Emulsions including a peg-derivative of tocopherol

[0001] Compositions and methods for making food and beverage products that contain additives such as nutraceuticals, pharmaceuticals and supplements are provided.

BACKGROUND OF THE INVENTION [0002] Non-polar compounds do not readily dissolve in aqueous solutions such as water or any other polar solvent. Many non-polar compounds are used in compositions for human consumption, for example pharmaceuticals, nutraceuticals and / or dietary supplements. Exemplary non-polar compounds used in such compositions are vitamins and minerals, fatty acids and other non-polar compounds, non-polar active agents and other non-polar active ingredients.

[0003] US Patent Document US2006 / 0088558 describes a liquid vitamin E TPGS concentrate composition, wherein said concentrate has a much higher percentage of TPGS per unit volume than traditional liquid TPGS formulations, providing TPGS in liquid form that can be used in soft gels or mixed with water to produce the desired concentrates for commercial use in supplements, beverages and pharmaceutical preparations, etc.

[0004] International Patent Application No. WO96 / 36316 describes self-emulsifying pre-concentrated pharmaceutical compositions comprising: (a) a lipophilic therapeutic compound, (b) 1000 d-alpha-tocopheryl polyethylene glycol succinate (TPGS) and (c) lipophilic phase as well pharmaceutical compositions containing such pre-concentrated concentrates in combination with a sufficient amount of water to form a stable emulsion.

[0005] US Patent Document US2005 / 208082 describes an aqueous emulsion in which the lipid phase of the emulsion contains a mixture of therapeutically effective lipophilic concentration, vitamin E concentration

TPGS and linoleic acid concentrations. The presence of linoleic acid increases the dissolution effect of vitamin E TPGS in lipophil and thus reduces the amount of vitamin E TPGS that would otherwise be required in the water emulsion.

[0006] Due to the poor water solubility, the inclusion of non-polar compounds in products for human consumption, for example, supplements, food products and beverages, is often a challenge. Available compositions containing non-polar compounds, in particular aqueous compositions containing non-polar compounds, and methods for formulating such compositions are limited. Thus, there is a need to improve compositions containing non-polar compounds and methods for making the compositions. Accordingly, one of the purposes here is to provide compositions, including aqueous compositions, containing non-polar compounds and methods for making the compositions.

EP 2 268 160 B1

SUMMARY [0007] The present invention provides a liquid nanoemulsion concentrate comprising:

vitamin E PEG derivative in an amount between 16% and 30% by weight concentrate;

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

a non-polar active ingredient comprising a compound or compounds selected from any or more 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 present in an amount between 5% and 10% by weight of the concentrate.

[0008] Primary compositions (concentrates) are provided that contain non-polar compounds, including liquid nanoemulsion concentrates. Also provided are methods that use such primary compositions to prepare other compositions, such as beverages and other hydrated liquids, in which the primary compositions are diluted to form liquid diluted compositions. Liquid dilution compositions are also provided comprising beverages or other hydrated liquid and diluted concentrate. Concentrates include dispersions, and / or can be used to prepare dispersions, effective amounts of additives, such as non-polar compounds, including non-polar active ingredients, such as nutraceuticals, pharmaceuticals, and supplements, such as fatty acids, including polyunsaturated fatty acids, such as acids 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 for supplementing the diet. The compositions provided herein are stable dispersions without phase separation and / or other changes.

[0009] For example, the compositions provided include concentrates containing non-polar active ingredients, surfactants and polar solvents in amounts by which the dilution of the concentrate in an aqueous medium, such as a drink, in a certain amount (e.g. any of the specified amounts, concentrations and dilutions of concentrates, and any of the amounts of non-polar active ingredients described herein below), provides a liquid diluted composition containing effective amounts of the non-polar active ingredient and having one or more desired properties. The compositions provided further include liquid diluted compositions, including liquid diluted compositions made from concentrates, containing an aqueous medium, non-polar active ingredients in effective amounts, and polar solvents that have the desired properties. The amount of concentrate and / or the amount of non-polar active ingredient can be determined. Desirable properties include the clarity of liquid dilute compositions, such as compositions that are clear or almost as clear as an aqueous medium in the absence of a concentrate and / or in the absence of a non-polar active ingredient; particle size, such as particle size smaller than 200 nm or smaller than about 200 nm, smaller than 100 nm or smaller than about 100 nm, smaller than 50 nm or smaller than about 50 nm or smaller than 25 nm or smaller than about 25 nm , at most or on average; turbidity, such as Nephelometric Turbidity Units (NTU,

Nephelometric Turbidity Units) 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 no visible particles, visible crystals, phase separation and / or ring formation.

[0010] The concentrates provided are liquid nanoemulsion concentrates that contain surfactants, a non-polar compound (s) (which is usually / constitutes a non-polar active ingredient that differs from the surfactant) and a polar solvent (e.g. water or edible hydrated liquid , such as a polar protic solvent such as dihydric or trihydric alcohol, e.g. propylene glycol or glycerol (glycerol)).

[0011] The amount of non-polar compound in the concentrate is between 5% or about 5% and 10% or about 10% by weight (w / w) of the concentrate, e.g. exactly or about 5, 5.2, 5.25, 6, 7, 8, 9 or 10% by weight of the concentrate.

[0012] In the concentrates provided, the surfactants have a Hydrophilic Lipophilic Balance (HLB) between 14 or about 14 and 20 or about 20, such as between 15 or about 15 and 18 or about 18, e.g. exactly or about 15, 16, 17 or 18.

[0013] Surfactants include vitamin E polyethylene glycol (PEG) derivatives as defined in the claims.

[0014] The amount of surfactant (s) in the concentrate is between 16% or about 16% and 30% or about 30% by weight of the concentrate, e.g. exactly or about 16, 17, 18, 19, 20, 21, 22, 23 , 24, 25, 26, 27, 28, 29 or 30% by weight (w / w) of the concentrate.

[0015] 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 (s) 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 composition, yes such as 17.75%, 20.25%, 20.5%, 22.7% or 25.2% (w / w) of the concentrate.

[0016] 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.

[0017] 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 may be at or about 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79 or 80% by weight (w / w wt.) of a concentrate, such as exactly or about 74.25%, exactly or about 75.8%, exactly or about 68.9%, exactly or about 71.74%, exactly or about 63.94%, exactly or about 68.79%, exactly or about 68.29%, exactly or about 69.02% or exactly or about 71.49% by weight (w / w) of the concentrate.

[0018] Among the surfactants obtained from PEG are PEG derivatives of vitamin E,

EP 2 268 160 B1 such as surfactants obtained from tocopherol or tocotrienol, in which the vitamin E derived unit represents the hydrophobic area of ​​the surfactant and is attached by means of a linker to another unit, such as a polyethylene glycol (PEG) unit. Exemplary surfactants derived from vitamin E include, but are not limited to, surfactants derived from tocopherol, including tocopherol polyalkylene glycol derivatives, typically tocopherol polyethylene glycol (PEG) derivatives, such as tocopherol polyethylene glycol (TPGD) diesters. tocopherol polyethylene glycol diesters), e.g. tocopherol polyethylene glycol succinate (TPGS), TPGS analogues, TPGS homologues and TPGS derivatives. Exemplary surfactants also include other PEG derivatives having similar properties, e.g., PEG sterol derivatives, e.g., cholesterol or sitosterol (including, for example, any of the PEG derivatives disclosed in US Patent No. 6,632,443) and PEG derivatives of other fat-soluble vitamins, on an example of certain forms of vitamin A (e.g. Retinol) or vitamin D (e.g. vitamins D1-D5).

[0019] Surfactants include vitamin E polyethylene glycol (PEG) derivatives, for example polyethylene glycol diester tocopherol (TPGD). In one example, TPGD is selected from tocopherol polyethylene glycol sebacate, tocopherol polyethylene glycol dodecanoate, tocopherol polyethylene glycol suberate, tocopherol polyethylene glycol azelaate, tocopherol polyethylene glycol citraconate, tocopherol methyl polyethylene glycolate, polyethylene glycol tocopherol, polyethylene glycol tocopherol, polyethylene glycol tocopherol glutaconate and tocopherol polyethylene glycol phthalate. In another example, the TPGD surfactant is tocopherol polyethylene glycol succinate (TPGS), such as TPGS-1000 and / or da TPGS. In another example, the surfactant is a TPGS analogue. In one aspect, the surfactant is a TPGS homologue, such as, for example, a TPGS homologue that differs from the parent TPGS compound by the addition or removal of one or more methylene units, e.g. - (CH2) n--.

[0020] PEG units in surfactants derived from PEG, including PEG units in PEG derivatives of vitamin E, include PEG units selected from any or more PEG-OH, PEG-NHS, aldehyde PEG, PEG-SH, PEG-NH2, PEGCO2H, methylated PEG (m-PEG) and branched PEG, and include PEG units having a molecular weight between 200 kDa or about 200 kDa to 20,000 kDa or about 20,000 kDa, usually between 200 kDa or about 200 kDa and 6,000 kDa or about 6,000 kDa . for example between 600 kDa or about 600 kDa and 6000 kDa or about 6000 kDa, usually 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.

In the provided compositions (including liquid nanoemulsion concentrates), exemplary non-polar compounds are non-polar active ingredients that include, but are not limited to, omega-3 fatty acids, omega-6 fatty acids, conjugated fatty acids, Coenzyme Q10 (e.g. ubidecarenone), phytosterols and sabal palm extracts. Non-polar active ingredients include, for example, non-polar compounds containing docosahexaenoic acid (DHA) and / or

EP 2 268 160 B1 eicosapentaenoic acid (EPA), alpha-linolenic acid (α-linolenic acid; ALA), conjugated linolenic acid (CLA) and gamma-linolenic acid (GLA), including, but not limited to, fish oil , algae oil, linseed oil, borage oil and sabal palm extracts.

[0022] Non-polar active ingredients include, but are not limited to, compounds containing any fat-soluble nutraceutical or pharmaceutical agent and / or oil, such as, for example, drugs, hormones, vitamins, nutrients, including any and other lipophilic compounds containing necessary unsaturated fatty acids, e.g. polyunsaturated fatty acids (PUFA), including, for example, omega-3 fatty acids, for example natural and synthetic omega-3 fatty acids, for example compounds containing polyunsaturated long chain omega-3 fatty acids, including eicosapentaenoic acid (EPA) (20: 5ω3), docosahexaenoic acid (DHA) (22: 6ω3), arachidonic acid (24 : 4ω3) docosapentaenoic acid (DPA, clupanodic acid (22: 5ω3), 16: 3 ω3; 24: 5 ω3 and / or nisin acid (24: 6ω3), which may contain, for example, fish oil, algae oil, krill oil, rapeseed oil, linseed oil, soybean oil and walnut oil; compounds containing short chain omega-3 fatty acids, e.g. alpha-linolenic acid (α-linolenic acid; ALA) (18: 3ω3) (e.g. linseed oil) and stearidic acid (18: 4ω3); esters of 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, e.g., EPA precursor, DHA precursor; derivatives such as polyglycolyzed derivatives or polyoxyethylene derivatives; oils containing omega-3 fatty acids, e.g. fish oil (e.g. marine oil), including, for example, high purity fish oil concentrates, perilla oil, krill oil and algae oil (e.g. oil from microalgae); compounds containing omega-6 fatty acids, for example compounds containing linoleic acid (18: 2ω6) (short chain fatty acid), gamma-linolenic acid (GLA) (18: 3ω6), dihomo-gammalinolenic acid (DGLA) (20: 3ω6) , eicosadienic acid (20: 2ω6), arachidonic acid (AA) (20: 4ω6), docosadienic acid (22: 2ω6), adrenic acid (22: 4ω6) and / or docosapentaenoic acid (22: 5ω6), for example oil from borage, corn oil, cottonseed oil, grape seed oil, peanut oil, evening primrose oil, for example evening primrose oil (Oenothera biennis), blackcurrant seed oil, hemp oil, spirulina extract, safflower oil, sesame oil and soybean oil;

compounds containing other fatty acids, e.g. triglycerides, including medium chain triglycerides, polar lipids, e.g. ether lipids, phosphoric acid, choline, fatty acids, glycerol, glycolipids, triglycerides and phospholipids (e.g. phosphatidylcholine (lecithin), phosphatidylethanolamine and phosphatidylinositol); sabal palm extract; and ethyl linolonate; and herb oils, for example garlic oils and scordinine; short chain saturated fatty acids (4: 010: 0), lauric acid (12: 0), myristic acid (14: 0), pentadecanoic acid (15: 0), palmitic acid (16: 0), palmitic acid (16: 1 ω7), heptadecanoic acid (17: 0), stearic acid (18: 0), oleic acid (18: 1 ω9), peanut acid (20: 0);

compounds containing trace elements, e.g. vitamins, minerals, cofactors, e.g. coenzymes such as coenzyme Q, e.g. coenzyme Q10 (CoQ10, also called ubiquinone, e.g. ubidecarenone or reduced form of CoQ10, e.g. ubiquinol), turmeric extract (e.g. curcuminoids),

Sabal palm lipid extract (e.g. sabal palm oil), echinacea extract, hawthorn extract, ginseng extract, lipoic acid (e.g. α-lipoic acid), ascorbyl palmitate, kava extract, St. John's wort (e.g. St. John's wort), quercitin extract, dehydroepiandrosterone, indole-3-carbinol; compounds containing carotenoids, including hydrocarbons and oxidized alcohol derivatives of hydrocarbons, e.g. beta-carotene, mixed carotenoids group, lutein, lycopene, zeaxanthin, cryptoxanthin, e.g. beta-cryptoxanthin, beta-carotene, mixed carotenoids group, astaxanthin, bixin, kantaxin , capsanthin, capsorubin, apo-carotenal, beta12'-apo-carotenal, beta-carotene, "Carotene" (a mixture of alpha and beta-carotene), gamma-carotene, betacryptoxanthin, ciolerythrin, zeaxanthin, their hydroxyl or carboxyl group containing esters;

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

compounds containing phytochemicals, including phytoestrogens, e.g. genistein and daidzin, e.g. isoflavones, e.g. soy isoflavones, flavonoids, phytoalexins, e.g. resveratol (3,5,4'-trihydroxystilbene), red clover extract and phytosterols; compounds containing fat-soluble drugs, including natural and synthetic forms of immunosuppressive drugs such as ciclosporin, protease inhibitors such as ritonavir, macrolide antibiotics and oil-soluble anesthetics such as propofol, natural and synthetic forms of steroid hormones, e.g. estrogens, estradiols, progesterone, testosterone, cortisone, phytoestrogens, dehydroepiandrosterone (DHEA), growth hormones and other hormones; compounds containing oil-soluble acids and alcohols, e.g. tartaric acid, lactylic acid, butylhydroxyanisole, butylhydroxytoluene, lignin, sterols, polyphenolic compounds, oryzanol, cholesterol, phytosterols, flavonoids, such as, but not limited to, quercetin and reservatol and diallyl disulfides.

[0023] Non-polar active ingredients include ingredients containing polyunsaturated fatty acids, such as compounds containing any or more omega-3 fatty acids, including docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA) and alpha-linolenic acid (ALA) (e.g., oils fish oils, krill oils, algae oils / or linseed oils), omega-6 fatty acids such as gamma-linolenic acid (GLA) (e.g. borage oils); conjugated fatty acids (e.g. conjugated linolenic acid (CLA)) and Sabal palm extracts; and ingredients containing coenzymes such as coenzymQ, for example coenzyme Q10 (e.g. ubidecarenone); and ingredients containing phytosterols and combinations thereof.

[0024] In one example, the non-polar active ingredient comprises eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA) or a combination thereof. In one example, the non-polar 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 comprises 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) 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 algae oil containing any such percentages of EPA and / or DHA.

[0025] In another example, the non-polar active ingredient contains alpha linolenic acid (ALA). In one example, the non-polar active ingredient contains ALA in an amount of at least 50% or about 50% by weight (w / w) of the non-polar 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 containing any such percentage of ALA.

[0026] In another example, the non-polar active ingredient contains gamma-linolenic acid (GLA). In one example, the non-polar active ingredient contains GLA in an amount of at least 22% or about 22% by weight (w / w) of the non-polar active ingredient. For example, non-polar active ingredients include borage oil containing GLA in an amount of at least 22% or about 22% by weight (w / w) of borage oil.

[0027] 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 of the non-polar compounds described herein. In one example, the total amount of non-polar active ingredient (s) is between exactly or about 5% and 10% by weight of the concentrate, for example wherein the combined weight of the non-polar active ingredient and the additional non-polar active ingredient (s) is no more than exactly or about 10 % w / w (w / w) concentrate.

[0028] Polar solvents contained in concentrates include polar protic solvents and polar aprotic solvents, and are usually 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 (e.g. exactly or about or at least exactly or 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 between exactly or about 3 and exactly or about 9 or a dipole moment between exactly or about 1.8 and exactly or about 2.8. Polar solvents include water and alcohols such as monohydric, dihydric, trihydric and other alcohols, and usually alcohols other than monohydric alcohols, alcohols having two or more hydroxyl groups, such as dihydric alcohols (two hydroxyl groups) and trihydroxy . 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

They may further include polyethylene glycols low molecular weight polyethylene glycols (PEGs) such as PEGs with molecular weights exactly or about, or less than exactly or about 600, 400 or 200 kDa. In some examples, the polar solvent is water, glycerin or propylene glycol.

[0029] The concentrates provided may contain one or more additional ingredients. In one example, the concentrate further contains a co-surfactant in an amount sufficient to stabilize the concentrate compared to the lack of a co-surfactant. In one aspect, the co-surfactant is a phospholipid such as, but not limited to, phosphatidylcholine. In one example, the amount of co-surfactant, e.g. phospholipid is between 0.1% or about 0.1% and 1% or about 1% by weight (w / w) of the concentrate.

[0030] In another example, the concentrate further contains a preservative in an amount sufficient to preserve the concentrate, compared to the lack of a preservative. Exemplary preservatives are natural preservatives such as benzyl alcohol and benzyl alcohol containing preservatives. 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 exactly 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.

[0031] In another example, the concentrate contains a non-polar solvent, for example a non-polar solvent, which dissolves and differs from the active ingredient. Typically, the amount of non-polar solvent is sufficient to dissolve the non-polar active ingredient and may be, for example, between 1% or about 1% and 6% or about 6%, for example exactly or about 1, 2, 3, 4, 5 or 6% by weight ( w / w) of the concentrate. The nonpolar solvent is usually an oil, such as any oil suitable for dissolving the nonpolar component. Examples of non-polar solvents are vitamin E oil, linseed oil, sunflower oil and any vegetable oils and other oils.

[0032] In another example, the concentrate contains an emulsion stabilizer. Usually, the emulsion stabilizer is contained in the concentrate in an amount sufficient to stabilize the concentrate. Emulsion stabilizers include, but are not limited to, compositions containing gum blends such as the Saladizer® trademark stabilizer. In one example, the emulsion stabilizer contains one or more of guar gum, xanthan gum and sodium alginate. In one example, the emulsion stabilizer contains guar gum, xanthan gum and sodium alginate.

[0033] In one example, the concentrates contain flavors. Usually, the agent (s) is contained in an amount sufficient to enhance the flavor of the concentrate, the aroma of the concentrate, or a combination thereof, compared to the lack of 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.

[0034] In another example, when the concentrates contain water as the polar solvent, for example, the concentrates contain a regulator. Typically, the pH regulator contains an acid or base in an amount sufficient to affect the pH of the concentrate, compared to the absence of a pH regulator. PH regulators include, but are not limited to, citric acid and phosphoric acid.

[0035] The concentrates provided include, but are not limited to, concentrates containing non-polar active ingredients containing PUFA, such as omega-3 fatty acid, omega-6 fatty acid, conjugated fatty acid and sabal palm oil containing non-polar active ingredients.

[0036] In one example, the concentrate provided contains a non-polar active ingredient that is fish oil containing eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) (e.g. fish oil containing 10% EPA and 70% DHA; fish oil containing about 13 % EPA and about 13% DHA; or fish oil containing 40% EPA and 20% DHA), a surfactant that is tocopherol polyethylene glycol succinate (TPGS) or an analogue TPGS (e.g. TPGS homolog) and polar solvent, which is water. In one aspect of the present invention, the amount of fish oil non-polar active ingredient is at or about 5% by weight (w / w) of the concentrate; the amount of TPGS or analog surfactant is exactly or about 18% by weight (w / w) of the concentrate and the amount of water is exactly or about 74.25% by weight (w / w) of the concentrate. In another aspect of this example, the amount of fish oil non-polar active ingredient is at or about 5% by weight (w / w) of the concentrate; the amount of TPGS surfactant or TPGS analogue is exactly or about 25.2% by weight (w / w) of the concentrate, the amount of water is exactly or about 68.79% by weight (w / w) of the concentrate. In another aspect of this example, the amount of fish oil active ingredient is at or about 10% by weight (w / w) of the concentrate; the amount of TPGS surfactant or TPGS analogue is exactly or about 20.2% by weight (w / w) of the concentrate, the amount of water is exactly or about 68.79% by weight (w / w) of the concentrate. In one aspect, the concentrate further comprises an emulsion stabilizer that is a blend of xanthan gum, guar gum and / or sodium alginate; a benzyl alcohol preservative; a pH regulator that is citric acid. In another aspect, the concentrate further contains D-limonene and / or lemon oil flavors. In another aspect, the concentrate further contains a phosphatidylcholine co-surfactant.

[0037] In another example, the concentrate provided contains a non-polar active ingredient that is an algae oil, such as algae oil containing 35% DHA, a surfactant that is TPGS or a TPGS analogue (e.g. TPGS homolog) and a polar solvent which is water. In one 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 surfactant or TPGS analogue is exactly or about 18% by weight (w / w) of the concentrate, the amount of water is exactly 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; quantity

The TPGS surfactant or TPGS analogue is exactly or about 25.2% by weight (w / w) of the concentrate and the amount of water is exactly or about 68.79% by weight (w / w) concentrate. In a further aspect of this example, the amount of the algal oil non-polar active ingredient is at or about 10% by weight (w / w) of the concentrate; the amount of TPGS surfactant or TPGS analogue is exactly or about 25.2% by weight (w / w) of the concentrate, the amount of water is exactly or about 63.94% by weight (w / w) of the concentrate. In one aspect of this example, the concentrate further comprises an emulsion stabilizer that is a blend of xanthan gum, guar gum and / or sodium alginate; a benzyl alcohol preservative; and a pH regulator that is citric acid. In another aspect, the concentrate further contains D-limonene and lemon oil flavors.

[0038] In another example, the concentrate contains a non-polar active ingredient, which is linseed oil (e.g. one containing 50% or 55% omega-3 fatty acids, e.g. 50% or 55% alpha-linolenic acid (ALA)), a surfactant that is TPGS or a TPGS analogue (e.g. TPGS homolog) and a polar solvent that is water. In one aspect of this example, the amount of linseed oil non-polar active ingredient is at or about 5% by weight (w / w) of the concentrate; the amount of TPGS surfactant or TPGS analogue is exactly or about 17.75% by weight (w / w) of the concentrate and the amount of water is exactly or about 71.74% by weight (w / w) of the concentrate. In another aspect of this example, the amount of linseed oil non-polar active ingredient is at or about 5% by weight (w / w) of the concentrate; the amount of TPGS surfactant or TPGS analogue is exactly or about 25.2% by weight (w / w) of the concentrate and the amount of water is exactly or about 68.79% by weight (w / w) of the concentrate. In another aspect of this example, the amount of linseed oil non-polar active ingredient is at or about 10% by weight (w / w) of the concentrate; the amount of TPGS surfactant or TPGS analogue is exactly or about 20.2% by weight (w / w) of the concentrate and the amount of water is exactly or about 68.79% by weight (w / w) of the concentrate. In one aspect, the concentrate further comprises an emulsion stabilizer that is a blend of xanthan gum, guar gum and / or sodium alginate; a benzyl alcohol preservative and a pH regulator that is citric acid. In another aspect, it further comprises D-limonene and lemon oil flavors. In another aspect, the concentrate further contains a phosphatidylcholine co-surfactant. In another aspect, the concentrate further contains a non-polar solvent that is vitamin E oil or other oil.

[0039] In another example, the concentrate contains a non-polar active ingredient, which is linseed oil (e.g., one containing 50% or 55% omega-3 fatty acids, e.g., 50% or 55% ALA), a surfactant that is TPGS or TPGS analogue (e.g. TPGS homolog) and polar solvent, which is glycerin. In one aspect of this example, the amount of linseed oil non-polar active ingredient is at or about 5% by weight (w / w) of the concentrate; the amount of TPGS surfactant or TPGS analogue is exactly or about 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 further contains a benzyl alcohol preservative.

[0040] In another example, the concentrate contains a non-polar active ingredient, which is linseed oil (e.g., one containing 50% or 55% omega-3 fatty acids, e.g., 50% or 55% ALA), a surfactant that is TPGS or TPGS analogue (e.g. TPGS homolog) and polar solvent, which is propylene glycol. In one aspect of this example, the amount of linseed oil non-polar active ingredient is at or about 5% by weight (w / w) of the concentrate; the amount of TPGS surfactant or TPGS analogue is exactly or about 25.2% by weight (w / w) of the concentrate and the amount of propylene glycol is exactly or about 69.02% by weight (w / w) of the concentrate. In one aspect, the concentrate further contains a benzyl alcohol preservative.

[0041] In another example, the concentrate contains a non-polar active ingredient that contains gamma-linolenic acid (GLA) (e.g. GLA-containing borage oil, e.g. 22% GLA), a surfactant that is TPGS or a TPGS analogue (e.g., TPGS homolog) and 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 analog 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 GLA-containing non-polar active ingredient is at or about 5% by weight (w / w) of the concentrate; the amount of TPGS surfactant or TPGS analogue is exactly or about 25.2% by weight (w / w) of the concentrate and the amount of water is exactly or about 68.79% by weight (w / w) of the concentrate. 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 surfactant or TPGS analogue is exactly or about 20.2% by weight (w / w) of the concentrate and the amount of water is exactly or about 68.79% by weight (w / w) of the concentrate. In one aspect, the concentrate further contains an emulsion stabilizer that is a blend of xanthan gum, guar gum and / or sodium alginate, a benzyl alcohol preservative, and a pH regulator that is citric acid. in another aspect, the concentrate further contains D-limonene and lemon oil. In another aspect, the concentrate further contains a phosphatidylcholine co-surfactant. In another aspect, the concentrate further comprises a non-polar solvent that is vitamin E oil or other oil.

In another example, the concentrate contains a non-polar active ingredient that contains gamma-linolenic acid (GLA) (e.g. GLA-containing borage oil, e.g. 22%), a surfactant that is TPGS or an analogue TPGS (e.g., TPGS homologue), 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 surfactant or TPGS analogue is exactly or about 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 further contains a benzyl alcohol preservative.

In another example, the concentrate contains a non-polar active ingredient that contains gamma-linolenic acid (GLA) (e.g. GLA-containing borage oil, e.g. 22%), a surfactant that is TPGS or an analogue TPGS (e.g. TPGS homolog) and 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 surfactant or TPGS analogue is exactly or about 25.2% by weight (w / w) of the concentrate and the amount of propylene glycol is exactly or about 69.02% by weight (w / w) of the concentrate. In one aspect, the concentrate further contains a benzyl alcohol preservative.

[0044] In another example, the concentrate contains a non-polar active ingredient that contains conjugated linoleic acid (CLA) (e.g. 80% CLA), a surfactant that is TPGS or a TPGS analogue (e.g. a TPGS homolog), and a polar solvent, which is water. In one aspect of this example, the amount of CLA-containing non-polar active ingredient is at or about 5% by weight (w / w) of the concentrate; the amount of TPGS surfactant or TPGS analogue is exactly or about 17.75% by weight (w / w) of the concentrate and the amount of water is exactly or about 71.74% by weight (w / w) of the concentrate. In another aspect of this example, the amount of the CLA-containing polar active ingredient is at or about 5% by weight (w / w) of the concentrate; the amount of TPGS surfactant or TPGS analogue is exactly or about 25.2% by weight (w / w) of the concentrate and the amount of water is exactly or about 68.79% by weight (w / w) of the concentrate. In another aspect of this example, the amount of CLA-containing non-polar active ingredient is at or about 10% by weight (w / w) of the concentrate; the amount of TPGS surfactant or TPGS analogue is exactly or about 20.2% by weight (w / w) of the concentrate and the amount of water is exactly or about 68.79% by weight (w / w) of the concentrate. In one aspect, the concentrate further contains an emulsion stabilizer that is a blend of xanthan gum, guar gum and / or sodium alginate, a benzyl alcohol preservative, and a pH regulator that is citric acid. In another aspect, the concentrate further contains D-limonene and lemon oil flavors. In another aspect, the concentrate further contains a phosphatidylcholine co-surfactant. In another aspect, the concentrate further comprises a non-polar solvent that is vitamin E oil or other oil.

[0045] In another example, the concentrate contains a non-polar active ingredient that contains conjugated linoleic acid (CLA) (e.g. 80% CLA), a surfactant that is TPGS or a TPGS analogue (e.g. a TPGS homolog) and a polar solvent that is glycerin. In one aspect of this example, the amount of CLA-containing non-polar active ingredient is at or about 5% by weight (w / w) of the concentrate; the amount of TPGS surfactant or TPGS analogue is exactly or about 25.2% by weight (w / w) of the concentrate and the amount of glycerin is exactly or about 69.02% by weight (w / w) of the concentrate.

EP 2 268 160 B1

In one aspect, the concentrate further contains a benzyl alcohol containing preservative.

[0046] In another example, the concentrate contains a non-polar active ingredient that is conjugated linoleic acid (CLA) (e.g. 80% CLA), a surfactant that is TPGS or a TPGS analogue (e.g. a TPGS homolog), and a polar solvent which is propylene glycol. In one aspect of this example, the amount of CLA-containing non-polar active ingredient is at or about 5% by weight (w / w) of the concentrate; the amount of TPGS surfactant or TPGS analogue is exactly or about 25.2% by weight (w / w) of the concentrate and the amount of propylene glycol is exactly or about 69.02% by weight (w / w) of the concentrate. In one aspect, the concentrate further contains a benzyl alcohol preservative.

[0047] In another example, the concentrate contains a non-polar active ingredient which is a sabal palm extract (e.g. one containing between exactly or about 85% and exactly or about 90% fatty acids), a surfactant that is TPGS or an analogue TPGS (e.g. TPGS homolog) and polar solvent, which is water. In one aspect of this example, the amount of non-polar active ingredient sabal palm extract is at or about 5% by weight (w / w) of the concentrate; the amount of TPGS or analog 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 non-polar active ingredient sabal palm extract is at or about 5% by weight (w / w) of the concentrate; the amount of TPGS surfactant or TPGS analogue is exactly or about 25.2% by weight (w / w) of the concentrate and the amount of water is exactly or about 68.79% by weight (w / w) of the concentrate. In another aspect of this example, the amount of non-polar active ingredient sabal palm extract is at or about 10% by weight (w / w) of the concentrate; the amount of TPGS surfactant or TPGS analogue is exactly or about 20.2% by weight (w / w) of the concentrate and the amount of water is exactly or about 68.79% by weight (w / w) of the concentrate. In one aspect, the concentrate further contains an emulsion stabilizer that is a blend of xanthan gum, guar gum and / or sodium alginate, a benzyl alcohol preservative, and a pH regulator that is citric acid. In another aspect, the concentrate further contains D-limonene and lemon oil flavors. In another aspect, the concentrate further contains a phosphatidylcholine co-surfactant. In another aspect, the concentrate further comprises a non-polar solvent that is vitamin E oil or other oil.

[0048] The concentrates provided further include concentrates having non-polar active ingredients coenzyme Q.

[0049] In one example, the concentrate contains a co-active Q10 (CoQ10) non-polar active ingredient (for example, the compound sold under the name Kaneka Q10 ™ (USP Ubidecarenone)), a TPGS surfactant or TPGS analogue (e.g. homologue), and a solvent polar, which is water. In one aspect of this example, quantity

The CoQ10 non-polar active ingredient is exactly or about 5% by weight (w / w) of the concentrate, the amount of TPGS or TPGS analogue is exactly or about 17.75% by weight (w / w) of the concentrate, and the amount of water is exactly or about 71.74% by weight (w / w) of the concentrate. In one aspect, the concentrate further contains an emulsion stabilizer that is a blend of xanthan gum, guar gum and / or sodium alginate, a benzyl alcohol preservative, and a pH regulator that is citric acid. In another aspect, the concentrate further contains D-limonene and lemon oil flavors. In another aspect, the concentrate further contains a phosphatidylcholine co-surfactant. In another aspect, the concentrate further comprises a nonpolar solvent, which is Vitamin E oil or other oil.

