valves and transfer tubes, for example food grade tubes.

As used herein, a mixer is any piece of equipment or combination of equipment that can be used to mix ingredients in the methods provided to prepare the compositions, eg, conventional mixers and homogenizers (shears). For example, mixers can be used to mix the ingredients of the water phase, the oil phase, and / or mix the additional ingredients.

As used herein, conventional mixers are mixers that are used to combine a group of ingredients, for example, the oil phase ingredients or the water phase ingredients, or to mix one or more ingredients with a liquid, eg with an emulsion, eg to mix additional ingredients with the emulsion. Conventional mixers can be any mixer that moves the material, for example, the ingredients during heating, for example, to promote dissolution of the ingredients.

As used herein, homogenizer and shear are used to refer to mixers that typically have high shear, which can be used, for example, to form an emulsion, for example, to emulsify the water phase and the oil phase in methods provided. Homogenizers are typically capable of high shear mixing, which emulsifies the phases.

As used herein, a cooling apparatus is any piece of equipment or combination of equipment that can be used with the methods provided to cool the compositions and phases and ingredients thereof, for example, during mixing and / or homogenization, for example, to cool the mixture while emulsifying the oil and water phases. Exemplary of the cooling apparatus are chillers (coolers), for example, recirculating coolers that can be attached, for example, to the tanks used in the methods provided, for example, remotely or by means of a tank mounted on the refrigerator, for recirculation the fluid from the tank, through the cooler and back to the tank, to rapidly cool and maintain mix temperature during mixing. Typically, the cooling apparatus can be used to cool the liquid to between 25 ° C or about 25 ° C and 45 ° C or about 45 ° C, for example, 25, 26, 27, 28, 29, 30, 31, 32 , 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44 or 45 ° C, typically between 25 ° C and 43 ° C, typically between 35 ° C and 43 ° C, for example, 26.5 ° C. The desired temperature, for example between 25 ° C or about 25 ° C and 45 ° C or about 45 ° C, typically between 35 ° C and 43 ° C, for example 26.5 ° C, in less than 2 hours or about 2 hours, typically less than 1 hour or about 1 hour, for example, in at least between 30 minutes or about 30 minutes and 60 minutes or about 60 minutes, for example, 30, 31, 32, 33, 34, 35 , 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59 or 60 minutes.

As used herein, a low heat refers to a temperature between 45 ° C or about 45 ° C and 85 ° C or about 85 ° C, for example, 45, 46, 47, 48, 49, 50, 51 , 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76 , 77, 78, 79, 80, 81, 82, 83, 84 or 85 ° C, for example, no more than 85 ° C or about 85 ° C, typically no more than 60 ° C or about 60 ° C, typically 60 ° C or 60 ° C. In the methods provided for preparing the liquid nanoemulsion concentrates, the ingredients of the oil phase and the water phase are typically heated, using low heat, to preserve the ingredients, for example, to prevent oxidation of the ingredients, for example, non-polar active ingredients, eg omega-3 containing compounds, eg DHA.

As used herein, "consisting essentially of" means that it contains the following list of ingredient (s) and does not include any additional active ingredients, for example, that it does not include any additional active drugs or pharmaceuticals. For example, a composition, eg, a liquid nanoemulsion, consisting essentially of an indicated plurality of ingredients contains those particular ingredients that do not contain any additional active drugs or pharmaceuticals.

As used herein, Ci-Cx includes C1-C2, C1-C3, ... Cx.

As used herein, the term "alkyl" and "alkyl group" refer to straight or branched chain substituted or unsubstituted hydrocarbon groups having any number of carbon atoms; the number of carbon atoms can be specified, for example, 1 to 30 carbon atoms, 8 to 28 carbon atoms, 7 to 27 carbon atoms, 8 to 22 carbon atoms, 8 to 20 atoms carbon of 8 to 18 carbon atoms and 12 to 18 carbon atoms. An alkyl group can be a saturated alkyl which means that it does not contain any alkene or alkylene groups or an unsaturated alkyl which means that it contains at least one alkene or alkylene group and can optionally be substituted. An alkyl group that includes at least one carbon-carbon double bond (C = C) is also referred to by the term alkenyl; alkenyl groups can optionally be substituted. An alkyl group that includes at least one carbon-carbon triple bond (C ° C) is also referred to by the term alkynyl; alkynyl groups can optionally be substituted.

B. COMPOSITIONS CONTAINING NON-POLAR COMPOUNDS

ES 2 396 946 T3

Provided herein are compositions containing nonpolar compounds and methods for preparing the compositions. Nonpolar compounds are poorly soluble in water (for example, they have low water solubility or are insoluble in water). Thus, it can generally be difficult to formulate nonpolar compounds into compositions for human consumption, particularly aqueous compositions, eg, food and beverages. The poor water solubility of nonpolar compounds can also contribute to their poor bioavailability. Improved methods and compositions for formulating nonpolar compounds are provided herein.

In general, emulsions (eg, oil-in-water emulsions) are colloidal dispersions of two immiscible liquids (eg, oil and water or other aqueous liquid), containing a continuous and a dispersed phase. Emulsions can be used to disperse nonpolar compounds in aqueous liquids. In an oil-in-water emulsion, the dispersed phase is the oil phase and the continuous phase is an aqueous phase (water). There remains a need for improved emulsions (eg, oil-in-water emulsions) containing nonpolar compounds in aqueous liquids and methods and compositions for generating the improved emulsions. In particular, there is a need for emulsions that are more suitable and desirable for human consumption than nonpolar compounds, for example in food and beverages. For example, emulsions having improved transparency (eg, small particle size, low turbidity), stability (eg, no separation), taste and odor, are necessary.

Among the compositions provided are such improved emulsions. For example, emulsions are provided that contain the nonpolar compounds dispersed in an aqueous liquid and have desirable properties, including improved transparency, stability, odor, and taste. The compositions provided (and methods for preparing the compositions) can be used to formulate any non-polar compound in aqueous compositions, including the non-polar compounds (eg, non-polar active ingredients) described herein and other known non-polar compounds.

Typically, the emulsions provided containing the non-polar compounds are nanoemulsions, which are emulsions having dispersed droplets (particles) with diameters less than 1000 nm or less than about 1000 nm, typically less than 500 nm or less than about 500 nm, typically less than 300 nm or about 300 nm, typically less than 250 or less than about 250 nm, typically less than 200 nm or less than about 200 nm, for example, less than or less than about 5, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, or 200 nm. Typically, the nanoemulsion compositions provided are oil-in-water nanoemulsions, containing the nonpolar compounds dispersed in an aqueous liquid.

The emulsion compositions provided are stabilized by one or more surfactants and / or cosurfactants and / or emulsion stabilizers. Surfactants form an interfacial film in the emulsion, between the oil and water phase, providing stability. Typically, the nanoemulsions of the provided compositions contain micelles, in which one or more surfactants surround the nonpolar active compound. The micelles are dispersed in the aqueous phase.

The emulsion compositions provided include liquid nanoemulsion concentrates containing the nonpolar compounds that can be diluted to provide nonpolar compounds in aqueous compositions such as beverages. Liquid nanoemulsion concentrates, which can be diluted to provide nonpolar compounds in aqueous compositions such as beverages. Liquid nanoemulsion concentrates can be diluted in a medium, eg, an aqueous medium, eg, a beverage, to form a liquid dilution composition (eg, an aqueous liquid dilution composition) containing the nonpolar compound. Also exemplary of the compositions provided are liquid dilution compositions (eg, aqueous liquid dilution compositions, which may be clear), prepared by diluting the liquid nanoemulsion concentrates in medium.

Compositions can be prepared using any non-polar compound. Nonpolar compounds are typically nonpolar active ingredients, for example pharmaceuticals, nutraceuticals, vitamins and minerals. Non-polar active ingredients include, but are not limited to, compounds that contain Polyunsaturated Fatty Acids (PUFAs), for example, active ingredients that contain omega-3s, for example, compounds that contain ALA, DHA and / or ePa, for example oils derivatives of fish and microalgae, krill and / or flaxseed extract, and non-polar active ingredients containing omega-6, for example, compounds containing gamma-linolenic acid (GLA), for example, borage oil; compounds containing saw palmetto oil; ingredients containing conjugated fatty acid, for example, compounds containing Conjugated Linoleic Acid (CLA); active ingredients containing coenzyme Q, eg, Coenzyme Q10 (CoQ10), typically compounds containing oxidized CoQ10 (ubidecarenone); and compounds containing phytosterols (plant sterols). Additional exemplary non-polar active ingredients are described therein. Any non-polar compound can be used in the compositions provided.

ES 2 396 946 T3

1. Liquid nanoemulsion concentrates containing the non-polar compounds

Liquid nanoemulsion concentrates (also referred to as liquid concentrates or concentrates) containing one or more nonpolar compounds are provided. The concentrates can be diluted in an aqueous medium to form aqueous dilution liquid compositions containing the nonpolar compounds. Liquid concentrates are formulated based on one or more desirable properties, eg, transfer; security; flavor; odor, stability, eg, absence of phase separation, ring formation and / or precipitation over time, and / or bioavailability of the concentrate and / or aqueous liquid dilution compositions containing the concentrate. In one example, the desirable property is the ability of the concentrate provided to produce a clear or partially clear dilution aqueous liquid composition when diluted in an aqueous medium, eg, a beverage such as water. In another example, the desirable property relates to the safety of the concentrates and / or the suitability of the concentrates for human consumption, eg, in food and beverages. In another example, it may be desirable for the concentrate to contain less than or equal to a particular concentration of the one or more ingredients. In another example, it may be desirable for the concentrate to contain more than or equal to a particular concentration of one or more ingredients.

In addition to the nonpolar compounds, the concentrates contain at least one surfactant. Typically, the surfactant has an HLB value of between 14 or about 14 and 20 or about 20, for example 14, 15, 16, 17, 18, 19, 20, about 14, about 15, about 16, about 17, about 18, about 19, or about 20. Exemplary of suitable surfactants are surfactants derived from Vitamin E, surfactants derived from polyethylene glycol (PEG) such as polyethylene glycol tocopherol succinate (TPGS), particularly those having an HLB value between or about 14 and of or about 20, and surfactants with similar properties such as hLb values. Typically, the surfactant is a natural surfactant, for example a surfactant that is certified for GRAS (generally recognized as safe) by the FDA and / or certified for Kosher, for example TPGS.

Liquid concentrates additionally contain a polar solvent, such as water (e.g. filtered water) or other edible aqueous liquid (e.g. propylene glycol or glycerin), or a combination thereof, typically a high amount of the polar solvent, e.g. , between 60% or about 60% and 80% or about 80% by weight (w / w) of the concentrate, typically between or about 60% and at or about 79% by weight of the concentrate.

Typically, concentrates additionally contain one or more additional ingredients. Exemplary of additional ingredients that may be included in concentrates are preservatives, non-polar solvents, cosurfactants, emulsion stabilizers, pH adjusters, and flavoring agents.

The nonpolar compounds in the concentrates and dilution compositions are contained in micelles. These micelles contain the nonpolar compound surrounded by one or more surfactants, allow dispersion of the polar compound between polar solvents, for example, when concentrates are divided to form aqueous liquid dilution compositions. Micelles containing the nonpolar compounds typically have a small or relatively small particle size, for example, less than 1000 nm or about 1000 nm, less than 500 nm or about 500 nm, typically less than 300 nm or about 300 nm, typically less than 200 nm or about 200 nm, for example 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18 , 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 125, 150 or 200 nm. A smaller particle size is correlated with the transparency of the aqueous liquid dilution compositions containing the dilute concentrates. For example, a liquid with a smaller particle size is more transparent than a liquid with a larger particle size. The smaller particle size can also contribute to other desirable properties, eg, stability.

A number of factors, including ingredients, their relative concentrations, and methods for preparing the concentrates, affect the particle size of the compositions and other desirable properties of the compositions, such as transparency. In particular, the nature of the surfactant, particularly the HLB of the surfactant, and the relative concentrations of the polar solvent (eg, water), surfactant, and the nonpolar compound, contribute to the small particle size in the transparency of aqueous dilution liquid compositions. . Typically, several of these parameters and properties are related to each other. For example, several of the parameters contribute to particle size, typically a small particle size of the compositions. Particle size contributes directly to the transparency of aqueous dilution liquid compositions containing concentrates. The particle size may also be related to other properties, for example, stability, absence of ring formation and / or formation of a precipitate of the aqueous liquid dilution compositions containing the concentrates.

Consequently, the properties of the ingredients and their relative concentrations in the concentrates are important to the ability of the concentrate to produce desirable dilution compositions. Methods are provided for formulating the liquid nanoemulsion concentrates. Determination of the appropriate ingredients, and the relative concentrations thereof, that will produce dilution compositions having desirable properties, is accomplished using the methods provided for formulating the liquid concentrates.

ES 2 396 946 T3

to. Formulation of liquid concentrates

In the provided formulation methods, concentrates are formulated by selecting the ingredients and the concentration ratios of the ingredients that produce compositions having one or more desired properties. When the concentrates are formulated, the selected ingredients and starting concentrations are used to prepare the initial concentrates, which are evaluated and modified, if necessary.

In a first step in formulating the provided concentrates, one or more of the initial concentrates are prepared and evaluated for the desired properties. For this step, ingredients are selected, for example, from the ingredients described in this document. The ingredients generally include surfactants, polar solvents, non-polar active ingredients, and other ingredients. A starting concentration (weight percent) of each selected ingredient is selected within the appropriate concentration range for that ingredient or category of ingredient, eg, the appropriate concentration range for the surfactant. In some cases, the initial concentrate is formulated based on the ingredients and concentrations thereof in an existing concentrate that has one or more desired properties.

The starting concentrate or concentrates are then prepared, using the concentrate preparation methods provided below, adding each ingredient to its starting concentration in the appropriate step. In one example, more than one starter concentrate, eg, multiple starter concentrates, each of which has a different concentration of one or more ingredients, are prepared and compared. In one example, multiple initial concentrates are produced to test various representative concentrations within an appropriate concentration range for one or more particular ingredients.

In a typical example, the initial concentrate is prepared by including at least one surfactant, such as among the surfactants described herein, having an HLB value between 14 or about 14 and 20 or about 20, at a starting concentration within the concentration range of between 16% or about 16% and 30% or about 30% by weight (w / w) of the concentrate; at least one non-polar compound, at a starting concentration within the concentration range of between 5% or about 5% and 10% or about 10%; and a polar solvent at a starting concentration of between 60% or about 60% and 80% or about 80% and typically between me and about 60% and or about 79% by weight. In one example the initial concentrate further includes other ingredients, eg, a preservative (s), co-surfactant (s), and / or other ingredients as described herein.

After preparing the initial concentrate (s), the concentrate (s) are evaluated for one or more desired properties, for example, the ability to form dilution compositions (eg, clear dilution compositions or dilution compositions that have a haze value). particle size, or other property). The ability to form dilution compositions having one or more properties is evaluated by diluting the concentrate in an aqueous medium, for example, diluting the concentrate in the aqueous medium at a dilution factor of between 1:10 or about 1:10 and 1 : 1000 or about 1: 1000 or greater, typically between 1:10 or about 1:10 and 1: 500 or about 1: 500 or greater, for example, does not dilute more than 1:10 or about 1:10, 1 : 20 or about 1:20, 1:25 or about 1:25, 1:50 or about 1:50, 1: 100 or about 1: 100, 1: 200 or about 1: 200, 1: 250 or about 1: 250, 1: 300 or about 1: 300, .1: 400 or about 1: 400, 1: 500 or about 1: 500, for example, 1:10, 1:20, 1:25, 1:30, 1:35, 1:40, 1:50, 1:55, 1:60, 1:65, 1:70, 1:75, 1:80, 1:90, 1: 100, 1: 110, 1: 120, 1: 130, 1: 140, 1: 150, 1: 160, 1: 170, 1: 180, 1: 190, 1: 200, 1: 210, 1: 220, 1: 230, 1: 235, 1: 240, 1: 250, 1: 260, 1: 270, 1: 280, 1: 290, 1: 300, 1: 350, 1: 400, 1: 450, 1: 500 or greater, or according to other dilutions provided in this document.

After evaluation, the ingredients and / or concentrations thereof can be adjusted to generate the desired properties in the final concentrate. Typically, the concentration of the non-polar compound, surfactant, and / or polar solvent is the concentration that is adjusted after evaluating the initial concentrate. Similarly, when multiple initial concentrates are formulated, one or more of the nonpolar compound, surfactant, and polar solvent concentration are varied among the multiple initial concentrates. In some cases, after evaluation, it may be determined that additional ingredients (not included in the initial formulation) are necessary or desirable to achieve the desired properties of a particular concentrate. This process can be repeated until a concentrate is generated that has the desired property (s).

i. Common Ingredients and Typical Concentration Ranges

Each of the concentrates provided contains a nonpolar compound such as, but not limited to, the exemplary nonpolar compounds described herein below. Typically, the non-polar compound is a non-polar active ingredient, for example, an oil-based active ingredient such as a polyunsaturated fatty acid (PUFA), a coenzyme Q, or a phytochemical. To formulate the initial concentrate, the starting concentration of the nonpolar compound is typically a concentration chosen from within a concentration range of between 5% or about 5% and 10% or about 10% (w / w) of the concentrate, such as a

ES 2 396 946 T3 starting concentration of 5% or about 5%, 6% or about 6%, 7% or about 7%, 8% or about 8%, 9% or about 9% or 10% or about 10% (w / w) of the concentrate. The nonpolar compound is typically added as part of an oil phase, according to the methods provided to prepare the concentrate.

The initial concentrate additionally contains at least one surfactant that can be added to either the water phase or the oil phase, and typically has an HLB value of between 14 or about 14 and 20 or about 20, eg, 14, 15, 16, 17 , 18, 19 or 20, or about 14, about 15, about 16, about 17, about 18, about 19, about 20, typically between or about 15 and or about 18, including but not limited to TPGS and analogs and derivatives thereof, typically a natural surfactant, which is safe and / or approved for human consumption.

Typically, the starting concentration of the surfactant is chosen from within a concentration range between 16% or about 16% and 30% or about 30% (w / w), for example 16% or

<td>about</td><td>the</td><td> 16%,</td><td> 17%</td><td>or</td><td>about</td><td>the</td><td> 17%,</td><td> 18%</td><td>or</td><td>about</td><td>the</td><td> 18%,</td><td> 19%</td><td>or</td>

<td>about</td><td>the</td><td> 19%,</td><td> 20%</td><td>or</td><td>about</td><td>the</td><td> 20%,</td><td> 21%</td><td>or</td><td>about</td><td>the</td><td> 21%,</td><td> 22%</td><td>or</td>

<td>about</td><td>the</td><td> 22%,</td><td> 23%</td><td>or</td><td>about</td><td>the</td><td> 23%,</td><td> 24%</td><td>or</td><td>about</td><td>the</td><td> 24%,</td><td> 25%</td><td>or</td>

<td>about</td><td>the</td><td> 25%,</td><td> 26%</td><td>or</td><td>about</td><td>the</td><td> 26%,</td><td> 27%</td><td>or</td><td>about</td><td>the</td><td> 27%,</td><td> 28%</td><td>or</td>

about 28%, 29% or about 29% or 30% or about 30% by weight (w / w) of the concentrate, such as, for example, 17.75%, 20.25%, 20.5%, 22.7% or 25.2% (w / w) of the concentrate.

In one example, the concentration range of the surfactant is between 16% or about 16% and 26% or about 26% by weight (w / w) of the concentrate, such as, for example, between 17% or about 17% and 25% or about 25% (w / w) of the concentrate; between 18% or about 18% and 26% or about 26% (w / w) of the concentrate; between 16% or about 16% and 18% or about 18% (w / w) of the concentrate; such as for example 18% or about 18% (w / w) of the concentrate, 20% or about 20% (w / w) of the concentrate, 23% or about 23% (w / w) of the concentrate, or 25% or about 25% (w / w) of the concentrate. In another example, the surfactant concentration range is between 17% or about 17% and 26% or about 26% (w / w) of the concentrate. In another example, the concentration range of the surfactant is between 18% or about 18% and 25% or about 25% (w / w) of the concentrate. In another example, the concentration range of the surfactant is between 18% or about 18% and 20% or about 20% (w / w) of the concentrate. In another example, the concentration range of the surfactant is between 17% or about 17% and 20% or about 20% (w / w) of the concentrate. In another example, the concentration range of the surfactant is between 16% or about 16% and 20% or about 20% (w / w) of the concentrate.

The concentrates additionally contain polar solvents (eg water or other edible polar solvent, eg propylene glycol and glycerin), typically a high concentration of polar solvent, which is added to the aqueous phase. Typically, the starting concentration of the polar solvent is chosen from within the range

<td>concentration</td><td>of</td><td colspan="5">between 60% or approximately</td><td colspan="11">60% and 80% or approximately 80% (w / w) of the</td>

<td>concentrated, by</td><td colspan="2">example,</td><td>the</td><td> 60%</td><td>or</td><td>about</td><td colspan="2">1 60%</td><td colspan="2"> , 61 %</td><td>or</td><td>about</td><td>the</td><td> 61%,</td><td>the</td><td> 62%</td><td>or</td>

<td>about</td><td>the</td><td> 62%,</td><td>the</td><td> 63%</td><td>or</td><td>about</td><td>the</td><td> 63%,</td><td>the</td><td> 64%</td><td>or</td><td>about</td><td>the</td><td> 64%,</td><td>the</td><td> 65%</td><td>or</td>

<td>about</td><td>the</td><td> 65%,</td><td>the</td><td> 66%</td><td>or</td><td>about</td><td>the</td><td> 66%,</td><td>the</td><td> 67%</td><td>or</td><td>about</td><td>the</td><td> 67%,</td><td>the</td><td> 68%</td><td>or</td>

<td>about</td><td>the</td><td> 68%,</td><td>the</td><td> 69%</td><td>or</td><td>about</td><td>the</td><td> 69%,</td><td>the</td><td> 70%</td><td>or</td><td>about</td><td>the</td><td> 70%,</td><td>the</td><td> 71%</td><td>or</td>

<td>about</td><td>the</td><td> 71%,</td><td>the</td><td> 72%</td><td>or</td><td>about</td><td>the</td><td> 72%,</td><td>the</td><td> 73%</td><td>or</td><td>about</td><td>the</td><td> 73%,</td><td>the</td><td> 74%</td><td>or</td>

<td>about</td><td>the</td><td> 74%,</td><td>the</td><td> 75%</td><td>or</td><td>about</td><td>the</td><td> 75%,</td><td>the</td><td> 76%</td><td>or</td><td>about</td><td>the</td><td> 76%,</td><td>the</td><td> 77%</td><td>or</td>

<td>about</td><td>the</td><td> 77%,</td><td>the</td><td> 78%</td><td>or</td><td>about</td><td>the</td><td> 78%,</td><td>the</td><td> 79%</td><td>or</td><td>about</td><td>the</td><td> 79%,</td><td>the</td><td> 80%</td><td>or</td>

<td>about</td><td>the</td><td colspan="3">80% (p / p) of</td><td colspan="3">concentrated, such as</td><td colspan="3">for instance,</td><td colspan="2"> 68,29%, 68,7865%,</td><td colspan="2"> 74,25%,</td><td colspan="2"> 71,74%,</td><td>or</td>

75.8165% (w / w) of the concentrate. In one example, the concentration range of the polar solvent is between 65% or about 65% and 80% or about 80% (w / w) of the concentrate. In another example, the concentration range of the polar solvent is between 65% or about 65% and 75% or about 75% (w / w) of the concentrate or between 65% or about 65% and 76 % or about 76% (w / w) of the concentrate.

One or more, typically more than one additional ingredient can be added to the initial concentrate. For example, concentrates typically contain at least one preservative, typically a natural preservative, eg, benzyl alcohol. Exemplary are other additional ingredients that can be added to concentrates, including emulsion stabilizing starting concentrates, eg, a combination of gums; a non-polar solvent for the non-polar compound, eg, an oil other than the non-polar compound, eg, a vitamin E oil or linseed oil; a pH adjuster, for example citric acid, phosphoric acid; one or more flavoring agents, for example, D-limonene or lemon oil; a co-surfactant, eg, a phospholipid, eg, phosphatidylcholine.

Appropriate concentration ranges for additional ingredients are described in the individual sections below. Typically, the concentration of additional ingredients depends, in part, on the

ES 2 396 946 T3 concentrations of the non-polar active ingredient, the surfactant and the polar solvent. Typically, the concentrations of these three ingredients (surfactant, polar solvent, and nonpolar compound) are focused on the formulation methods. For example, when it is determined that modifications should be made to ingredient concentrations in the initial concentrate, typically it is the concentrations of one or more of these three ingredients that are adjusted.

In one example, it may be desirable to add one or more of the additional ingredients after evaluation of the initial concentrate, for example, to improve the concentrate with respect to one or more desired properties.

ii. Initial concentrate evaluation

Formulation methods further include analysis of the initial concentrate based on one or more desired properties, eg, properties of an aqueous dilution liquid composition containing the dilute concentrate, eg, transparency, color, odor, taste, safety, stability, ring formation or formation of precipitates and / or the presence of crystals. For example, the methods typically include analyzing the initial concentrate's ability to form a clear liquid upon dilution in an aqueous medium, such as by analyzing the transparency / turbidity of the resulting dilution aqueous liquid composition containing the initial concentrate.

For evaluation of the properties of the aqueous liquid dilution composition, the initial concentrate is diluted in an aqueous medium, typically water or other polar solvent, for example, at a dilution factor of between 1:10 or about 1:10 and 1: 1000 or about 1: 1000, typically between 1:10 or about 1:10 and 1: 500 or about 1: 500, for example, no more than 1:10 or about 1:10 diluted, at least 1 : 20 or about 1:20, at least 1:25 or about 1:25, at least 1:50 or about 1:50, at least 1: 100 or about 1: 100, at least 1: 200 or about 1: 200, at least 1: 250 or about 1: 250, at least 1: 300, at least 1: 400 or at least 1: 500, for example, 1:10, 1:20, 1:25, 1:30, 1:35, 1:40, 1:50, 1:55, 1:60, 1:65, 1:70, 1:75, 1:80, 1:90, 1: 100, 1: 110, 1: 120, 1: 130, 1: 140, 1: 150, 1: 160, 1: 170, 1: 180, 1: 190, 1: 200, 1: 210, 1: 220, 1: 230, 1: 235, 1: 240, 1: 250, 1: 260, 1: 270, 1: 280, 1: 290, 1: 300, 1: 350, 1: 400, 1: 450, 1: 500, or any other dilution, such as others provided herein. Typically, the transparency of the aqueous liquid dilution composition containing the initial diluted concentrate is evaluated using one or more approaches. Additionally, other properties can be evaluated, for example odor and / or taste properties of the liquid, for example, when the nonpolar compound is a polyunsaturated fatty acid (PUFA), particularly fish oil or seaweed oil, if a liquid composition Aqueous fish-smelling dilution can be empirically evaluated.

(1) Transparency

In one example, dilution of the provided concentrates in aqueous medium produces clear liquids. The transparency of the aqueous dilution liquid composition containing the initial concentrate can be evaluated by one or more of a plurality of approaches, such as by empirical observation, measuring the particle size and / or measuring the haze value of the liquid.

In one example, concentrates can be diluted to form clear liquids (or liquids that are of equal transparency to known liquids), adding between 0.05 grams (g) or about 0.05 g and 10 g or about 10 g of the concentrate, typically between 0.05g and 5g, for example 0.05g, 0.06g, 0.07g, 0.08g, 0.09g, 0.1g, 0.2g, 0.3g, 0.4 g, 0.5 g, 0.6 g, 0.7 g, 0.8 g, 0.9 g, 1 g, 2 g, 3 g, 4 g, 5 g, 6 g, 7 g , 8 g, 9 g, or 10 g of the concentrate, in 8 fluid ounces (0.236588 liters), about 8 fluid ounces (0.236588 liters) or at least 8 fluid ounces (0.236588 liters) or at least about 8 fluid ounces (0.236588 liters), for example, 8 (0.236588 liters), 9 (0.26616 liters), 10 (0.29574 liters), 11 (0.32531 liters), 12 (0.35488 liters), 13 (0.38446 liters), 14 ( 0.41403 liters), 15 (0.44360 liters), 16 (0.47318 liters), 17 (0.50275 liters), 18 (0.53232 liters), 19 (0.56190 liters), 20 (0, 59147 liters), 25 (0.73934 liters), 30 (0.88721 liters), 35 (1.0351 liters), 40 (1.1829 liters), 45 (1.3308 liters), 50 (1.4787 liters), 100 (2.9574 liters), 200 (5 , 9147 liters) or more fluid ounces of aqueous medium, eg, water, forming a clear dilution aqueous liquid composition containing the nonpolar compound. In another example, concentrates can be diluted to form clear dilution aqueous liquid compositions by adding between 1 ml or about 1 ml and 10 ml or about 10 ml of the concentrate, for example, 1 ml, 2 ml, 3 ml, 4 ml, 5 ml, 6 ml, 7 ml, 8 ml, 9 ml, or 10 ml of the concentrate in 8 fluid ounces (0.236588 liters), approximately 8 fluid ounces (0.236588 liters), or at least 8 fluid ounces (0.236588 liters) or at least about 8 fluid ounces (0.236588 liters), for example, 8 (0.236588 liters), 9 (0.26616 liters), 10 (0.29574 liters), 11 (0.32531 liters), 12 (0.35488 liters), 13 (0.38446 liters) , 14 (0.41403 liters), 15 (0.44360 liters), 16 (0.47318 liters), 17 (0.50275 liters), 18 (0.53232 liters), 19 (0.56190 liters), 20 (0.59147 liters), 25 (0.73934 liters), 30 (0.88721 liters), 35 (1.0351 liters), 40 (1.1829 liters), 45 (1.3308 liters), 50 (1 , 4787 liters), 100 (2.9574 liters), 200 (5.9147 liters) or more fluid ounces of aqueous medium, for example, water, forming a clear dilution aqueous liquid composition containing the nonpolar compound.

In another example, the concentrate can be diluted in an aqueous medium to form a clear dilution aqueous liquid composition when at least 25 mg or about 25 mg, typically at least 35 mg, for example,

ES 2 396 946 T3

35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220,

230, 240, 250, 260, 270, 280, 290, 300, 325, 350, 375, 400, 425, 450, 475, 500, 550, 600, 700, 800, 900, 1000,

1500, 2000 mg, or more, of the nonpolar active ingredient is contained in at least 8 fluid ounces (0.236588 liters) or at least about 8 fluid ounces (0.236588 liters) of aqueous dilution liquid composition, for example, a drink for example, water.

In another example, the concentrate can be diluted in an aqueous medium to form a clear dilution aqueous liquid composition at a dilution factor of between 1:10 or about 1:10 and 1: 1000 or about 1: 1000, typically between 1: 10 or about 1:10 and 1: 500 or about 1: 500, for example, when diluted no more than 1:10 or about 1:10, 1:20 or about 1:20, 1:25 or about 1: 25, 1:50 or about 1:50, 1: 100 or about 1: 100, 1: 200 or about 1: 200, 1: 250 or about 1: 250, 1: 300 or about 1: 300, 1: 400 or about 1: 400, 1: 500 or about 1: 500, for example, 1: 10, 1:20, 1:25, 1:30, 1:35, 1:40, 1:50, 1:55, 1:60, 1: 65, 1:70, 1:75, 1:80, 1:90, 1: 100, 1: 110, 1: 120, 1: 130, 1: 140, 1: 150, 1: 160, 1: 170, 1: 180, 1: 190, 1: 200, 1: 210, 1: 220, 1: 230, 1: 235, 1: 240, 1: 250, 1: 260, 1: 270, 1: 280, 1: 290, 1: 300, 1: 350, 1: 400, 1: 450, 1: 500 or more. In another example, the clear liquid is formed in less dilute 1:10 dilutions of the concentrate.

The liquid nanoemulsion concentrates provided can be formulated using any non-polar compound for dilution in an aqueous medium. In one example, concentrates can be diluted in aqueous medium, for example, over a wide dilution range to form clear liquids, for example, at a dilution factor between 1:10 or about 1:10 and 1: 1000 or about 1 : 1000, typically between 1:10 or about 1:10 and 1: 500 or about 1: 500, for example when diluted no more than 1:10 or about 1:10, 1:20 or about 1:20, 1:25 or about 1:25, 1:50 or about 1:50, 1: 100 or about 1: 100,1: 200 or about 1: 200, 1: 250 or about 1: 250, 1: 300 or about 1: 300, 1: 400 or about 1: 400, 1: 500 or about 1: 500, for example, 1:10, 1:20, 1:25, 1:30, 1:35, 1:40, 1:50, 1:55, 1:60, 1:65, 1:70 , 1:75, 1:80, 1:90, 1: 100, 1: 110, 1: 120, 1: 130, 1: 140, 1: 150, 1: 160, 1: 170, 1: 180, 1 : 190, 1: 200, 1: 210, 1: 220, 1: 230, 1: 235, 1: 240, 1: 250, 1: 260, 1: 270, 1: 280, 1: 290, 1: 300 , 1: 350,1: 400,1: 450, 1: 500 or more. Typically, the transparency of the liquid is maintained with increasing dilutions, for example, to infinity.

The transparency of the aqueous liquid dilution composition can be evaluated using one or more of a plurality of approaches, for example, qualitatively, by empirical evaluation, or quantitatively, by measuring the particle size and / or measuring the haze value of the liquid. In one example, a particular quantitative or qualitative transparency value is desired. In another example, the aqueous dilution liquid composition is desired to be as clear, less clear or more transparent than another liquid, for example, an aqueous dilution liquid composition prepared according to the methods provided or a beverage, for example, a beverage or other aqueous medium that does not contain the concentrate. For example, an aqueous liquid dilution composition, which contains the liquid concentrate diluted in a beverage, may be as clear or about as clear as the beverage itself, which does not contain any concentrate. The evaluation can be carried out qualitatively, for example by empirical observation or quantitatively, for example by calculating the particle size and / or the turbidity value (NTU) for the liquid or liquids.

(2) Empirical evaluation

The relative transparency / turbidity of the aqueous liquid dilution composition containing the dilute concentrate (eg, initial concentrate) can be qualitatively evaluated by observation. In one example, a liquid is considered transparent if it does not have an opaque appearance and / or if no particles are visible when the liquid is viewed with the naked eye. Transparency can be empirically evaluated by comparison with other liquids, eg, water, fruit juice, soda, and / or milk. For example, it may be desirable for the liquid to be as clear or about as clear as water or another liquid, eg, a beverage. For example, the liquid (containing the liquid concentrate diluted in an aqueous medium, eg a beverage) is as clear or about as clear as the aqueous medium that does not contain the liquid concentrate. In a related example, it may be desired that there is no substantial difference, for example, that there is no observable difference between the aqueous liquid dilution composition containing the concentrate and the aqueous medium without the concentrate. A clear liquid is not necessarily colorless, for example a yellow liquid that does not contain visible particles or opacity can be considered transparent.

(3) Particle size

Alternatively, the transparency of the aqueous dilution liquid composition containing the dilute concentrate (eg, initial concentrate) can be evaluated by measuring the particle size of the liquid. Methods for measuring particle size are known and any method for measuring particle size that can measure particle sizes in the appropriate ranges can be used as described below.

Particle size can be analyzed by commercial services, for example, from Delta Analytical Instruments, Inc, such as using a light scattering analyzer, for example a dynamic light scattering analyzer, for

ES 2 396 946 T3 example, the Horiba® LB-550, which can measure particle sizes within a range of 0.001 microns to 6 microns and uses a Fourier Transform / Iterative Deconvolution technique to report data and can measure sample concentrations from ppm to 40% solids; the Horiba® IA-920, which is a laser light scattering instrument that has a He-Ne laser and a tungsten lamp and can determine particle sizes from 0.02 microns to 2000 microns using Mie's Theory; or other analyzers available from Delta Analytical Instruments, Inc.

Alternatively, the particle size can be measured microscopically, for example by viewing the liquid under a microscope, for example at 640X magnification. With this method, the particle size can be quantified by comparing with a measuring device, for example , a ruler that is visible when the liquid is viewed under the microscope. If any particles are observable at this magnification, they are measured by comparison with the measuring device. At 640X magnification, for example, any particle that is about 25nm, 25nm, or greater than 25nm is visible, while particle sizes less than 25nm are typically not visible.

Typically, aqueous liquid dilution compositions are desired to have a particle size less than 200 nm or less than about 200 nm, eg, 5, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190 or 200 nm. Typically, aqueous dilution liquid compositions are desired to have a particle size less than 100 nm or about 100 nm, less than 50 nm or about 50 nm, or less than 25 nm or about 25 nm. Typically, the particle size of the aqueous liquid dilution composition containing the concentrate is between 5 nm or about 5 nm and 200 nm or about 200 nm, typically between 5 nm or about 5 nm and 50 nm or about 50 nm.

(4) Turbidity measurement

Alternatively, the transparency of the liquid can be analyzed by taking an optical turbidity measurement, which indicates the level of opacity or turbidity of a liquid, which is correlated with the size / number of suspended particles in the liquid. The more transparent a particular liquid is, the lower its turbidity value.

Turbidity can be measured optically, for example, using a nephelometer, an instrument with a light, and a detector. The nephelometer measures turbidity by detecting scattered light resulting from exposure of the liquid to incident light. The amount of scattered light is correlated with the amount of particulate matter in the liquid. For example, a ray of light will pass through the sample with low turbidity with little alteration. Other methods for measuring turbidity are well known and can be used in the provided methods and compositions.

The units of a turbidity value measured with a nephelometer are Nephelometric Turbidity Units (NTU). In one example, the aqueous dilution liquid composition containing the dilute concentrate is desired to have a low haze, eg, a haze value (NTU) of 30 or about 30; or an NTU value less than 30 or about 30, for example, less than 29 or about 29, less than 28 or about

28, under 27 or about 27, under 26 or about 26, under 25 or about 25_;minor minor minor minor minor less than 9 or about 9_; or about 6, less than 5 or about 5, less than 4 or about 4, less than 3 or about 3, less than 2 or about 2, less than 1 or about 1; or 29 or approximately

29, 28 or about 28, 27 or about 27, 26 or about 26, 25 or about 25, 24 or about 24, 23 or about 23, 22 or about 22, 21 or about 21 about about about about about 0. In Another example, the haze value of the aqueous liquid dilution composition is less than 200 or less than about 200, eg, 200, 175, 150, 100, 50, 25 or less.

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In another example, it is desirable that the aqueous liquid dilution composition contain a haze value that is comparable to, for example, approximately equal to, the same as or less than or greater than the haze value of another liquid, for example, a Beverage that does not contain the liquid concentrate or an aqueous liquid dilution composition prepared by the methods provided.

ES 2 396 946 T3 iii. Selection of a formulation and modification of formulations

After evaluation of the initial concentrate (s), any particular formula is chosen or one or more modifications are made to the initial concentrate formula based on the results of the evaluation. When an initial concentrate does not show one or more desired properties, for example, a desired extent, based on the evaluation, the concentration of one or more ingredients can be adjusted and another initial concentrate can be prepared, repeating the process until a concentrate is prepared with the desired properties. For the modification of the initial concentrate, the amount of the polar solvent, surfactant and / or non-polar active ingredient can be adjusted, for example, to another concentration within the appropriate concentration range. Alternative ingredients can also be chosen. In one example, modifying the initial concentrate involves the addition of one or more additional ingredients. For example, if the evaluation reveals that the oil and water phases of the concentrate or the aqueous liquid dilution composition containing the dilute concentrate are separating, an emulsion stabilizer can be added to the formulation. In another example, a co-surfactant can be added to help emulsify the components of the concentrate. In another example, the phase (oil phase or aqueous phase) is modified, to which a particular ingredient is added. For example, the formulation can be modified to change whether the surfactant is added to the oil phase or the water phase.

In one example, when evaluation of the initial concentrate shows that it has the desired properties, no modifications are made. In this example, the starting concentrate formula is used to prepare the concentrate. When preparing two or more starting concentrates, for example with increasing concentrations of an ingredient, the formula of one or more starting concentrates can be chosen. Which formula is chosen can be based on which formula has the most desirable property. Alternatively, desirable properties can be balanced with relative amounts of ingredients. In one example, it is desirable to choose the formulation that uses the lowest or highest concentration of a particular ingredient but still provides a concentrate that produces a clear liquid upon dilution in an aqueous medium. In one example, the desired formulation is the formulation that has the lowest concentration of the surfactant, while still providing a concentrate that produces a clear liquid upon dilution in an aqueous medium. In another example, the desired formulation is the formulation that has the highest concentration of the non-polar active ingredient, while still providing a concentrate that provides a clear liquid upon dilution in an aqueous medium. In another example, the formulation that produces the most clear liquid is desired.

However, in another example, modifications are made to the formula even though the initial concentrate has the desired properties. For example, after determining that a particular concentrate formulation results in the desired properties, it may be desirable to modify the concentration of one or more ingredients to determine whether the same desired properties can be achieved if a higher or lower concentration of the ingredient (s) is used. . For example, it may be desirable to determine the lowest concentration of surfactant that can be used, while still generating a concentrate with a desired property, eg, the ability to form a clear liquid upon dilution in an aqueous medium. In another example, it may be desirable to determine the highest concentration of the nonpolar ingredient that can be incorporated into a concentrate, while still maintaining the desired property, eg, the ability of the concentrate to form a clear liquid upon dilution in an aqueous medium. In another example, one or more additional ingredients can be added after preparing an initial concentrate with the desired properties, for example, flavoring agents and / or pH adjusting agents.