[0050] In another example, the concentrate contains a co-active Q10 (CoQ10) non-polar active ingredient (for example, the compound sold under the name Kaneka Q10 ™ (USP Ubidecarenone)), a TPGS surfactant or TPGS analogue (e.g. homologue), and a solvent polar, which is glycerin. In one aspect of this example, the amount of CoQ10 non-polar 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 17.75% by weight (w / w) ) of the concentrate and the amount of glycerin is exactly or about 71.89% by weight (w / w) of the concentrate. In one aspect, the concentrate further contains a benzyl alcohol preservative. In another aspect, the concentrate further contains D-limonene and lemon oil flavors. In another aspect, the concentrate further contains a phosphatidylcholine co-surfactant. In another aspect, the concentrate further comprises a nonpolar solvent, which is Vitamin E oil or other oil.

[0051] In another example, the concentrate contains a co-active Q10 (CoQ10) non-polar active ingredient (for example, the compound sold under the name Kaneka Q10 ™ (USP Ubidecarenone)), a TPGS surfactant or TPGS analogue (e.g. homologue), and a solvent polar, which is propylene glycol. In one aspect of this example, the amount of CoQ10 non-polar 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 17.75% by weight (w / w) ) of the concentrate and the amount of propylene glycol is exactly or about 71.89% by weight (w / w) of the concentrate. In one aspect, the concentrate further contains a benzyl alcohol preservative. In another aspect, the concentrate further contains D-limonene and lemon oil flavors. In another aspect, the concentrate further contains a phosphatidylcholine co-surfactant. In another aspect, the concentrate further comprises a non-polar solvent that is a vitamin E oil or other oil.

[0052] In one example, the concentrate contains a non-polar phytosterol active ingredient (phytosterols), a TPGS surfactant or TPGS analogue (e.g. homologue), and a polar solvent that is water. In one aspect of this example, the amount of the non-polar phytosterol active ingredient (phytosterols) is exactly or about 5.25% by weight (w / w) of the concentrate, the amount of TPGS or TPGS analogue is exactly or about

EP 2 268 160 B1

20% w / w (w / w) of the concentrate and the amount of water is exactly or about 68.29% w / w (w / w) of the concentrate. In one aspect, the concentrate further contains a benzyl alcohol preservative and emulsion stabilizer which is a blend of xanthan gum, guar gum and / or sodium alginate. In another aspect, the concentrate further contains a phosphatidylcholine co-surfactant. In another aspect, the concentrate further comprises a non-polar solvent that is linseed oil or other oil.

[0053] In some examples, the concentrate is formulated based on the desired properties of the obtained diluted compositions prepared by diluting the concentrate in an aqueous liquid. Typically, the concentrate is prepared so that it can be diluted in an aqueous medium to produce a liquid diluted composition having one, more than one, all or any combination of the following:

[0054] In one example, the concentrate is formulated such that diluting a certain amount of the concentrate in a certain amount of an aqueous medium gives a liquid diluted composition having the desired particle size, in particular a particle size that is not larger than the specified particle size or not smaller than the specified particle size. The specified particle size can be expressed as the average particle size or largest particle size in the aqueous medium. For example, it may be desirable for the liquid dilution composition to contain an average or at most smaller than the specified particle size. For example, the concentrate may be formulated such that a 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 concentrate in exactly or about 8 ounces (0.236588 liter) of aqueous medium or dilution of the concentrate in aqueous medium at a dilution of not more than 1:10 or about 1: 10, not more than 1:25 or about 1:25, not more than 1:50 or about 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 about 1: 500; or diluting the concentrate in an aqueous medium to produce a liquid diluted 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 oz. volume of liquid diluted composition, gives a liquid diluted composition having a particle size of at most or on average 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.

[0055] In another example, the concentrate is formulated such that diluting a specific amount of the concentrate in an amount of aqueous medium gives a liquid diluted composition having the desired clarity, such as by providing a diluted composition having the desired NTU value, usually an NTU value that is not greater than or less than the given NTU value, or by providing a liquid diluted composition, which is as clear or almost as clear as the aqueous medium before adding the concentrate (i.e. without the presence of the concentrate). For example, the concentrate can be formulated such that a 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 or about 5 g or at least 10 g or about

G of concentrate in exactly or about 8 ounces (0.236588 liter) of aqueous medium; or diluting the concentrate in an aqueous medium at a dilution of not more than 1:10 or about 1:10, not more than 1:25 or about 1:25, not more than 1:50 or about 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 diluting the concentrate in an aqueous medium to produce 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 oz. volume of liquid diluted composition, gives a liquid diluted composition having 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, or provides a liquid dilute composition that is at least as clear or at least almost as clear as the aqueous medium containing no concentrate (i.e. compared to the clarity of the aqueous medium before adding the concentrate).

[0056] In another example, the concentrate is formulated such that upon dilution gives a stable liquid diluted composition, for example a composition that does not contain visible particles, does not contain visible crystals, does not show ring formation or a combination thereof. Stability may be for a specific period of time and / or when the concentrate or liquid dilution composition is stored at a specified temperature. For example, the concentrate can be formulated such that a 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 galbo at least 10 g or about 10 g of concentrate in 8 ounces (0.236588 liter) or about 8 ounces of aqueous medium; dilution of the concentrate in an aqueous medium at a dilution of not more than 1:10 or about 1:10, not more than 1:25 or about 1:25, not more than 1:50 or about 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; and / or diluting the concentrate in an aqueous medium to produce 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 oz volume of the diluted liquid composition, gives a diluted liquid composition that does not contain visible particles, it does not contain visible crystals, does not show phase separation and / or does not show ring formation and / or has a pleasant taste and / or smell. The concentrate can be formulated such that the liquid diluted composition is free of visible particles, is free of visible crystals, is free of phase separation, is free of ring formation, and / or has a pleasant taste and smell when the concentrate and / or liquid composition is diluted stored at room temperature, refrigeration temperature or freezing temperature. Storage may take place, for example, for at least one day, at least one week, at least thirty days, or at least one year.

[0057] The aqueous medium may be a drink such as, for example, water, juice, soda, tea, coffee, sports drinks, nutritional drinks, energy drinks, milk and other drinks, including those described herein.

[0058] Liquid dilution compositions are also described which contain concentrates diluted in an aqueous medium, e.g. a drink. Liquid dilution compositions may contain any of the concentrates provided. Thus, liquid diluted compositions contain non-polar active ingredients in an aqueous medium, such as beverages, which are indicated for human consumption. Liquid dilution compositions include those prepared by diluting concentrates, such as those having the properties as described above, such as desired particle size, clarity, NTU value, and / or stability, e.g., no ring formation, visible crystals, phase separation (or pleasant taste). odor, for example, in accordance with the specifications described above.

[0059] In one example, the aqueous medium contained in the liquid diluted composition is a drink, such as, for example, water, a carbonated drink, milk, tea, coffee, juice, energy drink or sports drink, or a nutritional drink. In one aspect, the liquid dilution composition is as clear or almost as clear as an aqueous medium, such as a drink, before adding the concentrate (e.g. compared to no concentrate), and / or remains as clear or almost as clear as a drink when stored at room temperature (e.g. 25 ° C or about 25 ° C) or at a cooling temperature (e.g. 0-10 ° C or about 0-10 ° C, e.g. exactly or about 4 ° C) or at freezing temperature (e.g. -20 ° C or about -20 ° C), where storage takes place for at least one day, at least one week, at least thirty days or at least one year.

[0060] The amount of concentrate in the liquid diluted composition can be determined. For example, liquid dilution compositions include those that contain 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 concentrate per 8 oz. Volume (0.236588 liter) of aqueous medium; or containing a concentrate with a dilution of not more than 1:10 or about 1:10, not more than 1:25 or about 1:25, not more than 1:50 or about 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 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 non-active ingredient per 8 oz. of volume of aqueous medium. Liquid dilution compositions typically have one or more desirable properties such as particle size, clarity, NTU value, stability, e.g. they are free of crystals, phase separation, ring formation or unpleasant taste / smell, as for at least a certain amount of time when stored under specified storage conditions.

[0061] For example, the compositions include liquid diluted compositions having a particle size on average or at most 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 50 nm or less than 25 or about 25 nm; those having 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 containing visible particles, not containing visible crystals, not showing ring formation and / or phase separation; and / or remaining free (or not) of visible particles, visible crystals, ring formation and / or phase separation, and / or an unpleasant taste / smell when stored in

At room temperature (e.g. 25 ° C or about 25 ° C) or at cooling temperature (e.g. 0-10 ° C or about 0-10 ° C, e.g. exactly 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 one week, at least thirty days, or at least one year.

[0062] Also described are methods for making concentrates and methods for making liquid diluted compositions. Generally, the methods for producing concentrates are carried out, producing the oil phase and the aqueous phase separately, and mixing the two phases, usually by emulsification, to produce a concentrate that is a liquid nanoemulsion concentrate. The oil phase ingredients are added to form the oil phase, and the water phase ingredients are added to form the water phase. The ingredients are selected from concentrate ingredients as described herein, which usually include a non-polar compound, a surfactant and a polar solvent as described herein. Typically, the oil phase ingredients include the non-polar compound (s), usually the non-polar active ingredient (s) of the concentrate, and the aqueous phase ingredients include the polar solvent. To provide concentrates as described herein, the ingredients are added in amounts in the range of the respective concentrations. In one example, the aqueous phase ingredients include a surfactant. In another example, the oil phase ingredients contain a surfactant. In one example, the aqueous phase ingredients and oil phase ingredients contain a surfactant.

[0063] The amounts of surfactant (s), non-polar active ingredient (s) and polar solvent are selected based on ranges of the respective concentrations of these ingredients in the resulting concentrate. For example, the non-polar active ingredient is included in an amount that is 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.

[0064] In one example, the oil phase ingredients further include a non-polar solvent (s). In one example, the concentrate is made from the first and second oil phase ingredients, and the first oil phase ingredients include a non-polar active ingredient and a solvent. In one example, the solvent contains an oil other than a non-polar active ingredient, such as, for example, vitamin E, linseed oil and / or safflower oil.

[0065] In one example, the oil phase ingredients and / or the water phase ingredients contain a co-surfactant, in an amount sufficient to stabilize the concentrate, such as a phospholipid, e.g., phosphatidylcholine. In one example, the amount of 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 oil phase ingredients and / or the water phase ingredients further contain at least one preservative in an amount sufficient to preserve the concentrate, such as, for example, a benzyl alcohol containing preservative. In one example, the amount of 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.

[0066] In another example, the oil phase ingredients and / or the water phase ingredients further contain an emulsion stabilizer in an amount sufficient to stabilize the concentrate, such as an emulsion stabilizer comprising a gum blend such as any or more of guar gum , xanthan gum and sodium alginate.

[0067] In an exemplary method of making the concentrate, the oil phase is prepared by mixing the oil phase ingredients in a first vessel and heating the oil phase ingredients; the water phase is prepared by mixing one or more water phase ingredients in a second vessel and heating the water phase ingredients; then the oil and aqueous phases are emulsified to form a concentrate.

[0068] In another exemplary method provided, the oil phase is prepared by mixing one or more ingredients of the first oil phase in a first vessel and heating the ingredients of the first oil phase at least until the ingredients of the first oil phase are dissolved; then adding one or more additional oil phase ingredients to the first vessel; and mixing and heating the first and additional oil phase ingredients; the aqueous phase is prepared by mixing one or more components of the aqueous phase in a second vessel and heating the aqueous phase component (s); then the aqueous and oily phases are emulsified to form a concentrate.

[0069] Heating and mixing of the water and oil phases can be carried out simultaneously or sequentially, in any order.

[0070] In any of the methods for making concentrates, mixing steps (e.g., mixing of oil and / or aqueous phases) can be carried out using a standard mixer, such as any of the standard mixers listed herein, or using any of the mixers described herein, such as using a homogenizer. In any of the methods provided, heating may be carried out using one or more heating devices, such as, for example, a hob, water jacket or any of the heating devices mentioned herein. In one example, the oil phase ingredients are heated using a first heating device and the water phase components are heated using a second heating device. In one example, heating requires heating the ingredients to 60 ° C or about 60 ° C, or to exactly or about 70 ° C, or to exactly or about 71 ° C. In one example, the components of the oil phase and / or the aqueous phase are heated to between between about 45 ° C or about 45 ° C and 85 ° C or about 85 ° C, for example; exactly or about 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.

[0071] In any of the methods, emulsification can be carried out using a homogenizer, such as a homogenizer as described herein. In one example, emulsification is carried out at between 850 rpm or about 850 rpm and 1200 rpm or about 1200 rpm. In another example, they are carried out at or about 30 revolutions per minute.

[0072] In some examples, the methods further include rapidly cooling the forming emulsion during the emulsifying step. In some examples, rapid cooling leads to cooling of the forming emulsion to between 25 ° C or about 25 ° C and 43 ° C or about 43 ° C, such as 25, 26, 27, 28,

EP 2 268 160 B1

29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42 or 44 ° C, e.g. between exactly or about 25 ° C and exactly or about 35 ° C, or between at or about 35 ° C and at or about 43 ° C. In one example, cooling of the emulsion is obtained in less than or about 60 minutes, in less than or about 30 minutes, or between exactly or about 30 and exactly or about 60 minutes. An exemplary means for carrying out rapid cooling involves repeatedly passing the phases through a cooling device attached to the vessel.

[0073] The methods may further comprise adding to the concentrate one or more flavors (e.g. lemon oil, D-limonene) and / or one or more pH regulators (e.g. citric acid, phosphoric acid), e.g. oil and water phase emulsification. The pH is usually measured simultaneously with, before and / or after the addition of the pH regulator, and the amount of the pH regulator is determined by the concentrate pH. Typically, the pH regulator includes an acid or base in an amount sufficient to affect the pH of the concentrate.

[0074] In one example of methods, the ingredients are added to the vessel (s) simultaneously or sequentially, in any order. In another example, the ingredients (e.g., the oil phase and / or water phase ingredients) are added to the vessels in a specific order, such as the specific order provided herein, for example in the particular examples provided. In one example, where the aqueous phase ingredients contain a polar solvent (e.g. water, propylene glycol, glycerin or other diol (such as another sugar alcohol) and emulsion stabilizer, the aqueous phase ingredients are added sequentially, in the following order: 1) a polar solvent (e.g. water, propylene glycol or glycerin); 2) emulsion stabilizer. In another example, where the oil phase ingredients contain a surfactant, a non-polar compound and a preservative, the oil phase ingredients are added sequentially, in the following order: 1) surfactant, 2) preservative; 3) non-polar compound (i.e. non-polar active ingredient). In another example, where the oil phase ingredients contain a surfactant, non-polar compound, preservative and emulsion stabilizer, the oil phase ingredients are added to the oil phase vessel in turn, in the following order: 1) surfactant; 2) preservative; 3) non-polar compound and 4) emulsion stabilizer. In another example, where the oil phase ingredients comprise a surfactant, non-polar compound, preservative, solvent and emulsion stabilizer, the oil phase ingredients are added to the oil phase vessel in turn in the following order: 1) surfactant; 2) preservative; 3) non-polar solvent; 4) non-polar compound and 5) emulsion stabilizer. In another example, where the oil phase ingredients contain a surfactant, a non-polar compound, a preservative, a non-polar solvent, a surfactant and an emulsion stabilizer, the oil phase ingredients are added to the oil phase vessel in turn in the following order: 1) agent surfactant; 2) preservative; 3) non-polar solvent; 4) surfactant; 5) non-polar compound and 6) emulsion stabilizer. In another example, where the aqueous phase comprises a polar solvent and a surfactant, the polar solvent and a surfactant are added sequentially, in that order. In another example, where the oil phase comprises a non-polar solvent, preservative, co-surfactant, surfactant and non-polar compound, the non-polar solvent, preservative and co-surfactant are added and mixed until the co-agent dissolves.

Surfactant; the surfactant is added as long as it dissolves; and the non-polar active ingredient is added in this order.

[0075] In one example, the methods for producing concentrates are carried out using a laboratory scale process as provided herein. In another example, in particular for large batches of concentrates, the methods are carried out using an enlarged scale process as described hereinbelow, such as the exemplary enlarged scale process shown in figure 1.

[0076] In one example, where the polar solvent is water, the water is first purified by passing through purifiers. In one example, water purification is carried out by passing through the purifiers, in order, in order: a carbon filter, ion exchange device, reverse osmosis device, 100 micron endpoint filter and 50 micron endpoint filter. In this example, after purification, water is added, along with the other components of the water phase, to the water phase tank. The water phase component (s) are then mixed using a standard agitator attached to the tank, for example mounted on top of the tank. A heating device (usually a water jacket on the water phase tank) is used to heat the water phase ingredients during the production of the water phase, usually for low heating (e.g. 60 ° C). To form the oil phase, the oil phase component (s) are weighed / metered and added to the oil phase tank. The oil phase ingredients are mixed using a standard agitator connected to the oil phase tank, for example mounted on the tank. A heating device (usually a water jacket on the oil phase tank) is used to heat the oil phase ingredients during the production of the aqueous phase, usually for low heating (e.g. 60 ° C). As soon as the water and oil phases reach 60 ° C, and after the oil phase ingredients have dissolved, the oil and water phases are combined by transferring the oil phase to the water phase vessel using a transfer medium. For this method, the homogenizer is mounted on an aqueous phase tank and switched on for example at 850-1200 rpm. Then the ball valves are opened and the transfer pump is turned on, thereby obtaining the transfer of the liquid oil phase to the water phase tank through the transfer hose (s). When the phases combine, the mixture is homogenized by continuing mixing using a homogenizer. The homogenizer can be adjusted, for example by adjusting the septum on the homogenizer, to obtain and preserve the emulsion, for example by moving the septum further into the forming emulsion and / or further from the forming emulsion. During the emulsifying step, the forming emulsion is cooled, usually quickly cooled, by repeatedly passing the forming emulsion through a recirculating cooling element that is attached to the water phase tank. The emulsion is transferred via a transfer medium to a storage / packaging tank where additional ingredients can be added and / or the mixture can be evaluated. Additional ingredients are mixed into the concentrate using a standard mixer. A final product filter is used to filter the concentrate before use.

[0077] To prepare any of the concentrates provided, any of the methods for producing concentrates may be used as described herein.

[0078] Also described are methods for making the supplied diluted liquid compositions

EP 2 268 160 B1 containing concentrates, such as beverages containing concentrates. These methods include methods of providing oil-based additives, for example in a food product and a drink. These methods include adding any of the concentrates provided, e.g., liquid nanoemulsion concentrates, to an aqueous medium such as a drink. Typically, the concentrate is added to a medium, e.g., a drink, such that the medium contains an effective amount of the additive (e.g., a non-polar active ingredient).

[0079] An effective amount of an additive, such as a non-polar active ingredient, is the amount and / or concentration of the ingredient required to prevent, treat, alleviate, inhibit or partially inhibit the symptom of the disease or disorder, or the amount and / or concentration indicated for consumption by the subject, such such as daily intake and / or supplementing nutritional deficiencies, for example enough to strengthen the nutritional, pharmaceutical, nutraceutical properties, health or energy food, drink or other consumer goods. In some examples, the concentrate is added to the aqueous medium such that the resulting diluted liquid composition contains an effective amount of a specific non-polar compound, for example a specific amount per volume or weight of the consumer product, such as for example 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 oz. volume of liquid diluted composition.

[0080] In one example, the effective amount is the concentration or amount of liquid nanoemulsion, wherein at least 25 mg or about 25 mg, usually 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 non-polar active ingredient is contained in at least 8 ounces of volume of aqueous medium.

[0081] US Provisional Application Serial Number: 61 / 070,381, filed March 20, 2008, entitled "COMPOSITIONS CONTAINING NON-POLAR COMPOUNDS;" and US Provisional Application Serial No. 61 / 132,424, filed June 16, 2008, entitled "COMPOSITIONS CONTAINING NON-POLAR COMPOUNDS," each by Philip Bromley, disclose, for example, compositions containing non-polar compounds and surfactants, PEG derivatives of vitamin E, such as tocopherol polyethylene glycol succinate ( TPGS) and TPGS analogues, including TPGS homologues, and methods of making the composition.

BRIEF DESCRIPTION OF THE DRAWINGS [0082]

Figure 1 shows an exemplary method on an enlarged scale of 100 provided methods for producing liquid nanoemulsion concentrates. This method is exemplary and its variations can be used. In this example, water 101 is used as the polar solvent and the water is first purified by passing it in succession through the following purifiers in the following order: carbon filter 105, ion exchange device 106, device for

Reverse osmosis 107, 100 micron endpoint filter 108 and 50 micron endpoint filter 109. After purification, water is added, along with the other components of the aqueous phase, to the aqueous phase tank 103. In other examples, the polar solvent is another a polar solvent such as glycerin or propylene glycol. In the example shown, the water phase component (s) are then mixed using a standard mixer 111 attached to the tank, for example mounted on top of the tank. To heat the components of the water phase during the production of the water phase, a heating device (usually a water jacket on the water phase tank) is used, usually for low heating (e.g. 60 ° C). To form the oil phase, the oil phase component (s) are weighed / metered and added to the oil phase tank 102. The oil phase ingredients are mixed using a standard mixer 111 connected to the oil phase tank, for example mounted on top of the tank. To heat the components of the oil phase during the formation of the aqueous phase, a heating device (usually a water jacket on the oil phase tank) is used, usually for low heating (e.g. 60 ° C). As soon as the oil and water phases reach 60 ° C, and after the oil phase components have dissolved, the water and oil phases are combined by transferring the oil phase to the water phase vessel through transfer medium 112. To this end, homogenizer 110 is mounted on the phase tank water, it turns on for example at 850-1200 revolutions per minute. Then the ball valves are opened and the transfer pump is turned on, thereby obtaining the transfer of the liquid oil phase to the water phase tank through the transfer hose (s). When the phases combine, the mixture is homogenized by continuing mixing using a homogenizer 110. The homogenizer can be adjusted, for example by adjusting the septum on the homogenizer, to obtain and preserve the emulsion, for example by moving the septum further into the forming emulsion and / or further from the forming emulsion. During the emulsification step, the forming emulsion is cooled, usually quickly cooled, by repeatedly passing the forming emulsion through a recirculating cooling element 115 (e.g. Model No. OC-1000 RO, sold by Turmoil, West Swanzey, NH), which is attached to the water phase tank. The emulsion is transferred via transfer medium 112 to storage / packaging tank 104 where additional ingredients can be added and / or the mixture can be evaluated. Additional ingredients are mixed into the concentrate using a standard mixer 111. To filter the concentrate, a final product filter 113 is used before use.

DETAILED DESCRIPTION

Outline [0083]

A. DEFINITIONS

B. COMPOSITIONS CONTAINING NON-POLAR COMPOUNDS

1. Liquid nanoemulsion concentrates containing non-polar compounds

a. Formulation of liquid concentrates

i. Common components and typical concentration ranges

EP2 268 160 B1 ii. Performing an initial concentrate assessment (1) Clarity (2) Empirical assessment (3) Particle size (5) Turbidity measurement iii. Choose a formulation and modify your formulation

b. Non-polar compounds

i. Active ingredients containing polyunsaturated fatty acids (PUFA) (1) Compounds containing omega-3 fatty acids (a) DHA / EPA (i) Fish oils (ii) Algae oil (b) Linseed oil-omega-3 (ALA) (2) Omega-6 compounds (a) Borage oil (gammalinolenic acid (GLA)) (3) Sabal palm extract (4) Conjugated linoleic acid (CLA) ii. Active ingredients containing coenzyme Q (1) Coenzyme Q10 iii. Active ingredients containing phytosterol

c. Surfactants

i. Surfactants derived from vitamin E (1) PEG derivatives of vitamin E (a) Tocopherols and tocotrienols (b) PEG units (c) Linkers (d) Polyethylene glycol tocopherol and tethotrienol polyethylene glycol diesters (vitamin E dicarboxylic acid esters attached to PEG) (e) Other vitamin E esters PEG (f) TPGS surfactants ii. Surfactant concentration iii. HLB

d. Surfactants (emulsifiers)

i. Phospholipids

e. Polar solvents

f. Preservatives and sterilizers

g. Emulsion stabilizers (co-emulsifiers)

h. Non-polar solvents

EP 2 268 160 B1

i. Flavorings

j. pH regulators

2. Composition powders

3. Liquid diluted compositions containing diluted concentrates

a. Clarity

i. Clarity as determined by empirical assessment ii. Clarity determined by particle size or number of particles iii. turbidity

b. Durability

c. Properties indicated for human consumption

d. Security

e. Oral bioavailability

C. WAYS OF MAKING LIQUID NANOEMULSION CONCENTRATES

CONTAINING NON-POLAR COMPOUNDS 1. Equipment for the production of concentrates

2. weights

a. Purifiers, including filters

b. Utensils for mixing ingredients

c. Agitators

d. Heating devices

e. Cooling instruments

f. Transfer measures

g. Equipment for performing the assessment

3. General methods for the preparation of liquid nanoemulsion concentrates

a. Preparation of the aqueous phase

i. Water phase ingredients

b. Preparation of the oil phase

i. Oil phase ingredients

c. Combining and emulsifying the oil and water phases i. Combining the oil and water phases ii. Oil and water phase emulsification iii. Cooling

d. Additional stages i. Additional components ii. Assessment of the concentrate iii. Concentrate filtration

4. Laboratory scale process

5. Enlarged processes

a. Water purification

b. Preparation of the water phase and the oil phase:

EP2 268 160 B1

c. Connecting and emulsifying the phases 147

d. Cooling 148

e. Additional stages 149

D. METHODS OF MANUFACTURING LIQUID DILUTED COMPOSITIONS 149

CONTAINING THINNESS CONCENTRATES

1. Dilutions 150

2. Analysis of aqueous liquid diluted compositions containing diluted 151 concentrates

a. Clarity / turbidity 152

i. Empirical assessment 152 ii. Particle size 153 iii. Turbidity measurement 154

E. EXAMPLES

A. DEFINITIONS [0084] Unless otherwise specified, all technical and scientific terms used herein have the same meanings as commonly understood by one of ordinary skill in the art to which the invention (s) belongs.

[0085] In the event that there are multiple definitions for the terms used herein, those in this section prevail. When reference is made to a URL or other identifier or solution, it will be understood that such identifiers may change and detailed information may appear and disappear from the internet, but equivalent information is known and can be easily accessed, such as by searching the internet and / or relevant databases. The reference to this proves the availability and dissemination of such information.

[0086] As used herein, the term colloid refers to a mixture comprising two phases, a dispersed phase and a dispersion phase, wherein said dispersed phase contains particles (droplets) dispersed in the dispersion phase. Colloidal mixtures include aerosols, foams and dispersions, e.g. emulsions, e.g. nanoemulsions. A liquid colloid, e.g. a nanoemulsion, may have a similar appearance, e.g. clarity, to a solution in which there is no dispersed phase.

[0087] As used herein, the term colloidal dissipation of two immiscible liquids, e.g., oil and water (or other aqueous liquid, e.g., polar solvent), one of which is part of the dispersion phase and the other part of the dispersed phase. The compositions provided include emulsions, usually oil-in-water nanoemulsions (which include any oil-soluble phase dispersed in any aqueous phase, also called the aqueous phase), in which the oil phase is a dispersed phase and the aqueous phase is a continuous phase. Emulsions are usually stabilized using one or more surfactants and / or co-surfactants and / or emulsion stabilizers. Surfactants form an interface between the oil and water phases of the emulsion, ensuring durability. Typically, nanoemulsions of the provided compositions contain micelles containing one or more surfactants surrounding a non-polar active ingredient that are dispersed in the aqueous phase. Examples

The emulsions provided are liquid nanoemulsion concentrates provided and liquid diluted compositions prepared by diluting the concentrates, usually in an aqueous medium.

[0088] The term nanoemulsion as used herein means 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, usually less than 500 nm or less than about 500 nm, usually less than 300 nm or about 300 nm, e.g. less than 250 nm or about 250 nm, e.g. less than 200 nm or less than about 200 nm, e.g. 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 nanoemulsions are provided liquid nanoemulsion concentrates and liquid diluted compositions, e.g., aqueous liquid diluted compositions containing diluted concentrates.

[0089] The terms "surfactant" and "surfactant" as used herein synonymously refer to synthetic and naturally occurring amphiphilic molecules that have a hydrophobic fragment (s) and a hydrophilic fragment (s). Due to their amphiphilic (amphipathic) nature, surfactants and co-surfactants can reduce the surface tension between two immiscible liquids, e.g. oil and water phases in the emulsion, stabilizing the emulsion. Various surfactants can be characterized based on their relative hydrophobicity and / or hydrophilicity. For example, relatively lipophilic surfactants are more soluble in fats, oils and waxes, usually having HLB values ​​less than 10 or about 10, while relatively hydrophilic surfactants are more soluble in aqueous compositions, e.g. water, and usually have HLB values ​​greater than 10 or about 10. Relatively amphiphilic surfactants are soluble in oil and water based liquids and usually have HLB values ​​close to 10 or about 10.

[0090] Surfactants include, for example, soaps, detergents, lipids, emulsifiers, dispersing agents and moisturizers, molecules that form liquid emulsions, for example by forming an emulsion in an aqueous medium or an aqueous liquid diluted composition, for example forming a colloidal dispersion of two immiscible liquids in the form of droplets, for example emulsions such as a microemulsion; and compounds that form various macromolecular structures, e.g., aggregates, in liquids, e.g., micelles, lipid bilayer structures, including liposomes and inverse micelles.

[0091] Typically, the surfactants used in the provided compositions have an HLB value between 14 or about 14 and 20 or about 20, for example exactly or about 14, 15, 16, 17, 18, 19 or 20, and usually between exactly or about 15 exactly or about 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, e.g., PEG tocotrienol or tocopherol diesters, such as TPGS ( for example. TPGS 1000) and TPGS analogues; and other known surfactants having HLB values ​​between 14 or about 14 and 20 or about 20, usually between exactly or about 15 and exactly or about 18, such

Like other surfactants described herein. Typically, the surfactant is a natural surfactant, for example a surfactant that is GRAS (widely considered safe) by the FDA and / or kosher certified.

[0092] The term PEG derivative of vitamin E as used herein means a compound containing one or more vitamin E units (e.g. tocopherol or tocotrienol) linked, for example via an ester, ether, amide or thioester linkage, to one or more polyethylene glycol (PEG) units ), via a linker, for example dicarboxylic or tricarboxylic acid. Exemplary PEG vitamin E derivatives are tocopherol polyethylene glycol succinate (TPGS), TPGS analogues, TPGS homologues, and TPGS derivatives.

[0093] The term tocopherol polyethylene glycol diester (TPGD) as used herein means a PEG derivative of tocopherol, wherein the linker is a dicarboxylic acid (carboxylic acid having two carboxyl groups, e.g. succinic acid) such as succinic acid. Exemplary dicarboxylic acids that can be used as linkers in these PEG tocopherol and tocotrienol diester surfactants are succinic, sebacic, dodecanedioic, suberic or azelaic acid, citraconic, methylcracraconic, itaconic, maleic, glutaric, glutaconic, and fumaric acid. Of many, exemplary TPGDs are tocopherol polyethylene glycol succinate (TPGS), tocopherol polyethylene glycol sebacate, tocopherol polyethylene glycol dodecanedione, polyethylene glycol tocopherol suberate, polyethylene glycol tocopherol azelaate, polyethylene glycol tocopherate, polyethylene glycol tocopherate, polyethylene glycol tocopherol, tocopherol polyethylene glycol glutarate, tocopherol polyethylene glycol glutaconate and tocopherol polyethylene glycol phthalate.