The following sections describe the ingredients used in the provided liquid nanoemulsion concentrates.

b. Non-Polar Compounds

Concentrates contain one or more nonpolar compounds. Nonpolar compounds include any soluble lipophilic or lipid compounds, for example active ingredients, that have greater solubility in organic solvents (for example, ethanol, methanol, ethyl ether, acetone, and benzene) and in fats and oils, than in liquid compositions. aqueous dilution solutions, eg water. Typically, nonpolar compounds are poorly soluble in water, or for example insoluble in water or compounds that have low solubility in water. Nonpolar compounds include, but are not limited to, drugs, hormones, vitamins, nutrients, and other lipophilic compounds. Exemplary nonpolar compounds are shown below in this document. The provided methods and compositions can be used to dilute (eg, dissolve / disperse) any non-polar compound in an aqueous medium. In one example, the nonpolar compound differs from the surfactant, for example, it is not polyethylene glycol tocopheryl succinate (TPGS). In another example, the nonpolar compound is not vitamin E. Exemplary nonpolar compounds that can be used in the concentrates provided are:

Non-polar ingredients containing essential fatty acids, for example, polyunsaturated fatty acids (PUFAs), for example, gamma-linolenic acid (GLA), for example, oil of borage or oil of evening primrose (Oenothera biennis), seed oil of black currant, hemp seed oil, spirulina extract; compounds containing omega-3 fatty acids, for example, natural and synthetic omega-3 fatty acids, for example, compounds containing omega-3 polyunsaturated long-chain fatty acids, including acid

ES 2 396 946 T3

Eicosapentaenoic (EPA) (20: 5w3); Docosahexaenoic acid (DHA) (22: 6cw3); Eicosatetraenoic acid (24: 4w3); Docosapentaenoic acid (DPA, Clupanodic acid) (22: 5w3); 16: 3 w3; 24: 5w3 and / or nisinic acid (24: 6w3) eg fish oil, seaweed oil, krill oil, canola oil, linseed oil, soybean oil and walnut oil; compounds containing short chain omega-3 fatty acids, for example, AlphaLinolenic acid (α-Linolenic acid; ALA) (18: 3w3) and Stearidonic acid (18: 4w3), esters of an omega-3 fatty acid and glycerol, for example, monoglycerides, diglycerides and triglycerides, esters of omega-3 fatty acid and a primary alcohol for example, fatty acid methyl esters and fatty acid esters, omega-3 fatty acid oil precursors, for example EPA precursor, DHA precursor, derivatives such as polyglycolized derivatives or polyoxyethylene derivatives, oils containing the omega-fatty acids 3, eg fish oil (marine oil), eg highly purified fish oil concentrates, perilla oil, krill oil and algae oils, eg microalgae oil; compounds containing omega-6 fatty acids, for example, compounds containing linoleic acid (18: 2w6) (a short chain fatty acid); Gammalinolenic acid (GLA) (18: 3w6); Dihomo gamma linolenic acid (DGLA) (20: 3w6); Eicosadienoic acid (20: 2w6); Arachidonic acid (AA) (20: 4w6); Docosadienoic acid (22: 2w6); Adenic acid (22: 4w6); and / or Docosapentaenoic acid (22: 5w6), for example, borage oil, corn oil, cottonseed oil, grapeseed oil, peanut oils, evening primrose oil for example, evening primrose oil Oenothera biennis), Blackcurrant seed oil, hemp seed oil, spirulina extract, safflower oil, sesame oil and soybean oil.

Other fatty acids, eg, triglycerides including medium chain triglycerides, polar lipids, eg, ether lipids, phosphoric acid, choline, fatty acids, glycerol, glycolipids, triglycerides, and phospholipids (eg, phosphedylcholine (lecithin), phosphatidylethanolamine and phosphatidylinositol); saw palmetto extract; and ethyl linoleate; and herbal oils such as garlic and scordinin oils; Short chain saturated fatty acids (4: 010: 0), Lauric acid (12: 0), Myristic acid (14: 0), Pentadecanoic acid (15: 0), Palmitic acid (16: 0), Palmitoleic acid (16 : 1 w7), Heptadecanoic acid (17: 0), Stearic acid (18: 0), Oleic acid (18: 1 w9), Arachidic acid (20: 0);

Micronutrients, eg vitamins, minerals, co-factors, eg Coenzyme Q10 (CoQ10, also known as ubiquinone), ubiquinol, tummeric extract (cucuminoids), lipid extract of saw palmetto (saw palmetto oil), echinacea extract , Hawthorne berry extract, ginsen extract, lipoic acid (thioctic acid), ascorbyl palmitate, kava extract, St. John's wort (St. John's wort, Klamath seed, horny goat weed), quercithin extract, dihydroepiandrosterone, indole-3-carbinol;

Carotenoids, including hydrocarbons and oxygenates, alcoholic derivatives of hydrocarbons, eg beta-carotene, mixed carotenoid complexes, lutein, lycopene, Zeaxanthin, Cryptoxanthin, eg, beta-cryptoxanthin, beta carotene, mixed carotenoid complexes, astaxanthin, bixubin, canthaxanthin , apo-carotene, beta-12'-apo-carotene, Carotene (mixture of alpha and beta-carotene), gamma carotene, cyolerritrine, zeaxanthin, members containing hydroxyl or carboxyl esters thereof;

Fat soluble vitamins, for example, Vitamins A, D, E and K, and corresponding provitamins and vitamin derivatives such as esters with an action similar to that of vitamin A, D, E or K, for example; retinol (vitamin A) and pharmaceutically acceptable derivatives thereof, for example, retinol palmitate ester and other retinol esters, and calciferol (vitamin D) and its pharmaceutically acceptable derivatives thereof and vitamin D precursors, d-alpha tocopherol (vitamin E) and derivatives thereof, including pharmaceutical derivatives thereof, for example, Tocotrienols, d-alpha tocopherol acetate and other esters of d-alpha tocopherol and ascorbyl palmitate, a fat-soluble version of vitamin C;

Phytochemicals, including phytoestrogens, eg, genistein and daidzein, eg, isoflavones, eg, soy isoflavones, flavonoids, phytoalexins, eg, Resveratrol (3,5,4'-trihydroxystilbene), red clove extract, and phytosterols;

Lipid-soluble drugs, including natural and synthetic forms of immunosuppressive drugs, such as Cyclosporine, protease inhibitors such as Ritonavir, oil-soluble anesthetics and macrolide antibiotics such as Propofol, natural and synthetic forms of steroid hormones, for example, estrogens, estradiols , progesterone, testosterone, cortisone, phytoestrogens, dehydroepiandrosterone (DhEa), growth hormones and other hormones;

Oil soluble acids and alcohols, eg, tartaric acid, lactic acid, butylated hydroxyanisole, butylated hydroxytoluene, lignin, sterols, polyphenolic compounds, oryzanol, cholesterol, phytosterols, flavonoids such as quercetin and reservoir and diallyl disulfides.

i. Active ingredients containing Polyunsaturated Fatty Acid (PUFA)

Exemplary nonpolar compounds contained in concentrates are fatty acid-containing compounds, eg, active ingredients containing polyunsaturated fatty acids (PUFAs). Fatty acids

ES 2 396 946 T3 are straight chain hydrocarbon molecules with a carboxyl group (COOH) at one end of the chain. The

PUFAs are fatty acids that contain more than one carbon-carbon double bond in the fatty acid's carbon chain. PUFAs, particularly essential fatty acids, are useful as supplements to the diet.

Different nomenclatures can be used to describe fatty acid molecules. The lipid nomenclature, for example 18: 3ω-3, indicates the length of the carbon chain, the number of double bonds, and the position along the carbon chain of the first carbon-carbon double bond in a fatty acid . Using this nomenclature, each carbon along the chain is marked according to its position relative to one end of the chain. For example, the first carbon away from the carboxylate end is named a, the second is named β, and so on. The last carbon in the molecule (the one farthest from the carboxy group) is always marked ω (or omega on). The number of carbons and the number of double bonds are shown first in the lipid name of a fatty acid, separated by a colon. For example, the name 18: 3 indicates that the molecule has eighteen (18) carbons and three (3) double bonds. Following these numbers, the position in which the first double bond appears, with respect to the last carbon (ω) is indicated. For example, the nomenclature 18: 3ω-3 (or 18: 3 omega-3; or 18: 3 n-3) describes a fatty acid with eighteen (18) carbons and three (3) double bonds, the first of which appears on the third carbon from the omega carbon.

Alternatively, chemical nomenclature can be used. The chemical name of a fatty acid describes the position of each double bond. In the chemical name, the carbons are listed, starting with 1, starting with the carbon that is part of the carboxy group (COOH). In this way, with this numbering system, the carbon becomes marked 2. The fatty acid's chemical name indicates the first carbon (from the COOH end) to participate in each double bond.

Certain PUFAs are called essential fatty acids because mammals, including humans, cannot synthesize them using any known chemical route, and must obtain them from the diet or by supplementation. (US Patent No. 6,870,077; Covington, American Family Physician (2004), 70 (1): 133-140). The essential PUFAs are omega-3 fatty acids (ω3; n-3) and omega-6 fatty acids (ω-6; n-6). Omega-3 and omega-6 fatty acids are methylene interrupted polyenes that have two or more cis double bonds, separated by a single methylene group. Exemplary Omega-3 fatty acids are Alpha-Linolenic acid (α-Linolenic acid; ALA) (18: 3ω3) (a short chain fatty acid); Stearidonic acid (18: 4-3) (a short chain fatty acid); Eicosapentaenoic acid (EPA) (20: 5w3); Docosahexaenoic acid (DHA) (22: 6w3); Eicosatetraenoic acid (24: 4w3); Docosapentaenoic acid (DPA, Clupanodic acid) (22: 5w3); 16: 3ω3; 24: 5ω3 and / or nisinic acid (24: 6ω3). Longer chain Omega-3 fatty acids can be synthesized from aLa (the short chain omega-3 fatty acid). Exemplary Omega-6 fatty acids are Linoleic acid (18: 2-6) (a short chain fatty acid); Gamma-linolenic acid (GLA) (18: 3w6); Dihomo gamma linolenic acid (DGLA) (20: 3w6); Eicosadienoic acid (20: 2w6); Arachidonic acid (AA) (20: 4w6); Docosadienoic acid (22: 2w6); Adenic acid (22: 4w6); and Docosapentaenoic acid (22: 5-6).

Although the longer chain Omega-3 and Omega-6 essential fatty acids can be synthesized from ALA (the short chain omega-3 fatty acid) and Linolenic acid (LA), respectively, evidence suggests that the conversion of these short-chain fatty acids in humans is slow. Thus, a major source of essential long-chain PUFAs is in the diet (for example, see Ross et al., Lipids in Health and Disease (2007), 6:21; Lands, The FASEB Journal (1992), 6 (8): 2530). Dietary supplements containing PUFAs, particularly essential PUFAs, are desirable for protection against cardiovascular disease, inflammation, and mental weakness (see for example, Ross et al., Lipids in Health and Disease (2007), 6:21; Lands, The FASEB Journal (1992), 6 (8): 2530; US Patent No. 6,870,077). Evidence suggests that essential fatty acids, particularly EPA and DHA, in the form of foods and nutritional supplements, play a role in the prevention of numerous pathologies, including cardiovascular disease, inflammation, mental health and behavioral diseases and disorders (see, for For example, Ross et al., Lipids in Health and Disease (2007), 6:21; Lands, The FASEB Journal (1992), 6 (8): 2530; US Patent No. 6,870,077; Covington, American Family Physician (2004), 70 (1): 133-140).

Omega-9 fatty acids are not essential PUFAs. Exemplary omega-9 fatty acids are Oleic acid (which is monounsaturated) (18: 1 w9); Eicosenoic acid (20: 1 w9); Meadic acid (20: 3 w9); Erucic acid (22: 1 w9); and Nervonic acid (24: 1 * 9).

Conjugated fatty acids are PUFAs with two or more conjugated double bonds. Conjugated fatty acids can be used as nutritional supplements. Exemplary conjugated fatty acids are conjugated linoleic acid (CLA), eg, 18: 2ω7, 18: 2ω6; Conjugated Linolenic Acid, eg, 18: 3-6,18: 3-5; and other conjugated fatty acids, for example, 18: 3ω3, 18: 4ω3, and 20: 5ω6.

(1) Omega-3 fatty acid compounds

Exemplary PUFA-containing active ingredients that can be used in the compositions provided are compounds containing one or more omega-3 (ω3; n-3) fatty acids, for example, compounds containing

ES 2 396 946 T3 DHA and / or EPA fatty acids, for example marine oils for example fish oil, krill oil and seaweed oil; and ALA fatty acid-containing compounds, for example flaxseed oil.

Typically, oily and aqueous compositions containing long chain polyunsaturated fatty acids (PUFAs) are susceptible to oxidation, rendering them unstable and giving them an unpleasant taste. The ingredients and relative concentrations thereof, as well as the methods for preparing the concentrates, contribute to the desirable properties of concentrates containing DHA / EPA. In one example, the ingredients and methods minimize the fishy odor and / or taste of DHA / EPA compositions and increase their stability over time. In one aspect, the compounds in the concentrates have low oxidation, contributing to these desirable properties.

(a) DHA / EPA

Exemplary non-polar active ingredients containing one or more omega-3 fatty acids, which can be used in the compositions provided, are compounds containing DHA and / or EPA, eg, marine oil, eg, fish oil, oil krill and seaweed oil. Any oil containing DHA and / or EPA can be used. In one example, the non-polar active ingredient contains between 20% or about 20% and 40% or about 40% DHA. In another example, the nonpolar active ingredient contains between 25% or about 25% and 35% or about 35% DHA. In another example, the non-polar active ingredient contains at least 70% or about 70% by weight (w / w), DHA, for example, at least 75% or about 75%, at least 80%. or about 80%, at least 85% or about 85%, or at least 90% or about 90% by weight (w / w), of DHA. In another example, the non-polar active ingredient contains between 5% or about 5% and 15% or about 15% EPA, for example 5, 6, 7, 8, 9, 10, 11, 12 , 13, 14 or 15% by weight (w / w), of EPA. In another example, the nonpolar active ingredient comprises no more than 10% or about 10% EPA or less than 10% or about 10% EPA. In another example, the nonpolar active ingredient contains DHA and EPA, for example, DHA representing at least 20% or about 20% by weight of the nonpolar active ingredient and EPA representing no more than 13% or about 13% of the nonpolar active ingredient, eg, no more than 10% or about 10% by weight of the nonpolar active ingredient. In another example, the nonpolar active ingredient contains DHA, representing at least 35% or about 35% of the nonpolar active ingredient and EPA representing no more than 13% or about 13% of the nonpolar active ingredient, for example, no more than 10% or about 10% of the non-polar active ingredient. In another example, the non-polar active ingredient contains dHa and EPA, for example, DHA representing at least 70% or about 70% of the non-polar active ingredient and EPA representing no more than 13% or about 13% of the active ingredient. non-polar, eg, no more than 10% or about 10% of the non-polar active ingredient.

(i) Fish oils

Exemplary PUFA-containing non-polar active ingredients that can be used in the provided compositions are fish derived oils containing DHA, EPA, or both DHA and EPA. In particular, cold-water marine fish is a known source of Omega-3 fatty acids (US Patent No. 4,670,285). Suitable fish oil containing DHA, EPA, or both DHA and EPA can be obtained from any of a number of commercial sources, for example, fish oils available from Jedwards International, Inc., any of which can be used with the compositions provided.

Fish oils are typically extracted from fish tissue, for example frozen fish tissue. In one example, the fish oil is an unflavored fish oil, for example cod liver oil, which has been isolated from fish, for example cod liver, and then refined and deodorized or has been treated in some other way so as to make its taste neutral, for example, as described in International Publication Nos. WO 00/23545 and WO 2004/098311. In one example, these fish oils are isolated from frozen fish tissue by a process that minimizes oxidation. Exemplary of such unflavored fish oils are Denomega ™ 100, Borregaard Ingredients, Sarpsborg, Norway; distributed by Denomega Nutritional Oils AS, Boulder, CO. Typically, unflavored fish oil, for example cod liver oil, contains between 25% or about 25% and 35% or about 35% Omega-3 fatty acids, for example 34% of Omega-3 fatty acids. In one example, fish oil, for example, Denomega ™ 100 oil contains 13% or about 13% DHA and 13% or about 13% EPA.

Fish oils containing high amounts of Omega-3 fatty acids, eg high amounts of DHA, are also exemplary of fish oils that can be included in the provided compositions. In one example of such fish oil they contain at least about 85% DHA, typically more than 85% DHA, and at least about 90% Omega-3 fatty acids, typically greater than 90% Omega-3 fatty acids. . In another example, fish oil may contain 98% PUFA, 89% Omega-3 fatty acids, approximately 70% DHA, approximately 10% EPA, 8.9% Omega-6 fatty acids, and 0.7 % Omega-9 fatty acids.

ES 2 396 946 T3

Exemplary of a fish oil containing high amounts of Omega-3 fatty acids that can be used as the non-polar compound in the provided compositions is Omega-3 Fish Oil EE (O3C Nutraceuticals, supplied by Jedwards International Inc., Quincy , MA), which contains 89% Omega-3 fatty acids, 8.9% Omega-6 fatty acids, 0.7% Omega-9 fatty acids, 0.1% saturated fatty acids, 1.0% of monounsaturated fatty acids, 74.5% Docosahexaenoic fatty acids (DHA), 9.3% Eicosapentaenoic fatty acids (EPA) and 98% polyunsaturated fatty acids (PUFA). This fish oil also contains 0.1% (16: 0) palmitic acid, 0.1% (16: 1 w7) palmitoleic acid, 0.1% (18: 0) stearic acid, 0.6 % (18: 1 ω 9) oleic acid, 0.1% (18: 1 ω 7) oleic acid, 0.3% (18: 2 w6) linoleic acid, 0.2% (18: 3 ω 3) linolenic acid, 0.2% (18: 4 ω 3) octadecatetraenoic acid, 0.1% (20: 1 ω 9) eicosanoic acid, 0.1% (20: 2 ω6) eicosadienoic acid, 0.2% (20: 3 ω6) Eicosatrienoic Acid, 2.4% (20: 4 ω6) Arachidonic Acid, 0.6% (20: 4 ω3) arachidonic acid, 0.1% (22: 1 ω11) erucic, 0.6% (21: 5 ω3) uncosapentaenoic acid, 0.5% (22: 4 ω6 ) of docosatetraenoic acid, 5.4% (22: 5 ω6) of docosapentaenoic acid, 3.6% (22: 5 ω3) of docosapentaenoic acid and 0.9% of other fatty acids.

Also exemplary of a fish oil containing high amounts of Omega-3 fatty acids that can be used in the provided compositions is Omega 85 DHA TG Ultra Concentrate (O3C Nutraceuticals AS, Oslo, Norway), which contains more than 85% DHA (C22: 6n-3) and more than 90% of total omega-3 fatty acids and is isolated from fatty fish species of the families Eugraulidae, Clupeidae and Scombridae. This fish oil is produced by purifying and concentrating the oils from these fish with moderate technologies to increase the concentration of the omega-3 fatty acid of DHA. Any fish oil containing DHA and / or EPA can be used as the nonpolar compound in the compositions provided. Exemplary of fish oils are also other fish oils manufactured by O3C Nutraceuticals, AS and other fish oils supplied by Jedwards, International, Inc.

Also exemplary of fish oils are krill oils, prepared according to International Publication No. WO 2007/080515.

(ii) Algae oil

Exemplary also of non-polar compounds containing Omega-3 PUFAs, particularly DHA (and optionally EPA), which can be used as the non-polar compound in the provided compositions are oils derived from microorganisms, for example oils derived from marine dinoflagellates, for example eg microalgae, eg Crypthecodinium sp, particularly Crypthecodinium cohnii. Microalgae oils, like fish oil, are an excellent source of omega-3 fatty acids, particularly DHA (US Patent No. 5,397,591, 5,407,957, 5,492,938, and 5,711,983). Exemplary oils derived from microalgae are the oils disclosed in (and oils prepared according to the methods described in) US Patent Nos. 5,397,591, 5,407,957, 5,492 938, and 5,711,983 and in US Publication No. United States number 2007/0166411, including DHASCO® and DHASCO-S® (Martek Biosciences Corporation).

For example, US Patent No. 5,397,591 describes, among others, single-celled edible oils (algae oils) (and a method for preparing the oils), which contain at least 70% triglycerides, which contain approximately one 20-35% DHA and lacking EPA, Crypthecodinium cohnii isolates, which preferably contain more than 70% triglycerides, which have 15-20% myristic acid; 20-2 5% palmitic acid; 10-15% oleic acid; 30-40% of DHA and 0-10% of other triglycerides. United States Patent No. 5,407,957 describes, among others, algal oils (and methods for the preparation of the oils) derived from Crypthecodinium cohnii, which preferably contain more than about 90% triglycerides, at least 35% of DHA by weight (w / w), in one example, having 15-20% myristic acid, 20-25% palmitic acid, 10-15% oleic acid, 40-45% DHA and 0-5% of other oils. United States Patent No. 492,938 describes, among others, single-cell edible oils (and method for preparing the oils) that contain at least 70% triglycerides, contain approximately 20-35% DHA, and are devoid of EPA, isolated from Crypthecodinium cohnii, in one example containing more than 70% triglycerides having 15-20% myristic acid; 20-25% palmitic acid; 10-15% oleic acid; 30-45% dHa; 0-10% of other triglycerides. US Patent No. 5,711,983 describes, among others, single-cell edible oils (and methods for preparing the oils) that contain at least 70% triglycerides, contain approximately 20-35% DHA, and which they lack EPA, isolated from Crypthecodinium cohnii, in one example containing more than 70% triglycerides, which has 15-20% myristic acid; 20-25% palmitic acid; 10-15% oleic acid; 30-40% of dHa and 0-10% of other triglycerides.

Also exemplary of suitable microalgae oils are those disclosed, for example, in US Patent No. 6,977,166 and US Publication Number US 2004/0072330. Any oil derived from dinoflagellates, eg microalgae, containing DHA and optionally EPA, is suitable as an algae oil for use with the compositions provided, eg V-Pure algae oil (Water4Life, Switzerland containing EPA and DHA.

(b) Flaxseed oil-omega 3 (ALA)

ES 2 396 946 T3

Also exemplary of non-polar Omega-3 containing compounds used in the provided compositions is linseed oil (linseed oil, flax seed oil). Flaxseed oils that are good sources of omega-3 fatty acids, particularly alpha-linolenic acid, have been used as nutritional supplements. Flaxseed oils are produced by pressing flax seed and refining the flax seed oil. Exemplary of linseed oil that can be used as the non-polar compound in the provided compositions is linseed oil derived from Linum usitatissimum L, for example, linseed oil supplied by Sanmark LLC, Greensboro, NC (Sanmark Limited, Dalian, Liaoning Province , China), which does not contain less than (NLT) 50% C18: 3 alpha-linolenic acid, and which additionally contains other fatty acids, for example 3-8% C16: 0 Palmitic acid, 2-8% of Stearic acid C18: 0, 11-24% C18: 1 Oleic acid, 11-24% C18: 2 linoleic acid and 0-3% other fatty acids. Also exemplary of the suitable linseed oil is a linseed oil containing 6% palmitic acid, 2.5% stearic acid, 0.5% arachidic acid, 19% oleic acid, 24.1% linoleic acid, 47.4% linolenic acid and 0.5% other fatty acids. The fatty acid composition of flaxseed oil can vary. Any linseed oil can be used as the nonpolar compound in the provided compositions. In one example, flaxseed oil contains at least 50% alpha-linolenic acid or at least about 50% alpha-linolenic acid. In another example, flaxseed oil contains at least 65% or about 65% or 70% or about 70% alpha-linolenic acid. Exemplary of a linseed containing a content greater than 65% linolenic acid (of the total fatty acid content), for example, 7080% or 70-75% is the linseed described in US Patent No. 6,870,077.

(2) Omega-6 compounds

Also exemplary of the non-polar compounds used in the provided compositions are compounds containing omega-6 PUFAs, for example, gamma-linolenic acid (GLA), for example, oil of borage and oil of evening primrose (Oenothera biennis), oil of blackcurrant seeds, hemp seed oil, fungal oil and spirulina extract. Any oil containing omega-6 fatty acids can be used in the compositions provided.

(a) Borage Oil (Gamma-Linolenic Acid (GLA))

Exemplary of the non-polar omega-6-containing compounds are GLA-containing compounds, for example, borage oil. GLA is an omega-6 PUFA, which is primarily derived from vegetable oils, for example evening primrose oil (Oenothera biennis), blackcurrant seed oil, hemp seed oil, and spirulina extract. GLA has been used as a nutritional supplement. GLA has been proposed to play a role in the treatment of various chronic diseases and in particular to have anti-inflammatory effects (Fan and Chapkin The Journal of Nutrition (1998), 1411-1414). In one example, the nonpolar active ingredient contains at least about 22% or about 22% by weight (w / w) of GLA, for example, a 22, 23, 24, 25, 26, 27, 28, 29 , 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 50, 60% or greater, by weight (w / w) of GLA.

Borage (Borago officinalis), also known as a flower with five pointed triangular petals, is an herb whose seeds contain high amounts of GLA. The exemplary borage oil that is used as a non-polar active ingredient in the provided compositions is borage oil supplied by Sanmark LLC, Greensboro, NC (Sanmark Limited, Dalian, Liaoning Province, China), which is obtained by pressing and isolating the oil from the seeds of Borago officinalis L. This oil does not contain less than (NLT) 22% C18: 3 gammalinolenic acid (GLA), between 9 and 12% C16: 0 Palmitic acid, between 3% and 5% C18: 0 Stearic acid, between 15% and 20% of Oleic acid C18: 1, between 35% and 42% of linoleic acid C18: 2, between 3% and 5% of Ocosenoic acid C20: 1, between 1% and 4% of Docosenoic acid C22: 1 and between 0% and 4% of other fatty acids. Other borage oils can be used. Other GLA-containing oils can also be used as the nonpolar compound.

(3) Saw Palmetto Extract

Also exemplary of the nonpolar compounds used in the provided compositions are saw palmetto extract, a lipophilic extract of ripe berries of the saw palmetto (also called Serenoa repens or Sabal serrulata), which has been used to treat genitourinary diseases and others. , and to enhance sperm production, breast size and libido, as a moderate diuretic, as a nerve sedative, as an expectorant and as a digestive tract tonic, and particularly to treat benign prostatic hyperplasia (BHP) (Ernst, Academia and Clinic (2002), 136; 42-53; Gordon and Shaughnessy, Complementary and Alternative Medicine (2003), 76 (6); 1281-1283). Saw palmetto extract is commercially available from numerous sources. Any lipid extract of saw palmetto can be used in the compositions provided. Exemplary of the saw palmetto extract used in the provided compositions is Saw Palmetto, Lipophilic Extract, commercially available from Natural Medicinals, Inc., Felda, fL. This Saw Palmetto Lipophilic Extract is extracted with Carbon Dioxide and, in one example, contains 85.9% total fatty acids, including 0.8% Caproic acid, 2% Caprylic acid, 2.4 % Capric acid, 27.1 Lauric acid, 10.3 Myristic acid, 8.1% Palmitic acid, 0.2% Palmitoleic acid, 2% Stearic acid, 26.7 Oleic acid, 4.9% Linoleic acid, 0.7% linolenic acid, 0.42%; 0.42% phytosterols, including 0.42% beta Sitosterol, 0.09% Campesterol, 0.03% Stigmasterol; and 0.2% humidity. Other sources of

ES 2 396 946 T3 saw palmetto extract.

(4) Conjugated Linoleic Acid (CLA) are also exemplary of the non-polar PUFA compounds that can be used in the provided compositions non-polar compounds containing conjugated fatty acids. Conjugated fatty acids are PUFAs with two or more conjugated double bonds. Conjugated fatty acids can be used as nutritional supplements. Exemplary of active ingredients containing conjugated fatty acids are compounds containing Conjugated Linoleic Acid (CLA), for example, 18: 2w7, 18: 2w6; Conjugated Linolenic Acid, eg, 18: 3w6, 18: 3w5; and other conjugated fatty acids, for example 18: 3w3, 18: 4w3 and 20: 5w6. CLA refers to a family of linoleic acid isomers found primarily in ruminant meat and dairy products. Typically, CLA compounds contain a mixture of different CLA isomers, for example, C18: 2 CLA c9, t11, CLA t10, c12, and other CLA isomers. Exemplary of CLA that can be used as an active ingredient in compositions provided is CLA (80%) commercially available from Sanmark, LTD (Dalian, Liaoning Province, China; product code 01057-A80). This CLA is a clear white to pale yellow oil and has the following fatty acid composition: NMT (not more than) 9.0% Palmitic acid C16: 0, NMT 4.0% Stearic acid, NMT 15.0% Oleic acid C18: 1, NMT 3.0% Linoleic acid C18: 2, NLT (not less than) 80% CLA C18: 2 (including the following isomers: NLT 37.5% C18: 2 CLA c9, t11, 37, 5% C18: 2 CLAt10, c12 and NMT 5.0% of other isomers of CLA); and NMT 5.0% of other fatty acids. Other CLA-containing compounds can be used.

ii. Active Ingredients of Coenzyme Q

Exemplary of the non-polar active ingredients are compounds containing Coenzyme Q, for example, Coenzyme Q10 (also referred to as CoQ10, ubiquinone, ubidecarenone, ubiquinol and vitamin Q10). Coenzyme Q compounds are benzoquinone compounds that contain isoprenyl units. The number of isoprenyl units in each of the different CoQ species is indicated by a number following the CoQ. For example CoQ10 contains 10 isoprenyl units. Coenzyme Q10 is a predominant Coenzyme Q species.

Coenzyme Q can exist in two different forms: an oxidized form and a reduced form. When the oxidized form of a Coenzyme Q species is reduced by one equivalent, it becomes ubisemiquinone, denoted QH, which contains a free radical in one of the oxygens on the benzene ring of benzoquinone. Both oxidized and reduced coenzyme Q-containing compounds can be used as active ingredients in the provided compositions.

(1) Coenzyme Q10

Exemplary of the non-polar active ingredients containing Coenzyme Q that can be used in the compositions provided active ingredients containing Coenzyme Q10. Coenzyme Q10 (also known as CoQ10, ubiquinone, ubidecarenone, ubiquinol, and vitamin Q10) is a benzoquinone compound that contains 10 isoprenoid units. The Q in the name refers to Quinone and the 10 refers to the number of isoprenoid units. CoQ10 typically refers to the oxidized form of CoQ10, which is also called ubidecarenone, as opposed to the reduced form of CoQ10. Both oxidized and reduced CoQ10 are exemplary of the coenzyme Q species that can be used as active ingredients in the compositions provided.

CoQ10 has electron transfer capabilities and is present in cell membranes such as those of the endoplasmic reticulum, peroxisomes, lysosomes, vesicles, and in mitochondria. A decrease in the synthesis of natural CoQ10 has been observed in sick and elderly people. Due to this observation and its powerful antioxidant properties, CoQ10 is used as a dietary supplement and a treatment for diseases such as cancer and heart disease. CoQ10, however, has a relatively poor bioavailability.

Compounds containing CoQ10 are commercially available. Any CoQ10 compound or reduced CoQ10 compound can be used with the provided composition. Exemplary of CoQ10 compounds that can be used as active ingredients are coenzyme Q10 compounds containing more than 98% or more than about 98% ubidecarenone, for example, the compound marketed under the name Kaneka Q10 ™ (Ubidecarenone USP) from Kaneka Nutrients, LP., Pasadena, TX. The compound marketed under the name Kaneka Q10 ™ is fermented entirely from yeast and is identical to the body's own CoQ10 and is free of the cis isomer found in some synthetically produced CoQ10 compounds. Any CoQ10 compound can be used in the compositions provided.

iii. Active Ingredients Containing Phytosterol

Exemplary of the nonpolar compounds used as active ingredients in the provided compositions are phytosterol-containing (plant sterol) compounds. Plant sterols are structurally similar to cholesterol and have been found to reduce the absorption of cholesterol from the diet, which can affect

ES 2 396 946 T3 serum cholesterol levels. According to the US Food and Drug Administration (FDA), two servings a day, each containing 0.4 grams of plant sterols, for a total daily intake of at least 0.8 grams, as part Eating a diet low in saturated fat and cholesterol can lower your risk of heart disease. In this way, plant sterols are used in nutritional supplements.

Any phytosterol-containing compound can be used as an active ingredient in the provided compositions. Exemplary of phytosterol-containing compounds that can be used as active ingredients in the provided compositions are compounds containing plant sterols, for example, the compound marketed under the name CardioAid ™, distributed by B&amp;D Nutrition and manufactured by ADM Natural Health and Nutrition, Decatur , IL. This compound contains Kosher, Pareve and Halal plant sterols that are produced according to the current food GPM. The sterols are PCR negative and the material is derived from genetically modified organisms (GMO). This phytosterol compound contains a minimum of 95% plant sterols, including up to 5 plant sterols. The compound may contain, for example, 40-58% Beta Sitosterol, 20-30% Campesterol, 14-22% Stigmasterol, 0-6% Brassicasterol, and 0-5% Sitostanol. The compound may additionally contain tocopherols, for example 0-15 mg / g tocopherols. The compound is tested and negative for Salmonella, E. coli, and Staphylococcus aureus.

c. Surfactants

The compositions provided contain surfactants. For example, in addition to the nonpolar compound (s), liquid concentrates contain one or more surfactants. In the provided methods for producing the concentrates, the surfactant is added to the water phase, the oil phase, or the water and oil phase. The compositions may additionally contain one or more co-surfactants or emulsifiers.

Surfactants are added to aqueous liquids, such as in provided compositions (eg, concentrates and aqueous liquid dilution compositions) to form micelles, which contain the nonpolar compound (s). The hydrophilic portion or portions of the surfactant molecules are oriented toward the outside of the micelle, in contact with the aqueous medium, while the hydrophobic portion or portions of the surfactant molecules are oriented toward the center of the micelle, in contact with the compound. or nonpolar compounds, which are contained in the center of the micelle. Micelles can contain more than one surfactant and / or co-surfactant. The properties of the compositions provided, eg, the particle size of the compositions and the desirable properties related to the particle size, are influenced by the choice of the surfactant (s) and the relative amount (concentration) of the surfactant. For example, the HLB of the surfactant (s) can affect the particle size, transparency, taste, odor, crystal formation, and other properties of the compositions provided. Similarly, the concentration of the surfactant compared to the concentration or concentrations or concentrations of other ingredients, particularly compared to the concentration of the polar solvent (s) and the concentration of the non-polar compound (s), can affect various desirable properties, for example, the ability to disperse or dissolve in aqueous media, for example, to form an aqueous liquid composition for dilution or with a pleasant taste and / or odor.

Surfactants (and co-surfactants) are molecules that contain hydrophobic and hydrophilic moieties. In one example, the hydrophobic portion is a hydrophobic tail and the hydrophilic portion is a hydrophilic head of the surfactant molecule.

Exemplary of the surfactants that can be used in the provided compositions and methods are surfactants having an HLB value of between 14 or about 14 and 20 or about 20, typically between 16 or about 16 and 18 or about 18. Exemplary suitable surfactants include, but are not limited to, surfactants derived from Vitamin E, such as surfactants derived from tocopherol and / or tocotrienol in which the Vitamin E moiety represents the hydrophobic region of the surfactant, and is attached, via linker to another moiety, such as a polyethylene glycol (PEG) moiety, which provides the hydrophilic portion of the surfactant. Vitamin-E derived surfactants include, but are not limited to, tocopherol derived surfactants, including polyalkylene glycol derivatives of tocopherol, typically polyethylene glycol (PEG) derivatives of tocopherol, such as polyethylene glycol tocopherol succinate (TPGS), TPGS analogs, TPGS homologs and TPGS derivatives. Alternatively, the surfactants can be other PEG derivatives that have similar properties, for example, PEG derivatives of sterols, for example a cholesterol or sitosterol (including, for example, any of the PEG derivatives disclosed in US Pat. Nos. 6,632,443) or PEG derivatives of other fat-soluble vitamins, eg, some forms of Vitamin A (eg, Retinol) or Vitamin D (eg, Vitamin D1-D5).

The HLB value of a surfactant comes from a semi-empirical formula; HLB values ​​are used to index surfactants according to their relative hydrophobicity and hydrophilicity. An HLB value is a numerical representation of the relative representation of hydrophilic groups and hydrophobic groups in a surfactant or a mixture of surfactants. The weight percent of these respective groups indicate properties of the molecular structure. See, for example, Griffin, WCJ Soc. Cos. Chem. 1: 311 (1949).

ES 2 396 946 T3

Surfactant HLB values ​​range from 1-45, while the range for nonionic surfactants is typically 1-20. The more lipophilic a surfactant is, the lower its HLB value. Conversely, the more hydrophilic a surfactant is, the higher its HLB value. Lipophilic surfactants have higher solubility in oil and lipophilic substances, while hydrophilic surfactants dissolve more easily in aqueous liquids. In general, surfactants with HLB values ​​greater than 10 or greater than about 10 are called hydrophilic surfactants, while surfactants having HLB values ​​less than 10 or less than about 10 are called hydrophobic surfactants. HLB values ​​are known for a number of surfactants. Table 1A shows HLB values ​​of exemplary surfactants and cosurfactants.

The surfactants in the compositions provided are typically nonionic surfactants and typically have an HLB value between and about 14 and about 20. Particular examples of suitable surfactants include PEG-derived surfactants, such as PEG derivatives of Vitamin E having appropriate HLB values, such as an HLB value between 14 or about 14 and 20 or about 20, eg, 14, 15, 16, 17, 18, 19, 20, about 14, about 15, about 16, about 17, about 18, about 19, or about 20. Typically, the surfactants are natural surfactants, for example, a surfactant that is GRAS (generally recognized as safe) by the FDA and / or certified Kosher.

i. Surfactants derived from vitamin E

Surfactants include, but are not limited to, Vitamin-E derived surfactants such as tocopherol and / or tocotrienol derived surfactants, in which the Vitamin E moiety represents the hydrophobic region of the surfactant, and is linked, through a linker, to other moieties, such as a polyethylene glycol (PEG) moiety, which provides the hydrophilic portion of the surfactant. Vitamin E-derived surfactants include, but are not limited to, tocopherol-derived surfactants, including polyethylene glycol derivatives of tocopherol, typically polyethylene glycol (PEG) derivatives of tocopherol, such as tocopherol polyethylene glycol succinate (TPGS), TPGS analogs, homologues of TPGS and derivatives of TPGS. Alternatively, the surfactants may be other PEG derivatives having similar properties as TPGS and TPGS analogs, for example, PEG derivatives of sterols, for example, a cholesterol or a sitosterol (including, for example, any of the derivatives of PEGs disclosed in U.S. Patent No. 6,632,443) or PEG derivatives of other fat-soluble vitamins, for example, some forms of Vitamin A (for example, Retinol) or Vitamin D (for example, Vitamin D1-D5).

(1) PEG derivatives of Vitamin E

Vitamin E derived surfactants (eg, tocopherol derived surfactants or tocotrienol derivatives) include polyalkylene glycol derivatives of Vitamin E, typically polyethylene glycol (PEG) derivatives of Vitamin E, eg, PEG derivatives of tocopherol and tocotrienol. Suitable PEG derivatives of Vitamin E typically contain one or more tocopherols or tocotrienols, linked (eg, via an ester, ether, amide, or thioester bond) to one or more PEG moieties, through a linker, eg, a dicarboxylic acid linker. An exemplary surfactant is schematically shown below:

<td>PEG -</td><td>- Crimping -</td><td>- Vitamin E</td>

where the line between the PEG and the Crimp; and the line between the Linker and Vitamin E each independently represent a covalent bond selected from an ester, ether, amide, or thioester.

Typically, PEG derivatives of Vitamin E are prepared by linking the PEG moiety, by esterification, to a vitamin E-linker conjugate (eg, a tocopherol-linker conjugate). In one example, the tocopherol-linker conjugate is formed first by covalently linking (by esterification) the hydroxyl moiety of tocopherol with a dicarboxylic acid to produce an ester bond. In this example, the tocopherol-linker conjugate is a tocopherol ester (such as tocopherol succinate). The esterification reaction can be carried out by any number of known methods (see, for example, US Patent Nos. 2,680,749, 4,665,204, 3,538,119, and 6,632,443). To prepare the tocopherol-PEG surfactant, the resulting tocopherol ester is then attached (via linker) to the PEG molecule, in another esterification reaction. In this example, the resulting surfactant is a polyethylene glycol tocopherol diester (TPGD).

Alternatively, PEG derivatives of a tocopherol-linker or tocotrienol-linker conjugate can be prepared by other methods. Various methods known in the art can be used to produce PEG derivatives to link a PEG molecule to tocopherol-linker or tocotrienol-linker compounds. For example, a tocopherol-linker conjugate can be covalently attached to the molecule through an amide, ether, or thioether bond. For example, a tocopherol-linker conjugate containing an amine group can be reacted with a PEG-NHS derivative to form an amide bond between the tocopherol-linker and the PEG molecule. A tocopherol-linker conjugate containing an amine group can be reacted with a PEG49 derivative.