[0094] As used herein, the terms "polyethylene glycol tocopherol succinate" "TPGS", "polyethylene glycol tocopherol surfactant succinate", and "TPGS surfactant" refer to polyethylene glycol (PEG) tocopherol diesters that are formed by attachment using esterification, tocopherol succinate, which itself is an ester produced by the esterification of tocopherol and succinic acid. The term tocopherol refers to any naturally occurring or synthetic form of vitamin E, and may refer to a single compound or mixture. Examples of tocopherols include, for example, α-tocopherol, D-tocopherol, β-tocopherol, γ-tocopherol and δ-tocopherol. The PEG unit of the TPGS surfactant may be any PEG unit, e.g. PEG units between 200 kDa or about 200 kDa and 20,000 kDa or about 20,000 kDa, usually between 200 kDa or about 200 kDa and 6,000 kDa or about 6,000 kDa, for example between 600 kDa or about 600 kDa and 6000 kDa or about 6000 kDa, usually 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, e.g. 200 kD 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 modified PEG units, e.g. methylated PEG (m-PEG) and / or PEG units including other PEG analogues, e.g. PEG-NHS, aldehyde PEG, PEG-SH, PEG-NH2, PEGCO2H and branched PEG.

[0095] An exemplary TPGS surfactant is TPGS-1000, which has a 1000 kDa PEG unit. TPGS may be any natural, water-soluble polyethylene glycol succinate tocopherol, for example food grade TPGS sold under the name Eastman Vitamin E TPGS®, food grade, by Eastman Chemical Company, Kingsport, TN. This TPGS is a water-soluble form of vitamin E of natural origin, which is produced by esterifying the carboxyl group of crystalline d-alpha-tocopheryl succinate with polyethylene glycol 1000 (PEG 1000) and contains between 260 and 300 mg / g of the total amount of tocopherol. A similar compound can be produced by esterifying the carboxyl group of the d, 1 synthetic vitamin E using PEG 1000. It forms a clear liquid when dissolved 20% in water. This tocopheryl and polyethylene glycol derivative is a water-soluble fat-soluble vitamin (vitamin E) preparation, e.g. as disclosed in US Patent Nos. 3,102,078, 2,680,749 and published US applications 2007/0184117 and 2007/0141203. Also, an exemplary TPGS surfactant that can be used in the provided compositions is water-soluble natural vitamin E TPGS), sold by ZMC-USA, The Woodlands, Texas. Any known TPGS source can be used. Usually, the TPGS surfactant is GRAS and is kosher certified. Usually TPGS has an HLB value between 16 or about 16 and 18 or about 18.

[0096] As used herein, the term analog refers to a chemical compound that is structurally similar to another compound (referred to as the parent compound here), but differs slightly in composition, e.g., by changing, adding or removing an atom, one or more units (e.g., units (units) methylene - (CH2) n--) or one or more functional groups. An analogue may have different chemical or physical properties compared to the original compound and / or may have improved biological and / or chemical activity. Optionally, the analogue may have similar or identical chemical or physical properties compared to the original 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. An analog may mimic the chemical and / or biological activity of the parent compound (i.e. it may have similar or identical activity) or, in some cases, may have increased or decreased activity. The analog may be a naturally occurring or unnaturally (e.g. synthetic) variant of the parent compound. Other types of analogs include isomers (e.g., enantiomers, diastereomers) and other types of chiral variants of the compound, as well as structural isomers. The analog may be a branched or cyclic variant of the linear compound. For example, a linear compound may have an analog that is branched or otherwise substituted to give some desired properties (e.g., improved hydrophobicity or bioavailability). Exemplary analogues used in the provided compositions and methods are TPGS analogs that can be used as surfactants instead of TPGS active in the provided compositions.

[0097] As used herein, the term homolog refers to an analogue that differs from the parent compound only by the presence or absence of a single unit, such as a methylene unit, or certain multiples of such units, e.g. - (CH2) n--. Usually, the homologue has similar chemical and physical properties as the parent compound. Examples of homologues used in the supplied

The compositions and methods are TPGS homologs.

[0098] As used herein, the terms "polyethylene glycol tocopherol succinate analog", "TPGS analog" and "TPGS analog surfactant" refer to compounds other than TPGS that are similar to the parent TPGS compound, but slightly differ in composition, e.g. , addition or removal of an atom, one or more units (e.g., units (units) - (CH2) n) or one or more functional groups. TPGS analogs include surfactants derived from vitamin E, including vitamin E PEG derivatives, including vitamin E PEG diesters such as, but not limited to, polyethylene glycol tocopherol sebacate (PTS), polyethylene glycol tocopherol dodecanedione (PTD) dodecanedione , tocopherol polyethylene glycol suberate (PTSr), tocopherol polyethylene glycol azelaate (PTAz) and polyoxyethanyl tocotrienyl sebacate (PTrienS), as well as other vitamin E PEG derivatives. In one example, the surfactant in the deposition compositions is a TPGS analogue.

[0099] Exemplary TPGS analogues shown in Scheme I:

are compounds other than TPGS compounds having formula

<img file="PL2268160T3_D0001.tif" />

where each R¹, R², R³ and R⁴ independently represents a hydrogen atom (H) or a methyl group (CH3); each dashed line is independently a single or double bond; n is an integer from 1 to 5000; each miq is independently 0 or 1; ap is an integer from 1 to 20.

[0100] For example, TPGS analogs include, but are not limited to, compounds having the formula shown in Scheme I, where when the dashed bonds designated "A" and "B" are single bonds and both m and q are 0, p is any integer from 2-10. TPGS analogs also include compounds where the dashed line at B or the dashed line at A, or both dashed lines, represent at least one double bond. For example, TPGS analogs include a compound as in Scheme I, where when the dashed line in A is only single bonds, the dashed line in "B" means one or more double bonds, e.g., PEG tocotrienol diesters. TPGS analogs also include compounds as in Scheme I, where when the dashed line marked "B" means only single bonds, the dashed line marked "A" means one or more double bonds; or when the dashed line marked "A" does not mean double bonds, and both m and q are zero, then p is greater than 1. For example, TPGS analogs include compounds where one or more dashed lines represent a double bond, e.g., PEG derivatives of tocotrienol esters (e.g., PTrienS).

[0101] Exemplary TPGS analogs are also compounds other than TPGS compounds having the formula shown in Scheme II:

EP 2 268 160 B1

<img file="PL2268160T3_D0002.tif" />

where each R¹, R², R³ and R⁴ independently represents a hydrogen atom (H) or a methyl group (CH3); the bond represented by the dashed line means either a single bond or a double bond; and m is an integer from 1 to 20, and n is an integer from 1 to 5000.

[0102] Exemplary TPGS analogs also include compounds other than TPGS having PEG units that differ in chain length according to the formula in Scheme III:

<img file="PL2268160T3_D0003.tif" />

where each R¹, R², R³ and R⁴ independently represents a hydrogen atom (H) or a methyl group (CH3), where n is an integer from 1 to 5000.

[0103] As used herein, the term TPGS-1000 analogs means compounds other than TPGS-1000 that are similar to the parent compound TPGS-1000, but slightly differ in composition, e.g., by changing, adding or removing an atom, one or more units (e.g. methylene (CH2) unit (s) n or one or more functional groups In one example, the surfactant in the compositions provided herein is an analog

TPGS-1000. Suitable TPGS-1000 analogs include, but are not limited to, other TPGS compounds having a PEG unit (s) that differ in chain length and molecular weight compared to TPGS-1000, including, for example, TPGS compounds having PEG units between 200 or about 200 kDa and 20,000 kDa or about 20,000 kDa, usually between 200 kDa or about 200 kDa and 6,000 kDa or about 6,000 kDa, for example between 600 kDa or about 600 kDa and 6,000 kDa or about 6,000 kDa, usually 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, but not limited to, 200, 300, 400, 500, 600 , 800 and 1000 kDa. Exemplary TPGS-1000 analogues are also TPGS compounds having modified PEG units, e.g. methylated PEG (m-PEG) and / or PEG units including other PEG analogues, e.g. PEG-NHS, PEG-aldehyde, PEG-SH, PEG-NH2 , PEG25 CO2H and branched PEG. Exemplary TPGS-1000 analogues are also any TPGS analogues, e.g. surfactants derived from vitamin E, including PEG vitamin E derivatives, including PEG vitamin E diesters such as, but not limited to, polyethylene glycol tocopherol sebacate (PTS), polyethylene glycol dodecanedione

Tocopherol (PTD), tocopherol polyethylene glycol suberate (PTSr), tocopherol polyethylene glycol azelaate (PTAz) and polyoxyethanyl tocotrienyl sebacate (PTrienS), as well as other PEG vitaminE derivatives.

[0104] As used herein, the term TPGS homologs means TPGS analogs that differ from the TPGS parent compound only by the presence or absence of a single unit, such as a methylene unit, or a certain multiple of such units, e.g. - (CH2) n--. In one aspect, TPGS homologs are used as surfactants in the provided compositions. Usually, suitable TPGS homologues have similar surfactant properties compared to the parent compound (TPGS), e.g. similar HLB properties, e.g. HLB properties between 14 or about 14 and 20 or about 20. Exemplary TPGS homologues are tocopherol polyethylene glycol sebacate (PTS) ), tocopherol polyethylene glycol dodecanedione (PTD), tocopherol polyethylene glycol suberate (PTSr), tocopherol polyethylene glycol azelaate (PTAz). Exemplary TPGS homologs are compounds having the formula shown in Scheme I (above), where neither dashed line A nor B represents a double bond, and where, when both m and q are 0, p is greater than 1.

[0105] As used herein, the term homologs TPGS-1000 means analogues TPGS-1000 that differ from the parent TPGS-1000 only by the presence or absence of a single unit, such as a methylene unit, or a multiple of such units, e.g. - (CH2) n -. Suitable TPGS-1000 homologues have similar surfactant properties compared to the parent compound (TPGS-1000), e.g. similar HLB properties, e.g. HLB properties between 14 or about 14 and 20 or about 20. Suitable TPGS-1000 homologues include homologues TPGS-1000 with minor changes in the PEG chain unit, and me-TPGS-1000, which is TPGS-1000 having a methyl end on the PEG unit.

[0106] As used herein, the term HLB refers to a value that is used to indicate and describe a surfactant according to its relative hydrophobicity / hydrophilicity with respect to other surfactants. The HLB value of the surfactant is an indication of the molecular balance of the hydrophobic and lipophilic fragments of the surfactant, which is an amphipathic molecule. Each surfactant or mixture of surfactants (and / or co-surfactants) has an HLB value, which is the relative weight percentage of hydrophobic and hydrophilic fragments of the surfactant molecule (s) represented by the number. HLB values ​​are obtained from the semi-empirical formula. The relative weight percentages of hydrophobic and hydrophilic groups indicate the properties of surfactants, including the structure of the molecule, for example, the types of aggregates that the surfactant will form and the solubility of the surfactant. See, for example, Griffin, WCJ Soc. Something. Chem. 1: 311 (1949).

[0107] HLB values ​​of surfactants range from 1-45, while the range for non-ionic surfactants is usually from 1-20. The more lipophilic the surfactant, the lower the HLB value. Conversely, the more hydrophilic the surfactant, the higher the HLB value. Lipophilic agents

Surfactants have greater solubility in oily and lipophilic substances, while hydrophilic surfactants dissolve more easily in aqueous media. Generally, 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 referred to as "hydrophobic surfactants" . HLB values ​​have been determined and are available for many surfactants (see, e.g., US Patent No. 6,267,985). It will be understood that the HLB values ​​for given surfactants and co-surfactants may vary depending on the empirical method used to determine the value. Thus, the HLB values ​​of surfactants and co-surfactants provide a preliminary guidance for formulating compositions based on relative hydrophobicity / hydrophilicity. For example, the surfactant is usually selected from surfactants having HLB values ​​in a specific surfactant or co-surfactant range that can be used for pilot formulation. Table 1A provides a list of HLB values ​​for exemplary surfactants and co-surfactants.

Table 1A: HLB values ​​of examples of surfactants and co-surfactants

<td>Surface measure active / co-surfactant active</td><td>HLB</td><td>Surfactant / co-surfactant</td><td>HLB</td>

<td>Hydrogenated castor oil PEG-2</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>Glycerol stearate</td><td> 3,5</td><td>Castor oil PEG-35</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>Hydrogenated castor oil PEG-40</td><td> 14</td>

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

<td>Hydrogenated castor oil PEG-7</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>Glycerol PEG-20 stearate</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>PEG-20 cetyl ether</td><td> 15,7</td>

<td>PEG-40 sorbitan peroxyisostearate</td><td> 10</td><td>Hexadecyl ether (PEG) 20</td><td> 15,7</td>

<td>PEG-10 olive glycerides</td><td> 10</td><td>Hydrogenated castor oil PEG-60</td><td> 16</td>

<td>PEG sorbitan 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>

EP 2 268 160 B1

<td>Surface measure active / co-surfactant active</td><td>HLB</td><td>Surfactant / co-surfactant</td><td>HLB</td>

<td>Hydrogenated castor oil PEG-25</td><td> 10,8</td><td>PEG-75 lanolin</td><td> 16,7</td>

<td>Polysorbate 85</td><td> 11</td><td>PEG-23 lauryl ether</td><td> 16,9</td>

<td>Glycerin ester of PEG-7 coconut oil fatty acids</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>PEG-40 isooctylphenyl ether</td><td> 17,9</td>

<td>PEG-15 glycerol isostearate</td><td> 12</td><td>PEG-100 stearate</td><td> 18,8</td>

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

<td>Tocopherol polyethylene glycol succinate (TPGS)</td><td> 16-18</td><td>phosphatidylcholine</td><td> 7,6</td>

[0108] The surfactants and HLB values ​​shown in Table 1 are exemplary. Any known surfactant or co-surfactant may be used with the provided compositions (see, e.g., US Patent No. 6,267,985). The surfactant (s) contained in the provided compositions typically have an HLB value between 14 or about 14 and 20 or about 20, e.g. 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.

[0109] The term micelle as used herein refers to aggregates formed by surfactants, which are usually formed when the surfactant is present in the aqueous composition, usually when the surfactant is used at a concentration above the critical micellar concentration (CMC). In micelles, the hydrophilic fragments of surfactant molecules contact the water or water phase (the original version uses the synonymous term, for which there is no equivalent in Polish, note the translator), while the hydrophobic fragments form the core of the micelle, which may enclose non-polar component (s), e.g., non-polar compounds in the provided compositions. Typically, the surfactants in the provided compositions form micelles containing the non-polar component in their center in aqueous liquid diluted compositions. Typically, micelles in the provided compositions have a particle size of about 1000 nm, usually less than 500 nm or less than about 500 nm, usually less than 300 or about 300 nm, e.g. less than 250 nm or about 250 nm, e.g. less than 200 nm or less than about 200 nm, e.g. 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.

[0110] The term reverse micelles as used herein means surfactant aggregates that typically form in a lipophilic solution, with the hydrophilic fragments forming the core. When the cross-sectional area of ​​the hydrophobic surfactant molecule is larger than that of the hydrophilic fragment of the molecule, the formation of micelles that can form hexagonal phase structures is favored.

[0111] The term liposomes as used herein means surfactant aggregates consisting of lipid bilayers, usually having an aqueous core. Liposomes are usually formed

By lipid surfactants, usually phospholipids, which are amphipathic phosphate-containing lipids, for example molecules containing one phosphate, glycerol and one or more fatty acids, and similar surfactants. Optionally, phospholipid surfactants can be used as co-surfactants that can be incorporated into aggregates of other surfactant (s), e.g. micelles. Lipid bilayers are two-dimensional layers in which all of the hydrophobic fragments, e.g. acyl side chains, are shielded from interaction with an aqueous liquid, except those at the ends of the layer. Adverse energy interactions of the acyl side chains with water result in the folding of the bilayers forming liposomes, three-dimensional vesicles of the lipid bilayer. In one example, the liposome is formed as a single bilayer enclosing a single water space (small single-plate vesicles; SUVS). In another example, the liposome is composed of concentric bilayers with a plurality of aqueous spaces alternating with bilayers (multi-platelet vesicles; MLVS). Liposomes can be used to enclose hydrophobic and hydrophilic active ingredients. In liposomes, non-polar active ingredients are usually divided into bilayers, while hydrophilic active ingredients are usually trapped in water compartments. In one example, liposomes can be used as a carrier / closing system because they are stable and can protect active ingredients from degradation, e.g. by oxidation and digestive enzymes.

[0112] The term "co-surfactant" as used herein is used to refer to a surfactant, usually a phospholipid, which is used in the compositions provided in combination with a surfactant (e.g., a major surfactant), for example to improve emulsification provided compositions and / or compounds, for example to emulsify the ingredients. In one example, the provided compositions contain at least one surfactant and at least one co-surfactant. Typically, the co-surfactant is a lipid, e.g., phospholipid, e.g., phosphatidylcholine. In one example, the co-surfactant has an HLB value between 7 or about 7 and 8 or about 8. Typically, compared to the surfactant, the co-surfactant is a lower weight (w / w) percentage of the provided compositions. Thus, the provided compositions usually have a lower concentration of surfactant (co-agents) than the surfactant.

[0113] As used herein, the term phospholipid means an amphipathic phosphate-containing lipid, for example a molecule containing one phosphate, glycerol and one or more fatty acids. In one example, one or more phospholipids are used as a co-surfactant in the provided compositions. Exemplary phospholipids used in the provided compositions are lecithin, including phosphatidylcholine (PC), phosphatidylethanolamine (PE), distearoylphosphatidylcholine (DSPC), phosphatidylserine (PS), phosphatidylglycerol (PG), phosphosphatidic acid (PA), phosphatidate or a combination thereof. Usually, the phospholipid is phosphatidylcholine (PC), which is sometimes referred to by the common name 'lecithin'. Exemplary phospholipids that can be used as co-surfactants in the provided compositions are phospholipids sold by Lipoid, LLC, Newark, NJ, on

For example, purified egg lecithins, purified soy lecithins, hydrogenated egg and soy lecithins, egg phospholipids, soybean phospholipids, hydrogenated egg and soybean phospholipids. Synthetic phospholipids, PEGylated phospholipids and phospholipid mixtures sold by Lipoid, LLC. An example of phosphatidylcholine that can be used as a surfactant in the provided compositions is the phosphatidylcholine composition sold by Lipoid, LLC under the name Lipoid S100, which is obtained from soy extract and contains more than 95% or more than about 95% phosphatidylcholine.

[0114] Typically, to form micelles, a surfactant (s) is used in which the cross-sectional area of ​​the hydrophilic fragment of the surfactant molecule is larger than that of the hydrophobic fragment of the molecule. For example, TPGS surfactants having HLB comprised between exactly or about 14 and exactly or about 20, usually between exactly or about 15 and exactly or about 18, are surfactants to stabilize oil-in-water emulsions containing non-polar ingredients active, e.g. nanometer-sized droplets suspended or dispersed in an aqueous phase or an aqueous liquid, e.g. an aqueous medium, such as spherical micelles, containing hydrophilic fragments of molecules (molecules) directed to the aqueous phase and hydrophobic fragments in the center of spherical micelles, for example, surrounding the non-polar active ingredient. Typically, surfactants and / or co-surfactants in the provided compositions aggregate in nanoemulsions and aqueous liquids to form micelles that contain the non-polar compound (s). The hydrophilic fragment (fragments) of the surfactant molecules is oriented outside the micelles in contact with the aqueous medium, while the hydrophobic fragment (fragments) of the surfactant molecules is oriented towards the center of the micelles, in contact with the non-polar compound (s) which is contained within micelles. Micelles may contain more than one surfactant.

[0115] As used herein, the terms "particle size" and "average particle size" synonymously refer to the average diameter of the particles in the supplied liquid, for example the diameter of the droplets or the diameter of the micelles in the emulsion. Typically, the supplied nanoemulsion concentrates, and liquids made from concentrates, have a particle size less than about 1000 nm, usually less than 500 nm or less than about 500 nm, usually less than 300 nm or about 300 nm, e.g., less than 250 nm or about 250 nm, e.g. less than 200 nm or less than about 200 nm, e.g. 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, the diluted compositions provided by diluting the nanoemulsion liquid concentrates with 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 more. Typically, supplied liquid nanoemulsion concentrates are formulated such that diluting the liquid nanoemulsion concentrates in an aqueous medium provides the liquid diluted composition having the appropriate particle size, for example between

Or about 15 nm to about 500 nm or about 500 nm. Optionally, information regarding the particles in liquid diluted compositions may be presented in the form of particle density, for example ppm (parts per million) or percent solids, in liquids.

[0116] The term visible particles as used herein means particles, e.g. in liquids, e.g. emulsions, which are visible when viewed with the naked eye (e.g., without magnification). In one example, visible particles are particles that are observed by a person skilled in the art of formulating compositions, for example concentrates or aqueous liquid diluted compositions containing diluted concentrates. In one example, the provided compositions do not contain visible particles. In another example, the compositions contain few visible particles, e.g. no more visible particles than another liquid, e.g. a beverage. The presence of visible particles and the number of visible particles is determined by empirical observations.

[0117] As used herein, the term visible crystals means crystals that are visible when viewed with the naked eye (e.g., without magnification). The presence or absence of visible crystals is usually determined by empirical observation and can be observed by a person skilled in the art of formulating compositions, for example concentrates or aqueous liquid diluted compositions containing diluted concentrates. In one example, the provided compositions do not contain visible crystals. In another example, the compositions contain few visible crystals, e.g. no more visible crystals than are contained in another liquid, e.g., a beverage.

[0118] The term "turbidity" as used herein is a measure of the turbidity or haze of liquids caused by particles in suspension in a liquid. Turbidity can be measured optically, for example using a nephelometer, an instrument with a light source and a detector. The nephelometer measures turbidity by detecting the scattered light obtained by irradiating the liquid with incident light. The amount of scattered light correlates with the amount of suspended solids in the liquid. For example, a light beam passes through a sample with low cloudiness with a small disturbance. Other methods of measuring turbidity are well known and can be used with the provided methods and compositions. Units of turbidity values, measured using a nephelometer, are nephelometric turbidity units (NTU). In one example, the provided compositions, e.g. aqueous liquid diluted compositions containing diluted liquid nanoemulsion concentrates have low turbidity, for example a turbidity value (NTU) of 30 or about 30; or an NTU value of less than 30 or about 30, e.g., 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

EP 2 268 160 B1

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 turbidity 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, e.g. 200, 175, 150, 100, 50, 25 or less.

[0119] The term "cloudy liquid" as used herein means a liquid that is thick and opaque with visible particles in suspension, for example a liquid that has a cloudy or muddy appearance.

[0120] The term "clear" as used herein may be used to describe a composition as provided herein, for example, hydrated liquid diluted compositions containing diluted nanoemulsion concentrates and / or emulsion concentrates as such. In one example, a clear liquid is one that does not look cloudy during empirical observation (e.g., to the naked eye) and / or does not contain particles or crystals that are visible to the naked eye, and does not exhibit "ring formation". In another example, a clear liquid is one that has a low or relatively low turbidity value, for example an NTU value that is less than or equal to the desired NTU value. In one example, the clear liquid has an NTU value of less than 300 or less than about 300, usually less than 250 or less than about 250, usually less than 200 or less than about 200, e.g. 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, e.g., 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 approximately 8, less than 7 or approximately 7, less than 6 or approximately 6, less than 5 or approximately 5, less than 4 or approximately 4, less than 3 or approximately 3, less than 2 or approximately 2, less than 1 or approximately 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 (e.g., less than 1000 nm or about 1000 nm, usually less than 500 nm or less than about 500 nm, usually less than 300 nm or about 300 nm , usually less than 250 nm or about 250 nm, usually less than 200 nm or about 200 nm, e.g. 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 (less than 10 nm or about 10 nm), e.g. less than or less than about 5,

EP 2 268 160 B1

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.

[0121] In another example, clarity is expressed in relative terms. For example, it may be desirable that a particular composition is equally clear, almost as clear or more clear than another liquid (as determined empirically or by measuring turbidity or particle size). For example, clarity can be estimated relative to another aqueous liquid diluted composition, for example a beverage. For example, in one example, the liquid is clear if in appearance it is similar to another clear liquid, e.g. a drink, e.g. water. For example, it may be desirable that the composition has a particle size that is less than or equal to another liquid, e.g., a beverage. In another example, it may be desirable that the composition has a turbidity value that is less than or equal to another liquid, e.g., a beverage. In another example, it may be desirable for the composition to look more clear or as clear as another liquid, for example a drink, for example by having no more visible particles, no more crystals formed, and / or no greater turbidity than another liquid. In one example, the provided compositions are clear. In another example, they are relatively clear or as clear as or almost as clear as another liquid, for example a beverage that does not contain a non-polar component or a liquid nanoemulsion composition.

[0122] As used herein, the terms "hydrophilic" and "polar" synonymously refer to components and / or compounds having greater solubility in aqueous liquids, e.g., water, than in fats, oils and / or organic solvents (e.g., methanol, ethanol, ethyl ether, acetone and benzene).

[0123] In the provided compositions, exemplary polar components are polar solvents, which are solvents that are more easily miscible with water and other polar components. Thus, polar components dissolve more easily in polar solvents than in non-polar solvents. Polar solvents are well known. Solvent polarity can be assessed by measuring a number of different parameters using well-known methods as described herein (see, e.g., Prizbytek, "High Purity Solvent Guide," Burdick and Jackson Laboratories, Inc., 1980). Polar solvents usually have high dielectric constants, usually dielectric constants greater than or about 15, such as 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 usually have high polarity indices, usually greater than exactly or about 3, such as exactly or about 3, 4, 5, 6, 7, 8 or 9, or greater than 9. Polar solvents have large dipole moments, usually greater than or about 1.4 debaja, such as or about 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0 , 2.1, 22, 2.3, 2.4, 2.5, 2.6, 3.0, 3.5, 4 or greater than 4 debates. Polar solvents include polar protic solvents and polar aprotic solvents.

[0124] As used herein, the term polar protic solvent means a polar solvent that contains a hydrogen atom attached to an electronegative atom such that hydrogen is proton-like and / or the bond between hydrogen and electronegative atom is polarized.

EP 2 268 160 B1

Exemplary polar solvents include, but are not limited to, water, alcohols, including monohydric, dihydric and trihydric alcohols, including, but not limited to, methanol, ethanol, glycerol, propylene glycol.

[0125] Dihydric alcohols are alcohols containing two hydroxyl groups. Exemplary dihydric alcohols include, but are not limited to, glycols, e.g., propylene glycol, ethylene glycol, tetraethylene glycol, triethylene glycol, and trimethylene glycol.

[0126] Trihydric alcohols are alcohols containing three hydroxyl groups. Exemplary trihydric alcohols include, but are not limited to, glycerin, butane1,2,3-triol, pentane-1,3,5-triol, 2-amino-2-hydroxymethyl-propane-1,3-diol.

[0127] Monohydric alcohols are alcohols containing 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 provided compositions is monohydric alcohol.

[0128] As used herein, the terms "non-polar", "lipophilic" and "fat-soluble" synonymously refer to compounds (e.g., non-polar compounds) and / or ingredients, e.g., non-polar active ingredients that have greater solubility in organic solvents (e.g. (ethanol, methanol, ethyl ether, acetone and benzene) and in fats and oils than in aqueous liquids, for example in water. Non-polar compounds include drugs, hormones, vitamins, nutrients and other lipophilic compounds. Typically, the non-polar compounds used in the provided compositions are sparingly water-soluble compounds, e.g., insoluble in water, or compounds having low water solubility. Exemplary non-polar compounds include non-polar active ingredients, e.g., fat-soluble drugs, hormones, exogenous amino acids, e.g., polyunsaturated fatty acids (PUFAs), e.g., omega-3 and omega-6 fatty acids, vitamins, nutrients, nutrients and other compounds . Additional exemplary non-polar compounds are described herein. The provided compositions can be formulated with any non-polar compound, for example, a non-polar active ingredient.

[0129] As used herein, the term non-polar active ingredient refers to a non-polar compound that when administered to an individual, for example a human, elicits or is claimed to elicit the desired response, such as altering the functioning of the organism at the cell, tissue, organ or other level and / or changing the appearance cosmetic or other property, or a non-polar compound that is taken to obtain the desired effect. The non-polar active ingredients can be any synthetic or natural non-polar ingredient or compound, including a pharmaceutical, drug, therapeutic agent, nutritional supplement, herb, hormone or other ingredient. Non-polar nutrients may include the non-polar active ingredients listed herein, as well as other pharmaceutically acceptable or food grade active derivatives of active ingredients, for example salts, esters, amides, prodrugs, active metabolites, isomers, fragments and analogs. Active ingredients may include compounds having documented possession of the desired effect, as well as compounds believed to cause such effects, for example compounds typically taken to supplement nutritional deficiencies.

[0130] As used herein, the subject includes an animal, usually a mammal, usually a human. [0131] As used herein, the term additives includes any that can be added to a food, beverage or other product suitable for human consumption to enhance one or more of its nutritional, pharmaceutical, dietary, health, nutraceutical, health beneficial, providing energy, healing, holistic or other properties. For example, compositions and methods for making food, drink and other aqueous products suitable for human consumption are provided herein that contain one or more additives, usually oil-based additives (e.g., non-polar compounds), such as nutraceuticals, agents pharmaceutical, vitamins, usually oil-soluble vitamins, e.g. vitamin D, E and A, minerals, fatty acids such as essential fatty acids, e.g. polyunsaturated fatty acids, e.g. omega-3 fatty acids and omega-6 fatty acids, e.g. alpha-linolenic acid (ALA), docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA), gamm-linolenic acid GLA, CLA, extract from Sabal palm, linseed oil, fish oil, algae oil, phytosterols and coenzymes, for example coenzyme Q10 and other additives.

[0132] As used herein, an effective amount of an additive such as a non-polar compound, such as a non-polar active ingredient, refers to the amount and / or concentration of the ingredient necessary to prevent, treat, alleviate or partially inhibit the symptom of the disease or disorder, and / or the amount or concentration indicated for individual consumption, such as daily intake, and / or supplementing nutritional deficiencies, for example, an amount sufficient to enhance the nutritional properties of a pharmaceutical, nutraceutical, health and energy product of a food, beverage or other product suitable for human consumption. In some examples, it is desirable that the provided compositions, e.g., liquid nanoemulsion concentrates and / or liquid diluted compositions, contain an effective amount of a specific non-polar compound, e.g., a specific amount per volume or weight of the composition.

[0133] In one example, the effective amount is the concentration or amount of the liquid nanoemulsion composition, wherein at least 25 mg or about 25 mg, usually 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 non-polar active ingredient is contained in at least 8 ounces of volume of an aqueous medium, e.g. a drink.

[0134] The term unit dosage form, as used herein, refers to physically discrete units suitable for human and animal subjects, and individually packaged, as is known in the art.

[0135] As used herein, the term "insoluble in water" refers to the properties of compounds, none of which dissolve when the compound is mixed with water, for example when mixed with water at room temperature, for example between 25 and 50 ° C or between about 25 and 50 ° C. In one example, the non-polar compounds in the provided compositions are poorly soluble in water, for example, have low water solubility.

[0136] As used herein, the term low water solubility refers to solubility in

Water less than 30 or about 30 mg / ml, usually less than 20 mg / ml or about 20 mg / ml, usually less than 10 mg / ml or about 10 mg / ml, usually less than 1 mg / ml or about 1 mg / ml, e.g. water solubility 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. The term poorly water-soluble as used herein can be used to refer to compounds, for example, non-polar compounds that are insoluble in water or have low water solubility.

[0137] As used herein, the terms concentrate, liquid concentrate and nanoemulsion liquid concentrate are used synonymously for compositions provided that contain non-polar compounds, are liquid at room temperature, for example at 25 ° C or about 25 ° C, or at a temperature between ° C or about 25 ° C and 50 ° C or about 50 ° C, and they can be diluted in an aqueous medium to make the provided aqueous liquid diluted compositions. Typically, the liquid nanoemulsion concentrate is a nanoemulsion concentrate that has a particle size (droplets) (or can be diluted to produce an aqueous liquid diluted composition having a particle size) that is less than 1000 or about 1000, usually less than 500 or about 500, usually less than 300 or about 300 nm, usually 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, 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.

[0138] As used herein, the term liquid composition is used to refer to any liquid, for example a composition that is liquid at room temperature, for example at 25 ° C or about 25 ° C, or at a temperature between 25 ° C or about 25 ° C and 50 ° C or about 50 ° C. Exemplary liquid dilution compositions provided are aqueous liquid dilution compositions in which one or more liquid nanoemulsion concentrates have been diluted, for example aqueous liquid dilution compositions containing diluted concentrates. In this example, the non-polar compound and other lipophilic compounds in the concentrate form a phase dispersed in the aqueous liquid, which is an emulsion (e.g., nanoemulsion).