ES 2 396 946 T3 aldehyde to form an amide bond between the tocopherol-linker and the PEG molecule. In another example, a tocopherol-linker containing a carboxylic acid can be activated to the corresponding acid halide and reacted with a derivative of PEG-SH to form a thioester bond between the tocopherol-linker and the molecule of

PEG.

(a) Tocopherols and Tocotrienols

The tocopherol (s) used to prepare the surfactant can be any natural or synthetic Vitamin E tocopherol including, but not limited to, alpha-tocopherols, beta-tocopherols, gamma-tocopherols, and delta-tocopherols, either in pure forms or in heterogeneous mixtures. in more ways than one. Exemplary tocopherols are da tocopherols and d, 1-tocopherols. To prepare the surfactant, tocopherol is typically esterified with a linker, eg, a dicarboxylic acid, to form a tocopherol ester, which is then attached to a PEG moiety.

The tocotrienol (s) used to prepare the surfactants can be any natural or synthetic Vitamin E tocotrienol including, but not limited to, alpha-tocotrienols, beta-tocotrienols, gamma-tocotrienols and delta tocotrienols, either in pure forms or in heterogeneous mixtures of more than one way. Mixtures of tocopherols and tocotrienols are contemplated for use in the provided methods and compositions. A tocotrienol can be esterified with a linker, such as a dicarboxylic acid, before linking it with a PEG moiety.

(b) PEG residues

The PEG used in the tocopherol-PEG derivative can be any of a plurality of known PEG moieties. Exemplary of suitable PEG residues are PEG residues having varying chain lengths, and varying molecular weights, for example, PEG 1000, PEG 200, PEG 500 and PEG 20,000. The numbers following the individual PEG residues indicate the molecular weight (in kilodaltons (kDa) of the PEG residues. The PEG moiety of the tocopherol-derived surfactant typically has a molecular weight between 200 kDa or about 200 kDa and 20,000 kDa or about 20,000 kDa, typically between 200 kDa or about 200 kDa and 6000 kDa or about 6000 kDa, for example, between 600 kDa. or about 600 kDa and 6000 kDa or about 6000 kDa, typically between 200 kDa or about 200 kDa and 2000 or about 2000 kDa, between 600 or about 600 kDa and 1500 kDa or about 1500 kDa, such as, but not limited to, 200, 300, 400, 500, 600, 800, and 1000 kDa. Exemplary of a PEG derivative of tocopherol ester having a PEG residue with 1000 kDa is TPGS-1000. Also exemplary of suitable PEG residues are PEG residues that are modified, for example, methylated PEG (m-PEG), which is a PEG chain protected with a methyl group. Other known PEG analogs can also be used. PEG moieties can be selected from any reactive PEG including, but not limited to, PEG-OH, PEG-NHS, PEG-aldehyde, PEG-SH, PEG-NH2, PEGCO2H, and branched PEG.

(c) Crimps

Typically, the PEG derivatives of Vitamin E are diesters or other esters, eg, triesters. When the PEG derivative is a diester, the linker that binds Vitamin E to PEG is typically a carboxylic acid, typically a dicarboxylic acid as for example in polyethylene glycol tocopherol succinate (TPGS), where the linker is a succinic acid, The surfactant is prepared by an esterification region linking a PEG moiety and a tocopherol ester of the dicarboxylic acid. In another example, the linker is another molecule, for example, an amino acid, such as glycine, alanine, 5-aminopentanoic acid, or 8-aminooctanoic acid; or an amino alcohol, such as ethanolamine.

(d) Polyethylene glycol diesters of Tocopherol and polyethylene glycol of Tocotrienol (Dicarboxylic acid esters of Vitamin E bound to PEG)

Typically, PEG derivatives of Vitamin E are polyethylene glycol diesters of Vitamin E, which are Vitamin E esters of PEG, prepared by linking a Vitamin E ester to one or more PEG moieties by esterification. Exemplary of the Vitamin E diesters are polyethylene glycol tocopherol diesters (TPGD) and polyethylene glycol tocotrienol diesters.

When the tocopherol or tocotrienol ester attached to the PEG moiety is a tocopherol ester of a dicarboxylic acid (for example, tocopherol succinate), the linker is a dicarboxylic acid (a carboxylic acid having two carboxy groups, for example, acid succinic). In this example, the PEG diester of tocopherol or tocotrienol is formed by an esterification reaction, in which PEG is bound to a tocopherol ester of a dicarboxylic acid.

Exemplary of dicarboxylic acids that can be used as linkers in these surfactants of PEG diester of tocopherol and tocotrienol are succinic acid, sebacic acid, dodecanedioic acid, suberic acid or azelaic acid, citraconic acid, methyl citraconic acid, itaconic acid, maleic acid, glutaric acid. , glutaconic acid, fumaric acids and phthalic acids. Accordingly, they are exemplary of the tocopherol esters that can be esterified to form the PEG derivatives tocopherol succinate, tocopherol sebacate, tocopherol dodecanedioate, tocopherol suberate, tocopherol azelaate, tocopherol citraconate, methylcitraconate, tocopherol.

ES 2 396 946 T3 tocopherol itaconate, tocopherol maleate, tocopherol glutarate, tocopherol glutaconate and tocopherol phthalate, among others.

Exemplary of vitamin E polyethylene glycol diesters prepared with dicarboxylic acids are compounds having the following formula shown in Scheme I below (and homologues, analogs, and derivatives thereof):

Scheme I

<img file="ES2396946T3_D0003.tif" />

in which each of R¹, R², R³ and R⁴ is, independently, H or Me; each dashed line is independently a single or double bond; n is an integer from 1-5000; each of myq is, independently, 0 or 1; and p is an integer from 1 to 20. In one example, the surfactant is a compound in which, when both m and q are 0, p is an integer between 2-20.

In one example, the surfactant has the following formula shown in Scheme II below (including homologues, analogs, and derivatives thereof):

Scheme II

<img file="ES2396946T3_D0004.tif" />

in which each of R¹, R², R³ and R⁴ is, independently, hydrogen (H) or methyl (CH2); the bond represented by the dashed line is a single or double bond, m is an integer from 1 to 20, and n is an integer from 1 to 5000.

In another example, the surfactant is an analog of TPGS, such as, but not limited to, a compound other than TPGS having the formula shown in SCHEME III:

<img file="ES2396946T3_D0005.tif" />

by the dashed line is a single or double bond, m is an integer from 1 to 20, and n is an integer from 1 to 5000.

ES 2 396 946 T3

Examples of PEG diesters of tocopherol and tocotrienol that can be used as surfactants in the compositions and methods provided include, but are not limited to: tocopherol polyethylene glycol succinates (TPGS; including Da TPGS and d, 1-TPGS; see for example, Patent US Patent No. 3,102,078), polyethylene glycol tocopherol sebacate (PTS; see, for example, US Patent No. 6,632,443), polyethylene glycol tocopherol dodecanedioate (PTD; see, for example, U.S. Patent No.

6,632,443), polyethylene glycol tocopherol suberate (PTSr; see for example US Patent No.

6,632,443), polyethylene glycol tocopherol azelaate (PTAz; see for example, U.S. Patent No.

6,632,443), tocotrienyl polyoxyethanyl sebacate (PTrienS, eg, PTrienS-600; see US Patent No. 6,632,443), as well as analogs, homologues and derivatives or any of the diesters of tocopherol.

(e) Other PEG Esters of Vitamin E

In another example, the tocopherol ester attached to the PEG to form the PEG diester of tocopherol is a tocopherol ester of a tricarboxylic acid, for example, Citric acid, Isocitric acid, Aconitic acid and Propane1,2,3-tricarboxylic acid ( tricarbalilic acid, carboxylic acid) or a carboxylic acid having three or more carboxy groups.

In another example, the PEG derivatives of tocopherol are polyethylene glycol tocopherol triesters (TPGT), for example, esters containing a tocopherol, a linker, a PEG moiety, and an additional moiety, for example an additional tocopherol, a second PEG moiety or a water soluble group, such as a quaternary amine. In one example, when the triester contains two PEG moieties, each PEG moiety has a shorter chain length (and lower molecular weight) than the PEG moiety in a PEG derivative of tocopherol, which has similar properties, containing only one PEG chain.

(f) TPGS surfactants

Exemplary are the tocopherol polyethylene glycol diester surfactants TPGS and analogs, homologues and derivatives thereof. TPGS is a natural surfactant that is GRAS and Kosher certified and thus desirable for use in products designed for human consumption, eg, beverages, food, and nutritional supplements. The TPGS typically has an HLB value between 16 or about 16 and 18 or about 18. Exemplary of the TPGS surfactants is TPGS-1000, which has a PEG residue of 1000 kDa. Exemplary of the TPGS surfactants that can be used in the provided compositions is the food grade TPGS surfactant marketed under the name Eastman Vitamin E Food grade TpGs® from Eastman Chemical Company, Kingsport, TN. This surfactant is a water-soluble form of vitamin E from a natural source, which is prepared by esterifying the carboxyl group of crystalline d-alpha-tocopheryl acid succinate with polyethylene glycol 1000 (PEG 1000) and contains between 260 and 300 mg / g of tocopherol. total. A similar compound can be prepared by esterifying the carboxyl group of synthetic Vitamin E form d, 1 with PEG 1000. This forms a clear liquid when dissolved 20% in water. This tocopheryl polyethylene glycol is a water-soluble preparation of a fat-soluble vitamin (vitamin E), for example, as disclosed in US Patent Nos. 3,102,078, 2,680,749 and US Published Applications N ° 2007/0184117 and 2007/0141203. The PEG moiety of alternative TPGS surfactants can have a molecular weight range between about 200 kDa or 200 kDa to 20,000 kDa or about 20,000 kDa, for example, between 600 kDa or about 600 kDa and 6000 kDa or about 6000 kDa, typically between 600 kDa or about 600 kDa and 1500 kDa or about 1500 kDa. Also exemplary of the TPGS surfactant that can be used in the provided compositions is Natural Water Soluble Vitamin E (TPGS) available from ZMC-USA, The Woodlands, Texas. Any known source of TPGS, or any analog, homologous or derivative thereof can be used.

Exemplary TPGS analogs are compounds other than TPGS, which are similar to a parent TPGS compound but differ slightly in composition, e.g., with the variation, addition, or removal of an atom, one or more units (e.g., a methylene unit (s) - (CH2) n) or one or more functional groups.

At room temperature, TPGS is typically a low melting waxy solid. In one example, the TPGS is heated before use, for example, to at least the melting temperature, for example, between 37 ° C or about 37 ° C and 41 ° C or about 41 ° C and the amount is poured desired. In another example, the TPGS can be added as a waxy solid to a container and heated with the heating apparatus.

They are also exemplary of the TPGS analog surfactants, including surfactants derived from Vitamin E, including PEG derivatives of Vitamin E, including PEG diesters of vitamin E, such as but not limited to, polyethylene glycol tocopherol (PTS) sebacate, dodecanediate polyethylene glycol tocopherol (PTD), polyethylene glycol tocopherol suberate (PTSr), Tocopherol polyethylene glycol azelaate (PTAz) and polyoxethanyl tocotrienyl sebacate (PTrienS) as well as other PEG derivatives of Vitamin E.

ii. Surfactant concentration

Typically, the concentration of the surfactant (s) in a particular concentrate composition is selected as described herein, by formulating an initial concentrate with a concentration of

ES 2 396 946 T3 surfactant (s) within a starting concentration range, followed by evaluation of the initial concentrate and, optionally, adjusting the concentration of the surfactant (s). Alternatively, the concentration of the surfactant can be chosen based on the concentration of the surfactant in one or more existing liquid concentrate formulas. Typically, the surfactant concentration is between 16% or about 16% and 30% or about 30% (w / w), for example, 16% or about 16%, 17% or about 17 %, 18% or about 18%, 19% or about 19%, the

20% or about 20%, 21% or about 21%, 22% or about 22%, 23% or about 23%, 24% or about 24%, 25% or about 25%, 26% or about 26%, 27% or about 27%, 28% or about 28%, 29% or about 29%, 30% or about 30% by weight ( w / w) of the concentrate. Exemplary of surfactant concentrations within the appropriate concentration range are 17.75% and 25.2% (w / w) of the concentrate. Typically, the surfactant concentration is less than or equal to 30% or about 30% (w / w) of the concentrate.

In one example, the surfactant concentration range is between 17% or about 17% and 25% or about 25% (w / w) of the concentrate. In another example, the concentration range of the surfactant is between 18% or about 18% and 25% or about 25% (w / w) of the concentrate. In another example, the concentration range of the surfactant is between 18% or about 18% and 20% or about 20% (w / w) of the concentrate. In another example, the concentration range of the surfactant is between 17% or about 17% and 20% or about 20% (w / w) of the concentrate. In another example, the concentration range of the surfactant is between 16% or about 16% and 20% or about 20% (w / w) of the concentrate.

iii. HLB

Exemplary of the properties of the surfactant (s) that contribute to the desirable properties of the compositions is the HLB (hydrophilic-lipophilic balance) of the surfactant (s). Generally, HLB is a value, derived from a semi-empirical formula, that is used to index surfactants according to their relative hydrophobicity / hydrophilicity. An HLB value is a numerical representation of the relative representation of hydrophilic groups and hydrophobic groups in a surfactant or mixture of surfactants. The weight percent of these respective groups indicates properties of the molecular structure. See, for example, Griffin, WCJ Soc. Cos. Chem. 1: 311 (1949).

Surfactant HLB values ​​range from 1-45, while the range for nonionic surfactants is typically 1-20. The more lipophilic a surfactant is, the lower its HLB value. Conversely, the more hydrophilic a surfactant is, the higher its HLB value. Lipophilic surfactants have higher solubility in oil and lipophilic substances while hydrophilic surfactants dissolve more easily in aqueous media. In general, surfactants with HLB values ​​greater than 10 or greater than about 10 are called hydrophilic surfactants, while surfactants having HLB values ​​less than 10 or less than about 10 are called hydrophobic surfactants.

HLB values ​​have been determined and are available for a plurality of surfactants (eg, see US Patent No. 6,267,985). It should be appreciated that the HLB values ​​for a given surfactant or co-surfactant can vary, depending on the empirical method used to determine the value. Thus, the HLB values ​​of surfactants and co-surfactants provide a rough guide for formulating compositions based on relative hydrophobicity / hydrophilicity. For example, a surfactant is typically selected from surfactants having HLB values ​​within a particular range of surfactant or co-surfactant that can be used for guide formulations. Table 1A shows the HLB values ​​of exemplary surfactants and cosurfactants.

The surfactants and HLB values ​​set forth in Table 1A are exemplary. Any known surfactant or co-surfactant can be used with the provided compositions (see, for example, US Patent No. 6,267,985), provided it has an appropriate HLB value, such as an HLB value between and about 14 and about 20. The surfactant (s) used in the concentrate provided typically has an HLB value between 14 or about 14 and 20 or about 20, eg, 14, 15, 16, 17, 18, 19, 20, about 14, about 15, about 16, about 17, about 18, about 19, or about 20. Exemplary surfactants include, but are not limited to, nonionic surfactants such as surfactants derived from polyethylene glycol (PEG), including, but not limited to, PEG derivatives of Vitamin E, such as PEG diesters of tocotrienol or tocopherol, such as TPGS (for eg, TPGS 1000) and TPGS analogs, homologues, and derivatives. Other known surfactants having HLB values ​​between 14 or about 14 and 20 or about 20, typically between about 15 and 18, may also be suitable. For example, surfactants having similar properties to TPGS can also be used. Typically the surfactant is a natural surfactant, for example a surfactant that is GRAS (generally recognized as safe) by the FDA and / or certified Kosher.

ES 2 396 946 T3

d. Co-surfactants (emulsifiers)

In one example, the liquid concentrate additionally contains one or more co-surfactants (emulsifiers). For example, a co-surfactant can be included to improve the emulsion of the active ingredient and / or the stability of the composition, for example by preventing or slowing the oxidation of the nonpolar compound. Exemplary of a cosurfactant used in the concentrates provided is a phospholipid, eg, phosphatidylcholine.

i. Phospholipids

Exemplary of the co-surfactants that can be used in the provided compositions are phospholipids. Phospholipids are amphipathic lipid-like molecules, typically containing a hydrophobic portion at one end of the molecule and a hydrophilic portion at the other end of the molecule. A number of phospholipids can be used as ingredients in the provided compositions for example, lecithin, including phosphatidylcholine (PC), phosphatidylethanolamine (PE), distearoylphosphatidylcholine (DSPC), phosphatidylserine (PS), phosphatidylglycerol (PG), phosphatidic acid (PA) (PI), sphingomyelin (PMS), or a combination thereof. Typically, the phospholipid is phosphatidylcholine (PC), which is sometimes referred to by the general name lecithin. Exemplary of the phospholipids that can be used as co-surfactants in the compositions provided are the phospholipids marketed by Lipoid, LLC, Newark, NJ, for example, Purified Egg Lecithins, Purified Soybean Lecithins, Egg Lecithins, and Soybean Seed. Hydrogenated, Egg Phospholipids, Soya Seed Phospholipids, Hydrogenated Egg Phospholipids and Soya Seed. Synthetic Phospholipids, PEGylated Phospholipids, and phospholipid combinations sold by Lipoid, LLC. Exemplary of phosphatidylcholine that can be used as a co-surfactant in the compositions provided is the phosphatidylcholine composition sold by Lipoid, LLC, under the name Lipoid S 100, which is derived from a soybean extract and contains more than 95% or more of about 95% phosphatidylcholine.

In one example, the phospholipid, eg, PC, represents less than or equal to 1% or about 1% by weight (w / w) of concentrate. In one example, phosphatidylcholine represents between 0.1% or about 0.1% and 1% or about 1%, for example 0.1, 0.15, 0.2, 0.25, 0.3, 0.35, 0.4, 0.45, 0.5, 0.6, 0.65, 0.66, 0.6690, 0.7, 0.75, 0.8, 0, 85, 0.9, 0.95 or 1% by weight (w / w) of the concentrate. In one example, the phospholipid represents between 0.15% or about 0.15% and 0.7% or about 0.7% by weight (w / w) of the concentrate.

and. Polar solvents

The compositions, including liquid nanoemulsion concentrates and liquid dilution compositions, additionally include polar solvents. Polar solvents are well known in the art. The polarity of a solvent generally indicates which compounds are soluble in the solvent and in which other solvents / liquids the solvent is miscible. Generally speaking, polar compounds are more easily solubilized in water and other polar solvents that are nonpolar compounds. Polar solvents are more likely to be miscible with water and other polar and liquid solvents.

The polarity of a solvent can be evaluated by measuring a number of different parameters according to well known methods (see for example, Prizbytek, High Purity Solvent Guide, Burdickand Jackson Laboratories, Inc., 1980), such as by determining a solvent property such as the dielectric constant, the dipole moment or the polarity index. For example, polar solvents generally have high dielectric constants, typically dielectric constants greater than or about 15 (see for example, Lowery et al., Mechanism and Theory in Organic Chemistry, Harper Collins Publishers, 3^to ed., 1987, p. 177), such as about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, or greater than 90. For example, the dielectric constant of water is at or about 80.10. Polar solvents generally have high polarity indices, typically greater than or about 3 (see, for example, Snyder, Classification of the solvent properties of common liquids, J. Chromatography A, 92: 223-230, 1974), such as of or about 3, 4, 5, 6, 7, 8 or 9 or greater than 9. Polar solvents generally have large dipole moments, typically greater than or about 1 , 4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6 , 3.0, 3.5, 4 or greater than 4 Debye (see, for example, CRC Handbook of Chemistry and Physics, Lide, ed., 82nd edition, CRC Press, 2001, p. 15 (14) 15 (18 )). Other methods for evaluating solvent polarity are known in the art including but not limited to the Kosower Z scale (Kosower, An introduction to physical organic chemistry, Wiley, 1969, p. 293), the donor number and donor acceptor scale ( Gutmann, Solvent effects on the reactivities of organometallic compounds, Coord. Chem. Rev., 18: 225-255, 1976) and the Hildebrand solubility parameters (see, for example, Giddings et al., High pressure gas chromatography of nonvolatile species. Compressed gas is used to cause migration of intractable solutes, Science, 162: 67-73, 1968).

Polar solvents include polar protic solvents and polar aprotic solvents. A polar protic solvent (eg, water, methanol, ethanol) contains a hydrogen atom attached to an electronegative atom, so that hydrogen has a proton-like character and / or the bond between hydrogen and an electronegative atom is polarized . Polar aprotic solvents on the other hand (eg acetone, acetonitrile), generally do not contain positively polarized hydrogen atoms.

ES 2 396 946 T3

Polar solvents in the compositions provided are typically polar protic solvents, including, but not limited to, water; alcohols, including but not limited to dihydric alcohols (eg glycols, eg propylene glycol, ethylene glycol, tetraethylene glycol, triethylene glycol, trimethylene glycol), containing two hydroxyl groups, trihydric alcohols (eg glycerin, butane-1,2,3 -triol, pentane-1,3,5-triol, 2-amino-2-hydroxymethyl-propane-1,3-diol), containing three hydroxyl groups, monohydric alcohols (for example, methanol, ethanol, propanol, isopropanol, n-butanol and t-butanol) and other alcohols; and acids, including but not limited to acetic acid and formic acid. Other polar solvents include, but are not limited to, acetone, acetonitrile, butyl acetate, dimethylformamide, dimethylsulfoxide, dioxane, ethyl acetate, tetrahydrofuran, and hexamethylphosphoric triamide. Typically, the polar solvent is water or is an alcohol that typically contains two or more hydroxyl groups, such as a trihydric or dihydric alcohol, such as, but not limited to, glycerol and propylene glycol. Polar solvents further include low molecular weight polyethylene glycols (PEGs), such as PEGs having a molecular weight of no greater than or about 600 kDa, such as between or about 200 kDa and or about 600 kDa, typically no greater than or about 400 kDa, eg, no more than 200 kDa.

In one example, the polar solvent has a dielectric constant greater than or about 15 and typically between or about 20 and or about 80, such as or about 80.1, 46.53, or 28.67. In another example, the polar solvent has a polarity index between or about 3 and about 9. In another example, the dipole moment of the polar solvent is between 1.5 and 3, and typically between or about 1.8 and 2.8, such as 1.9, 2.6 and 2.2 (for dielectric constants of solvents see, for example, Landolt-Bornstein, New Series IV / 17, Static Dielectric Constants of Pure Liquids and Binary Liquid Mixtures, Springer, 2008; and CRC Handbook of Chemistry and Physics, Lide, ed., 82^to edition, CRC Press, 2001; for the dipole moment of solvents, see, for example, CRC Handbook of Chemistry and Physics, Lide, ed., 82nd edition, CRC Press, 2001) and for polarity indices of solvents, see, for example, Snyder, Classification of the solvent properties of common liquids, J. Chromatography A, 92: 223-230, 1974).

The amount of the polar solvent is typically a high concentration, for example, within a concentration range of between 60% or about 60% and 80% or about 80% by weight (w / w) of the

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liquid concentrates provided 71.74%, 75.8165%, 74.25%, 68.7865% and 68.29% (w / w) of the concentrate. In one example, the concentration range of the polar solvent is between 65% or about 65% and 80% or about 80% (w / w) of the concentrate. In one example, the polar solvent concentration range is between 65% or about 65% and 75% or about 75% (w / w) of the concentrate, or between 65% or about 65% and the 76% or about 76% by weight (w / w) of the concentrate, or between 68% or about 68% and 75% or about 75% by weight (w / w) of the concentrate.

In the provided methods for preparing the concentrates, the polar solvent (eg, water, propylene glycol, or glycerin) is added to the aqueous phase. In one example, the polar solvent is water, for example, purified water, such as water that has been purified before adding to the concentrated formula, for example, by charcoal filter, ion exchange, reverse osmosis, sterilization by UV and / or filtrations using a filter, for example a 50-100 micron filter. Typically, when a filter is used, it is an end-of-use filter that filters the water before it reaches the tank in the process provided. Alternatively, pre-filtered water can be added to the concentrates.

F. Preservatives and Sterilizers

In one example, the liquid concentrate provided further comprises one or more preservatives (or preservatives) and / or sterilizers. The preservative (s) can be included to improve the stability of the concentrate and compositions prepared by diluting the concentrate over time. Preservatives, particularly preservatives for food and beverages are well known. Any known preservative can be used in the compositions provided. Exemplary of preservatives that can be used in the provided compositions are oil-soluble preservatives, eg, benzyl alcohol, Benzyl Benzoate, Methyl Paraben, Propyl Paraben, antioxidants, eg, Vitamin E, Vitamin A Palmitate and Beta Carotene. Typically, a preservative is selected so that it is safe for human consumption, eg, in food and beverages, eg, a GRAS-certified and / or Kosher-certified preservative, eg, benzyl alcohol.

The preservative typically represents less than 1%, less than about 1%, 1%, or about 1% by weight (w / w) of the nanoemulsion liquid concentrate or between 0.1% or about 0.1%. and 1% or

ES 2 396 946 T3 about 1% by weight (w / w) of the concentrate, for example 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0, 6%, 0.7%,

0.725%, 0.75%, 0.8%, 0.9%, 1%, about 0.1%, about 0.2%, about 0.3%, about 0.4%, about 0.5% , about 0.6%, about 0.7%, about 0.8%, about 0.9%, about 1% by weight (w / w) of the liquid concentrate.

g. Emulsion stabilizers (co-emulsifier)

In one example, the liquid concentrates provided additionally contain one or more emulsion stabilizers (co-emulsifiers), which can be used to stabilize the liquid nanoemulsion concentrate and / or the aqueous compositions containing the dilute concentrates. In one example, the emulsion stabilizer increases the viscosity of the liquid concentrate. In one example, one or more emulsion stabilizers are added during formulation, after evaluation of an initial concentrate, particularly if the oil and aqueous phases of the initial concentrate (or the aqueous liquid dilution composition resulting from dilution of the initial concentrate ) it seems that they are separating. The addition of the emulsion stabilizer can prevent the separation of the oil and water phases.

Exemplary of an emulsion stabilizer that can be used in the provided compositions is a composition containing combinations of gums, for example gums used as emulsifying agents, for example, a combination containing one or more of xanthan gum, guar gum and alginate Sodium, for example, the emulsion stabilizer marketed under the trade name SALADIZER®, available from TIC Gums, Inc. (Belcamp, MD). Other gums can be included in the emulsion stabilizer, for example gum arabic and sugar beet pectin. Other similar gum blends can be used as emulsion stabilizers.

The emulsion stabilizer can be added to the aqueous phase, the oil phase, and typically to the aqueous and oil phase during the formation of the liquid concentrates. In one example, the emulsion stabilizer is added to the aqueous phase at a concentration such that it represents less than 1% or about 1% w / w of liquid concentrate. In one example, the emulsion stabilizer is added to the aqueous phase to a final concentration of between 0.1% or about 0.1% and 1% or about 1%, eg 0.1%. 0.12%, 0.13%, 0.14%, 0.15%, 0.16%, 0.17%, 0.18%, 0.19%, 0.2%, 0.25% , 0.3%, 0.31%, 0.32%, 0.33%, 0.34%, 0.35%, 0.36%, 0.37%, 0.38%, 0.39% , 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9% or 1% w / w of the liquid concentrate. In one example, the emulsion stabilizer is added to the oil phase in such a way that it represents less than 0.1% or about 0.1%, for example, between 0.01% or about 0.01% and 0.1% or about 0.1%, for example, 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.061%, 0.062%, 0.063%, 0.0635%, 0.07%, 0.08%, 0.09% or 0.1% by weight (w / w) of the concentrate. In one example, the emulsion stabilizer is added to the water phase and oil phase, for example, at a concentration within the oil and water phase concentration ranges noted above. In one of these examples, the emulsion stabilizer represents less than 1%, for example, between 0.01% or about 0.01% and 1% or about 1% (w / w) of the emulsion stabilizer , for example, 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.061%, 0.062%, 0.063%, 0.0635%, 0 .07%, 0.08%, 0.09%, 0.1%, 0.12%, 0.13%, 0.14%, 0.15%, 0.16%, 0.17%, 0 , 18%, 0.19%, 0.2%, 0.25%, 0.3%, 0.31%, 0.32%, 0.33%, 0.34%, 0.35%, 0 , 36%, 0.37%, 0.38%, 0.39%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9% or 1% by weight (w / w) of the liquid concentrate.

h. Non-polar solvents

In one example, the liquid concentrates additionally contain a non-polar solvent, eg, an oil. Typically, the nonpolar solvent is included in the composition in addition to the nonpolar active ingredient, and is used to dissolve the nonpolar active ingredient. In one example, the solvent is an oil that is not contained in the nonpolar active ingredient. When a non-polar solvent is included in the concentrate, it is typically used to dissolve the non-polar compound before mixing it with the other ingredients, for example, before mixing it with the oil phase ingredients. In one example, the use of a non-polar solvent reduces the size of the crystal and / or increases the transparency of the aqueous liquid dilution composition containing the dilute concentrate. Exemplary of the non-polar solvents that can be used in the concentrates provided are oils (in addition to the non-polar active ingredient) eg Vitamin E oil, linseed oil, CLA, Borage Oil, Dlimonene, Canola oil, Canola oil. corn, MCT oil, and oat oil. Other oils can also be used. Exemplary of the Vitamin E oil, used as a non-polar solvent in the provided compositions, is the oil marketed by ADM Natural Health and Nutrition, Decatur, IL, under the name Novatol ™ 5-67 Vitamin E (D-alpha-Tocopherol; ADM product code 410217). This Vitamin E oil contains at least 67.2% Tocopherol and approximately 32.8% soybean oil.

In one example, the concentration of the non-polar solvent is within a concentration range of between 1% or about 1% and 10% or about 10%, for example, 1%, 2%, 3%, 3.25%, 3.5%, 3.75%, 4%, 5%, 5.25%, 5.5% or 5.75% w / w of the concentrate. In another example, the concentration is within the concentration range of between 3% or about 3% and 6% or about 6%, w / w of the liquid concentrate. In another example, it is between 3.75% and 5.25% w / w of the liquid concentrate.

ES 2 396 946 T3

1. Aromas

In one example, the concentrate additionally contains one or more flavorings or flavoring agents, for example, any compound for adding flavor to the concentrate and / or aqueous dilution liquid composition containing the dilute concentrate, for example, the food or beverage containing the concentrate. Various aromas are well known. Any flavor can be added to the concentrates, for example, any flavor marketed by Mission Flavors, Foothill Ranch, Ca. Examples of flavors that can be used are flavors of fruits such as guava, kiwi, peach, mango, papaya, pineapple, banana, strawberry, currant, blueberry, orange, grapefruit, tangerine, lemon, lime and lemon-lime; cola scents, tea scents, coffee scents, chocolate scents, dairy scents, sarsaparilla and birch soda scents, methyl salicylate (wintergreen oil, sweet birch oil), citrus oils and other scents. Typically the flavors are safe and / or desirable for human consumption, for example GRAS or Kosher certified flavors. Exemplary flavoring agents that can be used in the compositions are lemon oil, for example, lemon oil sold by Mission Flavors, Foothill Ranch, CA; and GRAS certified 99% D-limonene, available from Florida Chemical, Winter Haven, FL. Typically, the flavor is added, using the methods provided, to the nanoemulsion concentrates after combining the oil and water phases. Alternatively, the flavor (s) can be added to the aqueous and / or oily phase directly.

Typically, the concentration of the flavoring agent added to the concentrates provided is less than 5% or about 5%, typically less than 1% or about 1%, for example 0.1%, 0.2%, 0, 3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 0.37%, or 0.525%.

j. PH adjusters

In one example, one or more pH adjusters are added to the provided concentrates, typically to the emulsion that is formed after combining the water and oil phases according to the provided methods. In particular, the pH adjuster is used in compositions containing water. Alternatively, the pH adjuster can be added, at an appropriate concentration to achieve a desired pH, to the oil phase and / or the aqueous phase. Typically, the pH adjuster is added to adjust the pH of the concentrate within a range of 2.0 or about 2.0 to 4.0 or about 4.0. One or more of a plurality of pH adjusting agents can be used. Typically the pH adjusting agent is safe for human consumption, eg GRAS certified. An exemplary pH adjuster is citric acid, eg, citric acid available from Mitsubishi Chemical, Dublin, OH.

Typically, the concentration of the pH adjuster added to the concentrates provided is less than 5% or about 5%, typically less than 1% or about 1%, eg 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 0.28%, or 0.19%.

2. Powder forms of the compositions

The compositions can also be provided in powder form, ie, powder that is prepared by converting the provided nanoemulsion concentrates into a powder, using one of several well-known methods (eg, spray drying and / or milling). Powdered compositions include, but are not limited to, coated or uncoated swallowable or chewable tablets, dry powders in hard or soft gelatin capsules, and dry powders in single or multi-use containers for reconstituted suspensions or sprinkles. Preferable solid dosage forms are coated or uncoated swallowable or chewable tablets. Suitable methods for preparing powder compositions are well known in the art.

Additionally, the powder composition may further comprise at least one excipient. Excipients include, but are not limited to, diluents (sometimes referred to as fillers) including, for example, microcrystalline cellulose, mannitol, lactose, calcium phosphate, dextrates, maltodextrin, starch, sucrose, and pregelatinized starch; disintegrants including, for example, crospovidone, sodium starch glycolate, croscarmellose sodium, starch, pregelatinized starch, and sodium carboxymethyl cellulose; binders including, for example, starch, hydropropylcellulose, hydroxypropylmethylcellulose, pregelatinized starch, guar gum, alginic acid, acacia, sodium carboxymethylcellulose, and polyvinylpyrrolidone; emollients including, for example, colloidal silicon dioxide and talc; and lubricants / release agents including, for example, magnesium stearate, calcium stearate, stearic acid, sodium stearyl fumarate, glyceryl monostearate, hydrogenated vegetable oil, and talc. In a particular example, the excipients are selected from any one or more of maltodextrin and acacia.

The powder forms can be used for any convenient dosage amount of the nonpolar compound. Generally, the level of the nonpolar compound can be increased or decreased in accordance with the judgment of the physician, pharmacist, pharmaceutical scientist, or other person skilled in the art. The amount of the remaining non-active ingredients can be adjusted as needed.

ES 2 396 946 T3

Typically, the concentration of the excipients is within a concentration range of between 50% or about 50% and 85% or about 85%, for example, 50, 51, 52, 53, 54, 55, 56 , 57, 58, 59,

60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85 or greater,% by weight of the free-flowing powder.

The powder forms can be used for any convenient dosage amount of the nonpolar compound. Generally, the level of the nonpolar compound can be increased or decreased in accordance with the judgment of the physician, pharmacist, pharmaceutical scientist, or other person skilled in the art. The amount of remaining non-active ingredients can be adjusted as needed.

In one example, the powder form is a free-flowing powder. Free flowing powders can be obtained using techniques well known in the art, such that, but are not limited to, spray drying, freeze drying or absorption plating. In one example, to achieve a free flowing powder, the protein derivative is formulated with an excipient such as lactose or starch. For example, the formulation can be a spray-dried lactose formulation (see, for example, US Patent No. 4,916,163).

Methods for forming the powders include spray drying. Spray drying processes and spray drying equipment are generally described in Perry's Chemical Engineers' Handbook, pages 20-54 to 20-57 (Sixth Edition 1984). More details on spray drying processes and equipment are reviewed by Marshall, Atomization and Spray-Drying, 50 Chem. Eng. Prog. Monogr. Series 2 (1954), and Masters, Spray Drying Handbook (Fourth Edition 1985). Methods for spray drying are well known (see, for example, US Patent No. 5,430,021; 6,534,085 and US Application Publication No. US2007 / 0184117). In general, spray drying is used to dry a heated liquid by passing it through hot gas. One or more spray nozzles are used to atomize the liquid in a cooling tower or chamber. As the material is atomized (pulverized) the surface tension causes a uniform spherical particle to form, which is passed through the cooling chamber and hardens into a solid intact sphere. The spray-dried particles can be between or about 0.5 microns and or about 100 microns, and typically are less than or about 10 microns, typically less than or about 5 microns, and typically less than or about or or about 1 micrometer.

Methods are provided for spray drying liquid nanoemulsion compositions to form powder compositions. In spray drying methods, liquid nanoemulsion compositions can be heated, for example, to a temperature between or about 100 (37.777 ° C) and or about 150 ° F (65.555 ° C), typically between 110 ° F (43,333 ° C) and 140 ° F (60 ° C), for example, from or about 110 (43,333 ° C), 115 (46,111 ° C), 120 (48,888 ° C), 125 (51,666 ° C), 130 (54.444 ° C), 135 (57.222 ° C) or 140 ° F (60 ° C). The compositions can be mixed while heating, such as with any of the mixers described herein, eg, homogenizers (eg, reversible homogenizers and piston-actuated homogenization).

For spray drying, one or more excipients are mixed with a polar solvent, typically water, and heated, for example, to a temperature between or about 100 ° F (37.777 ° C) and or about 150 ° F ( 65,555 ° C), typically between 110 ° F (43,333 ° C) and 140 ° F (60 ° C), for example, from or about 110 (43,333 ° C), 115 (46,111 ° C), 120 (48,888 ° C ), 125 (51.666 ° C), 130 (54.444 ° C), 135 (57.222 ° C) or 140 ° F (60 ° C). In one example, the excipient is mixed with water in an amount of one part excipient (by weight) to two parts water (by weight). The excipient-solvent mixture (eg, water) can be mixed while heating, eg, using any of the mixers described herein, eg, homogenizers (eg, reversible homogenizers and piston-actuated homogenization) while heating during mixing. The heated liquid nanoemulsion composition and the heated water-excipient mixture are then mixed together, such as by transferring one mixture to the other, for example, by any of the transfer means provided herein. Typically, the two mixtures are homogenized, for example, with a reversible homogenizer or a piston-driven homogenizer or any other homogenizer. The homogenized mixture is then subjected to spray drying using a spray drier.

They are exemplary of the spray dryers cyclone spray dryers. During spray drying with cyclone spray dryers, the homogenized mixture is pumped into a spray device where it degrades into small droplets. Upon contact with a stream of hot air, moisture is very quickly removed from the droplets while they are still suspended in the drying air. Drying dust is separated from humid air in cyclones by centrifugal action. The centrifugal action is caused by the large increase in air velocity when the mixture of particles and air enters the cyclone system. The denser dust particles are forced toward the walls of the cyclone while the lighter, moist air is directed away through the exhaust pipes. The dust settles at the bottom of the cyclone where it is removed through a discharge device. Sometimes the air transport ducts for the dry powder are connected with cooling systems that admit cold air for the transport of the product through transport tubes. Cyclone dryers have been designed for production schemes capable of drying batches of tons of

ES 2 396 946 T3 dust per hour.

As will be appreciated by one of ordinary skill in the art, the inlet temperature and outlet temperature of the spray dryer are not critical but will be of a level such that it provides the desired particle size, less than or about 1 micron and results in a powder that has a desired property. Typically, the ability of the free-flowing powder to produce a clear (or relatively clear) dilution liquid composition upon dilution in an aqueous medium is the desired property that is evaluated. In this sense, the inlet and outlet temperatures are adjusted depending on the melting characteristics of the liquid nanoemulsion components and the composition of the homogenized mixture of liquid nanoemulsion / excipient concentrate. The inlet temperature is between or about 60 ° C and or about 170 ° C, with outlet temperatures of or about 40 ° C up to or about 120 ° C. Preferably the inlet temperatures are from or about 90 ° C to or about 120 ° C and outlet temperatures are from or about 60 ° C to or about 90 ° C. The flow rate used in spray drying equipment will generally be from or about 3 ml per minute to at or about 15 ml per minute. The air flow of the atomizer will vary between values ​​of or approximately 25 l per minute up to or approximately 50 l per minute. Commercially available spray dryers are well known to those of skill in the art and suitable accessories for any dispersion can be readily determined by one of ordinary skill in the art without undue experimentation. Operating conditions such as inlet temperature and outlet temperature, flow rate, atomization pressure, drying air flow rate, and nozzle configuration can be adjusted according to the manufacturer's guidelines.

In some examples, the dry powder is stored in a capsule form or pressed into a tablet form. For use as tablets, the compositions typically contain multiple other excipients. These excipients include tablet disintegrants, such as cornstarch, emollients, such as silicon dioxide, and lubricants, such as magnesium stearate. Ordinarily these compositions contain minor amounts by weight of emollients and lubricants, eg, every two percent (2%) or less by weight. Tablet disintegrators are optionally present and, if present, are included in amounts sufficient to ensure that the tablet disintegrates upon ingestion. Accordingly, materials such as cornstarch are employed at concentrations of from about zero to about 30 percent by weight of the composition.

Free-flowing powders can also be used to deliver the active agent by inhalation using a dry powder inhaler. Such dry powder inhalers typically deliver the active agent as a free-flowing powder that disperses in a patient's air stream during inspiration. To get the free flowing powder in the active agent it is typically formulated with a suitable excipient such as lactose or starch. For example, such a dry powder formulation can be prepared, for example, by combining the lactose with the active agent and then dry combining the components. Alternatively, if desired, the active agent can be formulated without an excipient. The pharmaceutical composition is then typically loaded into a dry powder dispenser, or inhalation cartridges or capsules for use with a dry powder delivery device. Examples of dry powder inhaler delivery devices include Diskhaler (GlaxoSmithKline, Research Triangle Park, NC) (see, for example, US Patent No. 5,035,237); Diskus (GlaxoSmithKline) (see, for example, US Patent No. 6,378,519; Turbuhaler (AstraZeneca, Wilmington, Del.) (See, for example, US Patent No. 4,524,769); Rotahaler (GlaxoSmithKline) (see, for example, US Patent No. 4,353,365) and Handihaler (Boehringer Ingelheim). Other examples of suitable DPI devices are described in US Patent Nos. 5,415,162, 5,239,993, and 5,715,810 and references cited therein.

3. Liquid dilution compositions containing the dilute concentrates

Among the compositions provided herein are liquid dilution compositions, typically aqueous liquid dilution compositions, containing the nonpolar compounds. Aqueous liquid dilution compositions are prepared by diluting the provided nanoemulsion liquid concentrates in aqueous media, eg beverages, eg water, flavored water, soda, milk, juices, including fruit juices, sauces, syrups, soups, beverages sports drinks, nutritional drinks, energy drinks, vitamin fortified drinks or any drink.

In one example, aqueous liquid dilution compositions contain between 0.05 grams (g) or about 0.05 g and 10 g or about 10 g, typically between 0.05 g and 5 g of the liquid concentrate per 8 fluid ounces (0.236588 liters) approximately 8 fluid ounces (0.236588 liters), at least 8 fluid ounces (0.236588 liters) or at least approximately 8 fluid ounces (0.236588 liters), or less than 8 fluid ounces (0.236588 liters) or less than about 8 fluid ounces (0.236588 liters) or per size of each feed, of the aqueous medium, for example, 0.05 g, 0.06 g, 0 .07 g, 0.08 g, 0.09 g, 0.1 g, 0.2 g, 0.3 g, 0.4 g, 0.5 g, 0.6 g, 0.7 g, 0 , 8 g, 0.9 g, 1 g, 2 g, 3 g, 4 g, 5 g, 6 g, 7 g, 8 g, 9 g or 10 g of the concentrate per 8 fluid ounces (0.236588 liters), approximately 8 fluid ounces (0.236588 liters), or at least 8 fluid ounces (0.236588 liters) or at least about 8 fluid ounces (0.236588 liters) of the aqueous medium, for example, 8 (0.236588 liters), 9 (0.26616 liters), 10 ( 0.29574 liters), 11 (0.32531 liters), 12 (0.35488

ES 2 396 946 T3 liters), 13 (0.38446 liters), 14 (0.41403 liters), 15 (0.44360 liters), 16 (0.47318 liters), 17 (0.50275 liters), 18 ( 0.53232 liters), 19 (0.56190 liters), 20 (0.59147 liters), 25 (0.73934 liters), 30 (0.88721 liters), 35 (1.0351 liters), 40 (1, 1829 liters), 45 (1.3308 liters), 50 (1.4787 liters), 100 (2.9574 liters), 200 (5.9147 liters) or more fluid ounces of aqueous medium.

In another example, the dilution liquid composition contains between 1 ml or about 1 ml and 10 ml or about 10 ml of the liquid concentrate, for example, 1 ml, 2 ml, 3 ml, 4 ml, 5 ml, 6 ml, 7 ml, 8 ml, 9 ml or 10 ml of the concentrate, per 8 fluid ounces (0.236588 liters), approximately 8 fluid ounces (0.236588 liters), at least 8 fluid ounces (0.236588 liters) or at least about 8 fluid ounces (0.236588 liters) or less than 8 fluid ounces (0.236588 liters) or less than about 8 fluid ounces (0.236588 liters) or per serving size of aqueous medium, for example, 8 (0.236588 liters), 9 (0.26616 liters), 10 (0.29574 liters), 11 (0.32531 liters), 12 (0.35488 liters) , 13 (0.38446 liters), 14 (0.41403 liters), 15 (0.44360 liters), 16 (0.47318 liters), 17 (0.50275 liters), 18 (0.53232 liters), 19 (0.56190 liters), 20 (0.59147 liters), 25 (0.73934 liters), 30 (0.88721 liters), 35 (1.0351 liters), 40 (1.1829 liters), 45 (1 , 3308 liters), 50 (1.4787 liters), 100 (2.9574 liters), 200 (5.9147 liters) or more fluid ounces of the aqueous medium.

In another example, the aqueous liquid dilution composition contains at least 10 mg or about 10 mg, typically at least 25 mg or about 25 mg, typically at least 35 mg of the non-polar compound, eg, the non-polar active ingredient, for example. 8 fluid ounces (0.236588 liters) or approximately 8 fluid ounces (0.236588 liters), at least 8 fluid ounces (0.236588 liters) or at least about 8 fluid ounces (0.236588 liters) of the aqueous medium or less than 8 fluid ounces (0.236588 liters) or less than about 8 fluid ounces (0.236588 liters) or by service size of the aqueous medium; for example, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 25, 25, 26, 27, 28, 29, 30, 31, 32, 33 , 34, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200 , 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 325, 350, 375, 400, 425, 450, 475, 500, 550, 600, 700, 800, 900, 1000, 1500 , 2000 mg, or more of the nonpolar compound per at least 8 fluid ounces (0.236588 liters) or at least about 8 fluid ounces (0.236588 liters) of aqueous medium.

In another example, the aqueous liquid dilution composition contains the concentrate diluted to a dilution factor of between 1:10 or about 1:10 and 1: 1000 or about 1: 1000 or greater, typically between 1:10 or about 1: 10 and 1: 500 or about 1: 500 or greater, for example, diluted no more than 1:10 or about 1:10, 1:20 or about 1:20, 1:25 or about 1:25, 1:50 or about 1:50, 1: 100 or about 1: 100,1: 200 or about 1: 200,1: 250 or about 1: 250,1: 300 or about 1: 300,1: 400 or about 1: 400, 1: 500 or about 1: 500, for example, 1:10, 1:20, 1:25, 1:30, 1:35, 1:40, 1:50, 1:55, 1:60, 1:65, 1:70 , 1:75, 1:80, 1:90, 1: 100, 1: 110, 1: 120, 1: 130, 1: 140, 1: 150, 1: 160, 1: 170, 1: 180, 1 : 190, 1: 200, 1: 210, 1: 220, 1: 230, 1: 235, 1: 240, 1: 250, 1: 260, 1: 270, 1: 280, 1: 290, 1: 300 , 1: 350, 1: 400, 1: 450, 1: 500 or greater. In another example, the aqueous liquid dilution compositions contain the liquid concentrate diluted in any amount. In another example, the dilution is less than 1:10 or about 1:10.

The properties of the provided liquid concentrates that are diluted in the aqueous medium contribute to various properties of the resulting aqueous liquid dilution compositions, eg, transparency; desirably for human consumption, eg, palatable taste and / or odor, eg, no fishy taste / odor, no ring formation, and no crystal formation; stability, eg, no oxidation, ring formation, and / or precipitation over time; and safety for human consumption. As described above, the liquid concentrates are formulated in accordance with the desired properties of the aqueous liquid dilution compositions containing the concentrates.

to. Transparency

In one example, aqueous dilution liquid compositions are clear dilution aqueous liquid compositions or non-turbid dilution aqueous liquid compositions, for example, as determined as described below, empirically or by measuring turbidity and / or particle size. . In another example, aqueous liquid dilution compositions are not transparent, or are not completely transparent. The liquids can be more or less transparent, or have the same transparency as another liquid, for example, an aqueous dilution liquid composition prepared according to the methods provided or a beverage, for example, a beverage that does not contain the diluted concentrate. The properties of liquid concentrates can affect the transparency of the liquid. A number of parameters can vary the transparency of the liquids, for example, the relative concentration of the surfactant, nonpolar compound and / or water; the type of non-polar ingredient; the concentration of the excipient (s) in the particular non-polar compound and the purity of the non-polar compound, for example, whether it has been standardized to high purity, or whether it is an extract or a filtered extract. For example, an aqueous liquid dilution composition prepared by diluting a concentrate containing a non-polar active ingredient that contains lecithin, for example, a high amount of lecithin, may be less transparent than one prepared with a concentrate containing a non-polar compound that does not contain lecithin. In another example, a liquid concentrate containing a nonpolar compound that is a filtered extract can produce a more transparent dilution aqueous liquid composition when diluted than a concentrate containing a crude extract.

ES 2 396 946 T3

i. Transparency determined by empirical evaluation

In one example, the transparency / turbidity of the dilution aqueous liquid composition containing the dilute concentrate is qualitatively evaluated, by observation. In one example, a liquid can be considered transparent if it does not have an opaque appearance and / or if there are no or very few particles visible when liquid is seen with the naked eye or if it is the same or substantially similar in transparency to another liquid, for example , a drink, for example water, fruit juice, soda or milk. In some cases, the aqueous liquid dilution composition is as clear or about as clear as water or another liquid, eg, a beverage. For example, the liquid (containing the liquid concentrate diluted in an aqueous medium, eg a beverage), may be as clear or about as clear as the aqueous medium that does not contain the liquid concentrate. In a related example, there is no substantial difference, for example, there is no observable difference between the aqueous liquid dilution composition containing the concentrate and the aqueous medium without the concentrate. A clear liquid is not necessarily colorless, for example, a yellow liquid that does not contain visible particles or opacity can be considered transparent. In another example, the liquid is transparent or partially transparent or substantially transparent if there are no visible crystals and / or if no ring formation is observed in the container containing the liquid.

ii. Transparency determined by particle size or number of particles

In another example, the transparency of the aqueous dilution liquid composition is evaluated by measuring the particle size and / or the number of particles in the liquid.

In one example, aqueous dilution liquid compositions have a particle size of less than 200 nm or less than about 200 nm, for example, 5, 10, 15, 20, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 60, 70, 80, 90, 100, 110 , 120, 130, 140, 150, 160, 170, 180, 190 or 200 nm. In another example, the aqueous liquid dilution composition has a particle size less than 100 nm or about 100 nm, less than 50 nm or about 50 nm, or less than 25 nm or about 25 nm. Typically, the particle size of the aqueous liquid dilution composition is between 5 nm or about 5 nm and 200 nm or about 200 nm, or between 5 nm or about 5 nm and 50 nm or about 50 nm.

Typically, the particle size of the provided aqueous dilution liquid composition containing the liquid concentrate containing the nonpolar compound is smaller than the particle size of a liquid containing the nonpolar compound (not formulated in a liquid concentrate ).

iii. Turbidity

In another example, the transparency of the liquid is evaluated and / or expressed using a turbidity measurement, eg, Nephelometric Turbidity Units (NTU), as measured using the provided methods, described below. In this example, the turbidity is measured optically, to obtain a value indicative of the opacity or turbidity of the liquid, which is correlated with the particles suspended in the liquid. The more transparent a liquid is, the lower its haze value.

In one example, the aqueous liquid dilution composition has a haze value (NTU) of 30 or about 30; or an NTU value of less than 30 or about 30, for example, less than 29 or about 29, less than 28 or about 28, less than 27 or about 27, less than 26 or about 26, less than 25 or about 25, less than 24 or about 24, less than 23 or about 23, less than 22 or about 22, less than 21 or about 21, less than 20 or about 20, less than 19 or about 19, less than 18 or about 18, less than 17 or about 17, less than 16 or about 16, less than 15 or about 15, less than 14 or about 14, less than 13 or about 13, less than 12 or about 12, less than 11 or about 11, less than 10 or about 10, less than 9 or about 9, less than 8 or about 8, less than 7 or about 7, less than 6 or about 6, less than 5 or about 5, less than 4 or about 4, less than 3 or about 3, less than 2 or about 2, less than 1 or about 1; or 29 or about 29, 28 or about 28, 27 or about 27, 26 or about 26, 25 or about 25, 24 or about 24, 23 or about 23, 22 or about 22, 21 or about 21, 20 or about 20 , 19 or about 19, 18 or about 18, 17 or about 17, 16 or about 16, 15 or about 15, 14 or about 14, 13 or about 13, 12 or about 12, 11 or about 11, 10 or about 10, 9 or about 9, 8 or about 8, 7 or about 7, 6 or about 6.5 or about 5, 4 or about 4, 3 or about 3, 2 or about 2, 1 or about 1, or 0 or about 0.

In another example, the haze value of the aqueous liquid dilution composition is less than 200 or less than about 200, eg, 200, 175, 150, 100, 50, 25 or less.

ES 2 396 946 T3

In another example, it is desirable that the aqueous dilution liquid composition contains a turbidity value that is comparable, for example, that is approximately equal to, equal to or less than or greater than the turbidity value of another liquid, for example, a beverage. It does not contain the liquid concentrate or an aqueous liquid dilution composition prepared by the methods provided.

b. Stability

Typically, the aqueous liquid dilution compositions provided containing the concentrates are stable, eg, free from one or more changes over a period of time, eg, 1, 2, 3, 4, 5, 6, 7, 8 , 9, 10, 11 or 12 months, 1, 2, 3, 4 or more years.

In one example, the compositions are stable in that they are free from oxidation or substantial oxidation over time. In another example, they are stable because they remain transparent over time. In another example, stable compositions remain safe and / or desirable for human consumption over time. In one example, stability refers to the absence of precipitates that form in the compositions over the period of time. In a related example, the compositions are stable in that they do not exhibit ring formation, off-white or opaque ring formation around the perimeter of the container containing the liquid, typically the surface of the liquid. Ring formation is typically undesirable, particularly in the case of a liquid for human consumption, for example a beverage.

In another example, the composition is stable if it does not exhibit any visible phase separation for a period of time, eg, after 24 hours, after a week, or after a month. In one example, compositions are stable if they exhibit one or more of these described characteristics, over time, when held at a particular temperature. In one example, the compositions remain stable at room temperature, eg, 25 ° C or about 25 ° C. In another example, the compositions remain stable between 19 ° C and 25 ° C. In another example, the compositions remain stable at refrigerated temperatures, eg, 4 ° C or about 4 ° C, or at a frozen temperature, eg, at -20 ° C or about 20 ° C.

Stability refers to a desirable property of the provided compositions, eg, the ability of the provided compositions to remain free from one or more changes over a period of time, eg, 1, 2, 3, 4, 5, 6. , 7, 8, 9, 10, 11 or 12 months, 1, 2, 3, 4 or more years. In one example, the composition is stable if it is formulated so that it remains free from oxidation or substantial oxidation over time. In another example, stable compositions remain transparent over time. In another example, stable compositions remain safe and / or desirable for human consumption over time. In one example, stability refers to the absence of precipitates that form in the compositions over the period of time. In a related example, stability refers to the absence of ring formation over the period of time. In another example, the composition is stable if it does not exhibit any visible phase separation over the period of time, eg, after 24 hours, after a week, or after a month. In one example, compositions are stable if they exhibit one or more of these described characteristics, over time, when held at a particular temperature.

In one example, the compositions are stable at room temperature, eg, 25 ° C or about 25 ° C. In another example, the compositions remain stable between 19 ° C and 25 ° C. In another example, the compositions remain stable at chilled temperatures, eg, 4 ° C or about 4 ° C, or at a frozen temperature, eg, at -20 ° C or about -20 ° C.

c. Desirable characteristics for human consumption

In one example, the liquid dilution composition is desirable for human consumption, eg, for use in a food or beverage. The different properties of the liquid dilution composition can contribute to the desirability as a consumable product. For example, taste, odor, transparency, color, crystal formation, precipitation, and ring formation can all be related to desirability.

In one example, the liquid dilution composition has a pleasant taste and / or odor, for example, due to one or more flavors added to the concentrate and / or aqueous medium. In another example, the liquid dilution composition containing the concentrate is free of an unpleasant taste or odor, for example, a fishy taste or odor. In one example, the concentrate smells or tastes less unpleasant, eg fishy, ​​compared to another aqueous liquid dilution composition.

In another example, the aqueous liquid dilution composition is desirable because it has no or fewer crystals compared to another aqueous liquid dilution composition. In another example, the aqueous liquid dilution composition is desirable in that it does not exhibit ring formation.

d. Security

ES 2 396 946 T3

Typically, the aqueous liquid dilution compositions containing the concentrates are safe for human consumption, eg, containing only FDA-approved ingredients for human consumption, eg, GRAS certified ingredients. In one example, one or more of the ingredients, for example, all ingredients are certified Kosher. The safety of the compositions also refers to stability over time. The absence of minimal oxidation of the compositions over time can contribute to the safety of the compositions.

and. Oral bioavailability

In one example, non-polar compounds, eg, non-polar active ingredients, contained in aqueous dilution liquid compositions exhibit high or relatively high bioavailability, eg, bioavailability that is greater than a liquid containing only the ingredient. nonpolar active (that is, it is not formulated in the liquid concentrate). Bioavailability refers to the body's ability to absorb the nonpolar active ingredient in a particular space, cell tissue, and / or cell compartment. Typically, non-polar active ingredients in liquids that have small particle sizes are better absorbed than those with larger particle sizes.

C. METHODS FOR PREPARING LIQUID NANOEMULSION CONCENTRATES CONTAINING NON-POLAR COMPOUNDS

Methods for preparing the liquid nanoemulsion concentrates are also described. The general equipment and the stages of the methods are detailed below. In one example, the general methods for preparing the concentrates are performed using an experimental scale manufacturing process that is used to prepare relatively smaller batches of the concentrates. In another example, the general methods for preparing the concentrates are performed using upscaling manufacturing processes, which are used to prepare relatively larger batches of the concentrates. The experimental scale process can be scaled up to large scale processes. Any concentrate prepared using the experimental method can be prepared using the large-scale process, increasing the scale of the method.

1. Equipment for preparing concentrates

Various equipment, for example, containers for mixing the oil phase, water phase and emulsion, for example, tanks; Balances; mixers including conventional mixers and homogenizers; heating and cooling apparatus, including jacketed water tanks, hot plates, water baths and coolers (coolers), including recirculating coolers; transfer apparatus, eg, transfer means, eg, pumps, hoses, sanitary fittings; Ball valves; purifiers eg filters eg carbon filters, ion exchange equipment, reverse osmosis equipment, end point filters and end product filters; evaluation means for example, pH and temperature meters; and other equipment is used in various stages of the methods provided to prepare the concentrates. The choice of equipment depends on a number of factors, including the batch size and the manufacturing process.

2. Balances

One or more balances are typically used to measure the ingredients before adding them to the appropriate container. Alternatively, the ingredients can be weighed into the container, for example in a tank on top of a balance.

Any of a plurality of well known commercially available balances can be used to weigh ingredients. The choice of balance (s) can depend on a number of factors, including the mass of the final concentrate that is prepared and the ingredient that is weighed. In one example, multiple balances are used to weigh the various ingredients of the concentrate. In general, a relatively larger capacity (weight) scale or scales are used in preparing larger batches of concentrate while a relatively smaller capacity scale or scales are used to prepare smaller batches.

Exemplary of balances used with the methods provided to weigh ingredients are the Toledo Balance (Model GD13x / USA), a Sartorius Basic Analytical Balance (Model BA11 OS) which is a basic series analytical balance with a capacity of 110 g and a resolution 0.1 mg; and an OHAUS Balance (Model CS2000), which is a compact portable digital scale that has a capacity of 2000 g and a resolution of 1 g.

to. Purifiers, including filters

Purifiers, typically more than one purifier, eg filters are used in the provided methods to remove impurities in the ingredients prior to their addition to the concentrate and / or from the final concentrate and / or an intermediate phase of the concentrate. For example, when the polar solvent is water, the water is typically purified water. In one example, one or more purifiers, for example carbon filters, ion exchange purifiers, reverse osmosis purifiers and / or end-point filters are used to filter water, for example water from

ES 2 396 946 T3 the city, before its addition to the aqueous phase, for example, to remove impurity, for example, sediments from the water.

Exemplary of the purifiers that can be used with the methods provided are filters, for example, 100 micron filters and carbon filters, which are filters that use activated carbon to remove impurities by chemical adsorption. Carbon filters are typically used for water purification and are particularly effective at filtering out chlorine, sediment, volatile organic compounds, and other impurities. Typically, the particles removed by the carbon filters are between about 0.5 microns and about 50 microns. Other filters are well known and can be used with the methods provided.

Also exemplary of the purifiers that can be used in the provided methods are reverse osmosis purifiers, which use mechanical pressure to purify liquids, eg, water. In one example, pressure forces water through a semi-permeable membrane to remove impurities.

Also exemplary of the purifiers that can be used in the methods provided are ion exchange purifiers, for example an ion exchange purifier using a resin bed, for example a zeolite resin bed, to replace salts. , for example, cations, for example, magnesium and calcium, with other cations, for example, sodium and potassium cations. Such purifiers can be purchased, for example, from Aquapure Filters, Clarkston, Ml.

In another example, a final product filter (for example, a 100 micron FSI filter, BPEM Product Number 100-5GP). This filter is used to filter out any impurities from the final product (for example, the final liquid nanoemulsion composition). Other filters are known and can be used with the methods provided.

b. Containers for mixing ingredients

One or more, typically two or more vessels, eg tanks, eg water jacketed tanks; jars; and / or beakers, eg, Pyrex® beakers, are used in the methods provided to contain the ingredient (s) of the liquid concentrates, eg, during mixing and / or heating or cooling. Typically, different vessels (an oil phase tank and a water phase tank) are used to mix and heat the ingredients of the oil phase and the water phase before combining the two phases to form an emulsion. In another example, an additional container, eg, a containment and / or packaging tank , is used to contain and / or package the emulsion and / or for adding / mixing the additional ingredients to the emulsion.

A number of containers are available for mixing the ingredients. Typically, containers are cleaned, eg, rinsed, soaped and / or sanitized according to known procedures, before use and between uses.

In one example, typically used with experimental scale processes, the container is a container, for example an experimental scale container, for example flasks, beakers, for example Pyrex® beakers, vials, containers of measuring, flasks and / or other containers of experimental scale.

In another example, typically used with the larger scale manufacturing process, the vessels are tanks, eg, water phase tanks, oil phase tanks, and containment / packaging tanks. Typically the tanks are equipped with one or more mixers, for example a conventional mixer and / or homogenizer, which are used to mix the ingredients added to the tank. In one example, the tank is additionally equipped with a heating and / or cooling device. For example, the tank can be a water jacketed tank. The temperature of the water jacketed tank is controlled through the water jacket for example to heat the contents, for example while mixing.

Examples of tanks that can be used with the methods are water-jacketed tanks, for example, the Overly 550-gallon (1,892-liter) Water-Jacketed Tank (Model 10576501G), which can have a capacity of 550 gallons (1,892 liters) and is typically used as a water phase tank, the Schweitzers 450 gallon (1,703.43 liter) tank (Model No. 5214-C), which has a 450 gallon (1,703.43 liter) capacity and is typically used as an oil phase tank and the 190 gallon (719.23 liter) Royal water jacket tank (Model 9977-5), which has a 190 gallon (719.23 liter) capacity and can be used as a water or oil phase tank when mixing small volumes. Other tanks are well known and can be used with the methods provided to mix the concentrates, for example the phases of the concentrates.

c. Mixers

Mixers are used in the methods for combining, mixing and / or emulsifying the liquid concentrates and / or various ingredients and / or phases of the liquid concentrates. In one example, mixers are used to keep ingredients and / or mix circulating to maintain temperature, viscosity, and / or other parameters of the

ES 2 396 946 T3 mixed. Exemplary of the mixers that can be used in the methods provided are conventional mixers, for example, conventional mixers that can be used, for example, to mix the ingredients in the aqueous and / or oil phases, to maintain a homogeneous mixture while heating. Exemplary of conventional mixes are the LIGHTNIN® mixer (LIGHTNIN, Rochester, NY), eg, Model Numbers XJC117 and ND-2. In one example, LIGHTNIN® mixers are fixed mount, gear driven, high flow mixers for use with closed tanks. Another example of the conventional mixer is a mixer marketed by IKA®, for example IKA® top head mixers, for example, models N ° RW-14 Basic and RE-165, which are laboratory stirrers and can be used for mixing ingredients, for example, to generate the oil and water phases. In one example, the mixer or mixers are attached to the vessels, eg, the tanks, eg, mounted or secured on the tanks, eg, on top of the tanks. In another example, the mixers are located in the vessels for mixing.

Also exemplary of mixers used with the methods are homogenizers (also called shears), which are typically used to form the emulsion by emulsifying the oil and water phases after they are combined. Homogenizers typically provide shear dispersion of solids and emulsion of immiscible liquids at high shear rates. Exemplary of the homogenizers that can be used in the methods provided are high-shear homogenizers, for example, reverse homogenizers sold by Arde Barinco, Inc., Norwood, NJ, for example, the Model CJ-50, which is a 3600 rpm mixer. which has a rotor diameter of 6 inches (15.24 cm), a top speed of 5575 ft / minute (1,699.26 m / min) and a depth of emergence of 33 inches (83.82 cm) and six separate openings at the bottom and top, which concentrate the liquid in six chambers, reducing surface volume and creating a shear effect, and the Model CJ-4E, which is a 10,000 rpm mixer with a fan-cooled motor, optimized for batch sizes of 1 to 5 gallons (3.79 to 18.93 liters) having a rotor diameter of 1.875 inches (4.76 cm), a top speed of 4920 rpm and a depth of immersion of 16 inches (40.64 cm). Other homogenizers, for example other reversible homogenizers available from Arde Barinco Inc., can be used with the methods.

In one example, the homogenizer is attached to the top of the container, for example, the tank, for example, by jaws or by channel locks and an electric lift. In another example, the homogenizer is located in the container. Arde Barinco reversible homogenizers contain axial flow impellers, which create two different mixing actions, depending on direction. Downward vortex flow pushes solids from top to bottom of the mix, while upward umbrella flow controls the mix at the highest shear rates and recirculation without splashing or entrainment of air. Reversible homogenizers are typically equipped with an adjustable baffle plate, which can be adjusted to control the type of mix, for example at different times during emulsification.

A number of additional mixers are well known and can be used with the methods. They are exemplary of mixers that can be used with the shear, in-line mixer / mixing methods. Forberg Belt, Plow &amp; Paddle Mixers, Conveyors, Bag Tipper &amp; Compactors, V-Shaped Mixers, Paddle Mixers, Double Cone Mixers, Continuous Mixers, Fast Flow Mixers, Discontinuous Mixers, Double Belt Mixers, Belt Mixers and bladed blades, Plow Mixers / Turbulence Mixers, Fluidizing Forberg Mixers, Air Mixers, Active Mixers, Passive Mixers, Top Entry Mixers, Side Entry Mixers, Static Mixers, Fixed Entry Mixers, Portable Mixers - Direct Drive and Gear Driven, Hygienic Mixers, Drum Mixers, IBC Bulk Container Mixers, Laboratory Agitators, Variable Speed ​​Mixers , kneader, vertical mixer, spiral mixer, double arm mixer, fork mixer, double spiral mixer, all agitators, Mixer-shaker, Banbury Mixers, Rubber Mixers, Blondheim Mixers, Butter Mixers, Conical Mixers, Continuous Mixers, Disperser Mixers, Concrete Mixer, Emulsifier Mixers, Hobart Mixers, Blender Mixer, Littleford Mixer, Meat Mixer, Littleford Mixer, Mincer Mixer Mixmuller, Nauta Mixers, Oakes Mixers, Planetary Mixers, Pony Mixers, PUG Mixers, Ribbon Mixers, Ross Mixer, Rotary Mixers, Sigma Mixers, Single Arm Mixers, Tote Bin Mixers, Turning Mixer, Suction Mixers, Turbine Mixer, Double Layer Mixers, V-Shaped Mixer Types, Zigzag Mixers, side arm mixers, portable mixers, stir bars, stir bars, magnetic mixers and top mixers, for example mechanical and / or electrical top mixers. .

d. Heating devices

One or more, typically more than one heating apparatus is used in the methods for controlling the temperature of the ingredients, phases and / or concentrate typically while mixing.

In one example, the heating appliances are water jackets. In this example, the vessels used to mix the ingredients and / or emulsify the phases are water jacketed tanks. The water jacket can be controlled, for example, using a control panel, to adjust the temperature of the contents of the container.

ES 2 396 946 T3

Alternatively, other heating apparatus can be used to heat the ingredients, phases, and / or concentrates. Exemplary heating appliances that can be used with the methods are immersion and / or submersible heaters, for example 12 KW or 13 KW sanitary heaters, which are food grade heaters that are immersed in tanks while mixing, typically for applications that require high heat, for example temperatures greater than approximately 60 ° C or 60 ° C or greater than 80 ° C or approximately 80 ° C. Also exemplary of the heating appliances are stoves, for example, propane stoves. Also exemplary of hot plate warming appliances are, for example, the Thermolyne hot plate, model number 846925 and model number SP46615. Typically the heater is capable of heating the mixture between 45 ° C or about 45 ° C and 85 ° C or about 85 ° C, for example 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84 or 85 ° C. Typically, the heater is capable of heating the mixture to 60 ° C or 60 ° C, for example, providing low heat.

and. Cooling appliances

One or more cooling apparatuses may be used with the methods, for example, to cool the ingredients during mixing, for example, to cool the mixture while emulsifying the oil and water phases. They are exemplary of chiller cooling apparatus, for example recirculating chillers, which can be attached to the vessel, for example remotely or via a tank mounted on the chiller, to recirculate fluid from the tank, through the chiller and back to the tank. to cool quickly and maintain mix temperature during mixing. They are exemplary of an open loop chiller that can be attached to the tank and used with the methods provided chillers marketed by Turmoil, West Swanzey, NH, for example open or closed loop chillers, for example Model No. OC-1000 RO . Other cooling apparatuses are well known and can be used with the provided methods.

Also exemplary of the cooling apparatus are water baths and ice baths, for example water baths and / or ice baths in which the container or containers are located, for example during homogenization.

Typically, cooling apparatus that can be used to cool the liquid to between 25 ° C or about 25 ° C and 45 ° C or about 45 ° C, for example 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44 or 45 ° C, typically between 25 ° C and 43 ° C; typically between 35 ° C and 43 ° C, for example 26.5 ° C. Typically, cooling is rapid cooling, for example, cooling between 25 ° C or about 25 ° C and 45 ° C or about 45 ° C, for example, between 35 ° C and 43 ° C, for example 26, 5 ° C, between 15 minutes or about 15 minutes and 2 hours or about 2 hours, typically between 30 minutes or about 30 minutes and 60 minutes or about 60 minutes, for example 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59 or 60 minutes.

F. Transfer media

Transfer means are used with the methods of transferring a liquid from one container to another container, for example, to transfer the contents of one or more containers to one or more other containers, for example, to transfer the aqueous phase to the container of the oil phase (for example, the oil phase tank) or to transfer the oil phase to the water phase container (for example, the water phase tank) to form the emulsion. Exemplary of the equipment used for the transfer media are transfer pumps and associated fittings, for example, ball valves, sanitary fittings (for example, sanitary fittings sold by Granger, Inc., Lake Forrest II), and transfer tubes (for example , tubes sold by Sani-Tech West, Oxnard, CA), for example, food grade hoses attached to transfer pumps. Exemplary transfer pumps that can be used with the methods provided are the Teel Pump (Model 2P377B), Granger, Inc. Lake Forrest I1, a self-priming pump having a power rating of 2 HP, a voltage of 60 Hz, 208 230/460 CA, a speed of 3450 rpm. Other pumps, for example other self-priming pumps from Grainger, Inc., can be used as part of the transfer media in the methods. Alternatively, the transfer means may include means for manually transferring the liquid to another container, for example, by pouring, pipetting and / or other well known methods of manual liquid transfer.

g. Evaluation team

The evaluation kit is used to evaluate one or more properties of the compositions, for example, the phases of the compositions and / or the final concentrates. For example, evaluation equipment can be used to measure one or more parameters of concentrates and / or phases, eg, temperature and pH of liquids. Exemplary of the evaluation equipment are pH meters and temperature meters. Exemplary of pH / temperature meters are the pH and temperature meter marketed by Hanna Instruments, (model number HI 8314), which can be used to measure the temperature and pH of the mixture or mixtures. Also exemplary of temperature gauges are temperature probes, for example, digital and / or waterproof temperature probes, for example, temperature probes sold by Cooper-Atkins, Middlefield, CT, for example, the waterproof temperature probe ( Cooper-Atkins Model No. DPP400W). Other evaluation team

ES 2 396 946 T3 for evaluating liquids and / or emulsions is well known and can be used with the methods provided.

3. General methods for preparing liquid nanoemulsion concentrates

In general, the methods for preparing the concentrates include the steps for generating phases (eg, the oil phase (s) and the aqueous phase (s)) and the steps for combining and emulsifying the phases, to form the liquid nanoemulsion concentrates. In some examples, the methods include additional steps, such as evaluation, addition of additional ingredients, packaging, and filtration. The methods provided can be performed using an experimental scale manufacturing process (typically for small batch sizes). Alternatively, the methods can be performed using a larger scale manufacturing process (typically for larger batch sizes). Each of the provided concentrates can be prepared using a large scale process or an experimental scale process. In one example, after the concentrate is first prepared using the experimental scale process, the method is scaled up to prepare larger amounts of the concentrate using the large scale processes. When concentrates are formulated according to the methods provided, the initial concentrate is typically prepared by an experimental scale procedure. In one example of the formulation methods, a selected formulation is then prepared using a large scale procedure. Any of the concentrates provided herein can be prepared by the methods provided using any of the manufacturing processes. Any method described in this document, when using an experimental scale method, can be scaled up for the production of the concentrates using the large scale processes.

In general, the methods for preparing liquid nanoemulsion concentrates include first generation steps, whereby one or more oil phases and one or more aqueous phases are produced. The generation of the aqueous phase and the generation of the oil phase are typically carried out in at least two different vessels, for example, an oil phase vessel and an aqueous phase vessel. Each of the generation steps typically includes a mixing step and a heating step, which can be performed simultaneously, sequentially in any order, or partially simultaneously.

To generate the aqueous phase, the aqueous phase ingredient (s) (eg, the polar solvent (eg, water, propylene glycol, glycerin, or other polar solvent) and, in some examples, additional aqueous phase ingredients) are added to an aqueous phase vessel. The ingredient (s) are mixed, typically using a conventional mixer, and heated, for example, using a heating apparatus. Typically, the ingredients of the aqueous phase are heated to a low heat temperature, for example, 60 ° C or about 60 ° C. To prepare the oil phase, the oil phase ingredients (eg, a nonpolar compound (s), surfactant (s), and, in some examples, another oil phase ingredient (s)) are added to an oil phase container. The oil phase ingredient (s) are mixed, typically using a conventional mixer, and heated, for example, using a heating apparatus. Typically the ingredients are heated to a low heat temperature, for example 60 ° C or about 60 ° C. The mixing / heating of the water and oil phase can be carried out simultaneously or sequentially in any order. In one example, the generation of the oil phase is carried out subsequent to the generation of the aqueous phase, for example, to preserve the non-polar active ingredient, for example, to prevent its oxidation. Typically both phases are heated to the desired temperature, eg, a low heat temperature, and / or until the ingredients dissolve, before combining the oil and water phases in a subsequent emulsion step.

In general, the methods additionally include an emulsion step. For the emulsion stage, the oil and water phases are combined, for example, using one or more transfer means. The oil and water phases are emulsified, typically with mixing, typically homogenizing, eg, using high shear to generate an emulsion (eg, the nanoemulsion liquid concentrate). The emulsion step can be carried out in the aqueous phase vessel, oil phase vessel or a different vessel.

Typically, during the emulsion step, the forming emulsion is cooled, eg, rapidly cooled, eg, using one or more cooling apparatus. Typically, the cooling stage is performed simultaneously with the emulsifying stage. In one example, cooling is carried out until the emulsion reaches a temperature between 25 ° C or about 25 ° C and 43 ° C or about 43 ° C.

The methods may include additional steps, eg, evaluation steps, steps to add additional ingredients, purification steps (eg, filtration), and / or packaging / containment steps, as detailed below.

to. Generation of the aqueous phase

Typically, the aqueous phase ingredients are weighed and / or measured, for example, using one or more balances (e.g., one or more of the balances described herein), prior to addition to the aqueous phase container (e.g. example, any container described in this document). In one example, the amount of each ingredient to be added to the aqueous phase container will be determined according to the methods provided for formulating the ingredients.

ES 2 396 946 T3 concentrates. Typically, the desired concentration by weight (w / w) of the final nanoemulsion concentrate is used to calculate the amount of each water phase ingredient that is added to the water phase container. Alternatively, desired volume by weight, volume by volume, or weight by volume can be used to calculate the correct amount of an ingredient to measure and add to the container.

In one example, when water is the polar solvent, impurities in the water, for example urban water, are removed using one or more purifiers (for example, one or more purifiers as described herein) above, prior to Add the water to the aqueous phase tank. In one example, the water is purified by passing through and using the following purifiers sequentially: a carbon filter, an ion exchange filter, a reverse osmosis purifier, and an end-point filter, for example, a 100 micron end point filter, prior to adding to aqueous phase vessel.

Typically, the aqueous phase ingredient (s) are mixed in the aqueous phase container using a conventional mixer (for example, any of the conventional mixers described herein), and heated, typically simultaneously or, in part, simultaneously, using a heating apparatus (eg, any of the heating apparatus described in this document). Typically, the aqueous phase is heated so that the ingredients in the aqueous phase reach a low heat temperature, for example, between about 45 ° C or about 45 ° C and 85 ° C or about 85 ° C, for example, 45 , 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70 , 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84 or 85 ° C, typically 60 ° C or 60 ° C, for example, to prevent oxidation of non-polar ingredients and / or maintain ingredient stability. Typically, mixing and / or heating the aqueous phase ingredients is continued in the aqueous phase vessel, for example, before combining the aqueous phase and the oil phase. In one example, the aqueous phase is mixed and / or heated until the ingredients dissolve in the aqueous phase. Typically, the temperature of the water phase is maintained by mixing before combining the oil and water phases.

i. Ingredients of the aqueous phase

The aqueous phase includes one or more polar solvents such as water, diols, such as propylene glycol, and sugar alcohols such as glycerin, and in some examples other aqueous phase ingredients are included. Typically, the aqueous phase ingredients are hydrophilic and / or amphipathic ingredients of the liquid nanoemulsion concentrate. For example, oils and other lipophilic ingredients are typically not added to the aqueous phase. Certain ingredients, eg ingredients having hydrophobic and hydrophilic moieties, eg surfactants and co-surfactants, can be added to either the oil phase or the water phase, or to the oil phase and the water phase. Exemplary aqueous phase ingredients include, but are not limited to, polar solvents, eg, water, typically filtered water, propylene glycol, glycerin, and other diols; emulsion stabilizers; pH adjusters, for example phosphoric acid and / or citric acid; aromas; surfactants, co-surfactants, eg, phosphatidylcholine and / or quillaja saponaria; and preservatives.

The ingredients of the aqueous phase can be added to the aqueous phase simultaneously and / or sequentially, in a specific order. In one example, one or more aqueous phase ingredients are added first and heated, prior to the addition of the additional ingredient (s). In one example, when the ingredients of the aqueous phase include a polar solvent and an emulsion stabilizer, these ingredients are added sequentially, in the following order: 1) polar solvent; 2) emulsion stabilizer. In one example, when the aqueous phase ingredients include water and an emulsion stabilizer, these ingredients are added sequentially, in the following order: 1) water, 2) emulsion stabilizer. In another example, when the aqueous phase ingredients include a surfactant, a polar solvent (eg, water), and an emulsion stabilizer, these ingredients are added to the aqueous phase container sequentially, in the following order: 1) surfactant; 2) polar solvent (eg water); 3) emulsion stabilizer. Alternatively, the ingredients of the aqueous phase can be added in any order. Typically, when the aqueous phase includes a surfactant, particularly when the surfactant is a surfactant that is solid at room temperature, for example, polyethylene glycol tocopherol succinate surfactant, the surfactant is the first ingredient in the aqueous phase added to the phase container. watery. Typically, when the aqueous phase ingredients include an emulsion stabilizer, the emulsion stabilizer is the last ingredient added to the aqueous phase container.

b. Generation of the oil phase

Typically the oil phase ingredient (s) are weighed and / or measured, for example, using one or more balances (e.g., one or more of the balances described herein), prior to addition to the oil phase container ( for example, any of the containers described in this document). In one example, the amount of each oil phase ingredient to add is determined according to the methods provided for formulating the concentrates. Typically, the desired concentration by weight (w / w) of the final nanoemulsion concentrate is used to calculate the amount of each oil phase ingredient that should be added to the oil phase container. Alternatively, the volume by weight, volume by volume or weight by volume can be used to calculate the correct amount of an ingredient to measure and add to the container.

ES 2 396 946 T3

Typically, the oil phase ingredients are mixed in the oil phase container using a conventional mixer (for example, any of the conventional mixers described herein) and heated, typically simultaneously, using a heating apparatus (for example, any of the heating devices described in this document). Typically, the oil phase is heated so that it reaches a low heat temperature for example between 45 ° C or about 45 ° C and 85 ° C or about 85 ° C, for example 45, 46, 47, 48, 49 , 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74 , 75, 76, 77, 78, 79, 80, 81, 82, 83, 84 or 85 ° C, typically 60 ° C or 60 ° C, for example, to avoid oxidation of the non-polar ingredients and / or maintain the stability of the ingredients. In one example, one or more of the oil phase ingredients are mixed and heated according to the methods provided prior to the addition of the remainder of the oil phase ingredients. For example, the nonpolar compound can be mixed and heated with one or more solvents, for example an oil, for example linseed oil and / or Vitamin E oil, until the nonpolar compound dissolves in the oil, before from the addition of the other oily ingredients. Typically, the oil phase ingredients are mixed in the oil phase container until dissolved. Typically, the temperature of the oil phase is maintained with the mixture before combining the oil and water phases.