[0139] As used herein, the terms "liquid diluted composition", "diluted composition" and "diluted liquid" are used synonymously with a composition that contains one or more supplied liquid nanoemulsion concentrates (e.g., liquid nanoemulsion concentrates containing a non-polar compound (s)) ), diluted in a liquid, for example in an aqueous medium. Examples of liquid dilution compositions provided are aqueous liquid dilution compositions, e.g., beverages or other liquids containing liquid nanoemulsion concentrates, e.g., water, sauces, soups, syrups, soda, juice, e.g. fruit juice, milk, coffee, tea, nutritional drinks , sports drinks, energy drinks, vitamin-enriched drinks, artificially colored and flavored water and other drinks containing diluted concentrates.

[0140] The term aqueous liquid dilution compositions as used herein means liquid

Diluted compositions which are primarily aqueous, for example compositions containing a liquid nanoemulsion concentrate diluted in an aqueous medium, for example water or other drink. It is not necessary for the aqueous liquid composition to be diluted completely aqueous. For example, the aqueous liquid diluted compositions may contain an aqueous part, e.g. an aqueous dispersion phase, as well as an additional part, e.g. a dispersed phase, e.g. a lipophilic dispersed phase. Typically, the lipophilic dispersed phase contains one or more lipophilic substances, e.g. one or more non-polar compounds, e.g. non-polar active ingredients. Examples of provided aqueous liquid diluted compositions are drinks containing active ingredients, e.g., water, carbonated drink, juice, e.g. fruit juice, milk, coffee, tea, nutritional drinks, sports drinks, energy drinks, vitamin-enriched drinks, artificial colored and flavored water and other drinks. Typically, aqueous liquid diluted compositions are non-polar containing beverages, e.g., beverages containing diluted concentrates.

[0141] The term "oil phase" as used herein is used to refer to a portion (or phase) of a composition such as that provided herein that contains one or more lipophilic components and / or amphiphilic components (oil phase components) and is typically a soluble phase. fats. In the emulsion compositions provided (e.g., nanoemulsion concentrates and diluted compositions), the oil phase usually means the dispersed phase. The term "oil phase" can also be used to refer to a liquid containing oil phase ingredients, which is usually produced in an oil phase vessel, when carrying out methods for making liquid nanoemulsion concentrates. For example, the oil phase may refer to a mixture of ingredients (oil phase ingredients) that are combined, mixed and heated, e.g. in an oil phase vessel (e.g. tank), prior to mixing with the aqueous phase. An "oil phase" may refer to an oil phase mixture that is formed after all ingredients have dissolved; optionally, it may refer to a forming mixture, for example when it is mixed / heated.

[0142] The term oil phase ingredient (s) as used herein refers to the ingredients of the provided compositions that are contained in the oil phase in the provided methods of making the compositions. Typically, the oil phase ingredients include non-polar compounds, e.g., non-polar active ingredients; surfactants; surfactants; oils such as non-polar solvents; preservatives and emulsion stabilizers. Other lipophilic and / or amphiphilic components may be included in the oil phase.

[0143] As used herein, the term "aqueous phase" is used to refer to a portion (phase) of a composition such as those provided herein that contains one or more hydrophilic components and / or amphiphilic components (aqueous phase components), and is typically a soluble phase. water. Typically, in the emulsion compositions provided, for example nanoemulsion concentrates and diluted concentrates, the aqueous phase is the dispersion phase. The term "hydrated phase" is also used to refer to a liquid containing components of the aqueous phase which is produced when carrying out processes for the preparation of liquid nanoemulsion concentrates. For example, the aqueous phase may refer to a mixture of ingredients (water phase ingredients) that are combined, mixed and heated, for example in a water phase vessel, before being mixed with the oil phase. The term 'phase

"Aqueous" may refer to a mixture of the aqueous phase which is formed after all ingredients have dissolved; optionally the term "aqueous phase" may refer to the mixture being formed, for example when it is stirred / heated.

[0144] As used herein, the term water phase component (s) refers to the components of the provided compositions that are contained in the aqueous phase (e.g., added to the water phase vessel) in the provided methods of making the compositions. Typical components of the aqueous phase include, but are not limited to, polar solvents, usually polar protic solvents, such as water and alcohols, usually alcohols having more than one hydroxyl group, such as dihydroxy or trihydroxy alcohols, e.g. glycerol and propylene glycol; surfactants; surfactants; preservatives and emulsion stabilizers. Other hydrophilic and / or amphiphilic components may be included in the aqueous phase.

[0145] The term initial concentrate as used herein means a concentrate (e.g., liquid nanoemulsion concentrate) that is prepared using the provided methods for formulating the provided concentrates, usually by selecting ingredients, e.g., surfactant (s), non-polar, polar compound (s) solvent and optionally other ingredients, and selection of initial ingredient concentrations from appropriate concentration ranges as described herein.

[0146] As used herein, stability refers to the desired properties of the provided compositions, e.g., the stability of the provided compositions 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 or for 1, 2, 3, 4 or more years. In one example, the composition is stable when formulated to remain free from oxidation or significant oxidation over time. In another example, the stable compositions remain clear over time. In another example, stable compositions remain safe and / or indicated for human consumption over time. In one example, the term stability refers to the absence of deposits forming in the compositions over a period of time. In a related example, the term persistence refers to the lack of "ring formation" over a period of time. In another example, the composition is stable if it shows no visible phase separation over a period of time, for example after 24 hours, after one week or after one week. In one example, the compositions are stable if they exhibit one or more of these described characteristics when stored at a particular temperature. In one example, the compositions remain stable at room temperature, for example 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 cooling temperatures, e.g., 4 ° C or about 4 ° C, or at freezing temperatures, e.g., at -20 ° C or about -20 ° C.

[0147] The term stabilize, as used herein, means increasing the stability of one of the provided compositions.

[0148] As used herein, the terms room temperature and ambient temperature are used to describe a temperature that is common in one or more enclosed

The space in which people usually reside or live. Room temperature may vary, but usually refers to temperatures between 19 ° C or about 19 ° C and 25 ° C or about 25 ° C. When the composition is stored at room temperature, it is understood that it is usually stored at a temperature in this range or at a temperature close to that range.

[0149] The term cooling temperature as used herein refers to a temperature that is common in refrigerators, e.g. household refrigerators or restaurants, e.g. a temperature that is below room temperature, but usually a few degrees higher than the freezing point of water (0 ° F or about 0 ° F, or -19 ° C or -20 ° C). Typically, the cooling temperatures are between 10 ° C or about 10 ° C and 0 ° C or about 0 ° C, for example 4 ° C or about 4 ° C. When the composition is stored at a cooling temperature, it is understood to be stored at a temperature common to household refrigerators and industrial refrigerators.

[0150] As used herein, the freezing temperature refers to a temperature of about or below the freezing point of water, e.g., the temperature commonly used in household freezers, e.g. 0 ° F or about 0 ° F, e.g. -19 ° C or about -19 ° C or -20 ° C or about -20 ° C or lower.

[0151] The singular forms "a", "an" and "the" used in the original description include plural references, unless the context clearly indicates otherwise. Thus, for example, reference to a compound comprising an "extracellular domain" includes compounds with one or more extracellular domains.

[0152] As used herein, ranges and amounts may be expressed as "about" a specified value or range. The term about also includes the exact amount. Hence, "about 5 grams" means "about 5 grams" and also "5 grams". It is also understood that the ranges expressed herein include integers in ranges and fractions thereof. For example, the range between 5 grams and 20 grams includes integer values ​​such as 5, 6, 7, 8, 9, 10, 11, 12, 13, .14, 15, 16, 17, 18, 19 and 20 grams and fractions from a range, for example, but not limited to, 5.25, 6.72, 8.5 and 11.95 grams.

[0153] As used herein, the terms "optional" and "optionally" mean that a later described event or situation occurs or does not occur, and that the description includes cases where said event or situation occurs and cases where it does not occur. For example, optionally a different part means that the part is different or not. In another example, an optional combining step means that the method includes the combining step or does not include the combining step.

[0154] The term "ring formation" as used herein 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 drink, for example a liquid diluted composition containing an emulsion or nanoemulsion. Typically, a ring is formed around the periphery of the container, usually at the level of the liquid surface in the container, e.g. at the neck of the container. Ring formation can occur over time, and if it occurs over a short period of time, it can be a sign of impermanence. Ring formation is usually undesirable, especially for liquids for human consumption, e.g., beverages. Usually the provided compositions show no "ring formation" or are stable without

Ring formation over a long period of time, for example days, weeks, months or years. In one example, the compositions are free of ring formation during storage, for example at room temperature, cooling and / or freezing.

[0155] The term fatty acid as used herein refers to straight chain hydrocarbon molecules with a carboxyl group (COOH) at one end of the chain.

[0156] As used herein, the terms polyunsaturated fatty acid and PUFA are synonymously used to refer to fatty acids that contain more than one carbon-carbon double bond in the fatty acid carbon chain. PUFAs, especially essential fatty acids, are useful as dietary supplements.

[0157] The term "essential fatty acids" as used herein means PUFAs which mammals, including humans, cannot synthesize using any known chemical transformation. Thus, essential fatty acids must be obtained from the diet or by supplementing the diet. Examples of necessary PUFA fatty acids are omega-3 (ω3; n-3) and omega-6 (ω-6; n-6) fatty acids.

[0158] The term omega-3 (ω3; n-3) fatty acids as used herein means polyenes interrupted by methylene groups that have two or more cis double bonds separated by a single methylene group and in which the first double bond occurs at the third carbon atom from the end ( ω). Omega-3 fatty acids are used as dietary supplements, for example for the treatment of the disease and prevention. In one example, the provided compositions 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: 3ω3) (short chain fatty acid); stearidonic acid (18: 4ω3) (short chain fatty acid); eicosapentaenoic acid (EPA) (20: 5ω3); docosahexaenoic acid (DHA) (22: 6ω3); eicosatetraenoic acid (24: 4ω3); docosapentaenoic acid (DPA, clupanodic acid) (22: 5ω3); 16: 3 ω3; 24: 5 ω3 and nisin acid (24: 6ω3). The longer chain omega-3 fatty acids can be synthesized from ALA (omega-3 short chain fatty acid). Exemplary non-polar active ingredients containing omega-3 fatty acids are non-polar active ingredients containing DHA and / or EPA, e.g. containing fish oil, krill oil and / or algae oil, e.g. microalgae oil, non-polar alpha-containing active ingredients linolenic (ALA), for example containing linseed oil.

[0159] The term omega-6 (ω-6; n-6) fatty acids as used herein means polyenes interrupted by methylene groups that have two or more cis double bonds separated by a single methylene group and in which the first double bond occurs at the sixth carbon atom from end (ω). In one example, the provided compositions contain non-polar active ingredients that contain at least one omega-3 fatty acid. Exemplary omega-6 fatty acids are linoleic acid (18: 2ω6) (short chain fatty acid); gamma-linolenic acid (GLA) (18: 3ω6); dihomo-gamma-linolenic acid (DGLA) (20: 3ω6); eicosadienic acid (20: 2ω6); arachidonic acid (AA) (20: 4ω6); docosadienoic acid (22: 2ω6); adren acid (22: 4ω6) and docosapentaenoic acid (22: 5ω6). Exemplary non-polar active ingredients containing omega-6 fatty acids are ingredients containing GLA, for example, oil from

EP 2 268 160 B1 borage. Also examples of non-polar active ingredients containing PUFAs are compounds containing conjugated fatty acids, e.g. conjugated linoleic acid (CLA) and compounds containing sabal palm extract.

[0160] The term algae oil as used herein refers to any oil obtained from sea furrows, for example Crypthecodinium sp, in particular Crypthecodinium cohnii. In one example, algae oil is used as a non-polar compound, for example as an active ingredient in the provided compositions. Algae oil usually contains DHA. In one example, algae oil is also a source of EPA.

[0161] The term fish oil as used herein refers to any oil obtained from any fish, usually cold water fish, e.g. fish tissue, from frozen fish tissue, e.g. cod liver. In one example, fish oil is used as a non-polar compound, for example, the active ingredient in the provided compositions. Fish oil usually contains DHA. In one example, fish oil also contains EPA.

[0162] As used herein, the terms preservative and preservative are synonymously used to refer to ingredients that can improve the stability of provided compositions. Preservatives, in particular preservatives for food and drink, are well known. Any known preservative can be used in the provided compositions. Exemplary preservatives that can be used in the provided compositions are oil-soluble preservatives, e.g., benzyl alcohol, benzyl benzoate, methylparaben, propylparaben, antioxidants, e.g. vitamin E, vitamin A palmitate and beta carotene. Usually, the preservative is selected so that it is safe for human consumption, for example in food and drink, e.g. preservative GRAS certified and / or kosher certified, e.g. benzyl alcohol.

[0163] As used herein, the term solvent means an ingredient that can be used to dissolve another ingredient. For example, solvents include polar and non-polar solvents. Non-polar solvents include oils and other non-polar components that dissolve non-polar compounds. In one example, the non-polar active ingredient is dissolved in the non-polar solvent in practical methods for preparing the provided compositions. In this example, the provided compositions contain polar solvents in amounts sufficient to dissolve the non-polar active ingredient. More than one polar solvent may be used. Typically, the non-polar solvent is an oil that is included in the composition in addition to the non-polar compound. For example, a non-polar solvent is usually not a non-polar compound as such, e.g. different from a non-polar solvent. Certain compounds, for example linseed oil and safflower oil, may be non-polar solvents and active non-polar ingredients. Typically, the non-polar solvent contains one or more oils, usually oils other than the non-polar active ingredient or the oil (s) contained in the active ingredient. Exemplary non-polar solvents include, but are not limited to, oils (in addition to the non-polar active ingredient), e.g., vitamin E oil, linseed oil, CLA, borage oil, Dlimonen, rapeseed oil, corn oil, MCT oil and oat oil . Sample Vitamin E Oil

EP 2 268 160 B1 is an 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). Vitamin E oil contains at least 67.2% tocopherol and about 32.8% soybean oil. In one example, a non-polar solvent is synonymously referred to as a "non-polar solubilizing agent".

[0164] As used herein, the terms "w / w", "weight by weight", "weight", "weight%" and "weight percent" are used synonymously to express the mass ratio of one component of the composition compared to the weight of the total composition . For example, when the amount of a particular ingredient constitutes 1% by weight (w / w) of a concentrate, the weight of this ingredient is 1% of the weight of the whole concentrate. Similarly, when the amount of the ingredient is 50% (w / w) of the concentrate, the weight of this ingredient is 50% of the total weight of the concentrate. Similarly, when the composition and / or compound contains 10% by weight of the component, the weight of the component is 10% of the total weight of the composition or compound. As soon as the concentration, amount or percentage (without units) is specified, it is understood that the concentration or percentage means the concentration or percentage by weight.

[0165] Similarly, the terms "v / v", "volume per volume", "volume percent" and "volume percent" as used herein are used synonymously to express the ratio of the volume of one component of the composition to the volume of the total composition.

[0166] As used herein, the term emulsion stabilizer refers to compounds that can be used to stabilize and / or emulsify and / or change the viscosity of the provided compositions, e.g., liquid nanoemusy concentrate and / or an aqueous composition 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 the initial concentrate, especially when the oil and water phases of the initial concentrate (or the aqueous diluted liquid composition obtained from the dilution of the initial concentrate) appear to want to separate. Addition of an emulsion stabilizer may prevent oil and water phases from separating.

[0167] An example of an emulsion stabilizer that can be used in the provided compositions is a composition containing a gum blend, for example, gums used as emulsifiers, for example a blend containing one or more of xanthan gum, guar gum and sodium alginate, for example an emulsion stabilizer sold under the brand name SALADIZER®, available from TIC Gums, Inc. (Belcamp, MD). Other gums may be included in the emulsion stabilizer, for example, acacia and sugar beet pectin. Other mixtures of similar gums may also be used as emulsion stabilizers.

[0168] The term pH regulator as used herein means any compound, usually an acid or base, that is capable of changing the pH of the provided compositions, for example, reducing the pH of the composition or increasing the pH of the composition, usually without changing other properties of the composition, or without significantly altering other properties . PH regulators are well known. Exemplary pH regulators are acids, e.g. citric and phosphoric acid, and bases.

[0169] The term flavoring as used herein means any ingredient that changes, usually improves, the taste and / or aroma of the provided composition, e.g., aqueous liquid dilute compositions, e.g., drinks.

[0170] As used herein, the terms "no more than" and "NMT" refer to an amount that is less than or equal to the specified amount. Similarly, the terms "not less than" and "NLT" refer to an amount that is greater than or equal to the specified amount.

[0171] As used herein, the term natural is used to refer to compositions and / or ingredients in the composition that can be found in nature and is not exclusively made by man. For example, benzyl alcohol is a natural preservative. Similarly, the derivative of polyethylene glycol and tocopheryl is a natural surfactant. In one example, the natural composition / ingredient is GRAS and / or is kosher certified. Usually, the compositions provided are natural, semi-natural and / or contain one or more natural ingredients.

[0172] As used herein, the terms "GRAS" and "GRAS" are used synonymously for compounds, compositions and ingredients that are "Generally Regarded as Safe" by USDA, FDA for use as additives, on for example in food, drink and / or other substances for human consumption, for example, any substance that meets the criteria of section 201 (s) and 409 of the US Federal Food, Drug and Cosmetic Act. Usually the compositions provided here are GRAS certified.

[0173] As used herein, the term kosher is used to refer to substances that comply with Jewish kosher dietary principles, for example, substances that do not contain ingredients derived from non-kosher animals or ingredients that were not produced according to kosher procedures. Usually the compositions provided here are kosher. [0174] As used herein, the term vessel refers to any container, for example, tanks, pots, vials, flasks, bottles, and beakers, which can be used to contain components and / or phases of the provided compositions, during methods of making the compositions. In one example (e.g. to provide larger-scale methods), the vessel is a reservoir that uses for mixing and / or heating one or more components and / or phases of the composition, e.g., water phase tanks and oil phase tanks. Usually, the oily and aqueous phases are mixed and heated in separate tanks before the phases are combined to form an emulsion. In another example, the reservoir is a packaging or storage reservoir that contains the compositions provided after making the composition, for example, an emulsion. Many containers are available for mixing ingredients. Usually, the tanks are cleaned, for example rinsed, washed with soap and / or disinfected according to known procedures, before use or between uses. Usually the tanks are equipped with one or more mixers, e.g. a standard stirrer and / or homogenizer, which are used to mix the ingredients added to the tank. In one example, the tank is further equipped with a heating and / or cooling device. For example, the tank may be a water-jacketed tank. The temperature of the water-jacketed tank is controlled by the water-jacket, for example to heat the contents, for example during mixing.

[0175] As used herein, the term aqueous phase vessel refers to a vessel used to mix and / or heat the components of the aqueous phase to form the aqueous phase of the provided composition. In one example (e.g. for enlarged scale methods), a water phase vessel

EP 2 268 160 B1 is a water phase tank. In one example, the water phase tank is a water jacketed tank that is equipped with a water jacket that can be used to heat the contents of the tank.

[0176] As used herein, the term oil phase vessel refers to the vessel used to mix and / or heat the oil phase ingredients to form the oil phase of the provided compositions. Usually the oil phase vessel is an oil phase tank. In one example, the oil phase tank is a water jacketed tank.

[0177] The term transfer medium as used herein refers to any equipment, combination of equipment and / or system that can be used to transfer a liquid, for example from one vessel to another vessel, in the provided methods of making the composition. Exemplary transfer means are a conveying pump and appropriate accessories, e.g. plumbing fittings, ball valves and transfer hoses, e.g. food grade hoses.

[0178] As used herein, the term agitator refers to any piece of equipment or combination of equipment that can be used to mix ingredients in the provided methods of making compositions, for example, standard agitators and (shear) homogenisers. For example, stirrers can be used to mix water phase, oil phase ingredients and / or to mix additional ingredients.

[0179] The term standard agitators as used herein means agitators that are used to combine a group of ingredients, e.g., oil phase ingredients or aqueous phase ingredients, and to mix one or more ingredients with a liquid, e.g., an emulsion, e.g., for mixing additional ingredients. with emulsion. Standard mixers can be any mixers that set the material in motion, e.g., ingredients, during heating, e.g., to promote dissolution of the ingredients.

[0180] As used herein, the terms "homogenizer" and "shear" are used to refer to mixers that typically have high shear, which can be used, for example, to form emulsions, e.g., emulsifying the aqueous phase and the oil phase, in the methods provided. Homogenizers are usually capable of high shear mixing which emulsifies the phases.

[0181] As used herein, the term "cooling device" means any piece of equipment or combination of equipment that can be used in the methods provided to cool compositions and phases and their components, e.g., during mixing and / or homogenizing, e.g., to cool the mixture during the oil phase emulsification and water. Exemplary cooling devices are coolers (coolers), e.g. recirculated coolers that can be attached, e.g., to tanks used in the provided methods, e.g. at a distance or by placing the tank in the cooler, to recirculate liquid from the tank through the cooler and back to tank to cool quickly and maintain the temperature of the mixture during mixing. Typically, a cooling device can be used to cool liquids 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, usually between 25 ° C and 43 ° C, usually between 35 ° C and 43 ° C, e.g. 26.5 ° C. Desired temperature, for example between 25 ° C or

About 25 ° C and 45 ° C or about 45 ° C, usually between 35 ° C and 43 ° C, for example 26.5 ° C, in less than 2 hours or about 2 hours, usually less than 1 an hour or about 1 hour, for example 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.

[0182] The term low heating as used herein 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 not more than 85 ° C or about 85 ° C, usually greater than 60 ° C or about 60 ° C, usually 60 ° C or 60 ° C. In the provided methods for producing liquid nanoemulsion concentrates, the oil phase and aqueous phase ingredients are typically heated using low heat to protect the ingredients, e.g., to prevent oxidation of ingredients, e.g., non-polar active ingredients, e.g., omega-3 containing compounds, on example of DHA. [0183] As used herein, the term "essentially consisting of" means comprising the list of ingredient (s) followed by it and not including any additional active ingredient, for example not including any additional active drug or pharmaceutical. For example, the composition, for example a liquid nanoemulsion essentially consisting of many of the components listed, contains these specific components and does not contain any additional active drug or pharmaceutical.

[0184] C here used₁-C_x includes C₁-C₂, C1-C3 ... C_x.

[0185] As used herein, the terms "alkyl" and "alkyl" refer to a straight or branched chain substituted or unsubstituted hydrocarbon groups having any number of carbon atoms; the number of carbon atoms can be determined, e.g. 1 to 30 carbon atoms, 8 to 28 carbon atoms, 7 to 27 carbon atoms, 8 to 22 carbon atoms, 8 to 20 carbon atoms, 8 to 18 carbon atoms and 12 to 18 carbon atoms . The alkyl group may be "saturated alkyl" meaning it does not contain any alkene or alkyne groups, or "unsaturated alkyl" meaning it contains at least one alkene or alkyne group, and may optionally be substituted. An alkyl group that contains at least one carbon-carbon double bond (C = C) is also termed "alkenyl"; alkenyl groups may be optionally substituted. An alkyl group that contains at least one carbon-carbon triple bond (C ° C) is also referred to by the term "alkynyl"; alkynyl groups may be optionally substituted.

B. COMPOSITIONS CONTAINING NON-POLAR COMPOUNDS [0186] Compositions containing non-polar compounds and methods of making the composition are provided herein. Non-polar compounds are sparingly soluble in water (e.g. have low water solubility or are insoluble in water). Thus, it can generally be difficult to formulate non-polar compounds in compositions for human consumption, in particular aqueous compositions, for example, food products and beverages. Poor water solubility of non-polar compounds may also contribute to their poor bioavailability. Improved methods and compositions for formulating non-polar compounds are provided herein.

[0187] In general, emulsions (e.g. oil-in-water emulsions) are colloidal dispersions of two immiscible liquids (e.g. oil and water or other aqueous liquid) containing a dispersion and a dispersed phase. Emulsions can be used to disperse non-polar compounds in aqueous liquids. In oil-in-water emulsions, the dispersed phase is the oil phase and the water phase disperse phase. There remains a need for improved emulsions (e.g. oil-in-water emulsions) containing non-polar compounds in aqueous liquids, and methods and compositions for making improved emulsions. In particular, emulsions are needed which are more suitable and indicated for human consumption of non-polar compounds, for example in food products and beverages. For example, emulsions having improved clarity (e.g., small particle size, low turbidity), stability (e.g., no separation), taste and smell are needed.

[0188] Among the provided compositions are such improved emulsions. For example, emulsions are provided that contain non-polar compounds dispersed in an aqueous liquid and have desirable properties, including improved clarity, stability, aroma and taste. The provided compositions (and methods for making the compositions) can be used to formulate any non-polar compound into aqueous compositions, including the non-polar compounds described herein (e.g. non-polar active ingredients) and other known non-polar compounds.

[0189] Usually provided emulsions containing non-polar compounds are nanoemulsions which are emulsions having dispersed droplets (particles) with a diameter less than 1000 nm or less than about 1000 nm, usually less than 500 nm or less than about 500 nm, usually less than 300 nm or about 300 nm, usually less than 250 or less than about 250 nm, usually 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. Commonly provided nanoemulsion compositions are oil-in-water nanoemulsions containing non-polar compounds dispersed in an aqueous liquid.

[0190] The provided emulsion compositions are stabilized using one or more surfactants and / or co-surfactants and / or emulsion stabilizers. Surfactants form an interfacial film in the emulsion, between the oil phase and the aqueous phase, providing durability. Typically, nanoemulsions of the provided compositions contain micelles in which one or more surfactants surrounds the non-polar active compound. The micelles are dispersed in the water phase.

[0191] The provided emulsion compositions include liquid nanoemulsion concentrates containing non-polar compounds that can be diluted to provide non-polar compounds in liquid compositions, such as beverages. Liquid nanoemulsion concentrates can be diluted in a medium, e.g., an aqueous medium, e.g., a beverage, to produce a liquid diluted composition (e.g., aqueous liquid diluted composition) containing a non-polar compound. Examples of provided compositions are also liquid diluted compositions (e.g., aqueous liquid diluted compositions that may be clear), prepared by diluting nanoemulsion concentrates in a medium.

[0192] The compositions can be prepared using any non-polar compound. compounds

Non-polar active compounds are usually non-polar active compounds, for example pharmaceuticals, nutraceuticals, vitamins and minerals. Non-polar active ingredients include, but are not limited to, polyunsaturated fatty acid (PUFA) containing compounds, e.g., omega-3 containing active ingredients, e.g., ALA, DHA and / or EPA containing compounds, e.g., oils obtained from fish and microalgae , krill and / or flax extract, and non-polar active ingredients containing omega-6, e.g. compounds containing gamma-linolenic acid (GLA), e.g. borage oil; compounds containing sabal palm oil; conjugated fatty acid containing ingredients, e.g. conjugated linoleic acid (CLA) containing compounds; active ingredients containing coenzyme Q, for example coenzyme Q10 (CoQ10), usually compounds containing oxidized CoQ10 (ubidecarenone) and compounds containing phytosterols (plant sterols). Additional exemplary non-polar active ingredients are described here. Any non-polar compound can be used in the provided compositions.

1. Liquid nanoemulsion concentrates containing non-polar compounds [0193] Liquid nanoemulsion concentrates (also called "concentrates" or "liquid concentrates") containing one or more non-polar compounds are provided. Concentrates can be diluted in an aqueous medium to produce aqueous diluted liquid compositions containing non-polar compounds. Liquid concentrates can be formulated based on one or more desired properties, for example, clarity; security; taste; smell; stability, e.g. no phase separation, "ring formation" and / or precipitation over time, and / or bioavailability of the concentrate and / or aqueous dilute liquid compositions containing the concentrate. In one example, a desirable property is the ability of the concentrate provided to provide a clear or partially clear aqueous liquid dilution composition when diluted in an aqueous medium, for example a drink such as water. In another example, the desired properties relate to the safety of the concentrates and / or the attractiveness of the concentrates for human consumption, for example in food and drink. In another example, it may be desirable for the concentrate to contain less than or equal to a particular concentration of one or more ingredients. In another example, it may be desirable for the concentrate to contain greater than or equal to a particular concentration of one or more ingredients.

[0194] In addition to non-polar compounds, the concentrates contain at least one surfactant. Typically, the surfactant has an HLB value between 14 or about 14 and 20 or about 20, e.g. 14, 15, 16, 17, 18, 19, 20, about 14, about 15, about 16, about 17, about 18, about 19 or about 20. Examples of suitable surfactants are surfactants derived from vitamin E, surfactants derived from polyethylene glycol (PEG), such as polyethylene glycol tocopherol succinate (TPGS), in particular those having an HLB value between exactly or about 14 and exactly or about 20, and surfactants with similar properties such as HLB values. Typically, the surfactant is a natural surfactant, for example a surfactant such as GRAS (widely considered safe) certified by the FDA and / or kosher certified, e.g. TPGS.

[0195] Liquid concentrates further 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, usually a large amount of polar solvent, e.g. between 60 % or about 60% and 80% or about 80% by weight (w / w) of the concentrate, usually between exactly or about 60% and exactly or about 79% by weight of the concentrate.

[0196] Typically, the concentrates further contain one or more additional ingredients. Exemplary additional ingredients that may be included in the concentrates are preservatives, non-polar solvents, surfactants, emulsion stabilizers, pH regulators, and flavoring agents.

[0197] Non-polar compounds in concentrates and in diluted compositions are contained in micelles. These micelles, containing a non-polar compound surrounded by one or more surfactants, allow the non-polar compound to be dispersed among polar solvents, for example when concentrates are diluted to form aqueous liquid diluted compositions. Micelles containing non-polar compounds usually have a small or relatively small particle size, e.g. smaller than 1000 nm or about 1000 nm, smaller than 500 nm or about 500 nm, usually smaller than 300 nm or about 300 nm, usually smaller 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. The smaller particle size is correlated with the clarity of the aqueous liquid diluted compositions containing the diluted concentrates. For example, a liquid with a smaller particle size is clearer than a liquid with a larger particle size. Small particle sizes can also contribute to other desirable properties, e.g. durability.

[0198] Many factors, including ingredients, their relative concentrations and methods for making concentrates, affect the particle size of the composition and other desirable composition properties, such as clarity. In particular, the nature of the surfactant, especially the HLB surfactant and the relative concentrations of polar solvent (e.g. water), a surfactant and a non-polar compound, contribute to the small particle size and clarity of the aqueous liquid diluted compositions. Usually several of these parameters and properties are related to each other. For example, several parameters contribute to the particle size, usually the small particle size of the composition. Particle size directly contributes to the clarity of the aqueous liquid dilute compositions containing concentrates. Particle size may also be related to other properties, for example stability, lack of "ring formation" and / or the formation of aqueous sediments of diluted liquid compositions containing concentrates.

[0199] Accordingly, the properties of the ingredients and their relative concentrations in the concentrates are important for the ability of the concentrate to give the desired diluted compositions. Methods for formulating liquid nanoemulsion concentrates are provided. Determination of the appropriate ingredients and their relative concentrations that dilute compositions having the desired properties will provide is accomplished using the liquid concentrate formulation methods provided.

a. Formulation of liquid concentrates

[0200] In the formulation methods provided, concentrates are formulated by choosing ingredients and concentration ratios of the ingredients that will give compositions having one or more desired properties. When concentrating, the selected ingredients and starting concentrations are used to make initial concentrates, which are evaluated and modified if necessary.

[0201] As a first step in the formulation of the concentrates provided, one or more initial concentrates are prepared and evaluated for initial properties. For this step, the ingredients are selected, for example, from the ingredients described herein. In general, the ingredients include surfactants, polar solvents, non-polar active ingredients and other ingredients. The starting concentration (weight percent) of each component selected is selected from the appropriate concentration range for such component or component category, for example, the appropriate concentration range for the surfactant. In some cases, the initial concentration is formulated based on the ingredients, and their concentration, of the existing concentrate. having one or more desired properties.

[0202] Next, the initial concentrate (s) are prepared using the methods below to prepare concentrates, adding each ingredient at its initial concentration in the appropriate stage. In one example, more than one initial concentrate is prepared and compared, e.g., multiple initial concentrates, each having a different concentration of one or more ingredients. In one example, multiple initial concentrates are prepared to test different representative concentrations in the appropriate concentration range for one or more specific ingredients.

[0203] In a typical example, the initial concentrate is prepared by incorporating at least one surfactant, such as selected from among the surfactants described herein, which has an HLB value between 14 or about 14 and 20 or about 20, at an initial concentration ranging from between 16% or about 16% and 30% or about 30% by weight (w / w) of the concentrate; at least one non-polar compound at an initial concentration ranging between 5% or about 5% and 10% or about 10%; and a polar solvent, at an initial concentration of between 60% or about 60% and 80% or about 80%, and usually between exactly or about 60% and exactly or about 79% by weight. In one example, the initial concentrate further includes other ingredients, e.g., preservative (s), co-agent (s), and / or other ingredients as described herein.

[0204] After the initial concentrate (s) are prepared, the concentrate (s) is evaluated for one or more desired properties, e.g., the ability to form dilute compositions (e.g., clear diluted compositions or diluted compositions having a specified turbidity, particle size, or other property). The ability to form diluted compositions having one or more properties is assessed by diluting the concentrate in an aqueous medium, for example, diluting the concentrate in an aqueous medium at a dilution factor between 1:10 or about 1:10 and 1: 1000 or about 1: 1000 or more, usually between 1:10 or about 1:10 and 1: 500 or about 1: 500 or more, for example a dilution not greater than 1:10 or about 1:10, 1:20 or about 1:20, 1:25 or around 1:25, 1:50 or

EP2 268160B1 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, e.g. 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, or as per other dilutions provided here.

[0205] After evaluation, the ingredients and / or their concentrations can be adjusted to produce the desired properties of the final concentrate. Typically, the concentration of the non-polar compound, surfactant and / or solvent is the concentration that is corrected after evaluation of the initial concentrate. Similarly, when multiple initial concentrates are formulated, one or more concentrations of non-polar compound, surfactant, and polar solvent vary among the multiple initial concentrates. In some cases, after evaluation, it can be determined that to obtain the desired properties of the particular concentrate, additional ingredients (not included in the initial formulation) are needed or desired. This process can be repeated until a concentrate having the appropriate property or properties is produced.

i. Common ingredients and typical concentration ranges [0206] Each of the concentrates provided contains a non-polar compound such as, but not limited to, the exemplary non-polar compound described below. Typically, the non-polar compound is a non-polar active ingredient, for example an oil-based active ingredient such as polyunsaturated fatty acid (PUFA), coenzyme Q or a phytochemical compound. For the formulation of the initial concentrate, the starting concentration of the non-polar compound is usually the concentration selected from a concentration range between 5% or about 5% and 10% or about 10% (w / w) of the concentrate, such as a 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 non-polar compound is usually added as part of the oil phase in accordance with the methods provided for making the concentrate.

[0207] The initial concentrate further contains at least one surfactant that can be added to the aqueous phase or oil phase, and usually has an HLB value between 14 or about 14 and 20 or about 20, for example 14, 15, 16, 17, 18, 19 or 20 or about 14, about 15, about 16, about 17, about 18, about 19, about 20, usually between exactly or about 15 and exactly or about 18, including, but not limited to, TPGS and its analogues and derivatives, usually a natural surfactant that is safe and / or approved for human consumption.

[0208] Typically, the starting concentration of the surfactant is selected from a concentration range between 16% or about 16% and 30% or about 30% (w / w), for example 16% or about

16%, 17% or approximately 17%, 18% or approximately 18%, 19% or approximately 19%, 20% or approximately 20%, 21% or approximately 21%, 22% or approximately 22%, 23% or approximately 23 %, 24% or approximately 24%, 25% or approximately 25%, 26% or approximately 26%, 27% or approximately 27%, 28% or approximately 28%, 29% or approximately 29% or 30% or approximately 30% by weight (w / w) concentrate, such as, for example, 17.75%, 20.25%, 20.5%, 22.7% or 25.2% (w / w) of the concentrate.

[0209] In one example, the surfactant concentration range 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) concentrate, 20% or about 20% (w / w) concentrate, 23% or about 23% (w / w) 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 surfactant concentration range is between 18% or about 18% and 25% or about 25% (w / w) of the concentrate. In another example, the surfactant concentration range is between 18% or about 18% and 20% or about 20% (w / w) of the concentrate. In another example, the surfactant concentration range is between 17% or about 17% and 20% or about 20% (w / w) of the concentrate. In another example, the surfactant concentration range is between 16% or about 16% and 20% or about 20% (w / w) of the concentrate.

[0210] The concentrates further contain polar solvents (e.g. water or other edible polar solvent, e.g. propylene glycol and glycerin), usually a high concentration of polar solvent which is added to the aqueous phase. Usually, the starting concentration of the polar solvent is selected from a concentration range between 60% or about 60% and 80% or about 80% (w / w) of the concentrate, e.g. 60% or about 60%, 61% or about 61%, 62 % or approximately 62%, 63% or approximately 63%, 64% or approximately 64%, 65% or approximately 65%, 66% or approximately 66%, 67% or approximately 67%, 68% or approximately 68%, 69% or about 69%, 70% or about 70%, 71% or about 71%, 72% or about 72%, 73% or about 73%, 74% or about 74%, 75% or about 75%, 76% or about 76% 77% or about 77%, 78% or about 78%, 79% or about 79%, 80% or about 80% (w / w) of the concentrate, such as for example 68.29%, 68.7865%, 74.25%, 71.74% or 75.8165% (w / w) of the concentrate. In one example, the polar solvent concentration range is between 65% or about 65% and 80% or about 80% (w / w) of the concentrate. In another 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 76% or about 76% (w / w ./wag.) of concentrate.

[0211] One or more, usually more than one, additional component may be added to the initial concentrate. For example, concentrates typically contain at least one preservative, usually a natural preservative, for example benzyl alcohol. Examples of other additive ingredients that can be added to concentrates, including initial concentrates, are emulsion stabilizers, for example, gum blends; a non-polar solvent for a non-polar compound, e.g., an oil other than a non-polar compound, e.g., vitamin E oil or linseed oil; pH adjuster, e.g. citric acid, phosphoric acid; one or more flavoring agents, for example D-limonene or lemon oil; a co-surfactant, e.g., phospholipid, e.g., phosphatidylcholine.

[0212] Suitable concentration ranges for additional ingredients are described in separate paragraphs below. Usually, the concentration of additional ingredients depends partly on the concentrations

EP 2 268 160 B1 of the non-polar active ingredient, surfactant and polar solvent. Usually, the concentrations of these three components (surfactant, polar solvent and non-polar compound) form the basis of the formulation methods. For example, when it is determined that the concentrations of the components in the initial concentrate need to be modified, they are usually the concentrations of one or more of the three components that are regulated.

[0213] In one example, after evaluating the initial concentrate, it may be desirable to add one or more additional ingredients to improve the concentrate with respect to one or more desired properties.

ii. Performing the evaluation of the initial concentrate [0214] Formulation methods further include concentrate analysis based on one or more desired properties, e.g., the properties of an aqueous liquid dilution composition containing the diluted concentrate, e.g., clarity, color, smell, taste, safety, stability, "ring formation 'Or the formation of deposits and / or the presence of crystals. For example, the methods typically involve analyzing the ability of the initial concentrate to form a clear liquid when diluted in an aqueous medium, such as analyzing the clarity / turbidity of the resulting aqueous diluted liquid composition containing the initial concentrate.

[0215] To assess the properties of the aqueous liquid dilution composition, the initial concentrate is diluted in an aqueous medium, usually water or another polar solvent, for example at a dilution ratio between 1:10 or about 1:10 and 1: 1000 or about 1: 1000, usually between 1:10 or about 1:10 and 1: 500 or about 1: 500, e.g. diluted not more than 1:10 or about 1:10, 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, e.g. 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 supplied here. Typically, the clarity of the aqueous diluted liquid composition containing the diluted initial concentrate is assessed using one or more approaches. In addition, other properties may be assessed, for example the aroma and / or taste properties of liquids, for example when the non-polar compound is polyunsaturated fatty acid (PUFA), especially fish oil or algae oil, it can be empirically assessed whether the aqueous diluted liquid composition has " smell.

(1) Clarity [0216] In one example, dilution of the supplied concentrates in an aqueous medium gives clear liquids. The clarity of the aqueous liquid diluted composition containing the initial concentrate can be assessed using one or more of many approaches, such as empirical observation, measurement of particle size, and / or measurement of liquid turbidity.

[0217] In one example, the concentrates can be diluted to form clear liquids (or liquids that are as clear as known liquids), by adding between 0.05 grams (g) or

About 0.05 g 10 g or about 10 g concentrate, usually between 0.05 g and 5 g, 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 concentrate to 8 ounces volume, about 8 ounces volume or at least 8 ounces volume or at least about 8 ounces volume e.g. 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 100, 200 or more ounces of volume of aqueous medium, e.g., water, to produce a clear liquid dilution composition that contains a non-polar compound. In another example, to make a clear aqueous liquid dilution composition, concentrates can be diluted by adding between 1 mL or about 1 mL 10 mL or about 10 mL concentrate, e.g. 1 mL, 2 mL, 3 mL, 4 mL, 5 mL , 6 ml, 7 ml, 8 ml, 9 ml or 10 ml concentrate to 8 ounces volume, about 8 ounces volume or at least 8 ounces volume or at least about 8 ounces volume, e.g. 8, 9, 10,11, 12 , 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 100, 200 or more ounces of volume of aqueous medium, e.g. water, to produce a clear aqueous diluted liquid composition that contains a non-polar compound.

[0218] In another example, to produce a clear aqueous liquid diluted composition, the concentrate may be diluted in an aqueous medium when at least 25 mg or about 25 mg, usually at least 35 mg, e.g. 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 or more non-polar active ingredient is contained in at least 8 ounces by volume or at least about 8 ounces by volume of aqueous diluted liquid composition, e.g., beverage, e.g.

[0219] In another example, to make a clear aqueous liquid dilution composition, the concentrate may be diluted in an aqueous medium at a dilution ratio between 1:10 or about 1:10 and 1: 1000 or about 1: 1000, usually 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, e.g. 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, a clear liquid is prepared at less diluted dilutions than 1:10 concentrate.

[0220] Diluted liquid nanoemulsion concentrates can be formulated using any non-polar compound for dilution in an aqueous medium. In one example, the concentrates can be diluted in an aqueous medium, for example, in a wide dilution range to produce clear liquids, for example at a dilution ratio between 1:10 or about 1:10 and 1: 1000 or about 1: 1000, usually 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, e.g. 1:10, 1:20, 1:25, 1:30, 1:35,

EP2 268160B1

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 clarity of the liquid is maintained with increasing dilutions, for example, to infinity.

[0221] The clarity of the aqueous liquid dilution composition can be assessed using one or more of many approaches, for example qualitatively by empirical evaluation or quantitative by measuring particle size and / or measuring the turbidity of a liquid. In one example, a particular qualitative or quantitative value is desired. In another example, it is desirable for the aqueous liquid dilution composition to be as clear as, less clear or more transparent than another liquid, e.g., an aqueous liquid dilution composition prepared according to the provided methods, or a beverage, e.g. a beverage or other aqueous medium. contains concentrate. For example, an aqueous liquid dilution composition containing a liquid concentrate diluted in a beverage may be as clear or nearly as clear as the same beverage without a concentrate. The assessment can be performed qualitatively, for example by empirical observation, or quantitatively, for example, by calculating particle size and / or turbidity (NTU) values ​​for one or more liquids.

(2) Empirical assessment [0222] The relative clarity / turbidity of the aqueous liquid diluted composition containing the diluted concentrate (e.g., initial concentrate) can be qualitatively assessed by observation. In one example, a liquid is considered clear if it does not appear cloudy in appearance and / or if there are no visible particles when looking at the liquid with the naked eye. Clarity can be determined empirically by comparison to other liquids, e.g. water, fruit juice, carbonated drink and / or milk. For example, it may be desirable for the liquid to be as clear or almost as clear as water or another liquid, such as a drink. For example, a liquid (containing a liquid concentrate diluted in an aqueous medium, e.g., a drink) is as clear or almost as clear as an aqueous medium not containing the diluted concentrate. In a related example, it may be desirable that there is no significant difference, e.g., no noticeable difference, between the aqueous diluted liquid 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 turbidity can be considered clear.

(3) Particle size [0223] Optionally, the clarity of the aqueous liquid dilution composition containing the diluted concentrate (e.g., initial concentrate) can be assessed by measuring the particle size of the liquid. Methods for measuring particle size are known and a method for measuring particle size can be used that can measure particle sizes in the appropriate ranges as described below.

[0224] Particle size can be analyzed using commercial devices, e.g., Delta Analytical Instruments, Inc., such as a light scattering analyzer, e.g., dynamic light scattering analyzer, e.g., Horiba® LB-550, which can measure particle sizes in the range of 0.001 micron up to 6 microns and uses the technique to present data

Fourier transformation / iterative deconvolution for the data obtained and can measure sample concentrations from ppm to 40% solids; Horiba® LA-920, which is a laser light scattering device having a He-Nu laser and tungsten lamp, can determine particle sizes from 0.02 microns to 2000 microns using Mie theory; or other analyzers available from Delta Analytical Instruments, Inc.

[0225] Optionally, the particle size can be measured microscopically, for example by observing the liquid under a microscope, for example at 640X magnification. Using this method, the particle size can be determined by comparison to a measuring instrument, for example a ruler, which is visible when looking at a liquid under a microscope. If any particles are observed at this magnification, they are measured by comparison with a measuring instrument. At 640X magnification, for example, any particle that is about 25 nm, 25 nm or more than 25 nm is visible, while particles smaller than 25 nm are usually not visible.

[0226] It is usually desirable that the aqueous liquid dilution compositions have a particle size less than 200 nm or less than about 200 nm, for example 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, SO, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190 or 200 nm. It is usually desirable that the aqueous liquid dilution compositions 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, usually between 5 nm or about 5 nm and 50 nm or about 50 nm.

(4) Turbidity measurement [0227] Optionally, liquid clarity can be analyzed by performing optical turbidity measurements that show the level of turbidity or haze that correlates with the size / number of particles in suspension in the liquid. The more clear the liquid is, the lower the turbidity value.

[0228] Turbidity can be measured optically, for example using a nephelometer, an instrument with a light source and a detector. The nephelometer measures turbidity by detecting the scattered light obtained by irradiating the liquid with incident light. The amount of scattered light is correlated with the amount of solid particles in the liquid. For example, a beam of light passes through a sample with low turbidity with little disturbance. Other methods of measuring turbidity are well known and can be used with the provided methods and compositions.

[0229] Units of turbidity values ​​measured using a nephelometer are nephelometric turbidity units (NTU). In one example, it is desirable that the aqueous liquid dilution composition containing the diluted concentrate has low turbidity, e.g., a turbidity value (NTU) of 30 or about 30; or an NTU value of less than 30 or about 30, e.g., 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 around 17, smaller

EP2 268 160 B1 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 approximately 9, less than 8 or approximately 8, less than 7 or approximately 7, less than 6 or approximately 6, less than 5 or approximately 5, less than 4 or approximately 4, less than 3 or approximately 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 turbidity value of the aqueous liquid dilution composition is less than 200 or less than about 200, for example 200, 175, 150, 100, 50, 25 or less.

[0230] In another example, it is desirable that the aqueous liquid dilution composition contain a turbidity value that is comparable, e.g., nearly the same as, same as or less than or greater than, the turbidity value of another liquid, e.g. a liquid concentrate or aqueous liquid diluted composition made using the methods provided.

iii. Choice of formulation and modification of the formulation [0231] After evaluating the initial concentrate (s), either a specific formula is selected or one or more modifications of the initial concentrate formula are made based on the results of the evaluation. When the initial concentrate does not have one or more desired properties, e.g. to the desired extent, based on evaluation, the concentration of one or more ingredients can be adjusted and another starting concentrate can be prepared to repeat the process until a concentrate with the desired properties is obtained. To modify the initial concentrate, the amount of polar solvent, surfactant and / or non-polar active ingredient can be adjusted, e.g. to a different concentration within the appropriate concentration range. Alternative ingredients can also be selected. In one example, modification of the initial concentrate requires the addition of one or more additional ingredients. For example, if the evaluation has shown that the oily and aqueous phases of the concentrate or aqueous diluted liquid composition containing the diluted concentrate separate, emulsion stabilizer may be added to the formulation. In another example, a co-surfactant may be added to facilitate the emulsification of the concentrate ingredients. In another example, the phase (oil phase or aqueous phase) to which a specific component is added is modified. For example, the formulation can be modified to change it depending on whether the surfactant is added to the oil phase or the aqueous phase.

[0232] In one example, when the evaluation of the initial concentrate has shown that it has the desired properties, no modifications are made. In this example, the initial concentrate formula is used to make the concentrate. When two or more initial concentrates are produced, for example with increasing concentrations of ingredients, the formula of one of the initial concentrates can be changed. Which formula is chosen can be done in

EP2 268 160 B1 based on which formula has the more desirable property. Optionally, the desired properties can be adjusted using relative amounts of ingredients. In one example, it is desirable to choose a formulation that uses the lowest and highest concentrations of a particular ingredient, but still provides a concentrate that produces a clear liquid when in an aqueous medium. In one example, the desired formulation is the formulation that has the lowest surfactant concentration while still providing a concentrate that produces a clear liquid when diluted in an aqueous medium. In another example, the desired formulation is the formulation that has the highest concentration of non-polar active ingredient while still providing a concentrate that gives a clear liquid when diluted in an aqueous medium. In another example, a formulation that provides the clearest liquid is desired.

[0233] However, in the next example, the formula is modified even if the initial concentrate has the desired properties. For example, when determining that a particular concentrate formulation gives the desired properties, it may be desirable to modify the concentration of one or more ingredients to determine if the same desired properties can be obtained if a higher or lower concentration of the ingredient (s) is used. For example, it may be desirable to determine the highest surfactant concentration that can be used while a concentrate will still be produced with the desired property, for example the ability to form a clear liquid when diluted in an aqueous medium. In another example, it may be desirable to determine the highest concentration of non-polar component that can be incorporated into the concentrate while still maintaining the desired property, e.g. the ability of the concentrate to form a clear liquid when diluted in an aqueous medium. In another example, one or more additional ingredients may be added after making an initial concentrate with the desired properties, e.g., flavoring agents and / or pH adjusting agents.

[0234] The following paragraphs describe the ingredients used in the supplied nanoemulsion liquid concentrates.

b. Non-polar compounds [0235] The concentrates contain one or more non-polar compounds. Non-polar compounds include any lipophilic or fat-soluble compounds, for example active ingredients that have greater solubility in organic solvents (e.g., ethanol, methanol, diethyl ether, acetone and benzene) and fats and oils, than in aqueous liquid diluted compositions on example in water. Usually, non-polar compounds are sparingly soluble in water, for example, insoluble in water or compounds having low water solubility. Non-polar compounds include, but are not limited to, drugs, hormones, vitamins, nutrients and other lipophilic compounds. Exemplary non-polar compounds are listed below. The provided methods and compositions can be diluted (e.g., dissolved / dispersed) any non-polar compound in an aqueous medium. In one example, the non-polar compound is different from the surfactant, e.g., it is not polyethylene glycol succinate tocopheryl (TPGS). In another example, the non-polar compound is not vitamin E. Examples of non-polar compounds that can be used in the provided concentrates are:

EP2 268160B1 [0236] Non-polar components containing essential fatty acids, for example polyunsaturated fatty acids (PUFA), for example gamma-linolenic acid (GLA), for example borage oil and evening primrose oil (Oenothera biennis), seed oil blackcurrant, hemp oil and spirulina extract; compounds containing omega-3 fatty acids, e.g. natural and synthetic omega-3 fatty acids, e.g. compounds containing polyunsaturated long chain omega-3 fatty acids, including eicosapentaenoic acid (EPA) (20: 5ω3); docosahexaenoic acid (DHA) (22: 6ω3); eicosatetraenoic acid (24: 4ω3); docosapentaenoic acid (DPA, clupanodic acid) (22: 5ω3); 16: 3 ω3; 24: 5 ω3 and / or nisin acid (24: 6ω3), for example fish oil, algae oil, krill oil, rapeseed oil, linseed oil, soybean oil and walnut oil; compounds containing short chain omega-3 fatty acids, for example alpha-linolenic acid (α-linolenic acid; ALA) (18: 3ω3) and stearidonic acid (18: 4ω3), omega-3 fatty acid and glycerol esters, e.g. monoglycerides, diglycerides and triglycerides, omega-3 fatty acid and primary alcohol esters, e.g. fatty acid methyl esters and fatty acid esters, omega-3 fatty oil precursors, for example EPA precursor, DHA precursor; derivatives such as polyglycolyzed derivatives or polyoxyethylene derivatives, oils containing omega-3 fatty acids, for example fish oil (marine oil), for example, high purity fish oil concentrates, perilla oil, krill oil and algae oil, for example microalgae oil; compounds containing omega-6 fatty acids, for example compounds containing linoleic acid (18: 2ω6) (short chain fatty acid); gammalinolenic acid (GLA) (18: 3ω6); dihomo-gamma-linolenic acid (DGLA) (20: 3ω6); eicosadienic acid (20: 2ω6); arachidonic acid (AA) (20: 4ω6); docosadienoic acid (22: 2ω6); adren acid (22: 4ω6) and / or docosapentaenoic acid (22: 5ω6), e.g. borage oil, corn oil, cottonseed oil, grape seed oil, peanut oil, evening primrose oil (Oenothera biennis), oil from black currant seeds, hemp oil, spirulina extract, safflower oil, sesame oil and soybean oil;

[0237] Other fatty acids, for example triglycerides including medium chain triglycerides, polar lipids, for example lipid ethers, phosphoric acid, choline, fatty acids, glycerol, glycolipids, triglycerides and phospholipids (e.g. phosphatidylcholine (lecithin), phosphatidylethanolamine and phosphatidylinositol); sabal palm extract; and ethyl linolonate; and herb oils, for example garlic oils and scordinine; 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), palmitolic acid (16 : 1 ω7), heptadecanoic acid (17: 0), stearic acid (18: 0), oleic acid (18: 1 ω9), peanut acid (20: 0);

[0238] Trace elements, for example vitamins, minerals, cofactors, for example coenzyme Q10 (CoQ10 also called ubiquinone), ubiquinol, turmeric extract (curcuminoids), sabal palm lipid extract (sabal palm oil), Echinacea extract, Hawthorn extract, ginseng extract, lipoic acid (α-lipoic acid), ascorbyl palmitate, kava extract, St. John's wort (St. John's wort, Klamath weed, goat weed), quercitin extract, dehydroepiandrosterone, indole-3-carbinol;

[0239] Carotenoids, including hydrocarbons and oxidized alcohol derivatives of hydrocarbons, on

Example beta-carotene, mixed carotenoids group, lutein, lycopene, zeaxanthin, cryptoxanthin, e.g. beta-cryptoxanthin, beta-carotene, mixed carotenoids group, astaxanthin, bixin, canthaxanthin, capsanthin, capsorotin, apo-carotin -12'-apokarotenal, "Carotene" (a mixture of alpha and beta-carotene), gamma-carotene, ciolerythrin, zeaxanthin, their hydroxyl or carboxyl group-containing esters;

[0240] Fat-soluble vitamins, for example vitamins A, D, E and K, and corresponding provitamins and vitamin derivatives, such as esters with effects similar to this of vitamin A, D, E or K, for example retinol (vitamin A) and its pharmaceutically acceptable derivatives, e.g. retinol palmitate ester and other retinol esters, and calciferol (vitamin D) and its pharmaceutically acceptable derivatives and precursors of vitamin D, d-alpha tocopherol (vitamin E) and its derivatives, including its pharmaceutical derivatives, for example tocotrienols, d-alpha tocopherol acetate and other d-alpha tocopherol esters, and ascorbyl palmitate, a fat-soluble version of vitamin C;

[0241] Phytochemicals, including phytoestrogens, e.g. genistein and daidzin, e.g. isoflavones, e.g. soy isoflavones, flavonoids, phytoalexins, e.g. resveratol (3,5,4'-trihydroxystilbene), red clover extract and phytosterols;

[0242] Fat-soluble drugs, including natural and synthetic forms of immunosuppressive drugs such as ciclosporin, protease inhibitors such as ritonavir, macrolide antibiotics and oil-soluble anesthetics 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;

[0243] Oil-soluble acids and alcohols, for example, tartaric acid, lactylic acid, butylhydroxyanisole, butylhydroxytoluene, lignin, sterols, polyphenolic compounds, oryzanol, cholesterol, phytosterols, flavonoids such as quercetin and reseratol and diallyl disulfides.

i. Active ingredients containing polyunsaturated fatty acids (PUFA) [0244] Exemplary non-polar compounds contained in concentrates are compounds containing fatty acids, for example active ingredients containing polyunsaturated fatty acids (PUFA). Fatty acids are straight chain hydrocarbon molecules with a carboxyl group (COOH) at one end of the chain. PUFAs are fatty acids that contain more than one carbon-carbon double bond in the fatty acid carbon chain. PUFAs, especially essential fatty acids, are useful as dietary supplements.

[0245] Various nomenclatures may be used to describe fatty acid molecules. The lipid nomenclature, for example 18: 3 ω-3, indicates the length of the carbon chain, the number of double bonds and the location along the carbon chain of the first double bond in the fatty acid. Using this nomenclature, each carbon atom along the chain is designated according to its location relative to one end of the chain. For example, the first carbon atom after the carboxylate terminus is called α, the second b and so on. The last carbon atom in the molecule (furthest from the carboxyl group) is always referred to as ω (or omega or n). The number of carbon atoms and the number of double bonds are listed first in the name of the fatty acid lipid,

EP 2 268 160 B1 separated by a colon. For example, the name "18: 3" indicates that the molecule has eighteen (18) carbon atoms and three (3) double bonds. By tracking these numbers, the position at which the first double bond appears in relation to the last (ω) carbon atom is listed. For example, the nomenclature 18: 3 ω-3 (or 18: 3 omega-3; or 18: 3 n-3) describes a fatty acid with eighteen (18) carbon atoms and three (3) double bonds, the first of which occurs at the third carbon atom after the omega carbon atom.

[0246] Optionally, a chemical nomenclature may be used. The chemical name for fatty acid describes the location of each double bond. In chemical terminology, carbon atoms are numbered starting with 1, starting with the carbon atom that is part of the carboxyl group (COOH). Thus, in this numbering system, the α carbon atom is designated "2". The chemical name of the fatty acid lists the first carbon atom (from the end with the COOH group) involved in the formation of each double bond.

[0247] Certain PUFAs are called essential fatty acids because mammals, including humans, cannot synthesize them using any known chemical transformation and must be obtained from the diet or by supplementing the diet. (US Patent No. 6,870,077; Covington, American Family Physician (2004), 70 (1): 133-140). Essential PUFAs are omega-3 (ω3; n-3) and omega-6 (ω-6; n-6) fatty acids. Omega-3 and omega-6 fatty acids are polyenes interrupted by methylene groups that have two or more cis double bonds separated by a single methylene group. Examples of omega-3 fatty acids are alphalinolenic acid (α-linolenic acid; ALA) (18: 3ω3) (short chain fatty acid); stearidic acid (18: 4ω3) (short chain fatty acid); eicosapentaenoic acid (EPA) (20: 5ω3); docosahexaenoic acid (DHA) (22: 6ω3); eicosatetraenoic acid (24: 4ω3); docosapentaenoic acid (DPA, clupanodic acid) (22: 5ω3); 16: 3 ω3; 24: 5 ω3 and nisin acid (24: 6ω3). The longer chain omega-3 fatty acids can be synthesized from ALA (omega-3 short chain fatty acid). Examples of omega-6 fatty acids are linoleic acid (18: 2ω6) (short chain fatty acid); gamma-linolenic acid (GLA) (18: 3ω6); dihomo-gamma-linolenic acid (DGLA) (20: 3ω6); eicosadienic acid (20: 2ω6); arachidonic acid (AA) (20: 4ω6); docosadienoic acid (22: 2ω6); adren acid (22: 4ω6) and docosapentaenoic acid (22: 5ω6).

[0248] Although the essential omega-3 and omega-6 longer chain fatty acids can be synthesized from ALA (short chain omega-3 fatty acid) and linolenic acid (LA), respectively, experimental data indicate that the conversion of these short chain fatty acids in humans is slow. Thus, the main source of essential PUFAs is diet (see, e.g., Ross et al., Lipids in Health and Disease (2007), 6:21; Lands, The FASEB Journal (1992), 6 (8): 2530). Dietary supplements containing PUFAs, especially the necessary PUFAs, are desirable to protect against cardiovascular disease, inflammatory and mental diseases (see e.g. 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). Experimental data indicate that essential fatty acids, especially EPA and DHA, in the form of food products and nutritional supplements, play a role in the prevention of many disease states, including cardiovascular, inflammatory,

Mental health and behavioral disorders and disorders (see, e.g., Ross et al., Lipids in Health and Disease (2007), 6:21; Lands, The FASEB Journal (1992), 6 (8): 2530; U.S. Patent No. 6,870,077; Covington, American Family Physician (2004), 70 (1): 133-140).

[0249] Omega-9 fatty acids are superfluous PUFAs. Examples of omega-9 fatty acids are oleic acid (which is monounsaturated acid) (18: 1 ω9); eicosenic acid (20: 1 ω9); honey acid (20: 3 ω9); erucic acid (22: 1 ω9) and neural acid (24: 1 ω9).

[0250] 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), for example 18: 2 ω7, 18: 2 ω6; conjugated linolenic acid, for example 18: 3ω6, 18: 3ω5; and other conjugated fatty acids, for example 18: 3 ω3, 18: 4 ω3 and 20: 5 ω6.

(1) Omega-3 fatty acid compounds [0251] Exemplary PUFA-containing active ingredients that can be used in the provided compositions are compounds that contain one or more omega-3 (ω3; n-3) fatty acids, for example compounds containing DHA and / or EPA fatty acids, e.g. marine oils, e.g. fish oil, krill oil and algae oil; and compounds containing ALA fatty acids, for example linseed oil.

[0252] Usually, oily and aqueous compositions containing long chain polyunsaturated fatty acids (PUFAs) are sensitive to oxidation, which makes them unstable and gives them an unpleasant odor. The ingredients and their relative concentrations, as well as methods for making concentrates, contribute to the desired properties of DHA / EPA containing concentrates. In one example, the ingredients and methods minimize the unpleasant "fishy" smell and / or taste of the DHA / EPA composition and increase their stability over time. In one aspect, the compounds in the concentrates have low levels of oxidation, contributing to their desired properties.

(a) DHA / EPA [0253] Exemplary non-polar active ingredients that contain one or more omega-3 fatty acids that can be used in the provided compositions. are DHA and / or EPA containing compounds, e.g. marine oil, e.g. fish oil, krill oil and algae 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 non-polar 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) EPA. In another example, the non-polar active ingredient comprises no more than 10% or about 10% EPA or less than 10% or about 10% EPA. In another example, non-polar

The active ingredient contains DHA and EPA, for example DHA represents at least 20% or about 20% by weight of the non-polar active ingredient, and EPA represents no more than 13% or about 13% of the non-polar active ingredient, for example no more than 10% or about 10% by weight of the non-polar active ingredient. In another example, the non-polar active ingredient contains DHA, representing at least 35% or about 35% of the non-polar active ingredient, and EPA, representing no more than 13% or about 13% of the non-polar active ingredient, for example no more than 10% or about 10 % non-polar active ingredient. In another example, the non-polar active ingredient contains DHA and EPA, e.g., DHA represents at least 70% or about 70% of the non-polar active ingredient, and EPA represents no more than 13% or about 13% of the non-polar active ingredient, e.g. no more than 10% or about 10% of the active ingredient.

(i) Fish oils [0254] Examples of non-polar active ingredients containing PUFA that can be used in the provided compositions are oils derived from fish that contain DHA, EPA or DHA as well as EPA. In particular, cold water marine fish are 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., all of which can be used in the compositions provided.

[0255] Fish oils are usually extracted from fish tissues, for example frozen fish tissues. In one example, fish oil is a tasteless fish oil, e.g., cod liver oil, which can be isolated from fish, e.g., cod liver, and then cleaned and deodorized, or otherwise treated as such, that its taste becomes neutral, for example as described in International Patent Publication No. WO 00/23545 and WO 2004/098311. In one example, these fish oils are isolated from frozen fish tissues using a method that minimizes oxidation. Examples of such tasteless fish oils are Denomega ™ 100, Borregaard Ingredients, Sarpsborg, Norway; distributed by Denomega Nutritional Oils AS, Boulder, CO. Usually tasteless fish oil, e.g. cod liver oil, contains between 25% or about 25% and 35% or 35% omega-3 fatty acids, e.g. 34% 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.