In some examples the oil and / or aqueous phase may be prepared in more than one container, for example, by mixing one or more of the aqueous phase ingredients in one container and mixing one or more other oil ingredients in another container. In this example, the mixed oil phase ingredients in the separate containers may be mixed together prior to emulsifying with the aqueous phase or, alternatively, they may be added separately during emulsification to the aqueous phase.

i. Oil phase ingredients

The oil phase includes the nonpolar compound, eg, the nonpolar active ingredient and, in some examples, other ingredients of the oil phase. Typically, the oil phase ingredients include one or more lipophilic and / or amphipathic ingredients of the liquid nanoemulsion concentrate. Oil phase ingredients typically do not include aqueous ingredients or hydrophilic ingredients. Certain ingredients, eg ingredients having hydrophobic and hydrophilic moieties, eg surfactants and co-surfactants, can be added to either the oil or water phase, or the oil and water phase. Exemplary of ingredients used in the oil phase of the concentrates provided are non-polar compounds, eg, non-polar active ingredients, including any of the non-polar active ingredients provided herein, emulsion stabilizers, pH adjusters, for example , phosphoric acid and / or citric acid; surfactants; co-surfactants, for example phosphatidylcholine and / or quillaja saponaria; preservatives and oils, eg solvents and other oil phase ingredients.

The oil phase ingredients can be added to the oil phase simultaneously and / or sequentially, for example, in any order or in a specific order. In one example, one or more oil phase ingredients are added first and heated, prior to the addition of an additional ingredient or ingredients. In one example, when the oil phase ingredients include a surfactant, a preservative, a solvent, a cosurfactant, and a nonpolar compound, these ingredients are added sequentially, in the following order: 1) surfactant; 2) preservative; 3) solvent; 4) co-surfactant; 5) nonpolar compound; and 6) emulsion stabilizer. In another example, when the oil phase ingredients include a surfactant, a preservative, and a nonpolar compound, the ingredients are added sequentially in the following order: 1) surfactant; 2) preservative; 3) nonpolar compound. In another example, when the oil phase ingredients include a surfactant, a preservative, a nonpolar compound, and an emulsion stabilizer, the ingredients are added sequentially in the following order: 1) surfactant, 2) preservative, 3) nonpolar compound. polar and 4) emulsion stabilizer. Alternatively, the oil phase ingredients can be added in a different order, eg any order. Two or more ingredients of the oil phase can be added simultaneously.

Typically, when the oil phase includes a surfactant, particularly when the surfactant is a surfactant that is solid at room temperature, for example, polyethylene glycol tocopherol succinate surfactant, the surfactant is the first oil phase ingredient added to the phase vessel. oily. Typically, when the oil phase ingredients include an emulsion stabilizer, the emulsion stabilizer is the last ingredient added to the oil phase container. Typically, the nonpolar compound is either of the last ingredient added to the oil phase, or is added immediately prior to the addition of the emulsion stabilizer, which is the last ingredient added to the oil phase container.

c. Combination and emulsion of the oil phase and the water phase

In general, in the methods, after generation of the oil phase and the water phase, the oil and water phases are combined, for example, using one or more transfer means (for example, any of the transfer means described in this document). The combined phases are emulsified, eg, by mixing, eg, homogenization, to form an emulsion (eg, a liquid nanoemulsion concentrate). Typically, the phases are mixed during the blending and emulsifying steps, eg, using a homogenizer (eg, any of the homogenizers described herein). In one example,

ES 2 396 946 T3 the oil and water phases (eg the emulsion being formed) are further cooled, eg rapidly cooled during the emulsion and / or blending steps.

i. Combination of the oil and water phases

To emulsify them, the oil and water phases are first combined, typically by transfer, using one or more transfer media (eg, any of the transfer media described herein). In one example, the oil phase is transferred to the water phase vessel. In another example, the aqueous phase is transferred to the oil phase vessel. In another example, a plurality of the oil phases or aqueous phases are transferred to an aqueous phase or oil phase vessel. In another example, the aqueous phase (s) and the oil phase (s) are transferred to another container, eg, an emulsion container.

Any transfer medium can be used to combine the phases. For example, any means for transferring the contents of one container to another container as described above, for example transfer pumps and associated equipment, for example sanitary fittings, hoses and / or ball valves; and manual transfer means, for example pouring and / or pipetting means or other known transfer means. In some examples, the phases are kept clean, eg, sterile during transfer, eg, using transfer media with sanitary fittings and / or by combining the phases in a sterile environment.

ii. Emulsion of the oil and water phases

Simultaneously with and / or subsequent to combining the phases, the phases are mixed (eg, homogenized), eg, using a homogenizer (eg, any of the homogenizers described) to form an emulsion. Typically, the emulsion is made in the container containing the combined liquids, eg, the oil phase or the aqueous phase container. For this emulsifying step, the oil and water phases are mixed, for example after the combining step, typically during and after the combining step, using a mixer that is capable of emulsifying liquids, for example a homogenizer, for example, a reversible homogenizer. Typically liquids are homogenized using the mixer (eg homogenizer) at a low speed, eg low rpm, eg between 850 rpm or about 850 rpm and 1200 rpm or about 1200 rpm, eg 850, 900, 950, 1000, 1050, 1100, 1150 or 1200 rpm.

The liquids are typically mixed, continuously or intermittently, until the liquids emulsify, for example, into a nanoemulsion. In one example, the mixing speed is maintained to emulsify the oil and water phases. In one example, the mixer baffle plate is adjusted, for example by moving the baffle plate further down into the mix or further up out of the mix, to control the type of mix, for example to change flow descending to ascending flow and vice versa, during mixing of the emulsion. In another example, the homogenizer can be adjusted to increase or decrease the shear or to maintain the shear at a particular speed. The methods for homogenizing the oil and water phases are well known and other methods can be used to homogenize the oil and water phases in the methods.

iii. Cooling

Typically, the emulsion is cooled during mixing, for example, by quenching. In one example, the emulsion is cooled to promote emulsion stability and phase emulsification, eg, avoiding or minimizing oxidation, eg, oxidation of the nonpolar compound. Cooling, eg quenching, is typically accomplished using one or more cooling apparatus, eg, any of the cooling apparatus described herein or any known cooling apparatus. In one example, the cooling apparatus is a recirculating cooler. In another example, the cooling apparatus is a water bath or an ice bath. In one example, when the apparatus is a recirculating cooler, the fluid in the container that is used for the emulsion stage is recirculated through the cooler, then back to the container, to rapidly cool and maintain the temperature of the mixture during mixing. . Typically, the forming emulsion is mixed and cooled until the phases emulsify and the temperature reaches between 25 ° C or about 25 ° C and 43 ° C or about 43 ° C, for example 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39; 40, 41, 42 or 43 ° C. Typically, when cooling is quenching, the temperature is reached in less than 2 hours or about 2 hours, typically less than 1 hour or about 1 hour, for example, in at least between 30 minutes or about 30 minutes and 60 minutes or about 60 minutes, for example 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51 , 52, 53, 54, 55, 56, 57, 58, 59 or 60 minutes.

Once the oil and water phases have been emulsified, thereby forming an emulsion, for example a liquid nanoemulsion concentrate, the emulsion can be used, for example, in the dilution methods provided to prepare a liquid dilution composition, for example, a drink, which contains the concentrate. Alternatively, one or more additional steps can be performed prior to using the concentrate.

ES 2 396 946 T3

d. Additional stages

Typically, one or more additional steps are performed, after emulsifying the phases, before using the concentrate. For example, the emulsion can be evaluated (eg, by measuring the pH and / or temperature of the concentrate). In another example, one or more additional ingredients can be added to the emulsion. In another example, the nanoemulsion concentrate is transferred to a containment container or packaging container, eg, a containment / packaging container, eg, a containment / packaging tank. In another example, the nanoemulsion is purified, eg filtered, before use. In one example, the addition of additional ingredients, evaluation, and / or purification can be done in the containment / packaging container. Other additional steps can be performed prior to use.

i. Additional ingredients

In one example, additional ingredients, for example pH adjusters and / or flavors, may be added to the emulsion after it is formed. In one example, citric acid and / or phosphoric acid is added to adjust the pH, for example, until the pH reaches a pH between 2.5 and 3.5, typically between 2.6 or about 2.6 and 3, 2 or about 3.2, for example 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, or 3.2. In another example, one or more flavors are added to the concentrate, for example, to enhance the taste and / or odor of the concentrate and / or beverages containing the concentrate. In another example, an additional polar solvent, eg water, may be added to the emulsion, eg in the case of evaporation to bring the concentrate to the appropriate volume. Other additional ingredients can also be added to the emulsion. Typically, additional ingredients are added to the container containing the emulsion, eg, the aqueous phase container, the oil phase container, the emulsion container, or another container, eg, a containment / packaging container. Typically, the emulsion is mixed (eg, using any of the described mixers, typically conventional mixers) while the additional ingredients are added.

ii. Concentrate evaluation

Typically, the concentrate is evaluated prior to use. Typically, pH and / or temperature are measured, for example, using a pH and temperature meter. In one example, pH and / or temperature are evaluated after additional ingredients have been added. In one example, additional ingredients can be added to adjust the parameters after evaluation.

iii. Concentrate filtration

In one example, the concentrate is purified (eg, with any of the purifiers described), eg, using a final product filter, before using the concentrate, eg, before diluting the concentrate in an aqueous medium.

Four. Experimental scale procedure

In one example of the methods for preparing the liquid nanoemulsion concentrates, the method steps are performed using an experimental scale manufacturing process that is performed on a bench, counter, table, and other surface. Typically, the experimental scale process is used to prepare emulsions that have volumes relatively smaller than those prepared with the large scale processes, for example, volumes less than 1 or about 1 or less than 1 gallon (3.79 liters) or about 1 gallon (3.79 liters), for example, less than about 500 ml, for example, 1000, 900, 800, 700, 600, 500, 450, 400, 350, 300, 250, 200, 150, 100, 50 or less.

For the experimental scale process, the equipment is typically compact enough to be used on top of a counter or other similar surface, typically compact enough to be moved, for example, to be moved, for example, lifted by the expert using the methods. . For example, the containers, for example, aqueous phase containers, oil phase containers, containment containers and packaging containers are typically experimental scale containers, for example, flasks, beakers, vials, measuring containers, flasks and / or other containers of experimental scale. In one example, the vessels in the experimental scale process are a Pyrex® beaker. Typically, the mixers are mixers that can be used in experimental scale vessels, for example, conventional mixers including hand mixers, stir bars, stir bars, magnetic mixers and top mixers, for example mechanical and / or electric top mixers and / or mixers. or other mixers that can be used in the containers. Exemplary of suitable experimental scale mixers are conventional mixers, for example conventional mixers marketed by IKA®, for example IKA® top mixers, for example models No. RW-14 Basic and RE-16S, which are laboratory mixers and which can be used to mix ingredients, for example to generate the oil and water phases. Also exemplary of suitable experimental scale mixers are homogenizers, eg, reversible homogenizers, including the Arde Barinco reversible homogenizer, Model No. CJ-4E, which can be used to emulsify the phases. Typically, heating appliances are those that can

ES 2 396 946 T3 be used with experimental scale containers, eg hot plates. Cooling apparatuses are typically apparatus suitable for use with smaller experimental scale vessels, eg ice baths and / or water baths in which the vessels can be placed, eg for rapid cooling. The evaluation means used in the experimental scale process, for example temperature and / or pH meters, typically may be capable of being put into the experimental scale vessels.

In general, for experimental scale processes, the oil phase and the water phase are generated by mixing and heating separately in experimental scale vessels, for example, flasks, beakers, vials, measuring containers, flasks and / or other containers. on an experimental scale. Mixing is typically performed using an appropriate experimental scale mixer, for example a conventional mixer, such as a hand mixer, stir bar, stir bar, magnetic mixer and / or top mixer, for example the commercially available IKA® mixer. For example, the IKA® top mixers, for example, models N ° RW-14 Basic and RE-16S, which are laboratory stirrers. Typically, the heating of the oil and water phases is carried out using a heating apparatus appropriate for the experimental scale method, for example, a heating apparatus on which one or more of the containers can be placed, for example, a hot plate. . To combine the oil phase and the aqueous phase, one or more phases, typically one phase, is typically transferred manually to another container eg, by pouring, pipetting, and / or other manual transfer means. To emulsify the oil and water phases, a reverse homogenizer is typically used. To cool the forming emulsion, for example to rapidly cool the emulsion, a cooling apparatus suitable for the experimental scale method is typically used, for example a cooling apparatus in which the container can be placed on top of or inside the container. For example, a water bath or an ice bath.

5. Large-scale manufacturing processes

The methods for preparing the liquid nanoemulsion concentrates can be carried out using a large-scale manufacturing process. A large-scale manufacturing process is typically used when the liquid nanoemulsion concentrate being prepared has a relatively larger volume than a concentrate being prepared by the experimental-scale process, for example, volumes greater than 1 or about 1 or more. greater than 1 gallon (3.79 liters) or about 1 gallon (3.79 liters), for example, greater than about 500 ml, for example, at least 0.5 l, 1 l, 2 l or 1 (3.79 liters), 2 (7.57 liters), 3 (11.36 liters), 4 (15.14 liters), 5 (18.93 liters), 6 (22.71 liters), 7 (26.0 liters), 8 (30 , 28 liters), 9 (34.07 liters), 10 (37.85 liters), 11 (41.64 liters), 12 (45.42 liters), 13 (49.21 liters), 14 (53.00 liters), 15 (56.78 liters), 16 (60.57 liters), 17 (64.35 liters), 18 (68.14 liters), 19 (71.92 liters), 20 (75.71 liters) , 21 (79.49 liters), 22 (83.28 liters), 23 (86.06 liters), 24 (90.85 liters), 25 (94.63 liters), 26 (98.42 liters), 27 (102.21 liters), 28 (105.99 liters), 29 (109.78 liters), 30 (113.56 liters), 40 (151.42 liters), 50 (189.27 liters), 60 (227.12 liters), 70 (264 , 98 liters), 80 (302.83 liters), 90 (340.69 liters), 100 (368.54 liters), 150 (567.81 liters), 200 (757.08 liters), 250 (946.35 liters), 300 (1135.62 liters), 350 (1324.89 liters), 400 (1514.16 liters), 450 (1703.43 liters), 500 (1892.70 liters), 550 (2081.97 liters) , 600 (2271.24 liters), 650 (2460.51 liters), 700 (2649.78 liters), 800 (3028.32 liters), 900 (3406.86 liters), 1000 (3785.40 liters) or more gallons.

In general, large-scale manufacturing processes are performed with equipment that is compatible with these higher volume batches (batch sizes). For example, vessels used in large-scale processes are typically tanks, e.g., water-jacketed tanks, which are equipped with water jackets that can be used as heating apparatus to heat the oil and water phase ingredients during the generation of the oil and water phases. Water jackets are typically controlled by control panels. Similarly, the transfer means used in the large-scale process typically include transfer pumps and associated accessories, eg, ball valves and hoses. Exemplary of mixers used in large-scale processes are conventional mixers (for example, mounted mixers, for example LIGHTNIN® mixers, for example, the Model XJC117 (a fixed-mount, gear-driven, high-flow mixer). and model ND2)). An exemplary large-scale process is set forth in Figure 1 and described in this section, below. The methods provided for preparing the concentrates can be performed using this exemplary large-scale process, or any variation of the large-scale process, for example, eliminating one or more steps from the exemplary process, adding one or more steps in accordance with the method and / or by substituting steps and / or equipment according to the methods described in this document.

Figure 1 sets forth an exemplary large-scale process 100 for preparing the liquid concentrate. In this example, the polar solvent is water. This exemplary large-scale process includes the following stages:

to. Water treatment

As indicated herein, the polar solvent can include water (including purified water) and other polar solvents, for example, glycerin and propylene glycol. In the example illustrated in Figure 1, the polar solvent is water 101 (e.g. urban water), which is purified prior to addition to the aqueous phase vessel by passing the water through the following purifiers, sequentially, in the following order: a carbon filter 105, an ion exchange kit 106, a reverse osmosis kit 107, a 100 micron end-point filter 108, and a 50 micron point-of-use filter 109.

ES 2 396 946 T3

b. Generation of the aqueous phase and the oil phase:

As described above, for the generation of the aqueous phase, the polar solvent and any other ingredients of the aqueous phase, typically are weighed and / or measured and added to the aqueous phase container and mixed, using a conventional mixer or another mixer, such as a homogenizer or other mixer described in this document and are typically heated during mixing, whereby the aqueous phase is generated with mixing and heating, typically at low heat (eg 60 ° C, 70 ° C, 71 ° C), according to the methods provided. In the example of the large-scale manufacturing process set forth in Figure 1, the aqueous phase vessel is an aqueous phase tank 103, which is a water jacketed tank. In the example illustrated in Figure 1, the ingredient (s) of the aqueous phase are mixed using a conventional mixer 111, for example, a LIGHTNIN® mixer (for example, model # XJC117, a gear-driven high flow mixer , fixed mounting), fixed to the tank, for example, mounted on the top of the tank. In the example illustrated in Figure 1, the heating apparatus used to heat the ingredients of the aqueous phase is the water jacket of the water jacket tank; the temperature in the water jacket is controlled through a control panel.

As described above, for the generation of the oil phase, the oil phase ingredients are typically weighed and / or measured, and added to the oil phase container and mixed, using a conventional mixer or other such mixer. as a homogenizer or other mixer described herein, and are typically heated during mixing, whereby the oil phase is generated with mixing and heating, typically with low heat (e.g. 60 ° C), according to the methods provided. In the example of the large scale manufacturing process set forth in Figure 1, the oil phase vessel is a water jacketed oil phase tank 102. In the example illustrated in Figure 1, the oil phase ingredients are mixed using a conventional 111 mixer, for example, a LIGHTNIN® mixer (for example, model ND2), attached to the oil phase tank, for example, mounted in the tank. In the example illustrated in Figure 1, the heating apparatus used to heat the oil phase ingredients is the water jacket of the water jacketed oil phase tank; the temperature in the water jacket is controlled by a control panel.

c. Combination and emulsion of the phases

As described in this document, once the oil and water phases reach the desired temperatures (for example, 60 ° C, 70 ° C, 71 ° C or other temperature), after the ingredients of the oil phase and / or aqueous phase, and optionally after cooling of one of the phases, for example cooling of the aqueous phase to 60 ° C according to the methods provided, the oil and water phases are combined by transfer and are typically emulsified by homogenization. In one example, the transfer is performed slowly to avoid build-up in the forming emulsion, such as by periodically stopping the transfer while continuing to mix the emulsion, or combining the phases slowly with the mixture. In the example of the large-scale manufacturing processes illustrated in Figure 1, phase combining is effected by transferring the oil phase to the aqueous phase container through transfer means 112, including a transfer pump (e.g., a Teel pump, model 2P377B, available from Granger, Inc.), sanitary fittings, a transfer tube or tubes (for example, food grade tubing marketed by Sani-Tech West) and a ball valve (s). Alternatively, the aqueous phase can be transferred to the oil phase. In the example set forth in Figure 1, to begin the blending / emulsifying steps, a homogenizer 110 (eg, an Arde Barinco, Inc. reversible homogenizer), mounted in the oil phase tank, is connected, for example, to 850-1200 rpm. The ball valves are then opened and the transfer pump is switched on, thus beginning the transfer of the liquid oil phase to the water phase tank through a transfer tube or tubes. As the phases are combined, the mixture is homogenized by continuous mixing with homogenizer 110.

In some examples of the large-scale manufacturing process, to avoid lumps, the pump is stopped periodically (eg by turning off the pump), while continuing to mix with the mixer, during emulsification. In one aspect of this example, this method is used to avoid lumps when the polar solvent is a solvent other than water, such as propylene glycol or glycerin. During mixing, the homogenizer can be adjusted, for example, by adjusting the baffle plate over the homogenizer to achieve and maintain an emulsion, for example, by moving the baffle plate further into the emulsion formation and / or further out of the emulsion in training. In one example, the shear rate is set to a rate where the oil phase can be seen exiting the top of the mixer. In one example, this setting is used when the polar solvent is a solvent other than water, such as propylene glycol or glycerin.

d. Cooling

As described herein, the forming emulsion typically cools, typically cools rapidly during the emulsion step. In the large-scale process, as shown in the example illustrated in Figure 1, quenching is typically accomplished by repeatedly passing the forming emulsion through a recirculating cooler 115 (for example, Model No. OC-1000 RO, marketed by Turmoil, West

ES 2 396 946 T3

Swanzey, NH), which is attached to the aqueous phase tank. Homogenization continues during the cooling stage, for example between 850 and 1200 rpm. Cooling continues, for example, until the temperature of the emulsion reaches between or about 25 ° C and about 43 ° C, such as between or about 25 ° C and about 35 ° C, between or about 35 ° C and about 43 ° C, or at about 40 ° C. Typically, the quench is done between or about 30 and in or about 60 minutes.

and. Additional stages

As described herein, additional steps can be performed after the emulsion is formed. In the example of the large-scale manufacturing process set forth in Figure 1, additional steps include transferring the emulsion, through a transfer means 112, which includes a transfer pump (e.g., a Teel pump, model 2P377B, sold by Granger, Inc.), sanitary fittings, a transfer tube or tubes (for example, food grade tubes sold by SaniTech West), and a ball valve or valves, to a containment / packaging tank 104. The transfer is done by connecting the transfer pump and opening the ball valves. Additional ingredients, eg, pH adjusters, may be added, eg, while controlling the pH, sufficient to bring the nanoemulsion to an appropriate pH, eg, between about 2.6 and 3.2. Flavors can also be added. The additional ingredients are mixed into a concentrate using a conventional mixer 111. The addition and mixing of additional ingredients and / or evaluation can be performed in the containment / packaging tank 104; alternatively, it can be done prior to transfer to the containment / packaging tank, eg, in tank 103 of the aqueous phase.

Variations of this exemplary large-scale process (Figure 1) can also be performed using the methods, including any of the variations described herein for preparing the concentrates. For example, by elimination and / or modification of one or more stages and / or equipment, according to the general methods described in this document.

D. Methods for preparing the liquid dilution compositions containing the dilute concentrates

Also described herein are methods for diluting liquid nanoemulsion concentrates to prepare liquid dilution compositions, typically aqueous liquid dilution compositions, containing nonpolar compounds. Generally, the nanoemulsion concentrate is diluted in an aqueous medium, for example, a beverage, for example, soda, water, milk, juice, health drinks, nutritional drinks, nutritional supplements, or other aqueous food or beverage. The concentrate and the aqueous medium can be mixed, for example, by stirring and / or combining or by any known mixing means. The concentrate is dispersed in the aqueous medium to form an aqueous dilution liquid composition, for example, a clear or partially clear aqueous dilution liquid composition. The aqueous liquid dilution composition can be evaluated, for example, to evaluate the transparency, taste, odor and / or stability of the liquid.

In one example, the liquid nanoemulsion concentrate is diluted in the aqueous medium, eg, water, with heating of the aqueous medium, eg, heating of the aqueous medium, eg, to at least 40 ° C or at least about 40 ° C. , for example, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50 or more ° C, for example, 48.9 ° C. In this example, the liquid nanoemulsion concentrate is added at an appropriate dilution, as described herein, to the heated aqueous medium, and stirred until dispersed or dissolved in the solution. The resulting liquid dilution composition can then be cooled, eg, to room temperature eg 25 ° C or about 25 ° C. After dilution, the aqueous liquid dilution composition can be packaged, for example, by transferring to containers, for example vials or beverage containers. In one example, a portion of the liquid dilution composition is transferred to vials for analysis, eg, evaluation of properties, such as transparency, turbidity, taste, odor, ring formation, crystal formation, and / or other properties.

They are exemplary of the equipment used to dilute the liquid nanoemulsion concentrates to form the liquid dilution compositions containing the diluted concentrates beakers, eg Pyrex® glass beakers, hot plates, eg Thermolyne hot plate, model number 846925 or model number SP46615, stir bars, temperature gauges, for example, temperature probes, for example, Cooper Temperature Probes (Model No. DPP400W) and Balances, for example, the 2.0 kg OHuAs Balance (Model No. CS2000) and / or the Sartorius Analytical Scale (Model BA 110S).

1. Dilutions

Typically, the concentrates provided can be diluted in an aqueous medium to form aqueous liquid dilution compositions over a wide range of dilutions. In one example, the concentrate can be diluted so that the aqueous liquid dilution composition contains between 0.05 g or about 0.05 g and 10 g or about 10 g, typically between 0.05 g and 5 g of the liquid concentrate per 8 fluid ounces. of the liquid (0.236588 liters), at least 8 fluid ounces (0.236588 liters) of the liquid or less than 8 fluid ounces (0.236588 liters)

ES 2 396 946 T3 of the liquid, or for a single service of the liquid. For example, the concentrate can be diluted so that the aqueous liquid dilution composition contains 0.05g, 0.06g, 0.07g, 0.08g, 0.09g, 0.1g, 0.2 g, 0.3 g, 0.4 g, 0.5 g, 0.6 g, 0.7 g, 0.8 g, 0.9 g, 1 g, 2 g, 3 g, 4 g, 5 g, 6 g, 7 g, 8 g, 9 g or 10 g of the concentrate per 8 fluid ounces (0.236588 liters), approximately 8 fluid ounces (0.236588 liters) or at least 8 fluid ounces (0.236588 liters) or at least about 8 fluid ounces (0.236588 liters) of the aqueous medium, for example, 8 (0.236588 liters), 9 (0.26616 liters), 10 (0.29574 liters), 11 (0.32531 liters), 12 (0.35488 liters), 13 (0.38446 liters) , 14 (0.41403 liters), 15 (0.44360 liters), 16 (0.47318 liters), 17 (0.50275 liters) , 18 (0.53232 liters), 19 (0.56190 liters), 20 (0.59147 liters), 25 (0.73934 liters), 30 (0.88721 liters), 35 (1.0351 liters), 40 (1.1829 liters), 45 (1.3308 liters), 50 (1 , 4787 liters), 100 (2.9574 liters), 200 (5.9147 liters) or more fluid ounces of the aqueous medium.

In another example, the concentrate is diluted such that the aqueous liquid dilution composition contains between 1 ml or about 1 ml and 10 ml or about 10 ml of the liquid concentrate, for example, 1 ml, 2 ml, 3 ml, 4 ml. , 5 ml, 6 ml, 7 ml, 8 ml, 9 ml or 10 ml of the concentrate, per 8 fluid ounces (0.236588 liters), approximately 8 fluid ounces (0.236588 liters), at least 8 fluid ounces (0.236588 liters) or at least about 8 fluid ounces (0.236588 liters) or less than 8 fluid ounces (0.236588 liters) or less than about 8 fluid ounces (0.236588 liters) or per serving size of the aqueous medium, for example, 8 (0.236588 liters), 9 (0.26616 liters), 10 (0.29574 liters), 11 (0.32531 liters), 12 (0.35488 liters) , 13 (0.38446 liters), 14 (0.41403 liters), 15 (0.44360 liters), 16 (0.47318 liters), 17 (0.50275 liters), 18 (0.53232 liters), 19 (0.56190 liters), 20 (0.59147 liters), 25 (0.73934 liters), 30 (0.88721 liters), 35 (1.0351 liters), 40 (1.1829 liters), 45 (1 , 3308 liters), 50 (1.4787 liters), 100 (2.9574 liters), 200 (5.9147 liters) or more fluid ounces of aqueous medium.

In another example, the liquid concentrate is diluted so that the aqueous liquid dilution composition contains at least 10 mg or about 10 mg, typically at least 25 mg or about 25 mg, typically at least 35 mg of the nonpolar compound, for example , the non-polar active ingredient per 8 fluid ounces (0.236588 liters) or approximately 8 fluid ounces (0.236588 liters), at least 8 fluid ounces (0.236588 liters) or at least about 8 fluid ounces (0.236588 liters) of the aqueous medium, or less than 8 ounces (0.236588 liters) or less than about 8 ounces (0.236588 liters) ), or by serving size, of the aqueous medium; for example, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 25, 25, 26, 27, 28, 29, 30, 31, 32, 33 , 34, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200 , 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 325, 350, 375, 400, 425, 450, 475, 500, 550, 600, 700, 800, 900, 1000, 1500 , 2000 mg or more of the nonpolar compound per at least 8 fluid ounces (0.236588 liters) or at least about 8 fluid ounces (0.236588 liters) of aqueous medium.

2. Analysis of Aqueous Liquid Dilution Compositions Containing Liquid Concentrates

The properties of the aqueous liquid dilution compositions containing the liquid concentrates can be evaluated using a number of different evaluation means. For example, transparency can be assessed; desirability for human consumption, eg, pleasant taste and / or odor, eg, no fishy taste / odor, no ring formation, and no crystal formation; stability, for example, absence of oxidation, ring formation, precipitation and / or overcoming of visible phases, with time; and safety for human consumption. Several of these properties can be empirically evaluated, for example, by observing liquids immediately or over time, or by smelling and / or tasting liquids. In one example, after evaluation of the aqueous dilution liquid compositions, the concentrates are reformulated to fit one or more parameters. In another example, the dilution factor can be adjusted.

to. Transparency / turbidity

The transparency of aqueous liquid dilution compositions can be evaluated using one or more of several approaches or for example, empirical observation, particle size measurement, and / or measurement of a haze value. Measurement can be qualitative or quantitative. In one example, a particular quantitative or qualitative transparency value is specified. In another example, the transparency of one liquid can be expressed relative to the transparency of another liquid, for example, an aqueous dilution liquid composition prepared according to the methods provided, or a beverage, for example, a beverage that does not contain the liquid concentrate. In this example, the liquid can be as clear as, less clear, or more clear than another liquid. For example, an aqueous liquid dilution composition containing the liquid concentrate diluted in a beverage may be as clear or about as clear as the same beverage that does not contain the concentrate. Any type of evaluation can be carried out qualitatively, for example by empirical evaluation or quantitatively, for example by taking a measurement of particle size or turbidity.

i. Empirical evaluation

In one example, the transparency / haze of the aqueous liquid dilution composition is qualitatively evaluated, eg, by observation. In one example, a liquid is considered transparent if it does not have an opaque appearance and / or if it contains no or few particles that are visible to the naked eye. In another example, the liquid can be considered relatively clear or relatively cloudy based on a comparison with other liquids, eg, water, fruit juice, soda and / or milk and / or other aqueous dilution liquid composition (s).

ES 2 396 946 T3 prepared according to the methods provided. For example, the aqueous liquid dilution composition can be as clear or about as clear as water or another liquid, eg, a beverage. For example, the liquid that contains the liquid concentrate diluted in a beverage may be as clear or about as clear as the beverage that does not contain the liquid concentrate. In a related example, the liquid can be clear or partially clear when there is no substantial difference, for example, there is no observable difference between the aqueous liquid dilution composition containing the concentrate and the aqueous medium that does not contain the concentrate. A clear liquid is not necessarily colorless. For example, a yellow liquid that does not contain or contains (few) visible particles or opacity can be transparent. In another example, the absence of crystal formation or ring formation may be indicative of a clear liquid.

ii. Particle size

In another example, transparency / haze is evaluated by quantitatively measuring particle size and / or number of particles, in the aqueous liquid dilution composition. In this example, transparency can be expressed as a numerical representation of the particle size, or as a comparison of the particle size of another liquid.

Methods for measuring the particle size of liquids are well known. Any method for measuring particle size can be used, provided it is sensitive to particle size in the expected and / or appropriate ranges of the provided aqueous liquid dilution compositions. For example, particle size analysis is commercially available, for example, from Delta Analytical Instruments, Inc., North Huntingdon, PA. In one example, the particle size of the aqueous liquid dilution composition is measured, for example, by Delta Analytical Instruments, Inc., using a light scattering analyzer, for example, a dynamic light scattering analyzer, for example , The Horiba® LB-550 which can measure particle sizes within a range of 0.001 microns to 6 microns and uses an iterative deconvolution / Fourier transform technique to present data and can measure sample concentrations from ppm to 4.0% of solid; the Horiba® LA-920, which is a laser light scattering instrument having a HeNe laser and a tungsten lamp that can determine particle sizes from 0.02 microns to 2000 microns using Mie's Theory; and other analyzers available from Delta Analytical Instruments, Inc.

Alternatively, the particle size can be measured by viewing the liquid under a microscope at magnification, eg, a 640X magnification. The particle size can then be measured by comparison with a measurement standard, for example a ruler, which is also viewed under magnification. In one example, particles of about 25 nm or greater than about 25 nm are visible, while particles less than 25 nm are not visible, for example, at 640X magnification.

iii. Turbidity measurement

In another example, the transparency / turbidity of the liquid is evaluated and / or expressed using a turbidity measurement, eg, Nephelometric Turbidity Units (NTU). In this example, the turbidity is measured optically, to obtain a value that indicates the opacity or turbidity of the liquid, which is related to the number and size of particles suspended in the liquid. The more transparent the liquid, the lower its turbidity value. Turbidity can be measured optically, for example, using a nephelometer, an instrument with a light, and a detector. The nephelometer measures turbidity by detecting scattered light resulting from exposure of the aqueous liquid dilution composition to incident light. The amount of scattered light is correlated with the amount and size of the particulate matter in the liquid and, thus, with the transparency. For example, a beam of light will pass through a sample that has low turbidity with little alteration, creating very little scattered light, resulting in a low turbidity value (NTU) reading. Other methods for measuring turbidity may be used, including commercial services for measuring turbidity, for example, services available from ACZ Laboratories, Inc., Steamboat Springs, CO.

The following examples are included for illustrative purposes only and are not intended to limit the scope of the invention.

E. Examples

Example 1: General Procedure Used to Prepare the Liquid Nanoemulsion Concentrates in Examples 2-4

Tables 2A (i) to 4A, below, set forth ingredients that were used to prepare a plurality of liquid nanoemulsion concentrates, described in greater detail in Examples 2 to 4, according to the methods provided. Each of these nanoemulsion concentrates contained one or more non-polar active ingredients, and contained water as the polar solvent, and a tocopherol polyethylene glycol succinate (TPGS) surfactant (the TPGS surfactant marketed under the name Vitamin E TPGS® from Eastman Chemical Company).

ES 2 396 946 T3

Each of Tables 2A (i) to 4A shows the milligrams (mg) for each serving (serving size is indicated) of each ingredient in the concentrate, the percentage, by weight (of the total concentrate), for each ingredient and the quantity (g) of each ingredient per batch of the indicated batch size (g). It is also indicated in each table, in the phase column, if each ingredient was added to the aqueous phase (water), the oil phase (oil) or was subsequently added to the emulsion that was formed by emulsifying the oil and aqueous phases (emulsion) .

Each of the liquid nanoemulsion concentrates set forth in Examples 2-4 was prepared using an experimental scale process according to the methods provided. Each of the concentrates could alternatively be prepared by scaling up the experimental scale process to prepare concentrates using a large scale manufacturing process of the methods provided, for example, to prepare larger batch sizes of the concentrates in the following Examples.

The experimental scale process to prepare the concentrates in Examples 2-4 was carried out using the following general steps (details are indicated in the specific examples).

To prepare each of the liquid nanoemulsion concentrates set forth in Examples 2-4 below, the indicated amount of each ingredient was weighed using a Toledo Balance (Model GD13x / USA), a Sartorius Basic Analytical Balance (Model BA110S), or a OHAUS Balance (Model CS2000). The selection of the scale (s) depended on the weight of the particular ingredient (s).

To prepare the aqueous phase, the ingredients of the aqueous phase (indicated as aqueous in each table in the phase column) were added in the indicated amounts (g / batch) to the aqueous phase container (a Pyrex® beaker), and were mixed using a conventional mixer (IKA® model No. RE-161S, which is a top mixer (laboratory shaker) compatible with the experimental scale process). While mixing, the aqueous phase ingredients were heated using a heating apparatus. The heating apparatus was a hot plate (a Thermolyne Hot Plate Model No. SP46615, Bamstead International, Dubuque, Iowa). Except where noted, when the aqueous phase included water and an emulsion stabilizer, those ingredients were added sequentially in the following order: 1) water; 2) emulsion stabilizer. The ingredients of the aqueous phase were heated with the hot plate until the temperature reached 60 ° C. The aqueous phase was kept at 60 ° C before combining and emulsifying the aqueous and oil phases. A temperature meter (temperature probe (Model No. DPP400W, Cooper-Atkins)) was used to evaluate (measure) the temperature of the aqueous phase.

The oil phase ingredients (indicated as oil in each table in the phase column) were added to an oil phase container (a Pyrex® beaker), and mixed using a conventional mixer (IKA® model No. RE -16 1S, which is a top mixer (laboratory shaker) compatible with the experimental scale process). In general, unless otherwise indicated, when the oil phase included two or more surfactants, preservative, nonpolar solvent, cosurfactant, nonpolar compound, and emulsion stabilizer, these ingredients were added sequentially, in the following order : 1) surfactant; 2) preservative; 3) non-polar solvent; 4) co-surfactant; 5) nonpolar compound; and 6) emulsion stabilizer. For example, when the oil phase included surfactant, preservative, and nonpolar compound, these ingredients were added in the following order (unless otherwise indicated): 1) surfactant, 2) preservative; 3) nonpolar compound.

As the oil phase ingredients were mixed, they were heated using a heating apparatus. The heating apparatus was a hot plate (Thermolyne Hot Plate Model No. SP46615, Bamstead International, Dubuque, Iowa). The oil phase ingredients were heated until the mixture reached 60 ° C. The oil phase was mixed at that temperature until all ingredients had dissolved, and was kept at 60 ° C before mixing with the aqueous phase. A temperature meter (temperature probe (Model No. DPP400W, Cooper-Atkins)) was used to evaluate (measure) the temperature of the oil phase.

After both phases had reached 60 ° C and the oil phase components had dissolved, the phases were combined and emulsified using the following steps. A reversible homogenizer (Arde Barinco, Inc .; Model CJ-4E) was placed in an aqueous phase vessel and turned on at 850-1200 RPM. The oil phase was then transferred to the water phase container by pouring the oil phase from the oil phase container into the water phase container. Mixing with the homogenizer continued, with the adjustment of the baffle plate in the homogenizer to achieve and maintain an emulsion, for example, by moving the baffle plate further into the forming emulsion and / or out of the forming emulsion.

Homogenization of the forming emulsion continued at between 850 and 1200 rpm, with rapid cooling. Rapid cooling was effected by placing the aqueous phase container (beaker), containing the forming emulsion, in a water bath, to cool the forming emulsion until the liquid temperature reached between 25 ° C and 43 ° C. C (specified specific temperatures; typically takes about 30 to about 60 minutes).

After emulsifying and quenching, additional ingredients were added, as indicated in the individual Examples / Tables. For example, any ingredient that was added later to the

ES 2 396 946 T3 emulsion (instead of to one of the individual phases) is indicated by the word emulsion in the phase column. The emulsion was mixed while adding any additional ingredients, using a conventional mixer (IKA® model No. RE-16 1S). Exemplary of the additional ingredients that were added in the following examples are flavors (D-limonene and lemon oil) and pH adjusters (eg, citric acid). In several examples (where indicated), the pH of the emulsion was measured using a pH and temperature meter (Hanna Instruments, model HI 8314). When necessary, the pH was adjusted with the appropriate amount of a pH adjuster (amount indicated in the tables), for example citric acid or phosphoric acid, until the emulsion reached a pH between 2.6 and 3.2. . Each of the concentrates produced in the following Examples had a pH of between about

2.6 and 3.2.

As a final step, the concentrates were filtered using a 100 micron end product filter, prior to further evaluation, dilution and / or use.

Example 2: Liquid Nanoemulsion Concentrates with Non-Polar Compounds containing PUFA

Examples 2A-2D set forth the details of nanoemulsion liquid concentrates containing nonpolar compounds (nonpolar active ingredients) including polyunsaturated fatty acids (PUFAs) (e.g. nonpolar active ingredients containing omega-3 fatty acids, omega-3 fatty acids, omega-6 fatty acids, conjugated fatty acids and other fatty acids). These concentrates were prepared using the general procedure set forth in Example 1, above.

Example 2A: Liquid Nanoemulsion Concentrates with Non-Polar Compounds containing Omega-3

Examples 2A (i) -2A (xii) set forth the details of liquid nanoemulsion concentrates containing nonpolar compounds including omega-3 fatty acids (eg DHA, EPA, ALA). These concentrates were prepared using the general procedure set forth in Example 1, above.

Example 2A (i): Liquid Nanoemulsion Concentrate with 5% of a Non-Polar Compound containing DHA (Fish Oil) and 18% of TPGS Surfactant

Table 2A (i), below, sets forth the ingredients used to prepare a 500 g batch of a liquid nanoemulsion concentrate containing DHA (serving size 2 ml), which was prepared according to the procedure set forth in Example 1 above, with the following details and modifications:

To prepare the aqueous phase using the method described in Example 1 above, the following aqueous phase ingredients were added sequentially, in the following order, to the aqueous phase container: 1) Water; 2) emulsion stabilizer. The water was purified city water, which was purified according to the methods provided by passing it through and using the following purifiers, sequentially: a carbon filter, an ion exchange purifier, a reverse osmosis purifier and a filter of end point, for example a 100 micron end point filter, before adding to the aqueous phase tank. The emulsion stabilizer was SALADIZER® brand emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

To prepare the oil phase using the method described in Example 1 above, the following oil phase ingredients were added sequentially, in the following order, to the oil phase container: 1) surfactant, 2) preservative; 3) co-surfactant; 4) non-polar active ingredient; and 5) emulsion stabilizer.