[0256] Exemplary fish oils that may be included in the provided compositions are also fish oils containing large amounts of omega-3 fatty acids, for example large amounts of DHA. One example of such fish oil contains at least about 85% DHA, usually more than 85% DHA, and at least about 90% omega-3 fatty acids, usually more than 90% omega-3 fatty acids. In another example, fish oil may contain 98% PUFA, 89% omega-3 fatty acids, about 70% DHA, about 10% EPA, 8.9% omega-6 fatty acids and 0.7% omega-9 fatty acids .

[0257] An exemplary fish oil containing large amounts of omega-3 fatty acids that can be used as a non-polar compound in the provided compositions is Omega-3 Fish Oil EE

EP2 268160B1 (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, 0, 1% saturated fatty acids, 1.0% monounsaturated fatty acids, 74.5% docosahexaenoic acid (DHA), 9.3% eicosapentaenoic acid (EPA) fatty acid and 98% polyunsaturated fatty acids (PUFA). This fish oil also contains 0.1% (16: 0) palmitic acid, 0.1% palmitic acid (16: 1 ω7), 0.1% (18: 0) stearic acid, 0.6% oleic acid (18: 1 ω 9), 0.1% oleic acid (18: 1 ω 7), 0.3% linoleic acid (18: 2 ω6), 0.2% linolenic acid (18: 3 ω 3), 0.2% octadecatetraenoic acid (18: 4 ω 3), 0.1% arachidic acid (20: 1 ω9), 0.1% eicosadienic acid (20 : 2 ω6), 0.2% eicosatrienic acid (20: 3 ω6), 2.4% arachidonic acid (20: 4 ω6), 0.6% arachidonic acid (20: 4 ω3), 0.1% erucic acid (22: 1 ω11) ), 0.6% ankosapentaenoic acid (21: 5 ω3), 0.5% docosatetraenoic acid (22: 4 ω6), 5.4% (22: 5 ω6), 3.6% docosapentaenoic acid (22: 5 ω3) ) and 0.9% of other fatty acids.

[0258] An example of fish oil containing large amounts of omega-3 fatty acids that can be used in the provided compositions is also Omega Concentrate 85 DHA TG Ultra (O3C Nutraceuticals AS, Oslo, Norway), which contains more than 85% DHA (C22 : 6n-3) and more than 90% of the total amount of omega-3 fatty acids, and is isolated from fatty species of fish of the anchovy (Engraulidae), herring (Clupeidae) and mackerel (Scombridae) families. Fish oil is produced by purifying and concentrating the oils of these fish using mild technologies to increase the concentration of omega-3 DHA. Any fish oil containing DHA and / or EPA may be used as the non-polar compound in the provided compositions. Exemplary fish oils are also other fish oils manufactured by O3C Nutraceuticals, AS and other fish oils provided by Jedwards, International, Inc.

[0259] Examples of fish oils are also krill oils, prepared according to International Patent Application Publication No. WO2007 / 080515.

(ii) Algae oil [0260] Exemplary non-polar compounds containing omega-3 PUFA, especially DHA (and optionally EPA) that can be used as a non-polar compound in the provided compositions, are also oils obtained from microorganisms, for example oils obtained from marine furrows , for example microalgae, for example Crypthecodinium sp, in particular Crypthecodinium cohnii. Oils from microalgae, like fish oil, are an excellent source of omega-3 fatty acids, especially DHA (US patents 5,397,591, 5,407,957, 5,492,938 and 5,711,983). Exemplary oils obtained from microalgae are the oils disclosed in (and oils prepared according to the methods described therein) US Patent Nos. 5,397,591, 5,407,957, 5,492,938 and 5,711,983 and in US Publication No. 2007/0166411, including DHASCO® and DHASCO-S® (Martek Biosciences Corporation ).

[0261] For example, US Patent No. 5,397,591 describes, among others, unicellular (derived from unicellular organisms) edible oils (algae oils) (and methods of producing oils) that contain at least 70% triglycerides that contain about 20-35% DHA and do not contain EPA, isolated from Crypthecodinium cohnii, preferably contain more than 70% triglycerides, contain 15-20% myristic acid; 20-25% palmitic acid; 10-15% oleic acid; 30-40%

EP2 268 160 B1

DHA and 0-10% other triglycerides. US Patent No. 5,407,957 describes, among others, oils from algae (and methods for producing oils) obtained from Crypthecodinium cohnii, preferably containing more than about 90% triglycerides, at least 35% by weight (w / w) DHA, in one example, contain 15-20% myristic acid, 20-25% palmitic acid, 10-15% oleic acid, 40-45% DHA and 0-5% other oils. US Patent No. 4,922,938 describes, among others, single-cell edible oils (and methods for their preparation), containing at least 70% triglycerides, which contain about 20-35% DHA and does not contain EPA, isolated from Crypthecodinium cohnii, in one example they contain more than 70 % triglycerides, contain 15-20% myristic acid; 20-25% palmitic acid; 10-15% oleic acid; 30-40% DHA; 0-10% other triglycerides. US Patent No. 5,711,983 describes, inter alia, single-cell edible oils (and methods for their preparation), containing at least 70% triglycerides, which contain about 20-35% DHA and do not contain EPA, isolated from Crypthecodinium cohnii, in one example they contain more than 70 % triglycerides, contain 15-20% myristic acid; 20-25% palmitic acid; 10-15% oleic acid; 30-40% DHA and 0-10% other triglycerides.

[0262] Examples of suitable microalgae oils are also those disclosed, for example, in US Patent No. 6,977,166 and in US Publication No. US 2004/0072330. Any oil obtained from furrows, for example microalgae, which contains DHA and optionally EPA, is suitable as algae oil for use in the provided compositions, for example V-Pure algae oil (Water4Life, Switzerland, which contains EPA and DHA.

(b) Linseed oil-omega-3 (ALA) [0263] An example of non-polar omega-3 containing compounds used in the provided compositions is also linseed oil (linseed oil, linseed oil). Linseed oils, which are good sources of omega-3 acids, in particular alpha-linolenic acid, can be used as nutritional supplements. Linseed oils are produced by extruding flax seeds and purifying flax seed oil. An example of linseed oil that can be used as a non-polar compound in the provided compositions is linseed oil obtained from Linum usitatissimum, e.g. linseed oil provided by Sanmark LLC, Greensboro, NC (Sanmark Limited, Dalian, Liaoning Province, China), which contains no less than (NLT) 50% C18: 3 alpha-linolenic acid. and furthermore contains other fatty acids, e.g. 3-8% C16: 0 palmitic acid, 2-8% C18: 0 stearic acid, 11-24% C18: 1 oleic acid, 11-24% C18: 2 linolenic acid and 0- 3% other fatty acids. Examples of suitable linseed oil are linseed oil containing 6% palmitic acid, 2.5% stearic acid, 0.5% peanut acid, 19% oleic acid, 24.1% linoleic acid, 47.4% linolenic acid and 0.5 % other acids. The linseed oil ingredient kit may vary. Any linseed oil may be used as the non-polar compound in the provided compositions. In one example, linseed oil contains at least 50% alpha linolenic acid or at least about 50% alpha linolenic acid. In another example, linseed oil contains at least 65% or about 65% or 70% or about 70% alpha-linolenic acid. Exemplary flaxseeds containing more than 65% linolenic acid (total fatty acids), e.g., 70-80% or 70-75%, are flax seeds described in US Patent No. 6,870,077.

EP2 268 160 B1 (2) Omega-6 compounds [0264] Examples of non-polar compounds used in the provided compositions are also compounds containing omega-6 PUFA, e.g. gamma-linolenic acid (GLA), e.g. borage oil and oil from evening primrose (Oenothera biennis), blackcurrant seed oil, hemp oil, mushroom oil and spirulina extract. Any oil containing omega-6 fatty acids can be used in the provided compositions.

(a) Borage oil (gamma-linolenic acid (GLA)) [0265] Exemplary non-polar compounds containing omega-6 are GLA-containing compounds, for example borage oil. GLA is omega-6 PUFA, which is mainly obtained from vegetable oils, for example, evening primrose oil (Oenothera biennis), blackcurrant seed oil, hemp oil and spirulina extract. GLA is used as a nutritional supplement. It is suggested that GLA plays a role in the treatment of various chronic diseases, and especially that it has anti-inflammatory properties (Fan and Chapkin The Journal of Nutrition (1998), 1411-1414). In one example, the non-polar active ingredient contains at least about 22% or about 22% by weight (w / w) GLA, e.g. 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 50, 60% by weight (w / w) or more GLA.

[0266] Borage (Borago officinalis), also known as "seven," is an herb whose seeds contain large amounts of GLA. An example of borage oil that is used as the non-polar active ingredient in the provided compositions is borage oil supplied by Sanmark LLC, Greensboro, NC (Sanmark Limited, Dalian, Liaoning Province, China), obtained by extrusion and isolation from Borago seeds officinalis L. This oil contains not less than (NLT) 22% gamma-linolenic acid (GLA) C18: 3; between 9 and 12% C16: 0 palmitic acid, between 3% and 5% C18: 0 stearic acid, between 15% and 20% C18: 1 oleic acid, between 35% and 42% C18: 2 linoleic acid, between 3% and 5% C20: 1 docosenoic acid, between 1% and 4% C22: 1 erucic acid and between 0% and 4% other fatty acids. Other borage oils may be used. Other GLA-containing oils can also be used as a non-polar compound.

(3) Sabal palm extract [0267] An example of the non-polar compounds used in the provided compositions is also a sabal palm extract, a lipophilic extract of ripe American dwarf berries (also called Serenoa repens or Sabal serrulata), which can be used to treat urinary tract diseases sexual and other, and to increase sperm production, breast size and libido, as a mild diuretic, sedative, expectorant and gastrointestinal enhancer, in particular for the treatment of benign prostatic hyperplasia (BHP) (Ernst, Academia and Clinic (2002), 136; 42-53; Gordon and Shaughnessy, Complementary and Alternative Medicine (2003), 76 (6 ); 1281-1283). Sabal palm extract is available on the market from many sources. Any lipodic sabal palm extract can be used in the provided compositions. An example of a sabal palm extract that can be 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

EP2 268 160 B1 and, in one example, contains 85.9% of the total amount of fatty acids, including 0.8% caproic acid, 2% caprylic acid, 2.4% capric acid, 27.1 lauric acid, 10, Myristic acid, 8.1% palmitic acid, 0.2% palmitic 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% moisture. Other sources of sabal palm extract may be used.

(4) Conjugated linoleic acid (CLA) [0268] Exemplary non-polar PUFAs that can be used in the provided compositions are also 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 active ingredients containing conjugated fatty acids are compounds containing conjugated linoleic acid (CLA), for example 18: 2 ω7, 18: 2 ω6; conjugated linolenic acid, for example, 18: 3ω6, 18: 3ω5; and other conjugated fatty acids, for example 18: 3 ω3, 18: 4 ω3 and 20: 5 ω6. CLA refers to the family of linoleic acid isomers originally found in meat and dairy products of ruminants. Usually CLA compounds contain a mixture of different CLA isomers, for example C18: 2 CLA c9, t11, CLA t10, c12 and other CLA isomers. An example of CLA that can be used as the active ingredient in the provided compositions 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 (no more than) 9.0% C 16: 0 palmitic acid, NMT 4.0% stearic acid, NMT 15.0% C18: 1 oleic acid , NMT 3.0% C18: 2 linoleic acid, NLT (not less than) 80% CLA C18: 2 (including the following isomers: NLT 37.5% C18: 2 CLA c9, t11, 37.5% C18: 2 TLA, C12 and NMT 5.0% of other CLA isomers); and NMT 50% other fatty acids. Other CLA containing compounds may be used.

ii. Active ingredients containing coenzyme Q [0269] Exemplary non-polar active ingredients will substitute compounds containing coenzyme Q, for example coenzyme Q10 (also called CoQ10, ubiquinone, ubidecarenone, ubiquinol and vitamin Q10). Coenzyme Q10 compounds are benzoquinone compounds containing isoprenyl units. The number of isoprenyl units and each of the different types of CoQ is indicated by the number following CoQ. For example, CoQ10 contains 10 isoprenyl units. Coenzyme Q10 is the predominant type of coenzyme Q.

[0270] Coenzyme Q can exist in two different forms: the oxidized form and the reduced form. When the oxidized form of coenzyme Q is reduced using one equivalent, it turns into ubisquinone, designated QH, which contains a free radical on one of the oxygen atoms in the benzoquinone benzene ring. As active ingredients in the provided compositions, compounds containing both oxidized and reduced coenzyme Q may be used.

(1) Coenzyme Q10 [0271] Exemplary non-polar active ingredients containing coenzyme Q that can be used in

The compositions provided are active ingredients containing coenzyme Q10. Coenzyme Q10 (also called CoQ10, ubiquinone, ubidecarenone, ubiquinol and vitamin Q10) is a benzoquinone compound that contains 10 isoprenoid units. "Q" in the name refers to quinone, and 10 refers to the number of isoprenoid units. CoQ10 usually refers to the oxidized form of CoQ10, which also refers to ubidecarenone, as opposed to the reduced form of CoQIO. For both oxidized and reduced CoQ10, there are examples of coenzyme Q types that can be used as active ingredients in the provided compositions.

[0272] CoQ10 has the ability to carry electrons and is present in cell membranes such as those of endoplasmic reticulas, peroxisomes, lysosomes, vesicles and mitochondria. Reduction of natural CoQ10 synthesis was observed in sick and elderly people. Because of these observations and its oxidizing properties, CoQ10 is used as a dietary supplement and for the treatment of diseases such as cancer and heart disease. However, CoQ10 has relatively poor bioavailability.

[0273] CoQ10 containing compounds are commercially available. Any CoQ10 compound or reduced CoQ10 compound may be used in the provided compositions. Exemplary 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 sold under the name Kaneka Q10 ™ (USP Ubidecarenone) by Kaneka Nutrients, LP, Pasadena, TX . The compound sold under the name Kaneka Q10 ™ is exclusively produced by yeast fermentation and is identical to the body's CoQ10 and does not contain the cis isomer found in some synthetically produced CoQ10 compounds. Any CoQ10 compound can be used in the provided compositions.

iii. Active ingredients containing phytosterols [0274] Exemplary non-polar compounds used as active ingredients in the provided compositions are compounds containing phytosterols (plant sterols). Plant sterols are structurally similar to cholesterol and have been found to reduce the absorption of cholesterol from food, which may affect serum cholesterol levels. According to the US Federal Food and Drug Administration (FDA), two administrations per day, each containing 0.4 grams of plant sterols, for a total daily intake of at least 0.8 grams, as part of a diet low in saturated fat and cholesterol, may reduce the risk heart diseases. Thus, plant sterols can be used as nutritional supplements.

[0275] Any compound containing phytosterol may be used as the active ingredient in the provided compositions. Exemplary phytosterol-containing compounds that can be used as active ingredients in the provided compositions are plant sterol-containing compounds, for example, the compound sold under the name CardioAid ™, distributed by B&amp;D Nutrition, and manufactured by ADM Natural Health and Nutrition, Decatur, IL. This compound contains kosher, not containing meat and dairy substances, and plant sterols in accordance with Muslim law, which are produced in accordance with current GMP (good manufacturing practices) of food. Sterols are PCR negative and the material is obtained from genetically modified organisms (GMOs). This phytosterol compound contains at least 95% plant sterols, which include up to 5 plant sterols.

EP2 268160B1

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 further contain tocopherols, e.g. 0-15 mg / g tocopherols. The compound is tested and gives negative results for Salmonella, E. coli and

Staphylococcus aureus.

c. Surfactants [0276] The provided compositions contain surfactants. For example, in addition to the non-polar compound (s), liquid concentrates contain one or more surfactants. In the processes provided for making concentrates, the surfactant is added to the aqueous phase, the oil phase, or to the water and oil phase. The compositions may further comprise one or more co-surfactants or emulsifiers.

[0277] Surfactants aggregate in aqueous liquids, such as in the compositions provided (e.g., concentrates and aqueous liquid diluted compositions) to form micelles that contain the non-polar compound (s). The hydrophilic fragment (fragments) of the surfactant molecules is oriented outside the micelles in contact with the aqueous medium, while the hydrophobic fragment (fragments) of the surfactant molecules is oriented towards the center of the micelles, in contact with the non-polar compound (s) which is contained within micelles. Micelles may contain more than one surfactant and / or co-surfactant. The properties of the provided compositions, for example the particle size of the composition and the desired properties related to the size of the particles, are influenced by the choice of surfactant (s) and the relative amount (concentration) of the surfactant. For example, the HLB surfactant (s) may affect particle size, clarity, taste, smell, crystal formation, and other properties of the provided compositions. Similarly, the concentration of surfactant compared to the concentration (concentrations) of other ingredients, especially when compared to the concentration of the polar solvent (s) and the concentration of the non-polar compound (s), can affect various desirable properties, e.g. dispersibility or dissolution in the medium aqueous, e.g. to produce a clear aqueous liquid composition diluted or a pleasant taste and / or smell.

[0278] Surfactants (and co-surfactants) are molecules that contain hydrophobic and hydrophilic fragments. In one example, the hydrophobic fragment is the hydrobic end and the hydrophilic fragment is the head of the surfactant molecule.

[0279] Exemplary surfactants that can be used in the provided methods and compositions are surfactants having an HLB value between 14 or about 14 and 20 or about 20, usually 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 tocopherol and / or surfactants derived from tocotrienol, in which the vitamin E unit represents the hydrophobic area of ​​the surfactant and is attached, through a linker, to another entity such as

A polyethylene glycol (PEG) unit that provides a hydrophilic surfactant fragment. Surfactants derived from vitamin E include, but are not limited to, surfactants derived from tocopherol, including tocopherol polyalkylene glycol derivatives, typically tocopherol polyethylene glycol derivatives (PEG), such as tocopherol polyethylene glycol succinate (TPGS), TPGS analogues , TPGS homologues and TPGS derivatives. Optionally, the surfactants may be other PEG derivatives having similar properties, e.g. PEG sterol derivatives, e.g. cholesterol or sitosterol (including, for example, any of the PEG derivatives disclosed in US Patent No. 6,632,443) or PEG derivatives of other fat-soluble vitamins, on example, some forms of vitamin A (e.g. retinol) or vitamin D (e.g. vitamin D1-D5).

[0280] The HLB value of the surfactant is obtained from a semi-empirical formula; HLB values ​​are used to indicate surfactants according to their relative hydrophobicity and hydrophilicity. The HLB value is a relative representation of hydrophilic groups and hydrophobic groups in a surfactant or in a mixture of surfactants represented by a number. The weight percentage of these representative groups indicates the molecular structure properties. See, for example, Griffin, WCJ Soc. Something. Chem. 1: 311 (1949).

[0281] HLB values ​​of surfactants range from 1-45, while the range for non-ionic surfactants is usually from 1-20. The more lipophilic the surfactant is, the lower its HLB value. Conversely, the more hydrophilic a surfactant, the higher its HLB value. Lipophilic surfactants have greater solubility in oil and lipophilic substances, while hydrophilic surfactants dissolve more easily in aqueous media. Generally, 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 referred to as "hydrophobic agents surfactant ". HLB values ​​are known for many surfactants. Table 1 provides a list of HLB values ​​for exemplary surfactants and co-surfactants.

[0282] In the provided compositions, the surfactants are typically nonionic surfactants and usually have an HLB value between at or about 14 and at or about 20. Specific examples of suitable surfactants include surfactants derived from PEG, such as vitamin E PEG derivatives having appropriate HLA values, such as an HLB value between 14 or about 14 and 20 or about 20, e.g. 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 (widely considered safe) by the FDA and / or kosher certified.

i. Surfactants derived from vitamin E [0283] Surfactants include, but are not limited to, agents

EP 2 268 160 B1 surfactants obtained from vitamin E, such as surfactants obtained from tocopherol and / or tocotrienol, in which the vitamin E unit represents the hydrophobic area of ​​the surfactant and is attached, via a linker, to another unit, such as a glycol unit polyethylene (PEG), which provides a hydrophilic surfactant fragment. Surfactants derived from vitamin E include, but are not limited to, surfactants derived from tocopherol, including tocopherol polyalkylene glycol derivatives, typically tocopherol polyethylene glycol (PEG) derivatives such as tocopherol polyethylene glycol succinate (TPGS), TPGS analogues , TPGS homologues and TPGS derivatives. Optionally, the surfactants may be other PEG derivatives having similar properties to TPGS and TPGS analogues, e.g. PEG sterol derivatives, e.g. cholesterol or sitosterol (including, for example, any of the PEG derivatives disclosed in US Patent No. 6,632,443) or other PEG derivatives fat-soluble vitamins, e.g. certain forms of vitamin A (e.g. retinol) or vitamin D (e.g. vitamins D1-D5).

(1) Vitamin E PEG derivatives [0284] Surfactants derived from vitamin E (e.g. surfactants derived from tocopherol or tocotrienol) include vitamin E polyalkylene glycol derivatives, usually vitamin E polyethylene glycol (PEG) derivatives, for example PEG derivatives tocopherol or tocotrienol. Suitable PEG derivatives of vitamin E contain one or more tocopherols or tocotrienols linked (for example by ester, ether, amide or thioester linkage) to one or more PEG units via a linker, e.g. a dicarboxylic acid linker. An example surfactant is schematically illustrated below:

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

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

[0285] Usually, vitamin E PEG derivatives are produced by attaching the PEG unit, by esterification, to a vitamin E-linker conjugate (e.g., tocopherol-linker conjugate). In one example, the tocopherol-linker conjugate is first formed by covalently linking (by esterifying) a tocopherol hydroxyl unit to a dicarboxylic acid to form 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 using any of a variety of known methods (see, for example, US Patent Nos. 2,680,749, 4,665,204, 3,538,119 and 6,632,443). To produce the tocopherol-PEG surfactant, the tocopherol ester obtained is then combined (via a linker) to the PEG molecule in another esterification reaction. In this example, the surfactant obtained is a tocopherol polyethylene glycol diester (TPGD). [0286] Optionally, PEG derivatives of a tocopherol-linker or tocotrienol-linker conjugate can be prepared using other methods. For attaching PEG molecules to tocopherol-linker compounds or

Tocotrienol-linker, various methods known in the art can be used. For example, a tocopherol-linker conjugate may covalently attach to a PEG molecule through an amide, ether or thioether bond. For example, a tocopherol-linker conjugate that contains an amide group can be reacted with a PEG-NHS derivative to form an amide bond between the tocopherol linker and PEG. The tocopherol-linker conjugate, which contains an amine, can be reacted with a PEG-aldehyde derivative to form an amide bond between the tocopherol-linker molecule and PEG. In another example, a tocopherol-linker that contains a carboxylic acid can be activated into the appropriate halide and reacted with a PEG-SH derivative to form a thioester linkage between the tocopherol-linker molecule and PEG.

(a) Tocopherols and tocotrienols [0287] The tocopherols (tocopherols) used to make the surfactant may 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 form or in the form of heterogeneous mixtures of one or more forms. Examples of tocopherols are da-tocopherols id, 1-tocopherols. To produce a surfactant, tocopherol is usually subjected to esterification with a linker, e.g. dicarboxylic acid, to form a tocopherol ester which is then attached to the PEG unit.

[0288] The tocotrienol (tocotrienols) used in the preparation of surfactants may be any natural or synthetic tocotrienol of vitamin E including, but not limited to, alpha-tocotrienols, beta-tocotrienols, gamma-tocotrienols and delta-tocotrienols, or in the form pure forms or in the form of heterogeneous mixtures of one or more forms. Mixtures of tocopherols and tocotrienols in the provided methods and compositions are contemplated. Before linking to the PEG unit, tocotrienol may be esterified with a linker such as dicarboxylic acid.

(b) PEG units [0289] The PEG used in the tocopherol-PEG derivative may be any of many known PEG units. Exemplary suitable PEG units are PEG units having different chain lengths and different molecular weights, for example PEG 1000, PEG 200, PEG 500 and PEG 20,000. The number of the following individual PEG units indicates the molecular weight (in kilodaltons (kDa)) of PEG units. The PEG unit of the surfactant obtained from tocopherol usually has a molecular weight between 200 kDa or about 200 kDa and 20,000 kDa or about 20,000 kDa, usually between 200 kDa or about 200 kDa and 6,000 kDa or about 6,000 kDa, for example between 600 kDa or about 600 kDa and 6000 kDa or about 6000 kDa, usually 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. An example of a PEG derivative of a tocopherol ester having a PEG 1000 kDa unit is TPGS-1000. Exemplary suitable PEG units are also modified PEG units, e.g. methylated PEG (m-PEG), which is a methyl terminated PEG chain. Other known PEG analogs may also be used. PEG units can be selected from any reactive PEG including, but not limited to, PEG-OH, PEG-NHS, PEG-aldehyde, PEG-SH,

EP 2 268 160 B1

PEG-NH2, PEG-CO2H and branched PEG.

(c) Linkers [0290] Typically, PEG derivatives of vitamin E are diesters or other esters, e.g. triesters. When the PEG derivative is a diester, the linker connecting vitamin E with PEG is usually a carboxylic acid, usually dicarboxylic acid, as for example in succinate of polyethylene glycol tocopherol (TPGS), where the linker is succinic acid and the surfactant is produced by esterification reaction connecting the unit PEG and dicarboxylic acid tocopherol ester. 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 amino alcohol such as ethanolamine.

(d) Diesters of polyethylene glycol tocopherol and polyethylene glycol tocotrienol (vitamin E dicarboxylic acid esters attached to PEG) [0291] Usually PEG derivatives of vitamin E are vitamin E polyethylene glycol diesters, which are esters of vitamin E PEG, produced by attaching vitamin E ester to one or more PEG units using esterification. Exemplary vitamin E diesters are tocopherol polyethylene glycol diesters (TPGD) and tocotrienol polyethylene glycol diesters.

[0292] When the tocopherol or tocotrienol ester attached to the PEG unit is a dicarboxylic acid tocopherol ester (e.g., tocopherol succinate), the linker is dicarboxylic acid (a carboxylic acid having two carboxyl groups, e.g. succinic acid). In this example, PEG tocopherol or tocotrienol diesters are prepared using an esterification reaction in which PEG attaches to the dicarboxylic acid tocopherol ester.

[0293] Exemplary dicarboxylic acids that can be used as linkers in these tegopherol and tocotrienol diester surfactants are succinic, sebacic, dodecanedioic, suberic or azelaic, citraconic, methylcracraconic, itaconic, maleic, glutaric, fumaric, and glutaconic phthalic acid. Thus, exemplary tocopherol esters that can be esterified to produce PEG derivatives include, but are not limited to, tocopherol succinate, tocopherol sebacate, tocopherol dodecanedione, tocopherol suberate, tocopherol azelaate, tocopherol citraconate, tocopherol methyl toconate, tocopherol maleate, tocopherol glutonate and tocopherol phthalate.

[0294] Exemplary vitamin E polyethylene glycol diesters made from dicarboxylic acids are compounds having the following formula, shown in Scheme I below (and their homologues, analogs and derivatives):

<img file="PL2268160T3_D0004.tif" />

EP2 268 160 B1 where each R¹, R², R³ and R⁴ independently is H or Me; each dashed line is independently a single or double bond; n is an integer from 1-5000; miq are independently 0 or 1; ap is an integer from 1-20. In one example, the surfactant is a compound where when both m and q are 0, p is an integer between 2-20.

[0295] In one example, the surfactant has the following formula, shown in Scheme II below (including its homologs, analogs and derivatives):

<img file="PL2268160T3_D0005.tif" />

where each R¹, R², R³ and R⁴ independently represents hydrogen (H) or methyl (Me); the bond represented by the dashed line means either a single or double bond, m is an integer from 1 to 20, and n is an integer from 1 to 5000.

[0296] In another example, the surfactant is a TPGS analog such as, but not limited to, a compound other than TPGS with the formula shown in Scheme III:

<img file="PL2268160T3_D0006.tif" />

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

[0297] Exemplary PEG tocopherol and tocotrienol diesters that can be used as surfactants in the provided compositions and methods include, but are not limited to, tocopherol polyethylene glycol succinates (TPGS; including Da-TPGS id, 1-TPGS;

see for example US Patent No. 3,102,078), polyethylene glycol tocopherol sebacate (PTS; see for example US Patent No. 6,632,443), polyethylene glycol tocopherol dodecanedione (PTD; see for example US Patent No. 6,632,443), polyethylene glycol tocopherol subperate (PTSr; see Example US Patent No. 6,632,443), Tocopherol Polyethylene Glycol Azelaate (PTAz; see, for example, US Patent No. 6,632,443), polyoxyethanyl tocotrienyl sebacate (PTrienS, e.g.

PTrienS-600; see, for example, US Patent No. 6,632,443) as well as analogs, homologues and derivatives or any of thiocopherol esters.

EP2 268 160 B1 (e) Other Vitamin E PEG Esters [0298] In another example, the tocopherol ester attached to PEG to form the PEG tocopherol diester is a tocopherol ester of a tricarboxylic acid, for example citric, isocitric, aconitic and propanoic acid 1,2,3-tricarboxylic acid (tricarballylic acid, carballylic acid) or carboxylic acid having three or more carboxyl groups.

[0299] In another example, the PEG tocopherol derivatives are tocopherol polyethylene glycol (TPGT) triesters, for example esters containing tocopherol, a linker, a PEG unit and an additional unit, for example an additional tocopherol, a second PEG unit or a water-soluble group such as quaternary amine. In one example, when the triester contains two PEG units, each PEG unit has a smaller chain length (and lower molecular weight) than the PEG unit in the PEG tocopherol derivative, having similar properties that contains only one PEG chain.

(f) TPGS surfactants [0300] Exemplary surfactants that are polyethylene glycol diesters of tocopherol are TPGS, their analogs, homologues and derivatives. TPGS is a natural surfactant that is GRAS and is kosher certified, and is therefore desirable for use in products intended for human consumption, for example, beverages, food and nutritional supplements. TPGS usually have an HLB value between 16 or about 16 and 18 or about 18. An exemplary TPGS surfactant is TPGS-1000, which has a 1000 kDa PEG unit. An exemplary surfactant that can be used in the provided compositions is a food grade TPGS surfactant, sold under the name Eastman Vitamin E TPGS®, food grade, by Eastman Chemical Company, Kingsport, TN. This surfactant is a water-soluble form of vitamin E of natural origin, which is produced by esterifying the carboxyl group of crystalline d-alpha-tocopheryl succinate using polyethylene glycol 1000 (PEG 1000), and contains between 260 and 300 mg / g of total tocopherol . A similar compound can be produced by esterifying the carboxyl group of Form 1, d of synthetic vitamin E using PEG 1000. It creates a clear liquid when dissolved 20% in water. This polyethylene glycol and tocopheryl derivative is a water-soluble preparation of fat-soluble vitamin (Vitamin E), for example as disclosed in US Patent Nos. 3,102,078, 2,680,749 and in published US applications 2007/0184117 and 2007/0141203. The PEG unit of alternative TPGS surfactants may 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 6,000 kDa or about 6,000 kDa, usually between 600 kDa or about 600 kDa and 1500 kDa or about 1500 kDa. An example of a TPGS surfactant that can be used in the provided compositions is also Water Soluble Natural Vitamin E (TPGS), sold by ZMC-USA, The Woodlands, Texas. Any known TPGS source or any analogue, homologue or derivative thereof may be used.

[0301] Examples of TPGS analogues are compounds other than TPGS that are similar to

The parent TPGS compound, but differ slightly in composition, for example, by changing, adding or removing an atom, one or more units (e.g., methylene (CH2) n units (units)) or one or more functional groups.

[0302] At room temperature, TPGS is usually a waxy low melting solid. In one example, the TPGS is heated prior to use, for example at least to a melting point, for example between 37 ° C or about 37 ° C and 41 ° C or about 41 ° C, and poured the desired amount. In another example, TPGS can be added to the vessel in the form of a waxy solid and heated using a heating device.