The non-polar active ingredient was fish oil, which contained approximately 70% (74%) DHA and approximately 10% (9.3%) EPA (Omega-3 EE Fish Oil, prepared by O3C Nutraceuticals, supplied by Jedwards International Inc., Quincy, MA). The non-polar active ingredient was added in an amount such that the active ingredient was 5% by weight of the final concentrate. The cosurfactant was a phosphatidylcholine cosurfactant, marketed under the trade name S-100, by Lipoid, LLC, Newark, NJ. This phosphatidylcholine was obtained from soybean extract and contained more than 95% phosphatidylcholine. The preservative was a natural preservative (GRAS certified), benzyl alcohol. The emulsion stabilizer was SALADIZER® brand emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

The flavorings (lemon oil, marketed by Mission Flavors, Foothill Ranch, CA; and GRAS certified 99% D-limonene, marketed by Florida Chemical, Winter Haven, FL) were added after emulsification and rapid cooling of the oil phases and watery. After the emulsion was formed, an appropriate amount of citric acid (set forth in Table 2A (i)) was added to adjust the pH of the emulsion to between 2.6 and 3.4 as described in Example 1 above.

ES 2 396 946 T3

Table 2A (i): Nanoemulsion Liquid Concentrate with 5% Non-Polar Compound containing DHA (Fish Oil) and 18% TPGS Surfactant

<td>Ingredient</td><td>mg / 2 ml serving</td><td>Phase</td><td>Percentage (by weight) of the concentrate</td><td>g / lot</td>

<td>Fish Oil (10.0% EPA and 70% DHA) (non-polar active ingredient)</td><td> 100</td><td>Oily</td><td> 5</td><td> 25</td>

<td>Water</td><td> 1485,05</td><td>Watery</td><td> 74,2525</td><td> 371,2625</td>

<td>SALADIZER® Brand Emulsion Stabilizer (combination of xanthan gum, guar gum and sodium alginate)</td><td> 6,8</td><td>Watery</td><td> 0,34</td><td> 1,7</td>

<td>Tocopherol Polyethylene Glycol Succinate (surfactant)</td><td> 360</td><td>Oily</td><td> 18</td><td> 90</td>

<td>SALADIZER® Brand Emulsion Stabilizer (combination of xanthan gum, guar gum and sodium alginate)</td><td> 1,27</td><td>Oily</td><td> 0,0635</td><td> 0,3175</td>

<td>Benzyl alcohol (preservative)</td><td> 10</td><td>Oily</td><td> 0,5</td><td> 2,5</td>

<td>D-limonene (aroma)</td><td> 10,5</td><td>Emulsion</td><td> 0,525</td><td> 2,625</td>

<td>Lemon oil (aroma)</td><td> 7,4</td><td>Emulsion</td><td> 0,37</td><td> 1,85</td>

<td>Phosphatidylcholine S 100 (co-surfactant)</td><td> 13,38</td><td>Oily</td><td> 0,669</td><td> 3,345</td>

<td>Citric Acid (pH adjuster)</td><td> 5,6</td><td>Emulsion</td><td> 0,28</td><td> 1,4</td>

<td>Totals</td><td> 2000,000</td><td></td><td> 100,0000</td><td> 500</td>

Example 2A (ii): Nanoemulsion Liquid Concentrate with 5% Non-Polar Compound containing DHA (Algae Oil) and 18% TPGS Surfactant

Table 2A (ii), below, sets forth the ingredients used to prepare a 200 g batch of liquid nanoemulsion concentrate containing DHA (serving size 2 ml), which was prepared according to the procedure set forth in Example 1 above, with the following details and modifications:

To prepare the oil phase using the method described in Example 1 above, the following oil phase ingredients were added sequentially, in the following order, to the oil phase container: 1) surfactant, 2) preservative; 3) non-polar active ingredient; and 4) emulsion stabilizer.

The non-polar active ingredient was seaweed oil, which contained 35% DHA. The non-polar active ingredient was added in an amount such that the ingredient was 5% (w / w) of the final concentrate. The preservative was a natural preservative (GRAS certified), benzyl alcohol. The emulsion stabilizer was a SALADIZER® brand emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

To prepare the aqueous phase using the method described in Example 1, the following aqueous phase ingredients were added, in the following order, to the aqueous phase container: 1) water; 2) emulsion stabilizer. The water was purified urban water, which was purified according to the methods provided by passing it through and using the following purifiers, sequentially: a carbon filter, a water exchange purifier

ES 2 396 946 T3 ions, a reverse osmosis purifier and an end point filter, for example a 100 micron end point filter, before adding it to the aqueous phase tank. The emulsion stabilizer was SALADIZER® brand emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

After the emulsion was formed, an appropriate amount of citric acid (set forth in Table 2A (ii)) was added to adjust the pH of the emulsion to between 2.6 and 3.4 as described in Example 1 above.

Table 2A (ii): Nanoemulsion Liquid Concentrate with 5% Non-Polar Compound containing DHA (Algae Oil) and 18% TPGS Surfactant

<td>Ingredient</td><td>mg / 2 ml serving</td><td>Phase</td><td>Percentage (by weight) of the concentrate</td><td>g / lot</td>

<td>Seaweed Oil (35% DHA)</td><td> 100</td><td>Oily</td><td> 5</td><td> 10</td>

<td>Water</td><td> 1516,33</td><td>Watery</td><td> 75,8165</td><td> 151,633</td>

<td>SALADIZER® Brand Emulsion Stabilizer (combination of xanthan gum, guar gum and sodium alginate)</td><td> 6,8</td><td>Watery</td><td> 0,34</td><td> 0,68</td>

<td>Tocopherol Polyethylene Glycol Succinate (surfactant)</td><td> 360</td><td>Oily</td><td> 18</td><td> 36</td>

<td>SALADIZER® Brand Emulsion Stabilizer (combination of xanthan gum, guar gum and sodium alginate)</td><td> 1,27</td><td>Oily</td><td> 0,0635</td><td> 0,127</td>

<td>Benzyl alcohol (preservative)</td><td> 10</td><td>Oily</td><td> 0,5</td><td> 1</td>

<td>Citric Acid (pH adjuster)</td><td> 5,6</td><td>Emulsion</td><td> 0,28</td><td> 0,56</td>

<td>Totals</td><td> 2000,000</td><td></td><td> 100,0000</td><td> 200</td>

Example 2A (iii): Nanoemulsion Liquid Concentrate with 5% Non-Polar Compound containing DHA (Seaweed Oil) and 25.2% TPGS surfactant

Table 2A (iii), below, sets forth the ingredients used to prepare a 150 g batch of liquid nanoemulsion concentrate containing DHA (serving size 2 ml), which was prepared according to the procedure set forth in Example 1 above, with the following details and modifications:

To prepare the oil phase using the method described in Example 1 above, the following oil phase ingredients were added sequentially, in the following order, to the oil phase container: 1) surfactant, 2) preservative; 3) non-polar active ingredient; and 4) emulsion stabilizer. The non-polar active ingredient was seaweed oil, which contained 35% DHA. The non-polar active ingredient was added in an amount such that the active ingredient was 5% by weight (w / w) of the final concentrate. The preservative was a natural preservative (GRAS certified), benzyl alcohol. The emulsion stabilizer was SALADIZER® brand emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

To prepare the aqueous phase using the methods described in Example 1 above, the following aqueous phase ingredients were added sequentially, in the following order, to the aqueous phase container: 1) water, 2) emulsion stabilizer. The water was purified city water, which was purified according to the methods provided by passing it through and using the following purifiers, sequentially: a carbon filter, an ion exchange purifier, a reverse osmosis purifier and a filter of end point, for example a 100 micron end point filter, before adding to the aqueous phase tank. Stabilizer

ES 2 396 946 T3 emulsion was SALADIZER® brand emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

After the emulsion was formed, an appropriate amount of citric acid (set forth in Table 2A (iii)) was added to adjust the pH of the emulsion to between 2.6 and 3.4 as described in Example 1 above.

Table 2A (iii): Nanoemulsion Liquid Concentrate with 5% Non-Polar Compound containing DHA (Algae Oil) and 25.2% Surfactant TPGS

<td>Ingredient</td><td>mg / 2 ml serving</td><td>Phase</td><td>Percentage (by weight) of the concentrate</td><td>g / lot</td>

<td>Seaweed Oil (35% DHA)</td><td> 100</td><td>Oily</td><td> 5</td><td> 7,5</td>

<td>Water</td><td> 1375,73</td><td>Watery</td><td> 68,7865</td><td> 103,17975</td>

<td>SALADIZER® Brand Emulsion Stabilizer (combination of xanthan gum, guar gum and sodium alginate)</td><td> 3,4</td><td>Watery</td><td> 0,17</td><td> 0,255</td>

<td>Tocopherol Polyethylene Glycol Succinate (surfactant)</td><td> 504</td><td>Oily</td><td> 25,2</td><td> 37,8</td>

<td>SALADIZER® Brand Emulsion Stabilizer (combination of xanthan gum, guar gum and sodium alginate)</td><td> 1,27</td><td>Oily</td><td> 0,0635</td><td> 0,09525</td>

<td>Benzyl alcohol (preservative)</td><td> 10</td><td>Oily</td><td> 0,5</td><td> 0,75</td>

<td>Citric Acid (pH adjuster)</td><td> 5,6</td><td>Emulsion</td><td> 0,28</td><td> 0,42</td>

<td>Totals</td><td> 2000,000</td><td></td><td> 100,0000</td><td> 150</td>

Example 2A (iv): Nanoemulsion Liquid Concentrate with 5% Non-Polar Compound containing ALA (Linseed Oil) and 17.75% TPGS surfactant

Table 2A (iv), below, lists the ingredients used to prepare a 500 g batch of nanoemulsion liquid concentrate containing alpha-linolenic acid (ALA) (serving size 2 ml), which was prepared according to the procedure set forth in Example 1 above, with the following details and modifications:

To prepare the oil phase using the method described in Example 1 above, the following aqueous phase ingredients were added sequentially, in the following order, to the oil phase container: 1) surfactant, 2) preservative; 3) non-polar solvent; 4) co-surfactant; 5) non-polar active ingredient; and 6) emulsion stabilizer.

The non-polar active ingredient, added to the oil phase, was a linseed oil compound, obtained from Sanmark LLC, Greensboro, NC (Sanmark Limited, Dalian, Liaoning Province, China), containing not less than (NLT) 50% C18: 3 alpha-linolenic acid. The non-polar active ingredient was added in an amount such that the ingredient was 5% by weight (w / w) of the final concentrate.

The cosurfactant was a phosphatidylcholine cosurfactant, marketed under the trade name S-100, by Lipoid, LLC, Newark, NJ. This phosphatidylcholine co-surfactant was obtained from soybean extract and contained more than 95% phosphatidylcholine. The non-polar solvent was a Vitamin E oil, marketed by aDm Natural Health and Nutrition, Decatur, IL, under the name Novatol ™ 5-67 Vitamin E (D-alpha-Tocopherol; product code ADM 410217). This oil contained at least 67.2% Tocopherol and approximately 32.8% soybean oil. The preservative was a natural preservative (GRAS certified), benzyl alcohol. The emulsion stabilizer was SALADIZER® brand emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

ES 2 396 946 T3

To prepare the aqueous phase using the method described in Example 1 above, the following aqueous phase ingredients were added sequentially, in the following order, to the aqueous phase container: 1) water, 2) emulsion stabilizer.

The water was purified city water, which was purified according to the methods provided by passing it through and using the following purifiers, sequentially: a carbon filter, an ion exchange purifier, a reverse osmosis purifier and a filter of end point, for example a 100 micron end point filter, before adding to the aqueous phase tank. The emulsion stabilizer was SALADIZER® brand emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

After the emulsion was formed, an appropriate amount of citric acid (set forth in Table 2A (iv)) was added to adjust the pH of the emulsion to between 2.6 and 3.4 as described in Example 1 above.

Table 2A (iv): Nanoemulsion Liquid Concentrate with 5% Non-Polar Compound containing ALA (Linseed Oil) and 17.75% TPGS Surfactant

<td>Ingredient</td><td>mg / 2 ml serving</td><td>Phase</td><td>Percentage (by weight) of the concentrate</td><td>g / lot</td>

<td>Vitamin E Oil (5-67) (non-polar solvent)</td><td> 75,00</td><td>Oily</td><td> 3,750</td><td> 18,75</td>

<td>SALADIZER® Brand Emulsion Stabilizer (combination of xanthan gum, guar gum and sodium alginate)</td><td> 1,270</td><td>Oily</td><td> 0,06</td><td> 0,3</td>

<td>Water</td><td> 1435</td><td>Watery</td><td> 71,74</td><td> 358,7</td>

<td>SALADIZER® Brand Emulsion Stabilizer (combination of xanthan gum, guar gum and sodium alginate)</td><td> 6,800</td><td>Watery</td><td> 0,34</td><td> 1,7</td>

<td>Tocopherol Polyethylene Glycol Succinate (surfactant)</td><td> 355,0</td><td>Oily</td><td> 17,75</td><td> 88,75</td>

<td>Phosphatidylcholine S-100 (co-surfactant)</td><td> 13,38</td><td>Oily</td><td> 0,6690</td><td> 3,345</td>

<td>Benzyl alcohol (preservative)</td><td> 10,00</td><td>Oily</td><td> 0,5000</td><td> 2,5000</td>

<td>Linseed oil 50% Omegas</td><td> 100,0</td><td>Oily</td><td> 5,0000</td><td> 25,000</td>

<td>Citric Acid (pH adjuster)</td><td> 3,800</td><td>Emulsion</td><td> 0,1900</td><td> 0,9500</td>

<td>Totals</td><td> 2000,00</td><td></td><td> 100,000</td><td> 500</td>

Example 2A (v): Nanoemulsion Liquid Concentrate with 10% Non-Polar Compound containing DHA (Seaweed Oil) and 20.2% Surfactant TPGS

Table 2A (v), below, sets forth the ingredients used to prepare a 500 g batch of liquid nanoemulsion concentrate containing DHA (serving size 2 ml), which was prepared according to the procedure set forth in Example 1 above, with the following details and modifications:

To prepare the oil phase using the method described in Example 1 above, the following oil phase ingredients were added sequentially, in the following order, to the oil phase container: 1) surfactant, 2)

ES 2 396 946 T3 preservative; 3) non-polar active ingredient; and 4) emulsion stabilizer.

The non-polar active ingredient was seaweed oil containing 35% DHA. The non-polar active ingredient was added in an amount such that the ingredient was 10% by weight (w / w) of the final concentrate. The preservative was a natural preservative (GRAS certified), benzyl alcohol. The emulsion stabilizer was SALADIZER® brand emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

To prepare the aqueous phase using the method described in Example 1 above, the following aqueous phase ingredients were added sequentially, in the following order, to the aqueous phase container: 1) water; 2) emulsion stabilizer.

The water was purified city water, which was purified according to the methods provided by passing it through and using the following purifiers, sequentially: a carbon filter, an ion exchange purifier, a reverse osmosis purifier and a filter of end point, for example a 100 micron end point filter, before adding to the aqueous phase tank. The emulsion stabilizer was SALADIZER® brand emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

After formation of the emulsion, an appropriate amount of citric acid (set forth in Table 2A (v)) was added to adjust the pH of the emulsion to between 2.6 and 3.4 as described in Example 1 previous.

Table 2A (v): Nanoemulsion Liquid Concentrate with 10% Non-Polar Compound containing DHA (Algae Oil) and 25.2% Surfactant TPGS

<td>Ingredient</td><td>mg / 2 ml serving</td><td>Phase</td><td>Percentage (by weight) of the concentrate</td><td>g / lot</td>

<td>Seaweed Oil (35% DHA)</td><td> 200</td><td>Oily</td><td> 10</td><td> 50</td>

<td>Water</td><td> 1278,76</td><td>Watery</td><td> 63,938</td><td> 319,69</td>

<td>SALADIZER® Brand Emulsion Stabilizer (combination of xanthan gum, guar gum and sodium alginate)</td><td> 1,2</td><td>Watery</td><td> 0,06</td><td> 0,3</td>

<td>Tocopherol Polyethylene Glycol Succinate (surfactant)</td><td> 504</td><td>Oily</td><td> 25,2</td><td> 126</td>

<td>SALADIZER® Brand Emulsion Stabilizer (combination of xanthan gum, guar gum and sodium alginate)</td><td> 0,44</td><td>Oily</td><td> 0,022</td><td> 0,11</td>

<td>Benzyl alcohol (preservative)</td><td> 10</td><td>Oily</td><td> 0,5</td><td> 2,5</td>

<td>Citric Acid (pH adjuster)</td><td> 5,6</td><td>Emulsion</td><td> 0,28</td><td> 1,4</td>

<td>Totals</td><td> 2000,000</td><td></td><td> 100,0000</td><td> 500</td>

Example 2A (vi): Nanoemulsion Liquid Concentrate with 5% Non-Polar Compound containing DHA (Fish Oil) and 25.2% TPGS Surfactant

Table 2A (vi), below, sets forth the ingredients used to prepare a 250 g batch of a liquid nanoemulsion concentrate containing DHA (serving size 2 ml), which was prepared according to the procedure set forth in Example 1 above, with the following details and modifications:

To prepare the oil phase using the method described in Example 1 above, the following phase ingredients

ES 2 396 946 T3 oil were added sequentially, in the following order, to the oil phase vessel: 1) surfactant; 2) preservative; 3) non-polar active ingredient; and 4) emulsion stabilizer.

The non-polar active ingredient was Denomega ™ 100, fish oil, which contained approximately 13% DHA and approximately 13% EPA. The non-polar active ingredient was added in an amount such that the active ingredient was 5% by weight of the final concentrate. The preservative was a natural preservative (GRAS certified), benzyl alcohol. The emulsion stabilizer was SALADIZER® brand emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD. To prepare the aqueous phase using the method described in Example 1 above, the following aqueous phase ingredients were added sequentially, in the following order, to the aqueous phase container: 1) water; 2) emulsion stabilizer.

The water, added to the aqueous phase, was purified urban water, which was purified according to the methods provided by passing it through and using the following purifiers, sequentially: a carbon filter, an ion exchange purifier, a purifier reverse osmosis and an end point filter, for example a 100 micron end point filter, before adding it to the aqueous phase tank.

The emulsion stabilizer was SALADIZER® brand emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

After the emulsion was formed, an appropriate amount of citric acid (set forth in Table 2A (vi)) was added to adjust the pH of the emulsion to between 2.6 and 3.4 as described in Example 1 above.

Table 2A (vi): Nanoemulsion Liquid Concentrate with 5% Non-Polar Compound containing DHA (Fish Oil) and 25.2% TPGS Surfactant

<td>Ingredient</td><td>mg / 2 ml serving</td><td>Phase</td><td>Percentage (by weight) of the concentrate</td><td>g / lot</td>

<td>Fish Oil (approx. 13% EPA; 13% DHA)</td><td> 100</td><td>Oily</td><td> 5</td><td> 12,5</td>

<td>Water</td><td> 1375,73</td><td>Watery</td><td> 68,7865</td><td> 171,96625</td>

<td>SALADIZER® Brand Emulsion Stabilizer (combination of xanthan gum, guar gum and sodium alginate)</td><td> 3,4</td><td>Watery</td><td> 0,17</td><td> 0,425</td>

<td>Tocopherol Polyethylene Glycol Succinate (surfactant)</td><td> 504</td><td>Oily</td><td> 25,2</td><td> 63</td>

<td>SALADIZER® Brand Emulsion Stabilizer (combination of xanthan gum, guar gum and sodium alginate)</td><td> 1,27</td><td>Oily</td><td> 0,0635</td><td> 0,15875</td>

<td>Benzyl alcohol (preservative)</td><td> 10</td><td>Oily</td><td> 0,5</td><td> 1,25</td>

<td>Citric Acid (pH adjuster)</td><td> 5,6</td><td>Emulsion</td><td> 0,28</td><td> 0,7</td>

<td>Totals</td><td> 2000,000</td><td></td><td> 100,0000</td><td> 250</td>

Example 2A (vii): Nanoemulsion Liquid Concentrate with 5% Non-Polar Compound containing DHA (Fish Oil) and 25.2% TPGS Surfactant

Table 2A (vii), below, lists the ingredients used to prepare a 250 g batch of a liquid nanoemulsion concentrate containing DHA (serving size 2 ml), which was prepared according to the

ES 2 396 946 T3 procedure set forth in Example 1 above, with the following details and modifications:

To prepare the oil phase using the method described in Example 1 above, the following oil phase ingredients were added sequentially, in the following order, to the oil phase container: 1) surfactant; 2) preservative; 3) non-polar active ingredient; and 4) emulsion stabilizer.

The non-polar active ingredient was fish oil containing approximately 70% (74%) DHA and approximately 10% (9.3%) EPA (Omega-3 EE Fish Oil, prepared with O3C Nutraceuticals, supplied by Jedwards International Inc., Quincy, MA). The non-polar active ingredient was added in an amount such that the active ingredient was 5% by weight of the final concentrate. The preservative was a natural preservative (GRAS certified), benzyl alcohol. The emulsion stabilizer was SALADIZER® brand emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

To prepare the aqueous phase using the method described in Example 1 above, the following aqueous phase ingredients were added sequentially, in the following order, to the aqueous phase container: 1) water; 2) emulsion stabilizer.

The water was purified city water, which was purified according to the methods provided by passing it through and using the following purifiers, sequentially: a carbon filter, an ion exchange purifier, a reverse osmosis purifier and a filter of end point, for example a 100 micron end point filter, before adding to the aqueous phase tank. The emulsion stabilizer was SALADIZER® brand emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

After the emulsion was formed, an appropriate amount of citric acid (set forth in Table 2A (vii)) was added to adjust the pH of the emulsion to between 2.6 and 3.4 as described in Example 1 above.

Table 2A (vii): Nanoemulsion Liquid Concentrate with 5% Non-Polar Compound containing DHA (Fish Oil) and 25.2% TPGS Surfactant

<td>Ingredient</td><td>mg / 2 ml serving</td><td>Phase</td><td>Percentage (by weight) of the concentrate</td><td>g / lot</td>

<td>Fish Oil (10% EPA; 70% DHA)</td><td> 100</td><td>Oily</td><td> 5</td><td> 12,5</td>

<td>Water</td><td> 1375,73</td><td>Watery</td><td> 68,7865</td><td> 171,96625</td>

<td>SALADIZER® Brand Emulsion Stabilizer (combination of xanthan gum, guar gum and sodium alginate)</td><td> 3,4</td><td>Watery</td><td> 0,17</td><td> 0,425</td>

<td>Tocopherol Polyethylene Glycol Succinate (surfactant)</td><td> 504</td><td>Oily</td><td> 25,2</td><td> 63</td>

<td>SALADIZER® Brand Emulsion Stabilizer (combination of xanthan gum, guar gum and sodium alginate)</td><td> 1,27</td><td>Oily</td><td> 0,0635</td><td> 0,15875</td>

<td>Benzyl alcohol (preservative)</td><td> 10</td><td>Oily</td><td> 0,5</td><td> 1,25</td>

<td>Citric Acid (pH adjuster)</td><td> 5,6</td><td>Emulsion</td><td> 0,28</td><td> 0,7</td>

<td>Totals</td><td> 2000,000</td><td></td><td> 100,0000</td><td> 250</td>

ES 2 396 946 T3

Example 2A (viii): Liquid Nanoemulsion Concentrate with 10% Non-Polar Compound containing

DHA (Fish Oil) and 20.2% TPGS Surfactant

Table 2A (viii), below, sets forth the ingredients used to prepare a 250 g batch of a liquid nanoemulsion concentrate containing DHA (serving size 2 ml), which was prepared according to the procedure set forth in Example 1 above, with the following details and modifications:

To prepare the oil phase using the method described in Example 1 above, the following oil phase ingredients were added sequentially, in the following order, to the oil phase container: 1) surfactant; 2) preservative; 3) non-polar active ingredient; and 4) emulsion stabilizer.

The non-polar active ingredient was Denomega ™ 100, fish oil, which contained approximately 13% DHA and approximately 13% EPA. The nonpolar active ingredient was added in an amount such that the active ingredient was 10% by weight of the final concentrate. The preservative was a natural preservative (GRAS certified), benzyl alcohol. The emulsion stabilizer was SALADIZER® brand emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

To prepare the aqueous phase using the method described in Example 1 above, the following aqueous phase ingredients were added sequentially, in the following order, to the aqueous phase container: 1) water; 2) emulsion stabilizer.

The water was purified city water, which was purified according to the methods provided by passing it through and using the following purifiers, sequentially: a carbon filter, an ion exchange purifier, a reverse osmosis purifier and a filter of end point, for example a 100 micron end point filter, before adding to the aqueous phase tank. The emulsion stabilizer was SALADIZER® brand emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

After the emulsion was formed, an appropriate amount of citric acid (set forth in Table 2A (viii)) was added to adjust the pH of the emulsion to between 2.6 and 3.4 as described in Example 1 above.

Table 2A (viii): Nanoemulsion Liquid Concentrate with 10% of a Non-Polar Compound containing DHA (Fish Oil) and 20.2% of TPGS Surfactant

<td>Ingredient</td><td>mg / 2 ml serving</td><td>Phase</td><td>Percentage (by weight) of the concentrate</td><td>g / lot</td>

<td>Fish Oil (approx. 13% EPA; 13% DHA)</td><td> 200</td><td>Oily</td><td> 10</td><td> 25</td>

<td>Water</td><td> 1375,73</td><td>Watery</td><td> 68,7865</td><td> 171,96625</td>

<td>SALADIZER® Brand Emulsion Stabilizer (combination of xanthan gum, guar gum and sodium alginate)</td><td> 3,4</td><td>Watery</td><td> 0,17</td><td> 0,425</td>

<td>Tocopherol Polyethylene Glycol Succinate (surfactant)</td><td> 404</td><td>Oily</td><td> 20,2</td><td> 50,5</td>

<td>SALADIZER® Brand Emulsion Stabilizer (combination of xanthan gum, guar gum and sodium alginate)</td><td> 1,27</td><td>Oily</td><td> 0,0635</td><td> 0,15875</td>

<td>Benzyl alcohol (preservative)</td><td> 10</td><td>Oily</td><td> 0,5</td><td> 1,25</td>

<td>Citric Acid (pH adjuster)</td><td> 5,6</td><td>Emulsion</td><td> 0,28</td><td> 0,7</td>

ES 2 396 946 T3

<td>Ingredient</td><td>mg / 2 ml serving</td><td>Phase</td><td>Percentage (by weight) of the concentrate</td><td>g / lot</td>

<td>Totals</td><td> 2000,000</td><td></td><td> 100,0000</td><td> 250</td>

Example 2A (ix): Nanoemulsion Liquid Concentrate with 10% Non-Polar Compound containing DHA (Fish Oil) and 20.2% TPGS Surfactant

Table 2A (ix), below, sets forth the ingredients used to prepare a 250 g batch of a liquid nanoemulsion concentrate containing DHA (serving size 2 ml), which was prepared according to the procedure set forth in Example 1 above, with the following details and modifications:

To prepare the oil phase using the method described in Example 1 above, the following oil phase ingredients were added sequentially, in the following order, to the oil phase container: 1) surfactant; 2) preservative; 3) non-polar active ingredient; and 4) emulsion stabilizer.

The non-polar active ingredient was fish oil, which contained approximately 70% (74%) DHA and approximately 10% (9.3%) EPA (Omega-3 Fish Oil EE, prepared with O3C Nutraceuticals, supplied by Jedwards International Inc., Quincy, MA). The nonpolar active ingredient was added in an amount such that the active ingredient was 10% by weight of the final concentrate. The preservative was a natural preservative (GRAS certified), benzyl alcohol. The emulsion stabilizer was SALADIZER® brand emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

To prepare the aqueous phase using the method described in Example 1 above, the following aqueous phase ingredients were added sequentially, in the following order, to the aqueous phase container: 1) water; 2) emulsion stabilizer.

The water was purified city water, which was purified according to the methods provided by passing it through and using the following purifiers, sequentially: a carbon filter, an ion exchange purifier, a reverse osmosis purifier and a filter of end point, for example a 100 micron end point filter, before adding to the aqueous phase tank. The emulsion stabilizer was SALADIZER® brand emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

After the emulsion was formed, an appropriate amount of citric acid (set forth in Table 2A (ix)) was added to adjust the pH of the emulsion to between 2.6 and 3.4 as described in Example 1 above.

Table 2A (ix): Nanoemulsion Liquid Concentrate with 10% Non-Polar Compound containing DHA (Fish Oil) and 20.2% TPGS Surfactant

<td>Ingredient</td><td>mg / 2 ml serving</td><td>Phase</td><td>Percentage (by weight) of the concentrate</td><td>g / lot</td>

<td>Fish Oil (10% EPA; 70% DHA)</td><td> 200</td><td>Oily</td><td> 10</td><td> 25</td>

<td>Water</td><td> 1375,73</td><td>Watery</td><td> 68,7865</td><td> 171,96625</td>

<td>SALADIZER® Brand Emulsion Stabilizer (combination of xanthan gum, guar gum and sodium alginate)</td><td> 3,4</td><td>Watery</td><td> 0,17</td><td> 0,425</td>

<td>Tocopherol Polyethylene Glycol Succinate (surfactant)</td><td> 404</td><td>Oily</td><td> 20,2</td><td> 50,5</td>

ES 2 396 946 T3

<td>Ingredient</td><td>mg / 2 ml serving</td><td>Phase</td><td>Percentage (by weight) of the concentrate</td><td>g / lot</td>

<td>SALADIZER® Brand Emulsion Stabilizer (combination of xanthan gum, guar gum and sodium alginate)</td><td> 1,27</td><td>Oily</td><td> 0,0635</td><td> 0,15875</td>

<td>Benzyl alcohol (preservative)</td><td> 10</td><td>Oily</td><td> 0,5</td><td> 1,25</td>

<td>Citric Acid (pH adjuster)</td><td> 5,6</td><td>Emulsion</td><td> 0,28</td><td> 0,7</td>

<td>Totals</td><td> 2000,000</td><td></td><td> 100,0000</td><td> 250</td>

Example 2A (x): Nanoemulsion Liquid Concentrate with 5% Non-Polar Compound containing ALA (Linseed Oil) and 25.2% TPGS Surfactant

Table 2A (x), below, lists the ingredients used to prepare a 250 g batch of a nanoemulsion liquid concentrate containing alpha-linolenic acid (ALA) (serving size 2 ml), which was prepared according to the procedure set forth in Example 1 above, with the following details and modifications:

To prepare the oil phase using the method described in Example 1 above, the following oil phase ingredients were added sequentially, in the following order, to the oil phase container: 1) surfactant, 2) preservative; 3) non-polar active ingredient; and 4) emulsion stabilizer.

The non-polar active ingredient was a linseed oil compound, obtained from Sanmark LLC, Greensboro, NC (Sanmark Limited, Dalian, Liaoning Province, China), containing not less than (NLT) 50% C18 alpha-linolenic acid: 3. The non-polar active ingredient was added in an amount such that the ingredient was 5% by weight (w / w) of the final concentrate. The surfactant was also added to the oil phase. The preservative was a natural preservative (GRAS certified), benzyl alcohol. The emulsion stabilizer was SALADIZER® brand emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

To prepare the oil phase using the method described in Example 1 above, the following aqueous phase ingredients were added sequentially, in the following order, to the aqueous phase container: 1) water, 2) emulsion stabilizer. The water was purified city water, which was purified according to the methods provided by passing it through and using the following purifiers, sequentially: a carbon filter, an ion exchange purifier, a reverse osmosis purifier and a filter of end point, for example a 100 micron end point filter, before adding to the aqueous phase tank. The emulsion stabilizer was SALADIZER® Emulsion Stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

After the emulsion was formed, an appropriate amount of citric acid (set forth in Table 2A (x)) was added to adjust the pH of the emulsion to between 2.6 and 3.4 as described in Example 1 above.

Table 2A (x): Nanoemulsion Liquid Concentrate with 5% Non-Polar Compound containing ALA (Linseed Oil) and 25.2% TPGS Surfactant

<td>Ingredient</td><td>mg / 2 ml serving</td><td>Phase</td><td>Percentage (by weight) of the concentrate</td><td>g / lot</td>

<td>Linseed oil 50% Omegas</td><td> 100</td><td>Oily</td><td> 5</td><td> 12,5</td>

<td>Water</td><td> 1375,73</td><td>Watery</td><td> 68,7865</td><td> 171,96625</td>

<td>SALADIZER® Brand Emulsion Stabilizer (combination of xanthan gum, guar gum and sodium alginate)</td><td> 3,4</td><td>Watery</td><td> 0,17</td><td> 0,425</td>

ES 2 396 946 T3

<td>Ingredient</td><td>mg / 2 ml serving</td><td>Phase</td><td>Percentage (by weight) of the concentrate</td><td>g / lot</td>

<td>Tocopherol Polyethylene Glycol Succinate (surfactant)</td><td> 504</td><td>Oily</td><td> 25,2</td><td> 63</td>

<td>SALADIZER® Brand Emulsion Stabilizer (combination of xanthan gum, guar gum and sodium alginate)</td><td> 1,27</td><td>Oily</td><td> 0,0635</td><td> 0,15875</td>

<td>Benzyl alcohol (preservative)</td><td> 10</td><td>Oily</td><td> 0,5</td><td> 1,25</td>

<td>Citric Acid (pH adjuster)</td><td> 5,6</td><td>Emulsion</td><td> 0,28</td><td> 0,7</td>

<td>Totals</td><td> 2000,000</td><td></td><td> 100,0000</td><td> 250</td>

Example 2A (xi): Nanoemulsion Liquid Concentrate with 10% Non-Polar Compound containing ALA (Linseed Oil) and 20.2% TPGS Surfactant

Table 2A (xi), below, lists the ingredients used to prepare a 250 g batch of liquid nanoemulsion concentrate containing alpha-linolenic acid (ALA) (serving size 2 ml), which was prepared according to the procedure set forth in Example 1 above, with the following details and modifications:

To prepare the oil phase using the method described in Example 1 above, the following oil phase ingredients were added sequentially, in the following order, to the oil phase container: 1) surfactant, 2) preservative; 3) non-polar active ingredient; and 4) emulsion stabilizer.

The non-polar active ingredient was a linseed oil compound, obtained from Sanmark LLC, Greensboro, NC (Sanmark Limited, Dalian, Liaoning Province, China), containing not less than (NLT) 50% alpha-linolenic acid (ALA ) C18: 3. The non-polar active ingredient was added in an amount such that the ingredient was 10% by weight (w / w) of the final concentrate. The preservative was a natural preservative (GRAS certified), benzyl alcohol. The emulsion stabilizer was SALADIZER® brand emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

To prepare the aqueous phase using the method described in Example 1 above, the following aqueous phase ingredients were added sequentially, in the following order, to the aqueous phase container: 1) water; 2) emulsion stabilizer. The water was purified city water, which was purified according to the methods provided by passing it through and using the following purifiers, sequentially: a carbon filter, an ion exchange purifier, a reverse osmosis purifier and a filter of end point, for example a 100 micron end point filter, before adding to the aqueous phase tank. The emulsion stabilizer was SALADIZER® Emulsion Stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

After the emulsion was formed, an appropriate amount of citric acid (set forth in Table 2A (xi)) was added to adjust the pH of the emulsion to between 2.6 and 3.4 as described in Example 1 above.

Table 2A (xi): Liquid Nanoemulsion Concentrate with 10% Non-Polar Compound containing ALA (Linseed Oil) and 20.2% Surfactant TPGS

<td>Ingredient</td><td>mg / 2 ml serving</td><td>Phase</td><td>Percentage (by weight) of the concentrate</td><td>g / lot</td>

<td>Linseed oil 50% Omegas</td><td> 200</td><td>Oily</td><td> 10</td><td> 25</td>

<td>Water</td><td> 1375,73</td><td>Watery</td><td> 68,7865</td><td> 171,96625</td>

ES 2 396 946 T3

<td>Ingredient</td><td>mg / 2 ml serving</td><td>Phase</td><td>Percentage (by weight) of the concentrate</td><td>g / lot</td>

<td>SALADIZER® Brand Emulsion Stabilizer (combination of xanthan gum, guar gum and sodium alginate)</td><td> 3,4</td><td>Watery</td><td> 0,17</td><td> 0,425</td>

<td>Tocopherol Polyethylene Glycol Succinate (surfactant)</td><td> 404</td><td>Oily</td><td> 20,2</td><td> 50,5</td>

<td>SALADIZER® Brand Emulsion Stabilizer (combination of xanthan gum, guar gum and sodium alginate)</td><td> 1,27</td><td>Oily</td><td> 0,0635</td><td> 0,15875</td>

<td>Benzyl alcohol (preservative)</td><td> 10</td><td>Oily</td><td> 0,5</td><td> 1,25</td>

<td>Citric Acid (pH adjuster)</td><td> 5,6</td><td>Emulsion</td><td> 0,28</td><td> 0,7</td>

<td>Totals</td><td> 2000,000</td><td></td><td> 100,0000</td><td> 250</td>

Example 2A (xii): Nanoemulsion Liquid Concentrate with 5% Non-Polar Compound containing DHA (Fish Oil) and 25.2% Surfactant TPGS

Table 2A (xii), below, sets forth the ingredients used to prepare a 250 g batch of a liquid nanoemulsion concentrate containing DHA (serving size 2 ml), which was prepared according to the procedure set forth in Example 1 above, with the following details and modifications:

To prepare the oil phase using the method described in Example 1 above, the following oil phase ingredients were added sequentially, in the following order, to the oil phase container: 1) surfactant; 2) preservative; 3) non-polar active ingredient; and 4) emulsion stabilizer.

The non-polar active ingredient was a fish oil, containing approximately 20% DHA and approximately 40% EPA (prepared by O3C Nutraceuticals, supplied by Jedwards International Inc., Quincy, MA). The non-polar active ingredient was added in an amount such that the active ingredient was 5% by weight of the final concentrate. The preservative was a natural preservative (GRAS certified), benzyl alcohol. The emulsion stabilizer was SALADIZER® brand emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

To prepare the aqueous phase using the method described in Example 1 above, the following aqueous phase ingredients were added sequentially, in the following order, to the aqueous phase container: 1) water; 2) emulsion stabilizer.

The water was purified city water, which was purified according to the methods provided by passing it through and using the following purifiers, sequentially: a carbon filter, an ion exchange purifier, a reverse osmosis purifier and a filter of end point, for example a 100 micron end point filter, before adding to the aqueous phase tank. The emulsion stabilizer was SALADIZER® brand emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

After the emulsion was formed, an appropriate amount of citric acid (set forth in Table 2A (xii)) was added to adjust the pH of the emulsion to between 2.6 and 3.4 as described in Example 1 above.

ES 2 396 946 T3

Table 2A (xii): Nanoemulsion Liquid Concentrate with 5% Non-Polar Compound containing DHA (Fish Oil) and 25.2% TPGS Surfactant

<td>Ingredient</td><td>mg / 2 ml serving</td><td>Phase</td><td>Percentage (by weight) of the concentrate</td><td>g / lot</td>

<td>Fish Oil (40% EPA; 20% DHA)</td><td> 100</td><td>Oily</td><td> 5</td><td> 12,5</td>

<td>Water</td><td> 1375,73</td><td>Watery</td><td> 68,7865</td><td> 171,96625</td>

<td>SALADIZER® Brand Emulsion Stabilizer (combination of xanthan gum, guar gum and sodium alginate)</td><td> 3,4</td><td>Watery</td><td> 0,17</td><td> 0,425</td>

<td>Tocopherol Polyethylene Glycol Succinate (surfactant)</td><td> 504</td><td>Oily</td><td> 25,2</td><td> 63</td>

<td>SALADIZER® Brand Emulsion Stabilizer (combination of xanthan gum, guar gum and sodium alginate)</td><td> 1,27</td><td>Oily</td><td> 0,0635</td><td> 0,15875</td>

<td>Benzyl alcohol (preservative)</td><td> 10</td><td>Oily</td><td> 0,5</td><td> 1,25</td>

<td>Citric Acid (pH adjuster)</td><td> 5,6</td><td>Emulsion</td><td> 0,28</td><td> 0,7</td>

<td>Totals</td><td> 2000,000</td><td></td><td> 100,0000</td><td> 250</td>

Example 2B: Liquid Nanoemulsion Concentrates with Non-Polar Compounds containing Omega-6

Example 2B (i): Nanoemulsion Liquid Concentrate with 5% Non-Polar Compound containing GLA (Borage Oil) and 17.75% TPGS Surfactant

Table 2B (i), below, lists the ingredients used to prepare a 500 g batch of liquid nanoemulsion concentrate containing GLA (Gamma Linoleic Acid) (serving size 2 ml), which was prepared according to the procedure set forth in Example 1 above, with the following details and modifications:

To prepare the oil phase using the method described in Example 1 above, the following oil phase ingredients were added sequentially, in the following order, to the oil phase container: 1) surfactant, 2) preservative; 3) non-polar solvent; 4) co-surfactant; 5) non-polar active ingredient; and 6) emulsion stabilizer.

The non-polar active ingredient was a borage oil compound, obtained from Sanmark LLC, Greensboro, NC (Sanmark Limited, Dalian, Liaoning Province, China), which was obtained by pressing and isolating the oil from the seeds of Borago officinalis L. This Oil contained not less than (NLT) 22% gamma-linolenic acid (GLA) C18: 3. The non-polar active ingredient was added in an amount such that the ingredient was 5% by weight (w / w) of the final concentrate. The cosurfactant was a phosphatidylcholine cosurfactant sold under the name S-100 by Lipoid, LLC, Newark, NJ. This phosphatidylcholine co-surfactant was obtained from soybean extract and contained more than 95% phosphatidylcholine. The non-polar solvent was Vitamin E oil, available from aDm Natural Health and Nutrition, Decatur, IL, under the name Novatol ™ 5-67 Vitamin E (D-alpha-Tocopherol; product code ADM 410217). This oil contained at least 67.2% Tocopherol and approximately 32.8% soybean oil. The preservative was a natural preservative (GRAS certified), benzyl alcohol, and SALADIZER® brand emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

To prepare the oil phase using the method described in Example 1 above, the following aqueous phase ingredients were added sequentially, in the following order, to the aqueous phase container: 1) water, 2) stabilizer

ES 2 396 946 T3 of emulsion.

The water was purified city water, which was purified according to the methods provided by passing it through and using the following purifiers, sequentially: a carbon filter, an ion exchange purifier, a reverse osmosis purifier and a filter of end point, for example a 100 micron end point filter, before adding to the aqueous phase tank. The emulsion stabilizer was SALADIZER® brand emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

After the emulsion was formed, an appropriate amount of citric acid (set forth in Table 2B (i)) was added to adjust the pH of the emulsion to between 2.6 and 3.4 as described in Example 1 above.

Table 2B (i): Nanoemulsion Liquid Concentrate with 5% Non-Polar Compound containing GLA (Borage Oil) and 17.75% TPGS Surfactant

<td>Ingredient</td><td>mg / 2 ml serving</td><td>Phase</td><td>Percentage (by weight) of the concentrate</td><td>g / lot</td>

<td>Vitamin E Oil (5-67) (non-polar solvent)</td><td> 75,00</td><td>Oily</td><td> 3,750</td><td> 18,75</td>

<td>SALADIZER® Brand Emulsion Stabilizer (combination of xanthan gum, guar gum and sodium alginate)</td><td> 1,270</td><td>Oily</td><td> 0,06</td><td> 0,3</td>

<td>Water</td><td> 1435</td><td>Watery</td><td> 71,74</td><td> 358,7</td>

<td>SALADIZER® Brand Emulsion Stabilizer (combination of xanthan gum, guar gum and sodium alginate)</td><td> 6,800</td><td>Watery</td><td> 0,34</td><td> 1,7</td>

<td>Tocopherol Polyethylene Glycol Succinate (surfactant)</td><td> 355,0</td><td>Oily</td><td> 17,75</td><td> 88,75</td>

<td>Phosphatidylcholine S-100 (co-surfactant)</td><td> 13,38</td><td>Oily</td><td> 0,6690</td><td> 3,345</td>

<td>Benzyl alcohol (preservative)</td><td> 10,00</td><td>Oily</td><td> 0,5000</td><td> 2,5000</td>

<td>Borage Oil 22% GLA</td><td> 100,0</td><td>Oily</td><td> 5,0000</td><td> 25,000</td>

<td>Citric Acid (pH adjuster)</td><td> 3,800</td><td>Emulsion</td><td> 0,1900</td><td> 0,9500</td>

<td>Totals</td><td> 2000,00</td><td></td><td> 100,000</td><td> 500</td>

Example 2B (ii): Nanoemulsion Liquid Concentrate with 5% Non-Polar Compound containing GLA (Borage Oil) and 25.2% TPGS Surfactant

Table 2B (ii), below, lists the ingredients used to prepare a 250 g batch of liquid nanoemulsion concentrate containing GLA (Gamma Linoleic Acid) (serving size 2 ml), which was prepared according to the procedure set forth in Example 1 above, with the following details and modifications:

To prepare the oil phase using the method described in Example 1 above, the following oil phase ingredients were added sequentially, in the following order, to the oil phase container: 1) surfactant, 2) preservative; 3) non-polar active ingredient; and 4) emulsion stabilizer.

ES 2 396 946 T3

The non-polar active ingredient was a borage oil compound, obtained from Sanmark LLC, Greensboro, NC (Sanmark Limited, Dalian, Liaoning Province, China), which was obtained by pressing and isolating the oil from the seeds of Borago officinalis L. This Oil contained not less than (NLT) 22% gamma-linolenic acid (GLA) C18: 3. The non-polar active ingredient was added in an amount such that the ingredient was 5% by weight (w / w) of the final concentrate. The preservative was a natural preservative (GRAS certified), benzyl alcohol. The emulsion stabilizer was SALADIZER® brand emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

To prepare the aqueous phase using the method described in Example 1 above, the following aqueous phase ingredients were added sequentially, in the following order, to the aqueous phase container: 1) water, 2) emulsion stabilizer.

The water was purified city water, which was purified according to the methods provided by passing it through and using the following purifiers, sequentially: a carbon filter, an ion exchange purifier, a reverse osmosis purifier and a filter of end point, for example a 100 micron end point filter, before adding to the aqueous phase tank. The emulsion stabilizer was SALADIZER® brand emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

After the emulsion was formed, an appropriate amount of citric acid (set forth in Table 2B (ii)) was added to adjust the pH of the emulsion to between 2.6 and 3.4 as described in Example 1 above.

Table 2B (ii): Nanoemulsion Liquid Concentrate with 5% Non-Polar Compound containing GLA (Borage Oil) and 25.2% Surfactant TPGS

<td>Ingredient</td><td>mg / 2 ml serving</td><td>Phase</td><td>Percentage (by weight) of the concentrate</td><td>g / lot</td>

<td>Borage Oil (22% GLA)</td><td> 100</td><td>Oily</td><td> 5</td><td> 12,5</td>

<td>Water</td><td> 1375,73</td><td>Watery</td><td> 68,7865</td><td> 171,96625</td>

<td>SALADIZER® Brand Emulsion Stabilizer (combination of xanthan gum, guar gum and sodium alginate)</td><td> 3,4</td><td>Watery</td><td> 0,17</td><td> 0,425</td>

<td>Tocopherol Polyethylene Glycol Succinate (surfactant)</td><td> 504</td><td>Oily</td><td> 25,2</td><td> 63</td>

<td>SALADIZER® Brand Emulsion Stabilizer (combination of xanthan gum, guar gum and sodium alginate)</td><td> 1,27</td><td>Oily</td><td> 0,0635</td><td> 0,15875</td>

<td>Benzyl alcohol (preservative)</td><td> 10</td><td>Oily</td><td> 0,5</td><td> 1,25</td>

<td>Citric Acid (pH adjuster)</td><td> 5,6</td><td>Emulsion</td><td> 0,28</td><td> 0,7</td>

<td>Totals</td><td> 2000,000</td><td></td><td> 100,0000</td><td> 250</td>

Example 2B (iii): Nanoemulsion Liquid Concentrate with 10% Non-Polar Compound containing GLA (Borage Oil) and 20.2% TPGS Surfactant

Table 2B (iii), below, lists the ingredients used to prepare a 250 g batch of liquid nanoemulsion concentrate containing GLA (Gamma Linoleic Acid) (serving size 2 ml), which was prepared according to the procedure set forth in Example 1 above, with the following details and modifications:

ES 2 396 946 T3

To prepare the oil phase using the method described in Example 1 above, the following oil phase ingredients were added sequentially, in the following order, to the oil phase container: 1) surfactant, 2) preservative; 3) non-polar active ingredient; and 4) emulsion stabilizer.

The non-polar active ingredient was a borage oil compound, obtained from Sanmark LLC, Greensboro, NC (Sanmark Limited, Dalian, Liaoning Province, China), which was obtained by pressing and isolating the oil from the seeds of Borago officinalis L. This Oil contained not less than (NLT) 22% C18: 3 gamma-linolenic acid (GLA). The non-polar active ingredient was added in an amount such that the ingredient was 10% by weight (w / w) of the final concentrate. The preservative was a natural preservative (GRAS certified), benzyl alcohol. The emulsion stabilizer was SALADIZER® brand emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

To prepare the aqueous phase using the method described in Example 1 above, the following aqueous phase ingredients were added sequentially, in the following order, to the aqueous phase container: 1) water, 2) emulsion stabilizer.

The water was purified city water, which was purified according to the methods provided by passing it through and using the following purifiers, sequentially: a carbon filter, an ion exchange purifier, a reverse osmosis purifier and a filter of end point, for example a 100 micron end point filter, before adding to the aqueous phase tank. The emulsion stabilizer was a SALADIZER® brand emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

After the emulsion was formed, an appropriate amount of citric acid (set forth in Table 2B (ii)) was added to adjust the pH of the emulsion to between 2.6 and 3.4 as described in Example 1 above.

Table 2B (iii): Nanoemulsion Liquid Concentrate with 10% Non-Polar Compound containing GLA (Borage Oil) and 20.2% Surfactant TPGS

<td>Ingredient</td><td>mg / 2 ml serving</td><td>Phase</td><td>Percentage (by weight) of the concentrate</td><td>g / lot</td>

<td>Borage Oil (22% GLA)</td><td> 200</td><td>Oily</td><td> 10</td><td> 25</td>

<td>Water</td><td> 1375,73</td><td>Watery</td><td> 68,7865</td><td> 171,96625</td>

<td>SALADIZER® Brand Emulsion Stabilizer (combination of xanthan gum, guar gum and sodium alginate)</td><td> 3,4</td><td>Watery</td><td> 0,17</td><td> 0,425</td>

<td>Tocopherol Polyethylene Glycol Succinate (surfactant)</td><td> 404</td><td>Oily</td><td> 20,2</td><td> 50,5</td>

<td>SALADIZER® Brand Emulsion Stabilizer (combination of xanthan gum, guar gum and sodium alginate)</td><td> 1,27</td><td>Oily</td><td> 0,0635</td><td> 0,15875</td>

<td>Benzyl alcohol (preservative)</td><td> 10</td><td>Oily</td><td> 0,5</td><td> 1,25</td>

<td>Citric Acid (pH adjuster)</td><td> 5,6</td><td>Emulsion</td><td> 0,28</td><td> 0,7</td>

<td>Totals</td><td> 2000,000</td><td></td><td> 100,0000</td><td> 250</td>

Example 2C: Liquid Nanoemulsion Concentrates with Non-Polar Compounds containing Conjugated Fatty Acid

Example 2C (i): Nanoemulsion Liquid Concentrate with 5% Non-Polar Compound containing CLA

ES 2 396 946 T3 and 17.75% Surfactant from TPGS

Table 2C (i), below, sets forth the ingredients used to prepare a 500 g batch of a liquid nanoemulsion concentrate containing CLA (serving size 2 ml), which was prepared according to the procedure set forth in Example 1 above, with the following details and modifications:

To prepare the oil phase using the method described in Example 1 above, the following oil phase ingredients were added sequentially, in the following order, to the oil phase container: 1) surfactant, 2) preservative; 3) non-polar solvent; 4) co-surfactant; 5) non-polar active ingredient; and 6) emulsion stabilizer.

The non-polar active ingredient was a conjugated linoleic acid (CLA) compound, obtained from Sanmark LTD (Dalian, Liaoning Province, China; product code 01057-A80), containing 80% CLA. The non-polar active ingredient was added in an amount such that the ingredient was 5% by weight (w / w) of the final concentrate. The cosurfactant was a phosphatidylcholine cosurfactant sold under the name S-100 by Lipoid, LLC, Newark, NJ. This phosphatidylcholine co-surfactant was obtained from soybean extract and contained more than 95% phosphatidylcholine. The non-polar solvent was a Vitamin E oil, marketed by ADM Natural Health and Nutrition, Decatur, IL, under the name Novatol ™ 5-67 Vitamin E (D-alpha-Tocopherol; product code ADM 410217). This oil contained at least 67.2% Tocopherol and approximately 32.8% soybean oil. The preservative was a natural preservative (certified GrAs), benzyl alcohol. The emulsion stabilizer was SALADIZER® brand emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

To prepare the aqueous phase using the method described in Example 1 above, the following aqueous phase ingredients were added sequentially, in the following order, to the aqueous phase container: 1) water, 2) emulsion stabilizer.

The water was purified city water, which was purified according to the methods provided by passing it through and using the following purifiers, sequentially: a carbon filter, an ion exchange purifier, a reverse osmosis purifier and a filter of end point, for example a 100 micron end point filter, before adding to the aqueous phase tank. The emulsion stabilizer was a SALADIZER® brand emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

After the emulsion was formed, an appropriate amount of citric acid (set forth in Table 2B (i)) was added to adjust the pH of the emulsion to between 2.6 and 3.4 as described in Example 1 above.

Table 2C (i): Nanoemulsion Liquid Concentrate with 5% Non-Polar Compound containing CLA and 17.75% TPGS Surfactant

<td>Ingredient</td><td>mg / 2 ml serving</td><td>Phase</td><td>Percentage (by weight) of the concentrate</td><td>g / lot</td>

<td>Vitamin E Oil (5-67) (non-polar solvent)</td><td> 75,00</td><td>Oily</td><td> 3,750</td><td> 18,75</td>

<td>SALADIZER® Brand Emulsion Stabilizer (combination of xanthan gum, guar gum and sodium alginate)</td><td> 1,270</td><td>Oily</td><td> 0,06</td><td> 0,3</td>

<td>Water</td><td> 1435</td><td>Watery</td><td> 71,74</td><td> 358,7</td>

<td>SALADIZER® Brand Emulsion Stabilizer (combination of xanthan gum, guar gum and sodium alginate)</td><td> 6,800</td><td>Watery</td><td> 0,34</td><td> 1,7</td>

<td>Tocopherol Polyethylene Glycol Succinate (surfactant)</td><td> 355,0</td><td>Oily</td><td> 17,75</td><td> 88,75</td>

<td>Phosphatidylcholine S-100 (co-surfactant)</td><td> 13,38</td><td>Oily</td><td> 0,6690</td><td> 3,345</td>

ES 2 396 946 T3

<td>Ingredient</td><td>mg / 2 ml serving</td><td>Phase</td><td>Percentage (by weight) of the concentrate</td><td>g / lot</td>

<td>Benzyl alcohol (preservative)</td><td> 10,00</td><td>Oily</td><td> 0,5000</td><td> 2,5000</td>

<td>Conjugated Linoleic Acid Oil 80%</td><td> 100,0</td><td>Oily</td><td> 5,0000</td><td> 25,000</td>

<td>Citric Acid (pH adjuster)</td><td> 3,800</td><td>Emulsion</td><td> 0,1900</td><td> 0,9500</td>

<td>Totals</td><td> 2000,00</td><td></td><td> 100,000</td><td> 500</td>

Example 2C (ii): Nanoemulsion Liquid Concentrate with 5% Non-Polar Compound containing CLA and 25.2% TpGS Surfactant

Table 2C (ii), below, sets forth the ingredients used to prepare a 250 g batch of liquid nanoemulsion concentrate containing CLA (serving size 2 ml), which was prepared according to the procedure set forth in Example 1 above, with the following details and modifications:

To prepare the oil phase using the method described in Example 1 above, the following oil phase ingredients were added sequentially, in the following order, to the oil phase container: 1) surfactant, 2) preservative; 3) non-polar active ingredient; and 4) emulsion stabilizer.

The non-polar active ingredient was a conjugated linoleic acid (CLA) compound, obtained from Sanmark LTD (Dalian, Liaoning Province, China; product code 01057-A80), containing 80% CLA. The non-polar active ingredient was added in an amount such that the ingredient was 5% by weight (w / w) of the final concentrate. The preservative was a natural preservative (GRAS certified), benzyl alcohol, and SALADIZER® brand emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, Md.

To prepare the aqueous phase using the method described in Example 1 above, the following aqueous phase ingredients were added sequentially, in the following order, to the aqueous phase container: 1) water, 2) emulsion stabilizer.

The water was purified city water, which was purified according to the methods provided by passing it through and using the following purifiers, sequentially: a carbon filter, an ion exchange purifier, a reverse osmosis purifier and a filter of end point, for example a 100 micron end point filter, before adding to the aqueous phase tank. The emulsion stabilizer was a SALADIZER® brand emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

After the emulsion was formed, an appropriate amount of citric acid (set forth in Table 2C (ii)) was added to adjust the pH of the emulsion to between 2.6 and 3.4 as described in Example 1 above.

Table 2C (ii): Nanoemulsion Liquid Concentrate with 5% Non-Polar Compound containing CLA and 25.2% Surfactant of TPGS

<td>Ingredient</td><td>mg / 2 ml serving</td><td>Phase</td><td>Percentage (by weight) of the concentrate</td><td>g / lot</td>

<td>Conjugated Linoleic Acid Oil 80%</td><td> 100</td><td>Oily</td><td> 5</td><td> 12,5</td>

<td>Water</td><td> 1375,73</td><td>Watery</td><td> 68,7865</td><td> 171,96625</td>

<td>SALADIZER® Brand Emulsion Stabilizer (combination of xanthan gum, guar gum and sodium alginate)</td><td> 3,4</td><td>Watery</td><td> 0,17</td><td> 0,425</td>

ES 2 396 946 T3

<td>Ingredient</td><td>mg / 2 ml serving</td><td>Phase</td><td>Percentage (by weight) of the concentrate</td><td>g / lot</td>

<td>Tocopherol Polyethylene Glycol Succinate (surfactant)</td><td> 504</td><td>Oily</td><td> 25,2</td><td> 63</td>

<td>SALADIZER® Brand Emulsion Stabilizer (combination of xanthan gum, guar gum and sodium alginate)</td><td> 1,27</td><td>Oily</td><td> 0,0635</td><td> 0,15875</td>

<td>Benzyl alcohol (preservative)</td><td> 10</td><td>Oily</td><td> 0,5</td><td> 1,25</td>

<td>Citric Acid (pH adjuster)</td><td> 5,6</td><td>Emulsion</td><td> 0,28</td><td> 0,7</td>

<td>Totals</td><td> 2000,000</td><td></td><td> 100,0000</td><td> 250</td>

Example 2C (iii): Nanoemulsion Liquid Concentrate with 10% Non-Polar Compound containing CLA and 20.2% TPGS Surfactant

Table 2C (iii), below, sets forth the ingredients used to prepare a 250 g batch of a liquid nanoemulsion concentrate containing CLA (serving size 2 ml), which was prepared according to the procedure set forth in Example 1 above, with the following details and modifications:

To prepare the oil phase using the method described in Example 1 above, the following oil phase ingredients were added sequentially, in the following order, to the oil phase container: 1) surfactant, 2) preservative; 3) non-polar active ingredient; and 4) emulsion stabilizer.

The non-polar active ingredient was a conjugated linoleic acid (CLA) compound, obtained from Sanmark LTD (Dalian, Liaoning Province, China; product code 01057-A80), containing 80% CLA. The non-polar active ingredient was added in an amount such that the ingredient was 10% by weight (w / w) of the final concentrate. The preservative was a natural preservative (GRAS certified), benzyl alcohol. The emulsion stabilizer was SALADIZER® brand emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

To prepare the aqueous phase using the method described in Example 1 above, the following aqueous phase ingredients were added sequentially, in the following order, to the aqueous phase container: 1) water, 2) emulsion stabilizer.

The water was purified city water, which was purified according to the methods provided by passing it through and using the following purifiers, sequentially: a carbon filter, an ion exchange purifier, a reverse osmosis purifier and a filter of end point, for example a 100 micron end point filter, before adding to the aqueous phase tank. The emulsion stabilizer was SALADIZER® brand emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

After the emulsion was formed, an appropriate amount of citric acid (set forth in Table 2C (iii)) was added to adjust the pH of the emulsion to between 2.6 and 3.4 as described in Example 1 above.

Table 2C (iii): Nanoemulsion Liquid Concentrate with 10% Non-Polar Compound containing CLA and 20.2% TPGS Surfactant

<td>Ingredient</td><td>mg / 2 ml serving</td><td>Phase</td><td>Percentage (by weight) of the concentrate</td><td>g / lot</td>

<td>Conjugated Linoleic Acid Oil 80%</td><td> 200</td><td>Oily</td><td> 10</td><td> 25</td>

<td>Water</td><td> 1375,73</td><td>Watery</td><td> 68,7865</td><td> 171,96625</td>

ES 2 396 946 T3

<td>Ingredient</td><td>mg / 2 ml serving</td><td>Phase</td><td>Percentage (by weight) of the concentrate</td><td>g / lot</td>

<td>SALADIZER® Brand Emulsion Stabilizer (combination of xanthan gum, guar gum and sodium alginate)</td><td> 3,4</td><td>Watery</td><td> 0,17</td><td> 0,425</td>

<td>Tocopherol Polyethylene Glycol Succinate (surfactant)</td><td> 404</td><td>Oily</td><td> 20,2</td><td> 50,5</td>

<td>SALADIZER® Brand Emulsion Stabilizer (combination of xanthan gum, guar gum and sodium alginate)</td><td> 1,27</td><td>Oily</td><td> 0,0635</td><td> 0,15875</td>

<td>Benzyl alcohol (preservative)</td><td> 10</td><td>Oily</td><td> 0,5</td><td> 1,25</td>

<td>Citric Acid (pH adjuster)</td><td> 5,6</td><td>Emulsion</td><td> 0,28</td><td> 0,7</td>

<td>Totals</td><td> 2000,000</td><td></td><td> 100,0000</td><td> 250</td>

Example 2D: Nanoemulsion Liquid Concentrates with Non-Polar Compounds with Saw Palmetto Extract

Example 2D (i): Nanoemulsion Liquid Concentrate with 5% Non-Polar Compound with Saw Palmetto Extract and 17.75% TPGS Surfactant

Table 2D (i), below, sets forth the ingredients used to prepare a 250 g batch of nanoemulsion liquid concentrate containing Saw Palmetto extract (serving size 2 ml), which was prepared according to the procedure outlined in Example 1 above, with the following details and modifications:

To prepare the oil phase using the method described in Example 1 above, the following oil phase ingredients were added sequentially, in the following order, to the oil phase container: 1) surfactant, 2) preservative; 3) non-polar solvent; 4) co-surfactant; 5) non-polar active ingredient; and 6) emulsion stabilizer.

The non-polar active ingredient was a Saw Palmetto extract, the Saw Palmetto Lipophilic Extract commercially available from Natural Medicinals, Inc., Felda, FL, which contained approximately 90% total fatty acids, including 0.8% acid. Caproic, 2% Caprylic Acid, 2.4% Capric Acid, 27.1 Lauric Acid, 10.3 Myristic Acid, 8.1% Palmitic Acid, 0.2% Palmitoleic Acid, 2% Acid Stearic, Oleic Acid 26.7, Linoleic Acid 4.9%, 0.7% linolenic acid, 0.42%; 0.42% phytosterol, including 0.42% beta Sitosterol, 0.09% Campesterol, 0.03% Stigmasterol and 0.2% moisture. The non-polar active ingredient was added in an amount such that the ingredient was 5% by weight (w / w) of the final concentrate.

The cosurfactant was a phosphatidylcholine cosurfactant marketed under the trade name S-100, by Lipoid, LLC, Newark, NJ. This phosphatidylcholine co-surfactant was obtained from soybean extract and contained more than 95% phosphatidylcholine.

The non-polar solvent was a Vitamin E oil marketed by ADM Natural Health and Nutrition, Decatur, IL, under the name Novatol ™ 5-67 Vitamin E (D-alpha-Tocopherol; product code ADM 410217). This oil contained at least 67.2% Tocopherol and approximately 32.8% soybean oil. The preservative was a natural preservative (certified by GRAS), benzyl alcohol. The emulsion stabilizer was SALADIZER® brand emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

To prepare the aqueous phase using the method described in Example 1 above, the following aqueous phase ingredients were added sequentially, in the following order, to the aqueous phase container: 1) water, 2) emulsion stabilizer.

ES 2 396 946 T3

The water was purified city water, which was purified according to the methods provided by passing it through and using the following purifiers, sequentially: a carbon filter, an ion exchange purifier, a reverse osmosis purifier and a filter of end point, for example a 100 micron end point filter, before adding to the aqueous phase tank. The emulsion stabilizer was a SALADIZER® brand emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

After the emulsion was formed, an appropriate amount of citric acid (set forth in Table 2D (i)) was added to adjust the pH of the emulsion to between 2.6 and 3.4 as described in Example 1 above.

Table 2D (i): Nanoemulsion Liquid Concentrate with 5% Non-Polar Compound with Saw Palmetto Extract and 17.75% TPGS Surfactant

<td>Ingredient</td><td>mg / 2 ml serving</td><td>Phase</td><td>Percentage (by weight) of the concentrate</td><td>g / lot</td>

<td>Vitamin E Oil (5-67)</td><td> 75,00</td><td>Oily</td><td> 3,750</td><td> 9,375</td>

<td>SALADIZER® Brand Emulsion Stabilizer (combination of xanthan gum, guar gum and sodium alginate)</td><td> 1,270</td><td>Oily</td><td> 0,06</td><td> 0,2</td>

<td>Water</td><td> 1435</td><td>Watery</td><td> 71,74</td><td> 179,3</td>

<td>SALADIZER® Brand Emulsion Stabilizer (combination of xanthan gum, guar gum and sodium alginate)</td><td> 6,800</td><td>Watery</td><td> 0,34</td><td> 0,9</td>

<td>Tocopherol Polyethylene Glycol Succinate (surfactant)</td><td> 355,0</td><td>Oily</td><td> 17,75</td><td> 44,38</td>

<td>Phosphatidylcholine S-100 (co-surfactant)</td><td> 13,38</td><td>Oily</td><td> 0,6690</td><td> 1,673</td>

<td>Benzyl alcohol (preservative)</td><td> 10,00</td><td>Oily</td><td> 0,5000</td><td> 1,2500</td>

<td>Saw Oil Palmetto (90% acid Fatty)</td><td> 100,0</td><td>Oily</td><td> 5,0000</td><td> 12,500</td>

<td>Citric Acid (pH adjuster)</td><td> 3,800</td><td>Emulsion</td><td> 0,1900</td><td> 0,4750</td>

<td>Totals</td><td> 2000,00</td><td></td><td> 100,000</td><td> 250</td>

Example 2D (ii): Nanoemulsion Liquid Concentrate with 5% Non-Polar Compound with Saw Palmetto Extract and 25.2% TPGS Surfactant

Table 2D (ii), below, sets forth the ingredients used to prepare a 250 g batch of liquid nanoemulsion concentrate containing Saw Palmetto extract (serving size 2 ml), which was prepared according to the procedure set forth in Example 1 above, with the following details and modifications:

To prepare the oil phase using the method described in Example 1 above, the following oil phase ingredients were added sequentially, in the following order, to the oil phase container: 1) surfactant, 2) preservative; 3) non-polar active ingredient; and 4) emulsion stabilizer.

The non-polar active ingredient was a Saw Palmetto extract, the Saw Palmetto Lipophilic Extract commercially available from Natural Medicinals, Inc., Felda, FL, containing approximately 85-90% total fatty acids, including 0, 8% Caproic Acid, 2% Caprylic Acid, 2.4% Capric Acid, 27.1 Lauric Acid, 10.3 Myristic Acid, 8.1% Palmitic Acid, 0.2% Palmitoleic Acid, 2% Acid Stearic, 26.7 Oleic Acid, 4.9% Linoleic Acid, 0.7% Linolenic Acid, 0.42%; 0.42% phytosterols including 0.42% beta Sitosterol, 0.09% Campesterol, 0.03% Stigmasterol and 0.2% moisture. The nonpolar active ingredient was added in an amount such that the

ES 2 396 946 T3 ingredient out of 5% by weight (w / w) of the final concentrate. The preservative was a natural preservative (certified

GRAS), benzyl alcohol. The emulsion stabilizer was SALADIZER® brand emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

To prepare the aqueous phase using the method described in Example 1 above, the following aqueous phase ingredients were added sequentially, in the following order, to the aqueous phase container: 1) water, 2) emulsion stabilizer.

The water was purified city water, which was purified according to the methods provided by passing it through and using the following purifiers, sequentially: a carbon filter, an ion exchange purifier, a reverse osmosis purifier and a filter of end point, for example a 100 micron end point filter, before adding to the aqueous phase tank. The emulsion stabilizer was SALADIZER® brand emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

After the emulsion was formed, an appropriate amount of citric acid (set forth in Table 2D (ii)) was added to adjust the pH of the emulsion to between 2.6 and 3.4 as described in Example 1 above.

Table 2D (ii): Nanoemulsion Liquid Concentrate with 5% Non-Polar Compound with Saw Palmetto Extract and 25.2% TPGS Surfactant

<td>Ingredient</td><td>mg / 2 ml serving</td><td>Phase</td><td>Percentage (by weight) of concentrate</td><td>g / lot</td>

<td>Saw Oil Palmetto 85-90% Fatty Acids</td><td> 100</td><td>Oily</td><td> 5</td><td> 12,5</td>

<td>Water</td><td> 1375,73</td><td>Watery</td><td> 68,7865</td><td> 171,96625</td>

<td>SALADIZER® Brand Emulsion Stabilizer (combination of xanthan gum, guar gum and sodium alginate)</td><td> 3,4</td><td>Watery</td><td> 0,17</td><td> 0,425</td>

<td>Tocopherol Polyethylene Glycol Succinate (surfactant)</td><td> 504</td><td>Oily</td><td> 25,2</td><td> 63</td>

<td>SALADIZER® Brand Emulsion Stabilizer (combination of xanthan gum, guar gum and sodium alginate)</td><td> 1,27</td><td>Oily</td><td> 0,0635</td><td> 0,15875</td>

<td>Benzyl alcohol (preservative)</td><td> 10</td><td>Oily</td><td> 0,5</td><td> 1,25</td>

<td>Citric Acid (pH adjuster)</td><td> 5,6</td><td>Emulsion</td><td> 0,28</td><td> 0,7</td>

<td>Totals</td><td> 2000,000</td><td></td><td> 100,0000</td><td> 250</td>

Example 2D (iii): Nanoemulsion Liquid Concentrate with 10% Non-Polar Compound with Saw Palmetto Extract and 20.2% TPGS Surfactant

Table 2D (iii), below, sets forth the ingredients used to prepare a 250 g batch of liquid nanoemulsion concentrate containing Saw Palmetto extract (serving size 2 ml), which is prepared according to the procedure outlined in Example 1 above, with the following details and modifications:

To prepare the oil phase using the method described in Example 1 above, the following oil phase ingredients were added sequentially, in the following order, to the oil phase container: 1) surfactant, 2) preservative; 3) non-polar active ingredient; and 4) emulsion stabilizer.

100

ES 2 396 946 T3

The non-polar active ingredient was a Saw Palmetto extract, Saw Palmetto Lipophilic Extract, commercially available from Natural Medicinals, Inc., Felda, FL, which contained approximately 85-90% total fatty acids, including 0.8 % Caproic acid, 2% Caprylic acid, 2.4% Capric acid, 27.1 Lauric acid, 10.3 Myristic acid, 8.1% Palmitic acid, 0.2% Palmitoleic acid, 2% Stearic acid, 26.7 Oleic Acid, 4.9% Linoleic Acid, 0.7% Linolenic Acid, 0.42%; 0.42% phytosterols including 0.42% beta Sitosterol, 0.09% Campesterol, 0.03% Stigmasterol; and 0.2% humidity. The non-polar active ingredient was added in an amount such that the ingredient was 10% by weight (w / w) of the final concentrate. The preservative was a natural preservative (GRAS certified), benzyl alcohol. The emulsion stabilizer was SALADIZER® brand emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

To prepare the aqueous phase using the method described in Example 1 above, the following aqueous phase ingredients were added sequentially, in the following order, to the aqueous phase container: 1) water, 2) emulsion stabilizer.

The water was purified city water, which was purified according to the methods provided by passing it through and using the following purifiers, sequentially: a carbon filter, an ion exchange purifier, a reverse osmosis purifier and a filter of end point, for example a 100 micron end point filter, before adding to the aqueous phase tank. The emulsion stabilizer was SALADIZER® brand emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

After the emulsion was formed, an appropriate amount of citric acid (set forth in Table 2D (iii)) was added to adjust the pH of the emulsion to between 2.6 and 3.4 as described in Example 1 above.

Table 2D (iii): Nanoemulsion Liquid Concentrate with 10% Non-Polar Compound from Saw Palmetto Extract and 20.2% Surfactant from TPGS

<td>Ingredient</td><td>mg / 2 ml serving</td><td>Phase</td><td>Percentage (by weight) of concentrate</td><td>g / lot</td>

<td>Saw Palmetto Oil 85-90% Fatty Acids</td><td> 200</td><td>Oily</td><td> 10</td><td> 25</td>

<td>Water</td><td> 1375,73</td><td>Watery</td><td> 68,7865</td><td> 171,96625</td>

<td>SALADIZER® Brand Emulsion Stabilizer (combination of xanthan gum, guar gum and sodium alginate)</td><td> 3,4</td><td>Watery</td><td> 0,17</td><td> 0,425</td>

<td>Tocopherol Polyethylene Glycol Succinate (surfactant)</td><td> 404</td><td>Oily</td><td> 20,2</td><td> 50,5</td>

<td>SALADIZER® Brand Emulsion Stabilizer (combination of xanthan gum, guar gum and sodium alginate)</td><td> 1,27</td><td>Oily</td><td> 0,0635</td><td> 0,15875</td>

<td>Benzyl alcohol (preservative)</td><td> 10</td><td>Oily</td><td> 0,5</td><td> 1,25</td>

<td>Citric Acid (pH adjuster)</td><td> 5,6</td><td>Emulsion</td><td> 0,28</td><td> 0,7</td>

<td>Totals</td><td> 2000,000</td><td></td><td> 100,0000</td><td> 250</td>

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ES 2 396 946 T3

Example 3: Liquid Nanoemulsion Concentrates with Non-Polar Compounds containing Coenzyme

Q

Example 3A Nanoemulsion Liquid Concentrate with 5% Non-Polar Compound CoQ10 and 17.75% Surfactant TPGS

Table 3A below lists the ingredients used to prepare a 650 g batch of liquid nanoemulsion concentrate containing CoQ10 (serving size 2 ml), which was prepared according to the procedure set forth in Example 1 above, with the following following details and modifications:

To prepare the oil phase using the method described in Example 1 above, the following oil phase ingredients were added sequentially, in the following order, to the oil phase container: 1) surfactant, 2) preservative; 3) non-polar solvent; 4) co-surfactant; 5) non-polar active ingredient; and 6) emulsion stabilizer.

The non-polar active ingredient was a Coenzyme Q10 (compound CoQ10, marketed under the name Kaneka Q10 ™ (USP Ubidecarenone) by Kaneka Nutrients, LP, Pasadena, TX, which contains more than 98% ubidecarenone (ubiquinone). The active ingredient did not polar was added in an amount such that the ingredient was 5% by weight (w / w) of the final concentrate. The cosurfactant was a phosphatidylcholine cosurfactant, marketed under the trade name S-100 by Lipoid, LLC, Newark, NJ. This phosphatidylcholine co-surfactant was obtained from soybean extract and contained more than 95% phosphatidylcholine. The non-polar solvent was a Vitamin E oil available from ADM Natural Health and Nutrition, Decatur, IL, under the name Novatol ™ 5-67 Vitamin E (Dalpha-Tocopherol; product code ADM 410217). This oil contained at least 67.2% Tocopherol and approximately 32.8% soybean oil. The preservative was a natural preservative (GRAS certified), benzyl alcohol, and a SALADIZER® brand emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

To prepare the aqueous phase using the method described in Example 1 above, the following aqueous phase ingredients were added sequentially, in the following order to the aqueous phase container: 1) water, 2) emulsion stabilizer.

The water was purified city water, which was purified according to the methods provided by passing it through and using the following purifiers, sequentially: a carbon filter, an ion exchange purifier, a reverse osmosis purifier and a filter of end point, for example a 100 micron end point filter, before adding to the aqueous phase tank. The emulsion stabilizer was a SALADIZER® brand emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

After the emulsion was formed, an appropriate amount of citric acid (set forth in Table 3A) was added to adjust the pH of the emulsion to between 2.6 and 3.4 as described in Example 1 above.

Table 3A: Nanoemulsion Liquid Concentrate with 5% Non-Polar Compound containing Coenzyme Q (CoQ10) and 17.75% TPGS Surfactant

<td>Ingredient</td><td>mg / 2 ml serving</td><td>Phase</td><td>Percentage (by weight) of concentrate</td><td>g / lot</td>

<td>Vitamin E Oil (5-67) (non-polar solvent)</td><td> 75,00</td><td>Oily</td><td> 3,750</td><td> 24,375</td>

<td>SALADIZER® Brand Emulsion Stabilizer (combination of xanthan gum, guar gum and sodium alginate)</td><td> 1,270</td><td>Oily</td><td> 0,06</td><td> 0,4</td>

<td>Water</td><td> 1435</td><td>Watery</td><td> 71,74</td><td> 466,3</td>

<td>SALADIZER® Brand Emulsion Stabilizer (combination of xanthan gum, guar gum and sodium alginate)</td><td> 6,800</td><td>Watery</td><td> 0,34</td><td> 2,2</td>

<td>Tocopherol Polyethylene Glycol Succinate (surfactant)</td><td> 355,0</td><td>Oily</td><td> 17,75</td><td> 115,38</td>

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<td>Ingredient</td><td>mg / 2 ml serving</td><td>Phase</td><td>Percentage (by weight) of concentrate</td><td>g / lot</td>

<td>Phosphatidylcholine S-100 (co-surfactant)</td><td> 13,38</td><td>Oily</td><td> 0,6690</td><td> 4,349</td>

<td>Benzyl alcohol (preservative)</td><td> 10,00</td><td>Oily</td><td> 0,5000</td><td> 3,2500</td>

<td>CoQ10</td><td> 100,0</td><td>Oily</td><td> 5,0000</td><td> 32,500</td>

<td>Citric Acid (pH adjuster)</td><td> 3,800</td><td>Emulsion</td><td> 0,1900</td><td> 1,2350</td>

<td>Totals</td><td> 2000,00</td><td></td><td> 100,000</td><td> 650</td>

Example 4: Liquid Nanoemulsion Concentrates with Non-Polar Compounds containing Phytosterol

Example 4A Nanoemulsion Liquid Concentrate with 5.25% Non-Polar Phytosterols Compound and 20% TPGS Surfactant

Table 4A, below, lists the ingredients used to prepare a 250 g batch of liquid nanoemulsion concentrate containing phytosterols (serving size 2 ml), which was prepared according to the procedure set forth in Example 1 above, with the following details and modifications:

To prepare the oil phase using the method described in Example 1 above, the following oil phase ingredients were added sequentially, in the following order, to the oil phase container: 1) surfactant, 2) preservative; 3) non-polar solvent; 4) co-surfactant; 5) non-polar active ingredient; and 6) emulsion stabilizer.

The non-polar active ingredient was a Phytosterols compound, marketed under the name CardioAid ™, distributed by B&amp;D Nutrition and manufactured by ADM Natural Health and Nutrition, Decatur, IL. This compound contained Kosher, Pareve and Halal plant sterols that are produced according to current food GMPs and contained a minimum of 95% plant sterols. The non-polar active ingredient was added in an amount such that the ingredient was 5.25% by weight (w / w) of the final concentrate. The cosurfactant was a phosphatidylcholine cosurfactant, marketed under the trade name S-100, by Lipoid, LLC, Newark, NJ. This phosphatidylcholine co-surfactant was obtained from soybean extract and contained more than 95% phosphatidylcholine. The non-polar solvent was a Flaxseed oil, obtained from Sanmark LLC, Greensboro, NC (Sanmark Limited, Dalian, Liaoning Province, China), containing not less than (NLT) 50% C18: 3 alpha-linolenic acid. The preservative was a natural preservative (GRAS certified), benzyl alcohol. The emulsion stabilizer was SALADIZER® brand emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

To prepare the aqueous phase using the method described in Example 1 above, the following aqueous phase ingredients were added sequentially, in the following order, to the aqueous phase container: 1) water, 2) emulsion stabilizer; and 3) pH adjuster.

The water was purified city water, which was purified according to the methods provided by passing it through and using the following purifiers, sequentially: a carbon filter, an ion exchange purifier, a reverse osmosis purifier and a filter of end point, for example a 100 micron end point filter, before adding to the aqueous phase tank. The emulsion stabilizer was SALADIZER® brand emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD. The pH adjuster was phosphoric acid, which was added at the indicated concentration to ensure that the pH of the concentrate was between 2.6 and 3.4.

After the emulsion was formed, the pH of the concentrate was measured with a peachmeter, as described above, to ensure that it was between 2.6 and 3.4.