[0303] Exemplary surfactants are also TPGS analogs that include vitamin E surfactants, including vitamin E PEG derivatives, including vitamin E PEG diesters such as, but not limited to, polyethylene glycol tocopherol sebacate (PTS) , tocopherol polyethylene glycol dodecanedione (PTD), tocopherol polyethylene glycol suberate (PTSr), Tocopherol polyethylene glycol azelaate (PTAz) and polyoxyethanyl tocotrienyl sebacate (PTrienS), as well as other vitamin E PEG derivatives.

ii. Surfactant Concentration [0304] Typically, the concentration of surfactant (s) in a particular concentrate composition is selected, as described herein, by formulating the initial concentrate with a surfactant (s) concentration that falls within the starting concentration range, followed by concentrate assessment initial and optionally regulating the concentration of the surfactant (s). Optionally, the surfactant concentration can be selected based on the surfactant concentration in one or more existing liquid concentrate formulations. Typically, the surfactant concentration is between 16% or about 16% and 30% or about 30% (w / w), e.g. 16% or about 16%, 17% or about 17%, 18% or about 18% , 19% or approximately 19%, 20% or approximately 20%, 21% or approximately 21%, 22% or approximately 22%, 23% or approximately 23%, 24% or approximately 24%, 25% or approximately 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 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.

[0305] 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 surfactant concentration range is between 18% or about 18% and 25% or about 25% (w / w) of the concentrate. In another example, the surfactant concentration range is between 18% or about 18% and 20% or about 20% (w / w) of the concentrate. In another example, the surfactant concentration range is between 17% or about 17% and 20% or about 20% (w / w) of the concentrate. In another example, the surfactant concentration range is between 16% or about 16% and 20% or about 20% (w / w) of the concentrate.

EP2 268 160 B1

i. HLB [0306] An example of surfactant (s) properties that contribute to the desired properties of the composition is HLB (hydrophilic-lipophilic balance) of the surfactant (s). Usually HLB is a value obtained from a semi-empirical formula that is used to indicate surfactants according to their relative hydrophobicity and hydrophilicity. The HLB value is a relative representation of hydrophilic groups and hydrophobic groups in a surfactant or in a mixture of surfactants represented by a number. The weight percentage of these representative groups indicates the molecular structure properties. See, for example, Griffin, WCJ Soc. Something. Chem. 1: 311 (1949).

[0307] HLB values ​​of surfactants range from 1-45, while the range for non-ionic surfactants is usually from 1-20. The more lipophilic the surfactant is, the lower its HLB value. Conversely, the more hydrophilic a surfactant, the higher its HLB value. Lipophilic surfactants have greater solubility in oil and lipophilic substances, while hydrophilic surfactants dissolve more easily in aqueous media. Generally, 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 referred to as "hydrophobic agents surfactant ".

[0308] HLB values ​​have been determined and are available for many surfactants (eg, see US Patent No. 6,267,985). It is understood that HLB values ​​for the surfactant or co-surfactant may vary depending on the empirical method used to determine the value. Thus, the HLB values ​​of surfactants and co-surfactants provide initial guidance for formulating compositions based on relative hydrophobicity / hydrophilicity. For example, the surfactant is usually selected from surfactants having HLB values ​​within a specific range of surfactant or co-surfactant so that it can be used for pilot formulations. Table 1 provides a list of HLB values ​​for exemplary surfactants and co-surfactants.

[0309] Surfactants and HLB values ​​shown in Table 1A are exemplary. Any known surfactant or co-surfactant may be used in the provided compositions (e.g., see US Patent No. 6,267,985) provided that it has an appropriate HLB value, such as an HLB value between exactly or about 14 and exactly or about

twenty. The surfactant (s) used in the concentrate provided usually has an HLB value between 14 or about 14 and 20 or about 20, e.g. 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 polyethylene glycol (PEG) surfactants, including but not limited to

To this end, PEG derivatives of vitamin E, such as PEG tocotrienol or tocopherol diesters, such as TPGS (e.g. TPGS 1000) and TPGS analogues, homologues and their derivatives. Other known surfactants having HLB values ​​between 14 or about 14 and 20 or about 20, usually between about 15 and 18, will also be suitable. For example, surfactants having TPGS-like properties may also be used. Typically, the surfactant is a natural surfactant, for example a surfactant that is GRAS (widely considered safe) by the FDA and / or kosher certified.

d. Co-active surfactants (emulsifiers) [0310] In one example, the liquid concentrate further comprises one or more co-surfactants (emulsifiers). For example, the co-surfactant may be included to improve the emulsification of the active ingredient and / or the stability of the composition, for example by preventing or slowing the oxidation of the non-polar compound. An exemplary surfactant used in the provided concentrates is a phospholipid, e.g., phosphatidylcholine.

i. Phospholipids [0311] Exemplary surfactants that can be used in the provided compositions are phospholipids. Phospholipids are amphipathic lipid-like particles, usually containing a hydrophobic fragment at one end of the molecule and a hydrophilic fragment at the other end of the molecule. Many phospholipids can be used as ingredients in the provided compositions, e.g. lecithin, including phosphatidylcholine (PC), phosphatidylethanolamine (PE), distearoylphosphatidylcholine (DSPC), phosphatidylserine (PS), phosphatidylglycerol (PGf), phosphatidylglycerol (PG), ), sphingomelin (SPM) or a combination thereof. Usually, the phospholipid is phosphatidylcholine (PC), which is sometimes referred to by the common name 'lecithin'. Exemplary phospholipids that can be used as co-surfactants in the provided compositions are phospholipids sold by Lipoid, LLC, Newark, NJ, for example, purified egg lecithins, purified soy lecithins, hydrogenated egg lecithins and soy, egg phospholipids, soy phospholipids , hydrogenated egg and soy phospholipids. Synthetic phospholipids, PEGylated phospholipids and phospholipid mixtures sold by Lipoid, LLC. An example of phosphatidylcholine that can be used as a surfactant in the provided compositions is the phosphatidylcholine composition sold by Lipoid, LLC under the name Lipoid S 100, which is obtained from soy extract and contains more than 95% or more than about 95% phosphatidylcholine.

[0312] In one example, the phospholipid, for example PC, is less than or equal to 1% or about 1% by weight (w / w) of the concentrate. In one example, phosphatidylcholine is 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 concentrate. In one example, the phospholipid is between 0.15% or about 0.15% and 0.7% or about 0.7% by weight (w / w) of the concentrate.

EP 2 268 160 B1

e. Polar solvents [0313] Compositions, including liquid nanoemulsion concentrates and liquid diluted compositions, further include polar solvents. Polar solvents are well known in the art. The solvent polarity usually indicates which compounds are soluble in the solvent and which other solvents / liquids this solvent is mixed with. Generally speaking, polar compounds dissolve more easily in water and other polar solvents than non-polar compounds. Polar solvents are more likely to mix with water and other polar solvents and liquids.

[0314] Solvent polarity can be determined by measuring a wide variety of parameters using well-known methods (see, e.g., Prizbytek, "High Purity Solvent Guide," Burdick and Jackson Laboratories, Inc., 1980), such as by determining solvent properties such as dielectric constant, dipole moment or polarity index. For example, polar solvents usually have higher dielectric constants, usually dielectric constants greater than or about 15 (see e.g. Lowery et al., Mechanism and Theory in Organic Chemistry, Harper Collins Publishers, ed. 3, 1987, p. 177), such as exactly 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. For example, the dielectric constant of water is exactly or about 80.10. Polar solvents usually have high polarity indexes, usually greater than or about 3 (see e.g. Snyder, "Classification of the solvent properties of common liquids," J. Chromatography A, 92: 223-230, 1974), such as exactly or about 3, 4, 5, 6, 7, 8 or 9 or greater than 9. Polar solvents usually have large dipole moments, usually greater than or about 1.4 debaja, 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 debates (see e.g. "CRC Handbook of Chemistry and Physics," Lide, ed., 82nd edition, CRC Press, 2001, pp. 15 (14) -15 (18)). Other methods for determining the polarity of solvents are known in the art including, but not limited to to this, the Z Kosower scale (Kosower, "An introduction to physical organic chemistry," Wiley, 1969, p. 293), the donor number and the donor acceptor scale (Gutmann, "Solvent effects on the reactivities of organometallic compounds," Coord. Chem. Rev., 18: 225-255, 1976), and Hildebrand solubility parameters (see, e.g., 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).

[0315] Polar solvents include protic polar solvents and aprotic polar solvents. The protic polar solvent (e.g. water, methanol, ethanol) contains a hydrogen atom attached to an electronegative atom such that the hydrogen is proton-like and / or the bond between hydrogen and electronegative atom is polarized. On the other hand, polar protic solvents (e.g. acetone, acetonitrile) usually do not contain positively polarized hydrogen atoms.

[0316] Polar solvents in the provided compositions typically are polar protic solvents including, but not limited to, water; alcohols including, but not limited to, dihydric alcohols (e.g., glycols, e.g., propylene glycol, ethylene glycol, tetraethylene glycol, triethylene glycol, and trimethylene glycol) that contain two groups

Hydroxyl alcohols, trihydric alcohols (e.g. glycerin, butane-1,2,3-triol, pentane-1,3,5-triol, 2 amino-2-hydroxymethylpropane-1,3-diol), which contain three hydroxyl groups, monohydric alcohols (e.g. 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, dimethyl sulfoxide, dioxane, ethyl acetate, tetrahydrofuran and hexamethylphosphoric triamide. Typically, the polar solvent is water or an alcohol that typically contains two or more hydroxyl groups, such as trihydroxy or dihydroxy alcohol, such as, but not limited to, glycerol and propylene glycol. Polar solvents further include low molecular weight polyethylene glycols (PEG), such as PEG having a molecular weight not greater than or about 600 kDa, such as between exactly or about 200 kDa and exactly or about 600 kDa, usually not more than exactly or about 400 kDa, for example no more than 200 kDa.

[0317] In one example, the polar solvent has a dielectric constant of greater than or about 15, and usually between exactly or about 20 and exactly or about 80, such as exactly or about 80.1, 46.53 or 28.67. In another example, the polar solvent has a polarity index between at or about 3 and at or about 9. In another example, the dipole moment of the polar solvent is between 1.5 and 3, and usually between exactly or about 1.8 and 2.8, such as 1.9, 2.6 and 2.2 (for solid dielectric 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, 2nd Edition, 82nd 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 the solvent polarity index see for example, Snyder," Classification of the solvent properties of common liquids, " J. Chromatography A, 92: 223-230, 1974).

[0318] The amount of polar solvent is usually a high concentration, for example, in the range of concentration between 60% or about 60% and 80% or about 80% by weight (w / w) of the concentrate, e.g. 60% or about 60% , 61% or approximately 61%, 62% or approximately 62%, 63% or approximately 63%, 64% or approximately 64%, 65% or approximately 65%, 66% or approximately 66%, 67% or approximately 67%, 68% or approximately 68%, 69% or approximately 69%, 70% or approximately 70%, 71% or approximately 71%, 72% or approximately 72%, 73% or approximately 73%, 74% or approximately 74%, 75% or approximately 75%, 76% or approximately 76%, 77% or approximately 77%, 78% or approximately 78%, 79% or approximately 79%, or 80% or approximately 80% (w / w) of concentrate . Exemplary concentrations of polar solvents in the liquid concentrates provided are 71.74%, 75.8165%, 74.25%, 68.7865% and 68.29% (w / w) of the concentrate. In one example, the polar solvent concentration range 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 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.

[0319] In the processes provided for making concentrates, a polar solvent (e.g. water,

(Propylene glycol or glycerin) is added to the aqueous phase. In one example, the polar solvent is water, e.g., purified water, such as water, which is purified before it is added to the concentrate formula, for example using a carbon filter, ion exchange, reverse osmosis, ultraviolet sterilization and / or filtration using a filter, such as a 50-100 micron filter. Usually, when a filter is used, it is the end point of the filter that filters the water before it enters the tank in the supplied process. Optionally, pre-filtered water can be added to the concentrates.

f. Preservatives and sterilizers [0320] In one example, the liquid concentrate provided further contains one or more preservatives (or preservatives) and / or sterilizers. A preservative (s) may be included to improve the stability of the concentrate and compositions made by diluting the concentrate over time. Preservatives, in particular food and beverage preservatives, are well known. Any known preservative can be used in the provided compositions. Exemplary preservatives that can be used in the provided compositions are oil-soluble preservatives, e.g., benzyl alcohol, benzyl benzoate, methylparaben, propylparaben, antioxidants, e.g. vitamin E, vitamin A palmitate and beta carotene. Usually, the preservative is selected so that it is safe for human consumption, for example in food and drink, e.g. preservative GRAS certified and / or kosher certified, e.g. benzyl alcohol.

[0321] The preservative usually represents less than 1%, less than about 1%, 1% or about 1% by weight (w / w) of the liquid nanoemulsion concentrate either between 0.1% or about 0.1% and 1% or about 1% by weight (w / w) of the concentrate, e.g. 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-emulsifiers) [0322] In one example, the liquid concentrates provided further comprise one or more emulsion stabilizers (co-emulsifiers) that can be used to stabilize liquid nanoemulsion concentrate and / or aqueous compositions containing diluted concentrates. In one example, the emulsion stabilizer increases viscosity of the liquid concentrate. In one example, one or more emulsion stabilizers are added during formulation, after evaluating the initial concentrate, especially when the oil and water phase concentrate of the initial concentrate (or the aqueous diluted liquid composition obtained after diluting the initial concentrate appears to want to separate). Addition of an emulsion stabilizer may prevent oil and water phases from separating.

[0323] An example of an emulsion stabilizer that can be used in the provided compositions is a composition containing a gum blend, for example, gums used as emulsifiers, for example a blend containing one or more of xanthan gum, guar gum and sodium alginate, for example an emulsion stabilizer sold under the brand name SALADIZER®, available from TIC Gums, Inc. (Belcamp, MD). Other gums may be included in the emulsion stabilizer, on

EP 2 268 160 B1 example gum arabic and sugar beet pectin. Other mixtures of similar gums may also be used as emulsion stabilizers.

[0324] The emulsion stabilizer may be added to the aqueous phase, the oil phase, and usually to the aqueous phase and oil phase, during the preparation of 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. liquid concentrate. In one example, an emulsion stabilizer is added to the aqueous phase for a final concentrate concentration between 0.1% or about 0.1% and 1% or about 1%, e.g. 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 liquid concentrate. In one example, an emulsion stabilizer is added to the oil phase such 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%, e.g. 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 aqueous phase and to the oil phase, for example at a concentration in the oil phase and the aqueous phase within the concentration ranges specified above. In one such example, 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) liquid concentrate.

h. Non-polar solvents [0325] In one example, the liquid concentrates further contain a non-polar solvent, for example an oil. Typically, a non-polar solvent is included in the composition in addition to the non-polar active ingredient and is used to dissolve the non-polar active ingredient. In one example, the solvent is an oil that is not included in the non-polar active ingredient. When the non-polar solvent is included in the concentrate, it is usually used to dissolve the non-polar compound before mixing with other components, for example, before mixing with other components of the oil phase. In one example, the use of a non-polar solvent reduces the crystal size and / or increases the clarity of the aqueous diluted liquid composition containing the diluted concentrate. Examples of non-polar solvents that can be used in the provided concentrates are oils (in addition to the non-polar active ingredient), e.g. vitamin E oil, linseed oil, CLA, borage oil, D-limonene, rapeseed oil, corn oil, MCT oil and oil from oats. An example of a vitamin E oil used as a non-polar solvent in the provided compositions is an 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). Vitamin E oil contains at least 67.2% tocopherol and about 32.8% soybean oil.

[0326] In one example, the concentration of the non-polar solvent is in the range of concentration between 1% or about 1% and 10% or about 10%, for example 1%, 2%, 3%, 3.25%, 3.5 %

EP2 268 160 B1

3.75%, 4%, 5%, 5.25%, 5.5% or 5.75% w / w concentrate. In another example, the concentration is between 3% or about 3% and 6% or about 6% w / w. liquid concentrate. In another example, it is between 3.75% and 5.25% w / w. liquid concentrate.

i. Flavors [0327] In one example, the concentrate further contains one or more flavors or flavors, for example any flavor compound and / or aqueous dilute liquid composition containing the dilute concentrate, on example of a food or drink containing a concentrate. Many flavors are well known. Any flavor may be added to the concentrates, for example, any flavor sold by Mission Flavors, Foothill Ranch, CA. Exemplary flavors that can be used are fruit flavors such as guava, kiwi, peach, mango, papaya, pineapple, banana, strawberry, raspberry, blueberry, orange, grapefruit, mandarin, lemon, lime and lemon -limonkowy; cola flavoring agents, tea flavors, coffee flavors, chocolate flavors, dairy flavors, root and birch beer flavors and aromas, methyl salicylate (wintergreen oil, birch oil) sugar), citrus oil and other flavors. Usually flavors are safe and / or recommended for human consumption, e.g. GRAS flavors or kosher certified. 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 D-limonene, e.g. 99% GRAS-certified D-limonene sold by FloridaChemical, Winter Haven, FL. Typically, flavor is added using the provided methods to nanoemulsion concentrates after the oil and water phases are combined. Optionally, the flavoring agent (s) may be added directly to the aqueous and / or oily phase.

[0328] Usually the concentration of the flavoring agent added to the concentrates provided is less than 5% or about 5%, usually 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 regulators [0329] In one example, one or more pH regulators are added to the concentrates provided, usually to the emulsion that forms after the water and oil phases are combined in accordance with the provided methods. The pH regulator is especially used in compositions containing water. Optionally, the pH regulator can be added, in an appropriate concentration to obtain the desired pH, to the oil phase and / or the aqueous phase. Typically, the pH adjuster is added to the concentrate's pH settings in the range of 2.0 or about 2.0 to 4.0 or about 4.0. One or more of many pH adjusting agents can be used. Usually the pH adjusting agent is safe for human consumption, for example GRAS certified. An example of a pH regulator is citric acid, e.g. citric acid sold by Mitsubishi Chemical, Dublin, OH.

[0330] Typically, the concentration of the pH regulator added to the concentrates provided is less than 5% or about 5%, usually 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%, 0.28% or 0.19%.

2. Powder forms of the compositions [0331] The compositions may also be provided in the form of a powder, i.e. a powder that is made by converting the provided nanoemulsion concentrates into powder using one of many well known methods (e.g., spray drying and / or milling). Powder compositions include, but are not limited to, coated or uncoated swallowing or lozenges, dry powders in hard or soft gelatin capsules, and dry powders for single or multiple use packaging to reconstitute suspensions or sprinklers. Preferably the solid dosage forms are coated or uncoated swallowing or lozenges. Suitable methods for preparing powder compositions are well known in the art.

[0332] In addition, 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, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, pregelatinized starch, guar gum, alginic acid, gum arabic, sodium carboxymethyl cellulose and polyvinylpyrrolidone; glidants including, for example, colloidal silicon dioxide and talc; and moisturizing / anti-sticking substances, including, for example, magnesium stearate, calcium stearate, stearic acid, sodium stearyl fumarate, glyceryl monostearate, hydrogenated vegetable oil and talc. In one particular example, excipients are selected from one or more of maltodextrin and acacia.

[0333] Powder forms can be used for any dosage amount of the non-polar compound. Usually, the level of non-polar compound can be increased or decreased, at the discretion of the physician, pharmacist, pharmacy scientist or other specialist. The amount of remaining inactive ingredients can be adjusted as needed.

[0334] Usually, the concentration of excipients is in the range of concentrations 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 more wt% loose powder.

[0335] Powder forms can be used for any dosage amount of the non-polar compound. Usually, the level of non-polar compound can be increased or decreased, at the discretion of the physician, pharmacist, pharmacy scientist or other specialist. The amount of remaining inactive ingredients can be adjusted as needed.

[0336] In one example, the powder form is a free-flowing powder. Loose powders can be obtained using techniques well known in the art, such as, but not limited to, spray drying, freeze drying or absorption coating. In one example, in order

To obtain a free-flowing powder, the protein derivative is formulated with an excipient such as lactose or starch. For example, the formulation may be a dried lactose spray formulation (see, e.g., US Patent No. 4,916,163).

[0337] Methods for forming 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 about spray drying processes and equipment are discussed by Marshall, "Atomization and Spray-Drying," 50 Chem. Eng. Threshold. Monogr. Series 2 (1954), and Masters, Spray Drying Handbook (fourth edition 1985). Spray drying methods are well known (see, e.g., US Patent Nos. 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. To spray the liquid in the cooling tower or chamber, one or more spray nozzles are used. When the material is sprayed (sprayed), surface tension causes the formation of homogeneous spherical particles that pass through the cooling tower and harden to permanent intact balls. Spray dried particles can be between exactly or about 0.5 microns and exactly or about 100 microns, and are usually smaller than exactly or about 10 microns, usually smaller than exactly or about 5 microns, and usually smaller than exactly or about or exactly or about 1 micron.

[0338] Methods of spray drying liquid nanoemulsion compositions to provide powder compositions are provided. In spray drying methods, liquid nanoemulsion compositions may be heated, e.g., to a temperature between exactly or about 100 and exactly or about 150 ° F, usually between 110 ° F and 140 ° F, e.g. exactly or about 110, 115, 120, 125, 130, 135 or 140 ° F. The compositions may be mixed during heating, such as using any of the mixers described herein, e.g., homogenizers (e.g., reversible homogenizers and piston-driven homogenization).

[0339] For spray drying, one or more excipients are mixed with a polar solvent, usually water, and heated, e.g., to a temperature between exactly or about 100 ° F and exactly or about 150 ° F, usually between 110 ° F and 140 ° F, e.g. exactly or around 110, 115, 120, 125, 130, 135 or 140 ° F. In one example, the excipient is mixed with water in an amount of one part excipient (by weight) to two parts water (by weight). Excipient-solvent mixture (e.g. water) can be stirred during heating, e.g., using any of the mixers described herein, e.g., homogenizers (e.g., reversible homogenizers and piston-driven homogenization), by heating during mixing. The heated nanoemulsion liquid composition and the heated water-excipient mixture are then mixed together, such as by transferring one mixture to another, e.g., using any of the transfer means provided herein. Usually these two mixtures are homogenized, e.g. using a reversible homogenizer or piston-driven homogenization or any other homogenizer. The homogenised mixture is then spray dried using a spray dryer.

[0340] Exemplary spray dryers are cyclone spray dryers. During spray drying using cyclone spray driers, homogenised

The mixture is pumped into a spraying device, where it is comminuted into small droplets. When in contact with a stream of hot air, moisture is removed very quickly from the droplets while they are still suspended in the drying air. The dry powder is separated from the moist air in the cyclones by centrifugal action. Centrifugal action is caused by a large increase in air velocity when a mixture of particles and air enters the cyclone system. The dense powder of particles is shifted towards the walls of the cyclone, while lighter, moist air is removed through the exhaust ducts. The powder settles on the bottom of the cyclone, where it is removed by a discharge device. Sometimes the air supply lines for drying the powder are connected to cooling systems that supply cold air to carry the product through the transfer lines. Cyclone dryers designed for large production schedules, are capable of drying many tons of powder per hour.

[0341] It will be understood by those skilled in the art that the inlet temperature and outlet temperature of the spray dryer are not critical, but will be at a level to provide the desired particle size, less than or about 1 micron exactly, and provide a powder that has the desired properties. Usually, the desired property to be evaluated is the ability of the free flowing powder to provide a clear (or relatively clear) liquid composition diluted when diluted in an aqueous medium. In this regard, the inlet and outlet temperatures are controlled depending on the melting characteristics of the nanoemulsion components and the composition of the homogenised mixture of the nanoemulsion liquid concentrate / excipient. The inlet temperature is between exactly or about 60 ° C and exactly or about 170 ° C, with the outlet temperatures between exactly or about 40 ° C to exactly or about 120 ° C. Preferably, the inlet temperatures are from exactly or about 90 ° C to exactly or about 120 ° C and the outlet temperatures are from exactly or about 60 ° C to exactly or about 90 ° C. The flow rate that is used in the spray drying equipment is usually exactly or about 3 ml per minute to exactly or about 15 ml per minute. The atomizing air flow rate varies between exactly or about 25 ml per minute to exactly or about 50 ml per minute. Commercial spray driers are well known to the skilled person and the appropriate settings for any given dispersion can be easily determined by a specialist without undue experimentation. Operating conditions such as inlet and outlet temperature, feed rate, atomization pressure, drying air flow rate and nozzle system can be adjusted according to the manufacturer's guidelines.

[0342] In some examples, the dry powder is stored in the form of a capsule or compressed into a tablet. For use in tablet form, the compositions usually contain many other excipients. These excipients include tablet disintegrating agents such as corn starch, glidants such as silicon dioxide and moisturizing agents such as magnesium stearate. Typically, these compositions contain lower amounts by weight of glidants and moisturizers, e.g. two percent each (2%) or less by weight. Tablet disintegrants are optionally present and, if present, are present in amounts sufficient to ensure that the tablet will disintegrate when ingested. Suitable materials, such as corn starch, are used at concentrations from about zero to about 30 weight percent of the composition.

[0343] Loose powders are also used to administer the active ingredient by inhalation using a dry powder inhaler. Such dry powder inhalers usually administer the active ingredient in the form of a loose powder, which is dispersed in the patient's air stream when inhaled. To obtain a free-flowing powder, the active ingredient is usually formulated with a suitable excipient, such as lactose or starch. For example, such a dry powder formulation can be prepared, for example, by combining lactose with the active ingredient followed by drying the blend of ingredients. Optionally, if desired, the active ingredient can be formulated without excipient. The pharmaceutical composition is then usually placed in a dry powder dispenser or in inhalation sachets or capsules for use with a dry powder delivery device. Examples of dry powder inhaler devices include Diskhaler (GlaxoSmithKline, Research Triangle Park, NC) (see, e.g., US Patent No. 5,035,237); Diskus (GlaxoSmithKline) (see e.g. US Patent No. 6,378,519; Turbuhaler (AstraZeneca, Wilmington, Del.) (See e.g. US Patent No. 4,524,769); Rotahaler (GlaxoSmithKline) (see e.g. US Patent 4,353,365) and Handihaler (Boehringer Ingelheim) Further examples of suitable DPI instruments are described in US patents 5,415,162, 5,239,993 and 5,715,810 and references cited herein.

3. Liquid Diluted Compositions Containing Diluted Concentrates [0344] Among the compositions provided herein are liquid diluted compositions, usually aqueous liquid diluted compositions containing non-polar compounds. Aqueous liquid dilution compositions are prepared by diluting the supplied nanoemulsion liquid concentrates in an aqueous medium, e.g., beverages, e.g., water, artificially colored and flavored water, carbonated drink, milk, juice, including fruit juices, sauces, syrups, soups, sports drinks, nutritional drinks, energy drinks, vitamin-enriched drinks or any drink.

[0345] In one example, the aqueous liquid dilution compositions contain between 0.05 grams (g) or about 0.05 g and 10 g or about 10 g, usually between 0.05 g and 5 g of liquid concentrate per 8 oz volume or about 8 ounces by volume, at least 8 ounces by volume or at least about 8 ounces by volume, or less than 8 ounces by volume or less than about 8 ounces by volume, or per serving size, water medium, e.g. 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 concentrate per 8 ounces by volume, about 8 ounces by volume, or at least 8 ounces by volume or at least about 8 ounces by volume of an aqueous medium, for example 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 100, 200 or more ounces of volume of aqueous medium.

[0346] In another example, the aqueous liquid dilution composition contains between 1 ml or about 1 ml and 10 ml or about 10 ml of a 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 concentrate per 8 ounce volume, about 8 ounce volume, at least 8 ounce volume, or at least about 8 ounce volume, or less than 8 ounce volume or less than about 8 ounce volume, or per portion size, water medium, e.g. 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 100, 200 or more ounces of volume of aqueous medium.

[0347] In another example, the aqueous liquid dilution composition contains at least 10 mg or about 10 mg, usually at least 25 mg or about 25 mg, usually at least 35 mg of the compound

For a non-polar active ingredient, e.g., a non-polar active ingredient, for 8 ounces of volume or about 8 ounces of volume, at least 8 ounces of volume or at least about 8 ounces of aqueous medium, or less than 8 ounces of volume or less than about 8 ounces, or per serving size, water 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 non-polar compound in at least 8 ounces volume or at least about 8 ounces volume aqueous medium.

[0348] In another example, the aqueous liquid dilution composition contains a concentrate diluted at a dilution factor between 1:10 or about 1:10 and 1: 1000 or about 1: 1000 or more, usually between 1:10 or about 1:10 and 1: 500 or about 1: 500 or more, e.g. not 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: 400, e.g. 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 aqueous liquid dilution compositions contain a liquid concentrate diluted to any amount. In another example, the dilution is less than 1:10 or about 1:10.

[0349] The properties of the provided liquid concentrates that are diluted in aqueous media contribute to the different properties of the resulting aqueous diluted liquid compositions, for example clarity; indications for human consumption, for example, a pleasant taste and / or smell, for example, the lack of a "fish" taste / smell, the lack of "ring formation" and the lack of crystal formation; stability, for example lack of oxidation, "ring formation" and / or precipitation over time; and safety for human consumption. As described above, liquid concentrates are formulated according to the desired properties of aqueous diluted liquid compositions containing concentrates.

a. Clarity [0350] In one example, the aqueous liquid dilution compositions are clear aqueous liquid compositions diluted or having no turbidity, aqueous liquid dilution compositions, for example, as determined as described below, empirically or by measuring turbidity and / or particle size . In another example, the aqueous liquid dilution compositions are not clear or not completely clear. The liquid may be more or less clear, or have the same clarity as another liquid, e.g., an aqueous dilute liquid composition prepared according to the provided methods, or a beverage, e.g., a beverage that does not contain a dilute concentrate. Many parameters can change the clarity of a liquid, for example the relative concentration of surfactant, non-polar compound and / or water; type of non-polar component; excipient (s) concentration in the specified non-polar compound; and the purity of the non-polar compound, for example, whether it has been standardized for 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 large amount of lecithin, it may be less clear than that made from a concentrate containing a non-polar compound that does not contain lecithin. In another example, a liquid concentrate containing a non-polar compound that is a filtered extract, when diluted can give a clearer aqueous liquid diluted composition than a concentrate containing a crude extract.

i. Clarity determined by empirical evaluation [0351] In one example, the clarity / turbidity of the aqueous liquid dilution composition containing the diluted concentrate is qualitatively assessed by observation. In one example, a liquid can be considered clear if it does not appear cloudy in appearance and / or if there are no visible particles or if there are few visible particles when looking at the liquid with the naked eye, or if it is the same or substantially similar in clarity to other liquid, e.g. water, fruit juice, carbonated drink or milk. In some cases, the aqueous liquid dilution composition is as clear or almost as clear as water or another liquid, for example a drink. For example, a liquid (containing a liquid concentrate diluted in an aqueous medium, e.g., a drink) can be as clear or almost as clear as an aqueous medium that does not contain a liquid concentrate. In the related example, there is no substantial difference, e.g., no observable difference, between the aqueous diluted liquid 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 turbidity can be considered clear. In another example, the liquid is clear or partially clear or substantially clear if no visible crystals and / or "ring formation" are observed on the container containing the liquid.

ii. Clarity Determined by Particle Size or Particle Size [0352] In another example, the clarity of the aqueous liquid dilution composition is assessed by measuring the particle size and / or the number of liquid particles.

[0353] In one example, the aqueous liquid dilution compositions have a particle size 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 50 nm.

[0354] Typically, the particle size of the provided aqueous liquid dilution composition containing a liquid concentrate that contains a non-polar compound is smaller than the particle size of a liquid containing a non-polar compound (not formulated as a liquid concentrate).

iii. Turbidity [0355] In another example, liquid clarity is evaluated and / or expressed by measurement

Turbidity, e.g., nephelometric turbidity units (NTU), as measured using the methods provided, described below. In this example, turbidity is measured optically to give a value indicating the turbidity or haze of a liquid that is correlated with particles in a liquid. The more clear the liquid, the lower its turbidity value.

[0356] In one example, the clear aqueous liquid dilution composition has a turbidity (NTU) value of 30 or about 30; or an NTU value of less than 30 or about 30, e.g., 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 approximately 14, less than 13 or approximately 13, less than 12 or approximately 12, less than 11 or approximately 11, less than 10 or approximately 10, less than 9 or approximately 9, less than 8 or approximately 8, less than 7 or approximately 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.