Table 4A: Nanoemulsion Liquid Concentrate with 5.25% Non-Polar Phytosterols Compound and 20% TPGS Surfactant

<td>Ingredient</td><td>mg / 2 ml serving</td><td>Phase</td><td>Percentage (by weight) of concentrate</td><td>g / lot</td>

<td>Flaxseed Oil (50% TO)</td><td> 105,00</td><td>Oily</td><td> 5,25</td><td> 13,13</td>

<td>Water</td><td> 1365,70</td><td>Watery</td><td> 68,29</td><td> 170,71</td>

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<td>Ingredient</td><td>mg / 2 ml serving</td><td>Phase</td><td>Percentage (by weight) of concentrate</td><td>g / lot</td>

<td>SALADIZER® Brand Emulsion Stabilizer (combination of xanthan gum, guar gum and sodium alginate)</td><td> 6,800</td><td>Watery</td><td> 0,17</td><td> 0,41</td>

<td>Tocopherol Polyethylene Glycol Succinate (surfactant)</td><td> 400,00</td><td>Oily</td><td> 20,00</td><td> 50,00</td>

<td>Phosphatidylcholine (cosurfactant)</td><td> 3,00</td><td>Oily</td><td> 0,15</td><td> 0,38</td>

<td>Benzyl alcohol (preservative)</td><td> 10,00</td><td>Oily</td><td> 0,50</td><td> 1,25</td>

<td>Phytosterols</td><td> 105,00</td><td>Oily</td><td> 5,25</td><td> 13,13</td>

<td>Phosphoric Acid (pH adjuster)</td><td> 8,00</td><td>Watery</td><td> 0,40</td><td> 1,00</td>

<td>Totals</td><td> 2000,00</td><td></td><td> 100,00</td><td> 250,00</td>

Example 5: Liquid Nanoemulsion Concentrates with Various Non-Polar Active Ingredients

Examples 5A-B set forth the details of liquid nanoemulsion concentrates containing two or more non-polar compounds (non-polar active ingredients), selected from polyunsaturated fatty acids (PUFAs) (for example, non-polar active ingredients containing omega-fatty acids). 3 and omega-6 fatty acids), Coenzyme Q10 and vitamins. These concentrates were prepared using the general procedure set forth in Example 1 above.

Example 5A: Liquid Nanoemulsion Concentrate formulated for Women

Table 5A, below, lists the ingredients used to prepare a 250 g batch (serving size 2 ml) of a nanoemulsion liquid concentrate containing nonpolar compounds including omega-3 acids (e.g. DHA, GLA, ALA ), omega-6 fatty acids, Coenzyme Q10 and vitamins, which were prepared according to the procedure set forth in Example 1 above, with the following details and modifications:

To prepare the oil phase using the method described in Example 1 above, the following oil phase ingredients were added sequentially, in the following order, to the oil phase container: 1) surfactant, 2) preservative; 3) non-polar active ingredients; and 4) emulsion stabilizer.

The non-polar active ingredients of omega-3 fatty acid and omega-6 fatty acid included:

A compound of borage oil, obtained from Sanmark LLC, Greensboro, NC (Sanmark Limited, Dalian, Liaoning Province, China), which was obtained by pressing and isolating oil from the seeds of Borago officinalis L. This non-polar active ingredient from borage oil contained not less than (NLT) 22% gamma-linolenic acid (GLA) C18: 3 and was added in an amount of 4.55% by weight (w / w) of the final concentrate, whereby the concentrate contained 1 0% GLA;

A flaxseed oil compound, which was Fresh Flaxseed Oil, obtained from Barleans Organic Oils, LLC, Ferndale, WA, which contained not less than (NLT) 55% C18: 3 alpha-linolenic acid and was added in a 2.4% by weight (w / w) amount of the final concentrate, whereby the concentrate contained 1.2% ALA; Y

Fish oil, containing approximately 30% DHA / EPA (marketed under the name Omega 30 TG Food Grade (Non-GMO) MEG-3 ™ Fish Oil from Ocean Nutrition Canada Limited, Nova Scotia, MA). The non-polar fish oil active ingredient was added in an amount of 0.2% by weight of the final concentrate, whereby the concentrate contained 0.06% EPA + DHA.

The non-polar active ingredient that contained Coenzyme Q was a Coenzyme Q10 compound (CoQ10, marketed under the name Kaneka Q10 ™ (USP Ubidecarenone) from Kaneka Nutrients, LP, Pasadena, TX, which contained more than 98% ubidecarenone (ubiquinone) The non-polar active ingredient was added in an amount such that the ingredient was 0.5% by weight (w / w) of the final concentrate.

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Non-polar active ingredients (vitamin) included Vitamin E and Vitamin D3. The vitamins were added in amounts corresponding to the Dietary Reference Intakes (DRI) for women. Vitamin D3 was obtained from DSM Nutritional Products, Parsippany, NJ. Vitamin E, as a Vitamin E oil, was marketed by ADM Natural Health and Nutrition, Decatur, IL, under the name Novatol ™ 5-67 Vitamin E (D-alpha Tocopherol; product code ADM 410217). This oil contained at least 67.2% Tocopherol and approximately 32.8% soybean oil.

The preservative was a natural preservative (GRAS certified), benzyl alcohol. The emulsion stabilizer was SALADIZER® brand emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

To prepare the aqueous phase using the method described in Example 1 above, the following aqueous phase ingredients were added sequentially, in the following order, to the aqueous phase container: 1) water and 2) emulsion stabilizer.

The water was purified city water, which was purified according to the methods provided by passing it through and using the following purifiers, sequentially: a carbon filter, an ion exchange purifier, a reverse osmosis purifier and a filter of end point, for example a 100 micron end point filter, before adding to the aqueous phase tank. The emulsion stabilizer was SALADIZER® brand emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

After the emulsion was formed, an appropriate amount of citric acid (set forth in Table 5A) was added to adjust the pH of the emulsion to between 2.6 and 3.4 as described in Example 1 above.

Table 5A: Liquid Nanoemulsion Concentrate formulated for Women

<td>Ingredient</td><td>mg / 2 ml serving</td><td>Phase</td><td>Percentage (by weight) of concentrate</td><td>g / lot</td>

<td>Borage Oil (22% GLA) (non-polar active ingredient)</td><td> 91,00</td><td>Oily</td><td> 4,55</td><td> 11,38</td>

<td>Flaxseed Oil 50% Omegas (non-polar active ingredient)</td><td> 48,00</td><td>Oily</td><td> 2,40</td><td> 6,0</td>

<td>Omega 30 TG Food Grade (Non-GM) Fish Oil MEG-3 ™ (non-polar active ingredient)</td><td> 4,00</td><td>Oily</td><td> 0,20</td><td> 0,5</td>

<td>Kaneka Q10 ™ (non-polar active ingredient)</td><td> 10,000</td><td>Oily</td><td> 0,50</td><td> 1,25</td>

<td>Vitamin E (5-67) oil (non-polar active ingredient)</td><td> 15,000</td><td>Oily</td><td> 0,75</td><td> 1,875</td>

<td>Vitamin D3 (non-polar active ingredient)</td><td> 0,010</td><td>Oily</td><td>5.0x10-⁶</td><td> 0,00125</td>

<td>Water</td><td> 1310,75</td><td>Watery</td><td> 65,538</td><td> 163,844</td>

<td>SALADIZER® Brand Emulsion Stabilizer (combination of xanthan gum, guar gum and sodium alginate)</td><td> 1,200</td><td>Watery</td><td> 0,06</td><td> 0,15</td>

<td>Tocopherol Polyethylene Glycol Succinate (surfactant)</td><td> 504,0</td><td>Oily</td><td> 25,20</td><td> 63,00</td>

<td>SALADIZER® Brand Emulsion Stabilizer (combination of xanthan gum, guar gum and sodium alginate)</td><td> 0,44</td><td>Oily</td><td> 0,0220</td><td> 0,06</td>

<td>Benzyl alcohol (preservative)</td><td> 10,000</td><td>Oily</td><td> 0,50</td><td> 1,25</td>

<td>Citric Acid (pH adjuster)</td><td> 5,60</td><td>Emulsion</td><td> 0,2800</td><td> 0,700</td>

<td>Totals</td><td> 2000,000</td><td></td><td> 100,0000</td><td> 250</td>

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Example 5B: Liquid Nanoemulsion Concentrate formulated for Children

Table 5B below lists the ingredients used to prepare a 250 g batch (serving size 2 ml) of a liquid nanoemulsion concentrate containing nonpolar compounds including omega-3 fatty acids (e.g. dHa, ALA). and vitamins that were prepared according to the procedure set forth in Example 1 above, with the following details and modifications:

To prepare the oil phase using the method described in Example 1 above, the following oil phase ingredients were added sequentially, in the following order, to the oil phase container: 1) surfactant, 2) preservative; 3) non-polar active ingredients; and 4) emulsion stabilizer.

Non-polar omega-3 fatty acid active ingredients included:

A flaxseed oil compound, which was Fresh Flaxseed Oil obtained from Barleans Organic Oils, LLC, Ferndale, WA, containing not less than (NLT) 55% C18: 3 alpha-linolenic acid, and was added in a 4.2% by weight (w / w) amount of the final concentrate, whereby the concentrate contained 2.1% ALA; Y

Fish oil, containing approximately 30% DHA / EPA (marketed under the name Omega 30 TG Food Grade (Non-GMO) MEG-3 ™ Fish Oil from Ocean Nutrition Canada Limited, Nova Scotia, MA). The non-polar fish oil active ingredient was added in an amount of 0.2% by weight of the final concentrate, whereby the concentrate contained 0.06% EPA + DHA.

Non-polar active vitamin ingredients included Vitamin E, Vitamin D3, and Vitamin A Palmitate. Vitamins were added in amounts corresponding to the Dietary Reference Intakes (DRI) for children. Vitamin D3 and Vitamin A Palmitate were obtained from DSM Nutritional Products, Parsippany, NJ. Vitamin E, in the form of Vitamin E oil, was marketed by ADM Natural Health and Nutrition, Decatur, IL, under the name Novatol ™ 5-67 Vitamin E (D-alpha-Tocopherol; product code ADM 410217). This oil contained at least 67.2% Tocopherol and approximately 32.8% soybean oil.

The preservative was a natural preservative (GRAS certified), benzyl alcohol. The emulsion stabilizer was SALADIZER® brand emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

To prepare the aqueous phase using the method described in Example 1 above, the following aqueous phase ingredients were added sequentially, in the following order, to the aqueous phase container: 1) water and 2) emulsion stabilizer.

The water was purified city water, which was purified according to the methods provided by passing it through and using the following purifiers, sequentially: a carbon filter, an ion exchange purifier, a reverse osmosis purifier and a filter of end point, for example a 100 micron end point filter, before adding to the aqueous phase tank. The emulsion stabilizer was SALADIZER® brand emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

After the emulsion was formed, an appropriate amount of citric acid (set forth in Table 5B) was added to adjust the pH of the emulsion to between 2.6 and 3.4 as described in Example 1 above.

Table 5B: Liquid Nanoemulsion Concentrate formulated for Children

<td>Ingredient</td><td>mg / 2 ml serving</td><td>Phase</td><td>Percentage (by weight) of the concentrate</td><td>g / lot</td>

<td>Flaxseed Oil 50% Omegas (non-polar active ingredient)</td><td> 84,00</td><td>Oily</td><td> 4,20</td><td> 10,50</td>

<td>Omega 30 TG Food Grade (Non-GM) Fish Oil MEG-3 ™ (non-polar active ingredient)</td><td> 4,00</td><td>Oily</td><td> 0,20</td><td> 0,50</td>

<td>Vitamin E (5-67) oil (non-polar active ingredient)</td><td> 30,000</td><td>Oily</td><td> 1,50</td><td> 3,75</td>

<td>Vitamin D3 (non-polar active ingredient)</td><td> 0,800</td><td>Oily</td><td> 0,04</td><td> 0,1</td>

<td>Vitamin A Palmitate (non-polar active ingredient)</td><td> 2,800</td><td>Oily</td><td> 0,14</td><td> 0,35</td>

<td>Water</td><td> 1357,16</td><td>Watery</td><td> 67,858</td><td> 169,645</td>

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<td>Ingredient</td><td>mg / 2 ml serving</td><td>Phase</td><td>Percentage (by weight) of the concentrate</td><td>g / lot</td>

<td>SALADIZER® Brand Emulsion Stabilizer (combination of xanthan gum, guar gum and sodium alginate)</td><td> 1,200</td><td>Watery</td><td> 0,06</td><td> 0,15</td>

<td>Tocopherol Polyethylene Glycol Succinate (surfactant)</td><td> 504,0</td><td>Oily</td><td> 25,20</td><td> 63,00</td>

<td>SALADIZER® Brand Emulsion Stabilizer (combination of xanthan gum, guar gum and sodium alginate)</td><td> 0,44</td><td>Oily</td><td> 0,0220</td><td> 0,06</td>

<td>Benzyl alcohol (preservative)</td><td> 10,000</td><td>Oily</td><td> 0,50</td><td> 1,25</td>

<td>Citric Acid (pH adjuster)</td><td> 5,60</td><td>Emulsion</td><td> 0,280</td><td> 0,70</td>

<td>Totals</td><td> 2000,000</td><td></td><td> 100,0000</td><td> 250</td>

Example 6: Dilution of Liquid Nanoemulsion Concentrates and Evaluation of Liquid Dilution Compositions

For the evaluation of various properties, the selected liquid nanoemulsion concentrates, prepared in the previous Examples, were diluted according to the methods provided, in an aqueous medium to form aqueous liquid dilution compositions. The results are described in detail in Examples 6A-B below.

Example 6A: Dilution and Transparency Evaluation of Dilution Compositions: Turbidity Analysis

The concentrates prepared in Examples 2A (vi), 2A (xii), 2A (iii), 2A (v) and 3A were diluted in the aqueous medium, according to the methods provided to dilute the concentrates. The resulting aqueous liquid dilution compositions were then evaluated for transparency by measuring turbidity using a nephelometer. Dilution parameters and evaluation results are set forth in Table 6A below. For each sample indicated in Table 6A, the Example in which the concentrate was prepared is indicated.

Each of the liquid nanoemulsion concentrates shown in Table 6A was diluted by adding the amount of concentrate indicated in Table 6A to the amount of water (purified according to the methods provided) indicated in Table 6A. Approximate dilution factors are also shown. The concentrates were diluted in an aqueous medium according to the methods provided to dilute the concentrates, using the following steps:

The indicated amount of water was heated in a Pyrex® beaker, by placing the beaker in a Thermolyne hot plate (Model No. 846925), until the water reached 49.8 ° C. The indicated amount of liquid nanoemulsion concentrate (approximately 1 g) was then added to the heated water, and stirred with a stir bar until dispersed. The resulting aqueous liquid dilution composition was added to a screw cap amber glass vial (Alcon) for evaluation. Each of the liquid dilution compositions containing DHA prepared from the concentrates of Examples 2A (iii) and 2A (v) contained 17.5 mg of DHA (in 250 ml and 500 ml of water, respectively). The liquid dilution composition containing DHA prepared from the concentrate of Example 2A (xii) contained 10 mg of DHA and 20 mg of EPA in 250 ml of water. The liquid dilution composition containing DHA prepared from the concentrate of Example 2A (vi) contained

6.5 mg of DHA and 6.5 mg of EPA in 250 ml of water.

The vials containing the liquid dilution compositions were sent to ACZ Laboratories, Inc., Steamboat Springs, CO, for turbidity analysis using a nephelometer. The results are shown in the form of Nephelometric Turbidity Units (NTU) and are listed in Table 6A below. As shown in Table 6A, each of the aqueous liquid compositions containing the dilute concentrates had an NTU value of less than about 300. Several of the compositions had an NTU value of between about 10 and about 12.

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Table 6A: Turbidity (NTU) of Aqueous Liquid Compositions containing Liquid Nanoemulsion Concentrates

<td>Concentrate of:</td><td>Non-Polar Active Ingredient</td><td>Concentrate (grams)</td><td>Water (grams)</td><td>Dilution</td><td>NTU</td>

<td>Example 2A (iii)</td><td>What does DHA (Seaweed Oil) contain</td><td> 1,0085</td><td> 250</td><td> 1:250</td><td> 12,1</td>

<td>Example 2A (v)</td><td>What does DHA (Seaweed Oil) contain</td><td> 1,0153</td><td> 500</td><td> 1:500</td><td> 159</td>

<td>Example 2A (vi)</td><td>What does DHA (Fish Oil) contain</td><td> 1,0215</td><td> 250</td><td> 1:250</td><td> 10,7</td>

<td>Example 2A (xii)</td><td>What does DHA (Fish Oil) contain</td><td> 1,0013</td><td> 250</td><td> 1:250</td><td> 10,4</td>

<td>Example 3A</td><td>CoQ10</td><td> 1,0246</td><td> 250</td><td> 1:250</td><td> 239</td>

Example 6B: Dilution and Evaluation of Transparency of Dilution Compositions: Particle Size

The nanoemulsion liquid concentrate prepared in Example 3A above, which included a non-polar active ingredient CoQ10, was shipped to Delta Analytical Instruments, Inc. for particle size measurement, which was made by diluting the concentrate and then analyzing it using the Horiba® LB-550 light scattering. The liquid nanoemulsion concentrate from Example 3A was mixed well and three drops of the concentrate were added to 25 ml of water. The mixture was put into a cell, which was used to measure the mean particle size on the Horiba® LB-550 light scattering analyzer. The results included the measurement of the mean particle size in the dilution composition, which was measured three times in different tests. The measurement for each trial and the average of the three trials are listed in Table 6B below. As indicated in Table 6B, the particle size of the liquid dilution composition was less than 150 nm.

Table 6B: Particle Size of the Liquid Aqueous Dilution Composition containing a Liquid Concentrate of Coenzyme Q Nanoemulsion

<td></td><td>Average Particle Size (nm)</td>

<td>Essay 1</td><td> 147,5</td>

<td>Essay 2</td><td> 143,8</td>

<td>Essay 3</td><td> 144,0</td>

<td>Average</td><td> 145,1</td>

Example 7: General Procedure Used to Prepare Liquid Nanoemulsion Concentrates in Examples 8 and 9

Tables 8A (i) through 9C, below, set forth ingredients that were used to prepare the exemplary nanoemulsion liquid concentrates described in Examples 8 and 9 below. Each of these concentrates contained a non-polar active ingredient, a polar solvent, and a surfactant. Each concentrate additionally contained a certified natural preservative GRAS (benzyl alcohol) and was produced, according to this general method, in a batch of 1000 grams (g) or 500 g (batch sizes indicated in the Tables).

Each of Tables 8A (i) to 9C shows the milligrams (mg) per 2 ml serving of each ingredient in the exemplary concentrate, the percentage, by weight (of the total concentrate) for each ingredient, and the amount in grams (g ) of each ingredient per 1000 g batch. As also indicated in each table, in the phase column, it is said whether each ingredient is added to the aqueous phase (aqueous), to the oil phase (oily) or is subsequently added to the emulsion formed after combining the oil phases and aqueous in the emulsifying stage (emulsion).

Each of the liquid nanoemulsion concentrates set forth in Examples 8 and 9 was prepared using an experimental scale process of the methods provided. To prepare larger batch sizes, the experimental scale process was scaled up to prepare any of these exemplary concentrates in Examples 8-9 using a large scale manufacturing process of the methods provided as above.

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The experimental scale process to prepare the concentrates in Examples 8 and 9 was performed using the following general steps (additional details are provided in the individual examples):

To prepare the concentrates, the indicated amount of each ingredient was weighed using a Toledo Balance (Model GD13x / USA), a Sartorius Basic Analytical Balance (Model BA110S) or an OHAUS Balance (Model CS2000). The selection of a scale or scales depends on the weight of the particular ingredient or ingredients. To generate the aqueous phase, the ingredients of the aqueous phase (indicated by aqueous in each table in the phase column) were added in the indicated amount (g / batch) to an aqueous phase container (a Pyrex® beaker ), and mixed using a reversible homogenizer (Arde Barinco, Inc .; Model CJ-4E), at 30 RPM. During mixing, the aqueous phase ingredients were heated until the ingredients reached the desired temperature of 60 ° C, using a hot plate as a warming device (a Thermolyne Hot Plate Model No. SP46615, Bamstead International, Dubuque, Iowa) . The temperature of the water phase and the mixing rate were maintained until the water and oil phases were combined and emulsified. A temperature meter (temperature probe (Model No. DPP400W, Cooper-Atkins)) was used to evaluate (measure) the temperature of the aqueous phase. The aqueous phase ingredients included a polar solvent (water, glycerin, or propylene glycol) and additional aqueous phase ingredients, where indicated.

The oil phase ingredients (indicated by oil in each table in the phase column) were added to an oil phase container (a Pyrex® beaker) and mixed using a conventional mixer (IKA® model No.RE- 16 1S, which is a top mixer (laboratory mixer) compatible with the experimental scale process). The oil phase ingredients included non-polar active ingredients and other oil ingredients as indicated in the Examples.

As the oil phase ingredients were mixed, they were heated using a hot plate as a warming apparatus (a Thermolyne Hot Plate Model No. SP46615, Bamstead International, Dubuque, Iowa), to a desired temperature of 60 ° C and they were generally mixed at that temperature until the ingredients had dissolved, and were kept at this temperature before mixing with the aqueous phase. A temperature meter (temperature probe (Model No. DPP400W, Cooper-Atkins)) was used to evaluate (measure) the temperature of the oil phase.

After both phases had reached the appropriate temperatures and the oil phase components had dissolved, the phases were combined and emulsified. The emulsion was carried out with a reversible homogenizer (Arde Barinco, Inc .; Model CJ-4E). The reversible homogenizer that was used to mix the aqueous phase ingredients was held at 30 RPM for mixing during the emulsifying step. While mixing with the homogenizer at this speed, the oil phase was transferred to the water phase container by pouring it from the oil phase container into the water phase container. Mixing with the homogenizer continued, with the adjustment of the baffle plate in the homogenate to achieve and maintain an emulsion, for example by moving the baffle plate further into the forming emulsion and / or out of the forming emulsion. During emulsification, the forming emulsion was rapidly cooled by placing the aqueous phase container (beaker) in a water bath, until the temperature of the liquid reached a desired temperature, as indicated in the Examples, between 35 ° C and 43 ° C, (which typically takes between about 30 and about 60 minutes).

In some examples, after emulsification and quenching, additional ingredients were added, as indicated in the individual Examples / Tables. In some examples, citric acid was added after combining and emulsifying the oil and water phases (indicated by emulsion on the phase column), while mixing with the reversible homogenizer (Arde Barinco, Inc .; Model CJ-4E).

As a final step, the concentrates were filtered using a 100 micron end product filter, prior to further evaluation, dilution and / or use.

Example 8: Liquid Nanoemulsion Concentrates with Non-Polar Compounds containing PUFA

Examples 8A-8C set forth the details of liquid nanoemulsion concentrates containing non-polar compounds (non-polar active ingredients) containing polyunsaturated fatty acids (PUFAs). The non-polar PUFA-containing active ingredients in the exemplified compositions were omega-3 fatty acids, omega-6 fatty acids, and conjugated fatty acids including:

A compound of flaxseed oil, which was Fresh Flaxseed Oil, obtained from Barleans Organic Oils, LLC, Ferndale, WA, which contained not less than (NLT) 55% C18: 3 alpha-linolenic acid and which was added in an amount of 5% by weight (w / w) of the final concentrate, whereby the concentrate contained 2.5% ALA;

A compound of borage oil, obtained from Sanmark LLC, Greensboro, NC (Sanmark Limited, Dalian, Liaoning Province, China), which was obtained by pressing and isolating oil from the seeds of Borago officinalis L. This oil from

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A conjugated linolenic acid (CLA) compound, marketed under the trade name Tonalin®, from Cognis Corporation, Cincinnati, OH, containing 1.7% by weight (w / w) C16 Palmitic acid: 0.6% by weight (w / w) of Stearic acid C: 18, 13.00% of Oleic acid C9 C18: 1, 0.20% by weight (w / w) of Linoleic acid C9 C18: 2 and 81.00% in weight (w / w) of Conjugated Linoleic Acid (CLA), which included 39.70% Conjugated C9 T11 isomer and 39.50% Conjugated T10, C12 isomer. This CLA-containing non-polar active ingredient was added in an amount of 5% by weight (w / w) of the final concentrate; Y

Fish oil, containing approximately 30% DHA / EPA (marketed under the name Omega 30 TG Food Grade (Non-GMO) MEG-3 ™ Fish Oil from Ocean Nutrition Canada Limited, Nova Scotia, MA). The non-polar fish oil active ingredient was added in an amount of 5% by weight of the final concentrate, whereby the concentrate contained 1.5% EPA + DHA.

Each of the concentrates containing these non-polar active ingredients was prepared using the general procedure set forth in Example 7 above.

Example 8A: Liquid Nanoemulsion Concentrates with Non-Polar Compounds containing PUFA, TPGS surfactant and glycerin

Tables 8A (i) -8A (iv) set forth the ingredients and other details of the liquid nanoemulsion concentrates, each of which contains a nonpolar PUFA-containing compound described above, the polar solvent glycerin, and a TPGS surfactant ( the TPGS surfactant marketed under the name Vitamin E TPGS® from Eastman Chemical Company). The specific PUFA-containing non-polar active ingredient is listed in each table.

These concentrates were produced according to the general method described in Example 7. Glycerin was obtained from Pan Century Oleochemicals SDN, BHD, Johor, Malaysia. For each of the concentrates, to prepare the oil phase, the general method of Example 7 was used, with the following details: The surfactant and preservative were added, mixed and heated (60 ° C) until the surfactant had melted and dissolved. The nonpolar active ingredient was then added, with continuous mixing, and heated to 60 ° C until combined with the aqueous phase.

During the emulsification of the water and oil phases as described in Example 7, the emulsion was rapidly cooled as described in the general method to a temperature of 35-43 ° C.

Table 8A (i): Nanoemulsion Liquid Concentrate with 5% of a Non-Polar Compound containing ALA, 25.2% of TPGS Surfactant and Glycerin

<td>Ingredient</td><td>mg / 2 ml serving</td><td>Phase</td><td>Percentage (by weight) of the concentrate</td><td>g / lot</td>

<td>Flaxseed Oil (55% Omega-3) (non-polar active ingredient)</td><td> 100</td><td>Oily</td><td> 5</td><td> 50</td>

<td>Glycerin (polar solvent)</td><td> 1386</td><td>Watery</td><td> 69,3</td><td> 693</td>

<td>Tocopherol Polyethylene Glycol Succinate (surfactant)</td><td> 504</td><td>Oily</td><td> 25,2</td><td> 252</td>

<td>Benzyl alcohol (preservative)</td><td> 10</td><td>Oily</td><td> 0,5</td><td> 5</td>

<td>Totals</td><td> 2000,000</td><td></td><td> 100</td><td> 1000</td>

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Table 8A (ii): Nanoemulsion Liquid Concentrate with 5% Non-Polar Compound containing GLA (Borage Oil), 25.2% TPGS Surfactant and Glycerin

<td>Ingredient</td><td>mg / 2 ml serving</td><td>Phase</td><td>Percentage (by weight) of concentrate</td><td>g / lot</td>

<td>Borage Oil (22% GLA) (non-polar active ingredient)</td><td> 100</td><td>Oily</td><td> 5</td><td> 50</td>

<td>Glycerin (polar solvent)</td><td> 1386</td><td>Watery</td><td> 69,3</td><td> 693</td>

<td>Tocopherol Polyethylene Glycol Succinate (surfactant)</td><td> 504</td><td>Oily</td><td> 25,2</td><td> 252</td>

<td>Benzyl alcohol (preservative)</td><td> 10</td><td>Oily</td><td> 0,5</td><td> 5</td>

<td>Totals</td><td> 2000,000</td><td></td><td> 100</td><td> 1000</td>

Table 8A (iii): Nanoemulsion Liquid Concentrate with 5% Non-Polar Compound containing CLA, 25.2% TPGS Surfactant and Glycerin

<td>Ingredient</td><td>mg / 2 ml serving</td><td>Phase</td><td>Percentage (by weight) of concentrate</td><td>g / lot</td>

<td>CLA Tonalin® oil (non-polar active ingredient)</td><td> 100</td><td>Oily</td><td> 5</td><td> 50</td>

<td>Glycerin (polar solvent)</td><td> 1386</td><td>Watery</td><td> 69,3</td><td> 693</td>

<td>Tocopherol Polyethylene Glycol Succinate (surfactant)</td><td> 504</td><td>Oily</td><td> 25,2</td><td> 252</td>

<td>Benzyl alcohol (preservative)</td><td> 10</td><td>Oily</td><td> 0,5</td><td> 5</td>

<td>Totals</td><td> 2000,000</td><td></td><td> 100</td><td> 1000</td>

Table 8A (iv): Nanoemulsion Liquid Concentrate with 5% Non-Polar Compound containing DHA 10 (Fish Oil), 25.2% TpGS Surfactant and Glycerin

<td>Ingredient</td><td>mg / 2 ml serving</td><td>Phase</td><td>Percentage (by weight) of concentrate</td><td>g / lot</td>

<td>Omega 30 TG Food Grade (Non-GM) Oil Fish Me-3^TM(non-polar active ingredient)</td><td> 100</td><td>Oily</td><td> 5</td><td> 25,00</td>

<td>Glycerin (polar solvent)</td><td> 1386</td><td>Watery</td><td> 69,3</td><td> 346,50</td>

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<td>Ingredient</td><td>mg / 2 ml serving</td><td>Phase</td><td>Percentage (by weight) of concentrate</td><td>g / lot</td>

<td>Tocopherol Polyethylene Glycol Succinate (surfactant)</td><td> 504</td><td>Oily</td><td> 25,2</td><td> 126,00</td>

<td>Benzyl alcohol (preservative)</td><td> 10</td><td>Oily</td><td> 0,5</td><td> 2,50</td>

<td>Totals</td><td> 2000,000</td><td></td><td> 100</td><td> 500</td>

Example 8B: Liquid Nanoemulsion Concentrates with Non-Polar Compounds containing PUFA, TPGS Surfactant and Propylene Glycol

Tables 8B (i) -8B (iv) set forth the ingredients and other details of the liquid nanoemulsion concentrates, each of which contains the nonpolar PUFA-containing compounds described above, the polar solvent propylene glycol, and a TPGS surfactant ( the TPGS surfactant marketed under the name Vitamin E TPGS® from Eastman Chemical Company). The concentrates were produced according to the general method described in Example 7. Propylene glycol was produced by Shell Chemicals, Alberta, Canada, and obtained from the distributor Mitsubishi International Food Ingredients, Inc., Dublin, OH. Specific PUFA-containing non-polar active ingredients are listed in the tables.

For each of the concentrates, to prepare the oil phase, the general method of Example 7 was used, with the following details: The surfactant and preservative were added, mixed and heated (60 ° C) until the surfactant had melted and dissolved. The non-polar active ingredient was then added, with continuous mixing and heating at 60 ° C until combined with the aqueous phase.

During the emulsification of the oil and water phases according to the general method of Example 7, the emulsion was rapidly cooled as described in Example 7, to a temperature of 35-43 ° C.

Table 8B (i): Nanoemulsion Concentrate with 5% Non-Polar Compound containing ALA, 25.2% TPGS Surfactant and Propylene Glycol

<td>Ingredient</td><td>mg / 2 ml serving</td><td>Phase</td><td>Percentage (by weight) of concentrate</td><td>g / lot</td>

<td>Flaxseed Oil (55% Omega-3) (non-polar active ingredient)</td><td> 100</td><td>Oily</td><td> 5</td><td> 50</td>

<td>Propylene Glycol (Polar Solvent)</td><td> 1386</td><td>Watery</td><td> 69,3</td><td> 693</td>

<td>Tocopherol Polyethylene Glycol Succinate (surfactant)</td><td> 504</td><td>Oily</td><td> 25,2</td><td> 252</td>

<td>Benzyl alcohol (preservative)</td><td> 10</td><td>Oily</td><td> 0,5</td><td> 5</td>

<td>Totals</td><td> 2000,000</td><td></td><td> 100</td><td> 1000</td>

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Table 8B (ii): Nanoemulsion Liquid Concentrate with 5% Non-Polar Compound containing GLA (Borage Oil), 25.2% TPGS Surfactant and Propylene Glycol

<td>Ingredient</td><td>mg / 2 ml serving</td><td>Phase</td><td>Percentage (by weight) of concentrate</td><td>g / lot</td>

<td>Borage Oil (22% GLA) (non-polar active ingredient)</td><td> 100</td><td>Oily</td><td> 5</td><td> 50</td>

<td>Propylene Glycol (Polar Solvent)</td><td> 1386</td><td>Watery</td><td> 69,3</td><td> 693</td>

<td>Tocopherol Polyethylene Glycol Succinate (surfactant)</td><td> 504</td><td>Oily</td><td> 25,2</td><td> 252</td>

<td>Benzyl alcohol (preservative)</td><td> 10</td><td>Oily</td><td> 0,5</td><td> 5</td>

<td>Totals</td><td> 2000,000</td><td></td><td> 100</td><td> 1000</td>

Table 8B (iii): Nanoemulsion Liquid Concentrate with 5% Non-Polar Compound containing CLA, 25.2% TPGS Surfactant and Propylene Glycol

<td>Ingredient</td><td>mg / 2 ml serving</td><td>Phase</td><td>Percentage (by weight) of concentrate</td><td>g / lot</td>

<td>CLA Tonalin® oil (non-polar active ingredient)</td><td> 100</td><td>Oily</td><td> 5</td><td> 50</td>

<td>Propylene Glycol (Polar Solvent)</td><td> 1386</td><td>Watery</td><td> 69,3</td><td> 693</td>

<td>Tocopherol Polyethylene Glycol Succinate (surfactant)</td><td> 504</td><td>Oily</td><td> 25,2</td><td> 252</td>

<td>Benzyl alcohol (preservative)</td><td> 10</td><td>Oily</td><td> 0,5</td><td> 5</td>

<td>Totals</td><td> 2000,000</td><td></td><td> 100</td><td> 1000</td>

Table 8B (iv): Nanoemulsion Liquid Concentrate with 5% Non-Polar Compound containing DHA 10 (Fish Oil), 25.2% TPGS Surfactant and Propylene Glycol

<td>Ingredient</td><td>mg / 2 ml serving</td><td>Phase</td><td>Percentage (by weight) of concentrate</td><td>g / lot</td>

<td>Omega 30 TG Food Grade (Non-GM) Fish Oil MEG-3 ™ (non-polar active ingredient)</td><td> 100</td><td>Oily</td><td> 5</td><td> 25,00</td>

<td>Propylene Glycol (Polar Solvent)</td><td> 1386,00</td><td>Watery</td><td> 69,3</td><td> 346,50</td>

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<td>Ingredient</td><td>mg / 2 ml serving</td><td>Phase</td><td>Percentage (by weight) of concentrate</td><td>g / lot</td>

<td>Tocopherol Polyethylene Glycol Succinate (surfactant)</td><td> 504</td><td>Oily</td><td> 25,2</td><td> 126,00</td>

<td>Benzyl alcohol (preservative)</td><td> 10</td><td>Oily</td><td> 0,5</td><td> 2,50</td>

<td>Totals</td><td> 2000,000</td><td></td><td> 100</td><td> 500</td>

Example 9: Liquid Nanoemulsion Concentrates with Non-Polar Compounds containing Coenzyme Q

Examples 9A-9C set forth the details of liquid nanoemulsion concentrates, each prepared with a nonpolar compound (nonpolar active ingredient) containing coenzyme Q, using the general procedure set forth in Example 7 above. The non-polar active ingredients in each of these concentrates will be Coenzyme Q10 (CoQ10), marketed under the trade name Kaneka Q10 ™ (USP Ubidecarenone) from Kaneka Nutrients, L. P., Pasadena, TX, which contains more than 98% ubidecarenone (ubiquinone), and is added in an amount of 5.25% by weight (w / w) of the final concentrate. In addition to the nonpolar compound, surfactant, polar solvent and preservative (as described in Example 7), the concentrates in Examples 9A-9C additionally contained a nonpolar solvent (Vitamin E oil sold by ADM Natural Health and Nutrition, Decatur, IL, under the name Novatol ™ 5-67 Vitamin E (D-alpha-Tocopherol; product code ADM 410217), which contained at least 67.2% Tocopherol and approximately 32.8% seed oil soy); and a cosurfactant (a phosphatidylcholine cosurfactant marketed under the name S-100 by Lipoid, LLC, Newark, NJ, or a soybean extract extract containing greater than 95% phosphatidylcholine).

Example 9A: Nanoemulsion Liquid Concentrate with 5% Non-Polar Compound Containing Coenzyme Q, 17.75% TPGS Surfactant and Glycerin

Table 9A below sets forth ingredients and other details of a liquid nanoemulsion concentrate containing the non-polar active ingredient CoQ10, the polar solvent glycerin, and a TPGS surfactant (the TPGS surfactant marketed under the name Vitamin E TPGS® by Eastman Chemical Company). This concentrate was produced according to the general method in Example 7. Glycerin was obtained from Pan Century Oleochemicals SDN, BhD, Johor, Malaysia).

The oil phase was prepared using the general methods described in Example 7, with the following details: The oil phase ingredients were added to the oil phase container in the following order: 1) non-polar solvent; 2) preservative; and 3) co-surfactant and mixed with the conventional mixer and heated at a temperature of 60 ° C, until the surfactant had dissolved. The TPGS surfactant was then added and dissolved at 60 ° C. The non-polar active ingredient CoQ10 was then added and dissolved at 60 ° C during the emulsification of the oil and water phases, as described in Example 7, the mixture was rapidly cooled to a temperature of 35-43 ° C.

Table 9A: Nanoemulsion Liquid Concentrate with 5% Non-Polar Compound containing Coenzyme Q, 17.75% TPGS Surfactant and Glycerin

<td>Ingredient</td><td>mg / 2 ml serving</td><td>Phase</td><td>Percentage (by weight) of the concentrate</td><td>g / lot</td>

<td>Vitamin E Oil (5-67) (non-polar solvent)</td><td> 75</td><td>Oily</td><td> 3,75</td><td> 37,5</td>

<td>Glycerin (polar solvent)</td><td> 1442</td><td>Watery</td><td> 72,08</td><td> 720,8</td>

<td>Tocopherol Polyethylene Glycol Succinate (surfactant)</td><td> 355</td><td>Oily</td><td> 17,75</td><td> 177,5</td>

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<td>Ingredient</td><td>mg / 2 ml serving</td><td>Phase</td><td>Percentage (by weight) of the concentrate</td><td>g / lot</td>

<td>Phosphatidylcholine (Alcolec PC95) (cosurfactant)</td><td> 13,38</td><td>Oily</td><td> 0,669</td><td> 6,69</td>

<td>Benzyl alcohol (preservative)</td><td> 10</td><td>Oily</td><td> 0,5</td><td> 5</td>

<td>Kaneka Q10 ™ (non-polar active ingredient)</td><td> 105</td><td>Oily</td><td> 5,25</td><td> 52,5</td>

<td>Totals</td><td> 2000,000</td><td></td><td> 100,0000</td><td> 1000</td>

Example 9B: Nanoemulsion Liquid Concentrate with 5% Non-Polar Compound containing Coenzyme Q, 17.75% TPGS Surfactant and Propylene Glycol

Table 9B below sets forth ingredients and other details of a liquid nanoemulsion concentrate containing the non-polar active ingredient CoQ10, the polar solvent propylene glycol, and a TPGS surfactant (the TPGS surfactant marketed under the name Vitamin E TPGS® from Eastman Chemical Company). This concentrate was produced according to the general method in Example 7. Propylene glycol was produced by Shell Chemicals, Alberta, Canada and obtained through distributor Mitsubishi International Food Ingredients, Inc., Dublin, OH.

The oil phase was prepared using the general methods described in Example 7, with the following details: The oil phase ingredients were added to the oil phase container in the following order: 1) non-polar solvent; 2) preservative; and 3) co-surfactant and mixed with the conventional mixer and heated at a temperature of 60 ° C, until the co-surfactant had dissolved. The TPGS surfactant was then added and dissolved at 60 ° C. The non-polar active ingredient CoQ10 was then added and dissolved at 60 ° C. This temperature was maintained until mixing with the aqueous phase for emulsion.

During the emulsification of the oil and water phases, as described in Example 7, the mixture was rapidly cooled to a temperature of 35-43 ° C.

Table 9B: Nanoemulsion Liquid Concentrate with 5% Non-Polar Compound containing Coenzyme Q, 17.75% TPGS Surfactant and Propylene Glycol

<td>Ingredient</td><td>mg / 2 ml serving</td><td>Phase</td><td>Percentage (by weight) of concentrate</td><td>g / lot</td>

<td>Vitamin E Oil (5-67) (non-polar solvent)</td><td> 75</td><td>Oily</td><td> 3,75</td><td> 37,5</td>

<td>Propylene Glycol (Polar Solvent)</td><td> 1442</td><td>Watery</td><td> 72,08</td><td> 720,8</td>

<td>Tocopherol Polyethylene Glycol Succinate (surfactant)</td><td> 355</td><td>Oily</td><td> 17,75</td><td> 177,5</td>

<td>Phosphatidylcholine (Alcolec PC95) (cosurfactant)</td><td> 13,38</td><td>Oily</td><td> 0,669</td><td> 6,69</td>

<td>Benzyl alcohol (preservative)</td><td> 10</td><td>Oily</td><td> 0,5</td><td> 5</td>

<td>Kaneka Q10 ™ (non-polar active ingredient)</td><td> 105</td><td>Oily</td><td> 5,25</td><td> 52,5</td>

<td>Totals</td><td> 2000,000</td><td></td><td> 100,0000</td><td> 1000</td>

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Since the modifications were apparent to those skilled in the art, it is intended that this invention be limited only by the scope of the appended claims.

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