[0357] In another example, the turbidity value of the aqueous liquid dilution composition is less than 200 or less than about 200. For example, 200, 175, 150, 100, 50, 25 or less.

[0358] In another example, it is desirable that the aqueous liquid dilution composition have a turbidity value that is comparable, for example, almost the same as, same as, or less than or greater than the turbidity value of another liquid, e.g., a beverage containing no a liquid concentrate or aqueous liquid diluted composition made using the methods provided.

b. Stability [0359] Typically, provided aqueous liquid dilution compositions containing concentrates are stable, e.g., free of one or more changes over a period of time, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9 , 10, 11 or 12 months, 1, 2, 3, 4 or more years.

[0360] In one example, the compositions are stable because they are free from oxidation or significant oxidation over time. In another example, they are permanent because they remain clear over time. In another example, stable compositions remain safe and / or indicated for human consumption over time. In one example, stability refers to the lack of sediment formation in the compositions over a period of time. In a related example, the composition is considered stable because it has no "ring formation", no whitish or opaque ring formation around the periphery of the container containing the liquid, usually on the surface of the liquid. Ring formation is usually undesirable, especially for liquids for human consumption, for example, a beverage.

[0361] In another example, the composition is stable if it shows no visible phase separation over a period of time, for example after 24 hours, after one week or after one month. In one example, the compositions are stable if they do not exhibit one or more of these described properties when stored at a particular temperature. In one example of the compositions, the compositions remain stable at room temperature, for example 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 cooling temperatures, e.g., 4 ° C or about 4 ° C, or at freezing temperature, e.g., at -20 ° C or about -20 ° C.

[0362] Stability refers to the desired property of the provided compositions, e.g. the ability of the provided compositions to remain free of one or more changes over a period of time, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months, 1, 2, 3, 4 or more. In one example, the composition is stable if formulated to remain free from oxidation or significant oxidation over time. In another example, stable compositions remain clear over time. In another example, stable compositions remain safe and / or recommended for human consumption over time. In one example, stability refers to the lack of sediment formation in the compositions over a period of time. In a related example, persistence refers to the lack of "ring formation" over a period of time. In another example, a composition is stable if it shows no visible phase separation over a period of time, for example after 24 hours, after one week, or after one month. In one example, the compositions are stable if they exhibit one or more of these desirable properties, when stored at a particular temperature.

[0363] In one example, the compositions are stable at room temperature, for example 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 cooling temperatures, e.g., 4 ° C or about 4 ° C, or at freezing temperatures, e.g., at -20 ° C or about -20 ° C.

c. Properties indicated for human consumption [0364] In one example, the liquid dilution composition is indicated for human consumption, for example for use in a food or drink. The different properties of the liquid diluted composition may contribute to its benefits as a consumable product. For example, taste, smell, clarity, color, crystal formation, precipitation and "ring formation" can all have advantages.

[0365] In one example, the liquid dilution composition has a pleasant taste and / or aroma, 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 does not have an unpleasant taste or smell, such as a "fish" taste or smell. In one example, the concentrate has a less unpleasant smell or taste, for example fishy, ​​compared to another aqueous liquid diluted composition.

[0366] In another example, the aqueous liquid dilution composition is desirable because it contains no crystals or has few crystals compared to the other aqueous liquid dilution composition. In another example, an aqueous liquid dilution composition is desired,

EP2 268 160 B1 because it has no ring formation.

d. Safety [0367] Typically, aqueous liquid dilute compositions containing concentrates are safe for human consumption, e.g., they only contain FDA approved ingredients for human consumption, e.g., GRAS-certified ingredients. In one example, one or more ingredients, for example all ingredients, are kosher certified. Composition safety also applies to durability over time. The lack of minimal oxidation of the composition over time can contribute to the safety of the composition.

e. Oral bioavailability [0368] In one example, non-polar compounds, e.g., non-polar active ingredients, contained in aqueous dilute liquid compositions exhibit high or relatively high bioavailability, e.g. bioavailability, which is higher than liquid containing only non-polar active ingredient (i.e. not formulated in a liquid concentrate). Bioavailability refers to the body's ability to absorb a non-polar active ingredient in a specific space, tissue cell and / or cellular compartment. Usually, non-polar active ingredients in liquids having small particle sizes are better absorbed than those with larger particle sizes.

C. METHODS OF MAKING LIQUID NANOEMULSION CONCENTRATES CONTAINING NON-POLAR COMPOUNDS [0369] Methods for producing liquid nanoemulsion concentrates have also been described. Common equipment and method steps are described in detail below. In one example, general methods for producing concentrates were carried out using a laboratory scale production process that is used to make batches of concentrates of relatively smaller sizes. In one example, general methods for making concentrates are carried out using large scale production processes that are used to obtain relatively larger batches of concentrates. A laboratory-scale process can be increased to an enlarged process. Any concentrate produced using a laboratory-scale process can be made using an enlarged-scale process by changing the scale of the process.

1. Equipment for making concentrates [0370] Various equipment, for example, vessels for mixing the oil phase, the aqueous phase and the emulsion, for example tanks; weight; mixers including standard mixers and homogenizers; heating and cooling instruments, including water-jacketed tanks, hotplates, water baths and coolers (coolers) including recirculated coolers; conveying devices, e.g. conveying means, e.g. pumps, hoses, plumbing fittings; ball valves; purifiers, for example filters, for example carbon filters, ion exchange equipment, reverse osmosis equipment, endpoint filters and end product filters; evaluation equipment, e.g., pH and temperature measuring instruments; and other equipment is used at various stages of the provided methods for producing concentrates. The choice of equipment depends on many factors, including batch size and production process.

EP2 268 160 B1

2. Weights [0371] For measuring out ingredients before adding them to the appropriate vessel, usually one or more weights are used. Optionally, the ingredients can be weighed in a vessel, for example in a container placed on a scale.

[0372] For weighing the ingredients, any of the many well-known scales sold on the market can be used. The choice of weight (weights) depends on many factors, including the weight of the final concentrate produced and the weighted ingredients. In one example, multiple weights are used to weigh the various components of the concentrate. Generally, correspondingly higher load (load) weights (weights) are used to produce larger batches of concentrate, while correspondingly lower load (weight) weights are used to produce smaller batches.

[0373] Exemplary balances used in the methods of weighing ingredients provided are Toledo Scale (model GD13x / USA), Sartorius Basic Analytical Scale (model BA11 OS), which is the basic series of analytical balances with a 110 g capacity and a resolution of 0.1 mg; and OHAUS Scale (model CS2000), which is a compact portable digital scale, with a load capacity of 2000 g and a resolution of 1 g.

a. Purifiers, including filters [0374] Purifiers, usually more than one purifier, for example filters, are used in the provided methods to remove impurities in the ingredients before they are added to the concentrate and / or to form the final concentrate and / or concentrate intermediate. For example, when the polar solvent is water, the water is usually purified. In one example, one or more purifiers, e.g., carbon filters, ion exchange purifiers, reverse osmosis purifiers and / or endpoint filters, are used to filter water, e.g. municipal water, before adding it to the aqueous phase, e.g. to remove impurities, for example sediment from water.

[0375] Exemplary purifiers that can be used in the methods provided are filters, 100 micron filters and carbon filters that are filters that use activated carbon to remove contaminants by chemical absorption. Carbon filters are usually used for water purification and are particularly effective for filtering chlorine, sediment, volatile organic compounds and other impurities. Typically, the particles removed using carbon filters fall between about 0.5 microns and about 0.5 microns. Other filters are well known and can be used in the methods provided.

[0376] Exemplary purifiers that can be used in the methods provided are also reverse osmosis purifiers that use mechanical pressure to purify liquids, such as water. In one example, to remove impurities, pressure pushes water through the semipermeable membrane.

[0377] Examples of purifiers that can be used in the methods provided are ion exchange purifiers, e.g., an ion exchange purifier using a resin bed, e.g., a zeolite resin bed, to exchange salts, e.g., cations, e.g., magnesium and calcium, with other cations, for example sodium and potassium cations. Such purifiers can be purchased for example

EP2 268 160 B1 from Aquapure Filters, Clarkston, MI.

[0378] In another example, a final product filter (e.g., a 100 micron FSI filter, BPEM 100-5GP product number). This filter is used to filter out any impurities in the final product (e.g., final liquid nanoemulsion composition). Other filters are known and can be used in the methods provided.

b. Vessels for mixing ingredients [0379] One or more, usually two or more vessels, for example tanks, for example water-jacketed tanks; pots; and / or beakers, e.g., Pyrex® beakers, are used in the methods provided to contain liquid concentrate component (s), for example, during mixing and / or heating or cooling. Typically, separate vessels (oil phase tank and water phase tank) are used to mix and heat the oil phase and water phase ingredients before combining the two phases to form an emulsion. In another example, an additional vessel, e.g. a storage and / or packaging tank, is used to store and / or pack the emulsion and / or to add / mix additional ingredients to the emulsion.

[0380] Many vessels are available for mixing ingredients. Usually, the tanks are cleaned, for example rinsed, washed with soap and / or disinfected according to known procedures, before use or between uses.

[0381] In one example, the vessel typically used in a laboratory scale process is a container, e.g., a laboratory scale container, e.g., flasks, beakers, e.g., Pyrex® beakers, vials, measuring containers, bottles, and / or other scale containers. laboratory.

[0382] In another example, the vessels typically used in an enlarged scale manufacturing process are tanks, e.g., water phase tanks, oil phase tanks, storage / packaging tanks. Usually, tanks are equipped with one or more mixers, e.g. a standard stirrer and / or homogenizer, which are used to mix the ingredients added to the tank. In one example, the tank is further equipped with a heating and / or cooling device. For example, the tank may be a water-jacketed tank. The temperature of the water-jacketed tank is controlled by the water-jacket, for example to heat the contents, for example while mixing.

[0383] Exemplary tanks that can be used in the methods are water jacketed tanks, for example an Overly 550 Gallon water jacket tank (model 10576501G) which has a capacity of 550 gallons and is usually used as a water phase tank, Schweitzers 450 tank gallon (model # 5214-C), which has a capacity of 450 gallons and is usually used as an oil phase tank and a Royall 190 gallon water jacket tank (model 9977-5), which has a capacity of 190 gallons and can be used as a water phase or oil phase tank when smaller volumes are mixed. Other reservoirs are well known and can be used in the methods provided to mix concentrates, e.g. concentrate phases.

c. Agitators [0384] Agitators are used in methods for combining, mixing and / or emulsifying liquid concentrates and / or various components and / or liquid concentrate phases. In one example,

The stirrers are used to keep the ingredients and / or the mixture circulating to maintain the temperature, viscosity and / or other parameters of the mixture. Exemplary mixers that can be used in the methods provided are standard mixers, e.g., standard mixers, which can be used, for example, to mix ingredients in the water and / or oil phases to maintain a homogeneous mixture during heating. An example of a standard agitator is the LIGHTNIN® agitator (LIGHTNIN, Rochester, NY), for example, model numbers XJC117 and ND-2. In one example, LIGHTNING® mixers are permanently mounted mixers, driven by gears, with a high flow, for use in closed tanks. Another example of a standard mixer is the mixer sold by IKA®, e.g. IKA® tripod mixers, e.g. models No. RW-14 Basic and RE-165, which are laboratory mixers and can be used to mix ingredients, e.g. oil and water phases . In one example, the agitator (s) are attached to vessels, e.g. tanks, e.g. assembled or gripped with a laboratory paw on the tank, e.g. at the top of the tank. In another example, the stirrers are placed in mixing vessels.

[0385] Exemplary mixers used in the methods are also homogenizers (also known as shear), which are usually used to form emulsions by emulsifying the oil and water phases after they are combined. Homogenizers usually provide high shear dispersion of solids and emulsification of immiscible liquids at high shear rates. Exemplary homogenizers that can be used in the methods provided are high shear homogenizers, e.g., reversible homogenizers sold by Arde Barinco, Inc., Norwood, NJ, for example the CJ-50 model, which is a 3,600 rpm mixer having 6-inch diameter impeller, 5575 ft / min speed and 33-inch immersion depth, and has six separate holes at the top and bottom, which concentrate the liquid into six chambers, reducing the surface volume and creating a shearing effect; and the CJ-4E model, which is a 10,000-rpm mixer with a cooling fan motor, optimized for batch sizes of 1 to 5 gallons, having a rotor diameter of 1.875 inches, speed of 4920 revolutions per minute, and immersion depth of 16 inches. Other homogenizers can be used in the methods, for example, other reversible homogenizers sold by Arde Barinco Inc.

[0386] In one example, the homogenizer is attached to the top of a vessel, for example a reservoir, for example with lab feet or with pliers and an electric lift. In another example, the homogenizer is placed in a vessel. Arde Barinco reversible homogenizers incorporate an axial flow impeller that creates two distinct mixing actions, depending on the direction. A downward "swirl flow" pulls solids from above and below the mixture, while an upward "umbrella flow" controls mixing at highest shear and recirculation speeds without splashing or introducing air. Reversible homogenizers are usually equipped with adjustable baffles that can be adjusted to control the type of mixing, for example at different times during emulsification.

100

[0387] Many additional stirrers (mixers, mixers) are known in the art and can be used with methods. Examples of mixers (mixers, mixers) that can be used with the methods are shear mixers, built-in mixers. Ribbon, furrow / paddle for loose substances, Forberg mixers, portable, bag dumps &amp; compactors, type V mixers, turbine mixers, two-cone mixers, continuous mixers, high flow mixers, periodic mixers, double band mixers, blade and ribbon mixers with propellers, plow mixers / turbulent mixers, Forberg fluidized mixers, air mixers, active mixers, passive mixers, top mounted mixers, side-entry mixers, static mixers, fixed mixers, portable mixers - direct and toothed drive, sanitary mixers, drum mixers, mixers with collecting container (IBC), laboratory mixers, speed controlled mixers, dough mixer, vertical mixer , spiral mixer, twin arm mixer, fork mixer, double spiral mixer, all mixers (mixers, mixers), mixers with mixer, Banbury closed mixers, rubber mixers, Blondheim mixers, butter mixers, cone mixers, continuous mixers, dispersing mixers, mixing kneaders, emulsifying mixers, Hobart mixers, liquefying mixers, Littleford mixers, meat mixers, plow mixers, mixing crushers, Nauta mixers, Oakes mixers, planetary mixers, Pony mixers, PUG mixers, ribbon mixers, Ross mixers, rotary mixers, Sigma mixers, single arm mixers, Tote Bin mixers, drum mixers, vacuum mixers, turbulators, two-chamber mixers, V-type mixers, Zig-Zag mixers, side agitator mixers, hand mixers, mixing rods, mixers, magnetic mixers and tripod mixers, example of mechanical and / or electric tripod stirrers.

d. Heating devices [0388] The methods use one or more, usually more than one heating device to control the temperature of ingredients, phases and / or concentrate, usually while mixing.

[0389] In one example, the heating devices 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 control the temperature of the contents of the vessel.

[0390] Optionally, other heating devices may be used to heat the ingredients, phases and / or concentrates. Exemplary heating devices that can be used in the methods are immersion and / or immersion heaters, for example 12 KW and 13 KW sanitary heaters, which are food grade heaters that are immersed in a tank during mixing, usually to apply the required strong heating, for example, temperatures higher than about 60 ° C or 60 ° C, or higher than 80 ° C or about 80 ° C. Exemplary heating devices are also ovens, e.g. propane ovens. Exemplary heating devices are also hotplates, e.g. the Thermolyne hot plate, model number 846925 and model number SP46615. Usually, the preheater is able to heat the mixture to 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,

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67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84 or 85 ° C. Typically, the preheater is able to heat the mixture to 60 ° C or 60 ° C, for example, providing poor heating.

e. Cooling devices [0391] One or more cooling devices may be used in the methods, for example for cooling ingredients during mixing, for example for cooling the mixture during the emulsification of the oil and water phases. Exemplary cooling devices are refrigerators, e.g. recirculated coolers, which can be attached to a vessel, e.g. remotely, or by mounting the tank in a cooler, to recirculate liquid from the tank through the cooler and back to the tank, to quickly cool and maintain the temperature of the mixture during mixing. Examples of open system coolers that can be attached to the tank and used in the methods provided are those sold by Turmoil, West Swanzey, NH, for example open or closed system coolers, for example model No. OC-1000 RO. Other cooling devices are well known and can be used in the methods provided.

[0392] Exemplary cooling devices are also water baths and ice baths, for example water baths and / or ice baths in which the vessel (s) are placed, for example during homogenization.

[0393] Typically, cooling devices can be used to cool a liquid 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, usually between 25 ° C and 43 ° C; usually between 35 ° C and 43 ° C, for example 26.5 ° C. Typically, cooling is rapid cooling, e.g., cooling to between 25 ° C or about 25 ° C and 45 ° C or about 45 ° C, e.g. between 35 ° C and 43 ° C, e.g. 26.5 ° C, in between 15 minutes or about 15 minutes and 2 hours or about 2 hours, usually 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 means [0394] Transfer means are used in methods for transferring liquid from one vessel to another vessel, for example to transfer the contents of one or more vessels to one or more vessels, e.g., transferring the aqueous phase into the vessel an oil phase (e.g., oil phase tank) or transferring the oil phase to a water phase vessel (e.g., water phase tank) to form an emulsion. Exemplary equipment used for transfer means are conveying pumps and associated accessories, e.g., ball valves, plumbing fittings (e.g., plumbing fittings sold by Granger, Inc., Lake Forrest II) and transfer hoses (e.g., hoses sold by Sani-Tech West, Oxnard, CA), for example food grade hoses attached to conveying pumps. Exemplary conveying pumps that can be used in the methods provided are Teel Pump (model 2P377B), Granger, Inc. Lake Forrest I1, suction pump with 2 HP rated power, voltage 60 Hz 208-230 / 460 AC, speed 3450 rpm. Other pumps, e.g. other suction pumps from Grainger, Inc. may be used as part of the transfer means. Optionally, transfer means can

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Include means for manually transferring liquid to another vessel, e.g., pouring, pipetting and / or other well known methods of manually transferring liquid.

g. Assessment equipment [0395] Assessment equipment is used to evaluate one or more properties of the composition, for example composition phases and / or final concentrates. For example, assessment equipment may be used to measure one or more concentrate and / or phase parameters, e.g., liquid temperature and pH. Examples of evaluation equipment are pH meters and temperature measuring instruments. An example of a pH / temperature measuring instrument is a pH and temperature measuring instrument sold by Hanna Instruments (model number HI 8314) that can be used to measure and pH of a mixture (s). Exemplary temperature measuring instruments are also temperature sensors, e.g., digital and / or waterproof temperature sensors, e.g., temperature sensors sold by Cooper-Atkins, Middlefield, CT, e.g., digital waterproof temperature sensor (model # DPP400W) from Cooper-Atkins. Other evaluation equipment for assessing liquids and / or emulsions is well known and can be used in the methods provided.

3. General methods for preparing liquid nanoemulsion concentrates [0396] In general, to make liquid nanoemulsion concentrates, methods for producing concentrates include the steps for producing phases (e.g., oil phase (s) and aqueous phase (s)) as well as the steps for combining and emulsifying the phases. In some examples, the methods include additional steps such as evaluation, addition of further ingredients, packaging and filtration. The provided methods can be carried out using a laboratory scale production process (usually for small batches). Optionally, the methods can be carried out using a larger scale production process (usually for larger lots). Each of the concentrates provided can be produced either using an enlarged process scale or a laboratory scale process. In one example, after the primary concentrate has been prepared using a laboratory scale process, the method is scaled up to produce larger quantities of the concentrate using an enlarged scale process. When concentrates are formulated according to the methods provided, the primary concentrate is usually prepared using a laboratory scale process. In one example of the formulation methods, the selected formulation is then prepared using an enlarged scale process. Any of the concentrates provided herein can be made using the methods provided, using any of the manufacturing processes. Any method described herein where a laboratory scale process is used can be enlarged to produce concentrates using an enlarged scale process.

[0397] Typically, methods for producing liquid nanoemulsion concentrates include first production steps in which one or more oil phases and one or more aqueous phases are produced. The production of the oil phase and the production of the aqueous phase are usually carried out in at least two separate vessels, e.g. an oil phase vessel and an aqueous phase vessel. Each of the manufacturing steps usually includes a mixing step and a heating step that can be carried out simultaneously, sequentially in any order or partly simultaneously.

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[0398] To form the aqueous phase, the aqueous phase component (s) (e.g., polar solvent (e.g., water, propylene glycol, glycerin or other polar solvent) and, in some examples, additional components of the aqueous phase) is added to the water phase of the vessel. The component (s) are mixed, usually using a standard mixer, and heated, for example using heating devices. Typically, the aqueous phase ingredients are heated to a low heating temperature, e.g. to 60 ° C or about 60 ° C. To form the oil phase, the oil phase ingredients (e.g., non-polar compound (s), surfactant (s) and, in some examples, the other oil phase ingredient (s)) are added to the oil phase vessel. The oil phase component (s ) are mixed, usually using a standard mixer, and heated, for example using a heating device. Typically, the ingredients are heated to a low heating temperature, e.g. to 60 ° C or about 60 ° C. Mixing / heating of the oil and water phases can be carried out simultaneously or sequentially, in any order. In one example, the production of the oil phase is carried out simultaneously with the production of the aqueous phase, for example to protect the non-polar active ingredient, for example to prevent its oxidation. Usually, before the oil and water phases are combined in the next emulsification step, both phases are heated to the desired temperature, e.g. low heating temperature, and / or until the ingredients dissolve.

[0399] In general, the methods further include an emulsifying step. For the emulsifying step, the oil and aqueous phases are combined, for example, using one or more transfer means. The oily and aqueous phases are emulsified, usually with stirring, usually homogenized, using high shear, to form an emulsion (e.g., liquid nanoemulsion concentrate). The emulsification step can be carried out in an aqueous phase vessel, in an oil phase vessel or in a separate vessel.

[0400] Typically, during the emulsification step, the forming emulsion is cooled, for example, cooled rapidly, for example using one or more cooling devices. Usually, the cooling step is carried out simultaneously with the emulsifying step. 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.

[0401] The methods may include additional steps, e.g., evaluation steps, steps for adding additional ingredients, purification steps (e.g., filtration) and / or packaging / storage steps. as detailed below.

a. Preparation of the aqueous phase [0402] Typically, the aqueous phase ingredients are weighed and / or measured, for example, using one or more weights (e.g., one or more weights described herein), before adding the aqueous phase (e.g., any dish described here). In one example, the amount of each component added to the aqueous phase vessel is determined in accordance with the methods provided for making concentrates. Typically, the desired concentration, by weight (w / w), of the final nanoemulsion concentrate is used to calculate the amount of each component of the aqueous phase that is added to the aqueous phase vessel. Alternatively, the desired volume per mass, volume per volume and mass per volume may be used to calculate the correct amount of ingredient to be measured and added to the vessel.

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[0403] In one example, when the polar solvent is water, impurities in water, e.g., municipal water, are removed using one or more purifiers (e.g., one or more purifiers as described herein) above, before adding water to the water phase vessel. In one example, the water is purified by passing through, using the following purifiers, successively: a carbon filter, an ion exchange purifier, a reverse osmosis purifier and an endpoint filter, for example a 100 micron endpoint filter; before adding the water phase to the vessel.

[0404] Typically, the aqueous phase ingredient (s) are mixed in the aqueous phase vessel using a standard stirrer (e.g., any of the standard mixers described herein) and heated, usually simultaneously or partially simultaneously, using a heating device (e.g., any of those described herein heating devices). Typically, the aqueous phase is heated so that the components of the aqueous phase reach a low heating temperature, e.g. between about between 45 ° C or about 45 ° C and 85 ° C or about 85 ° C, e.g. 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, usually 60 ° C or 60 ° C, for example to prevent oxidation of non-polar components and / or to preserve the stability of the components. Typically, mixing and / or heating of the water phase ingredients in the water phase vessel is continued, for example, before combining the water phase and the oil phase. In one example, the aqueous phase is mixed and / or heated until the aqueous phase ingredients are dissolved. Typically, the temperature of the aqueous phase is kept stirring before the oil and water phases are combined.

i. Water phase components [0405] The water 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 includes other components of the water phase. Typically, the aqueous phase ingredients are the hydrophilic and / or amphipathic components of the liquid nanoemulsion concentrate. For example, oils and other lipophilic ingredients are usually not added to the aqueous phase. Certain ingredients, for example ingredients having hydrophobic and hydrophilic units, for example surfactants and co-surfactants, can be added to either the oil phase or the water phase or the oil and water phase. Exemplary components of the aqueous phase include, but are not limited to, polar solvents, e.g. water, usually filtered water, propylene glycol, glycerin and other diols; emulsion stabilizers; pH regulators, e.g., phosphoric acid and / or citric acid; flavors; surfactants; surfactants, e.g., phosphatidylcholine and / or specific quila; and preservatives.

[0406] The aqueous phase ingredients may be added to the aqueous phase simultaneously and / or sequentially in a specific order. In one example, before adding the next ingredient (s), one or more water phase ingredients are first added and heated. In one example, when the ingredients of the aqueous phase include a polar solvent and an emulsion stabilizer, these components are added sequentially, in the following order: 1) a 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 (e.g., water)

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An emulsion stabilizer, these components are added to the water phase vessel in turn, in the following order: 1) surfactant, 2) polar solvent (e.g. water); 3) emulsion stabilizer. Optionally, the aqueous phase ingredients can be added in any other order. Typically, when the aqueous phase includes a surfactant, especially when the surfactant is a surfactant that is solid at room temperature, for example a surfactant that is tocopherol polyethylene glycol succinate, the surfactant is the first component of the water phase added to water phase vessels. Typically, when the aqueous phase ingredients include an emulsion stabilizer, the emulsion stabilizer is the last ingredient added to the water phase vessel.

b. Oil phase preparation [0407] Typically, the oil phase ingredient (s) are weighed and / or metered, for example using one or more weights (e.g. one or more weights described herein), before adding the oil phase ( e.g., any vessel described herein). In one example, the amount of each oil phase component added is determined according to the formulated concentrate methods provided. Typically, the desired concentration, by weight (w / w), of the final nanoemulsion concentrate is used to calculate the amount of each oil phase component that should be added to the oil phase vessel. Alternatively, volume by mass, volume by volume or weight by volume can be used to calculate the correct amount of ingredient to be measured and added to the vessel.

[0408] Typically, the oil phase ingredients are mixed in an oil phase vessel using a standard stirrer (e.g., any of the standard stirrers described herein) and heated, usually simultaneously, using a heating device (e.g., any of the heating devices described herein). Usually the oil phase is heated so that it reaches a low heating temperature, e.g. between 45 ° C or about 45 ° C and 85 ° C or about 85 ° C, e.g. 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, usually 60 ° C or 60 ° C, for example to prevent oxidation of non-polar components and / or to preserve the stability of the components. In one example, one or more oil phase ingredients are mixed and heated according to the provided methods, before the remaining oil phase ingredients are added. For example, before adding the remaining components of the oil phase, the non-polar compound can be mixed and heated with one or more solvents, e.g., oil, e.g., linseed oil and / or vitamin E oil, until the non-polar compound is dissolved in the oil. Typically, the oil phase ingredients are mixed in an oil phase vessel until dissolved. Typically, the temperature of the oil phase is kept under stirring before the oil and water phases are combined.

[0409] In some examples, the oil phase and / or the aqueous phase may be prepared in more than one vessel, for example by mixing one or more oil phase ingredients in one vessel and mixing one or more other ingredients in another vessel. In this example, the oil phase ingredients mixed in separate vessels can either be mixed together prior to emulsification with the aqueous phase, or optionally, can be added separately, during emulsification, to the aqueous phase.

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i. Oil phase ingredients [0410] The oil phase includes a non-polar compound, for example a non-polar active ingredient, and, in some examples, other oil phase ingredients. Typically, the oil phase ingredients include one or more lipophilic and / or amphipathic ingredients of the liquid nanoemulsion concentrate. The oil phase ingredients usually do not include aqueous and hydrophilic ingredients. Certain ingredients, for example ingredients having hydrophobic and hydrophilic units, for example surfactants and co-surfactants, can be added to either the oil or water phase or the oil and water phase. Exemplary ingredients used in the oil phase of the provided concentrates are non-polar compounds, for example non-polar active ingredients, including any of the non-polar active ingredients provided herein; emulsion stabilizers, pH regulators, e.g., phosphoric acid and / or citric acid; surfactants; surfactants, e.g., phosphatidylcholine and / or specific quila; preservatives and oils, for example, solvents and other components of the oil phase.

[0411] The oil phase ingredients may be added to the oil phase simultaneously and / or sequentially, for example in any order or in a specific order. In one example, before adding the further ingredient (s), one or more oil phase ingredients are first added and heated. In one example, when the oil phase ingredients include a surfactant, a preservative, a solvent, a co-surfactant and a non-polar compound, these ingredients are added sequentially, in the following order: 1) a surfactant; 2) preservative; 3) solvent; 4) surfactant; 5) non-polar compound and 6) emulsion stabilizer. In another example, when the oil phase ingredients include a surfactant, a preservative and a non-polar compound, the ingredients are added sequentially, in the following order: 1) surfactant, 2) preservative; 3) non-polar relationship. In another example, when the oil phase ingredients include a surfactant, preservative, non-polar compound and emulsion stabilizer, the ingredients are added sequentially, in the following order: 1) surfactant, 2) preservative; 3) non-polar compound and 4) emulsion stabilizer. Optionally, the components of the oil phase and a different order, for example, in any order. Two or more oil phase ingredients can be added simultaneously.

[0412] Typically, when the oil phase comprises a surfactant, especially when the surfactant is a surfactant that is a solid at room temperature, for example a surfactant that is polyethylene glycol succinate tocopherol, the surfactant is the first component oil phase added to the oil phase vessel. Typically, when the oil phase ingredients include an emulsion stabilizer, the emulsion stabilizer is the last ingredient added to the oil phase vessel. Typically, the non-polar compound is either the last component added to the oil phase vessel, or is added just prior to the addition of the emulsion stabilizer that is the last component added to the oil phase vessel.

c. Combining and emulsifying the oil phase and the aqueous phase [0413] Generally, in the methods, after producing the oil phase and the aqueous phase, the oil and aqueous phases combine

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For example, one or more transfer means (e.g., any of the transfer means described herein) are used. The combined phases are emulsified, e.g., by mixing, e.g., homogenization, to form an emulsion (e.g., liquid nanoemulsion concentrate). Typically, the phases are mixed during the joining and emulsifying steps, for example using a homogenizer (e.g., any of the homogenizers described herein). In one example, the oil and water phases (e.g. formed emulsion) is further cooled, e.g. cooled rapidly, during the emulsification and / or joining steps.

i. Combining the oil and water phases [0414] To emulsify them, the oil and water phases are first combined, usually by transfer, using one or more transfer means (e.g., any of the transfer means described herein). In one example, the oil phase is transferred to a water phase vessel. In another example, the aqueous phase is transferred to an oil phase vessel. In another example, many oil or water phases are transferred to the water or oil phase vessel. In another example, the aqueous phase (s) and the oil phase (s) are transferred to another vessel, an emulsifying vessel.

[0415] Any transfer means can be used to connect the phases. For example, any means for transferring the contents of one vessel to another vessel as described above, e.g. conveying pumps and associated equipment, e.g. plumbing, hoses and / or ball valves; and manual transfer means, e.g., pipette pouring and / or piping means or other known transfer means. In some examples, the phases are kept clean, e.g., sterile during transfer, e.g., by using transfer means with plumbing fittings and / or joining the phases in a sterile environment.

ii. Emulsification of oil and aqueous phases [0416] Simultaneously and / or after the phases have been combined, the phases are mixed (e.g., homogenized), e.g. Typically, emulsification is carried out in a vessel containing combined liquids, e.g. an oil phase vessel or an aqueous phase vessel. For this emulsifying step, the oil and aqueous phases are mixed, for example after the joining step, usually during and after the joining step, using a mixer that is capable of emulsifying the liquid, e.g. a homogenizer, e.g. a reversible homogenizer. Typically, liquids are homogenized using a mixer (e.g. homogenizer) at low speed, e.g. at low rpm, e.g. between 850 rpm or about 850 rpm and 1200 rpm or about 1200 rpm, e.g. 850, 900, 950, 1000, 1050 , 1100, 1150 or 1200 revolutions per minute.

[0417] Liquids are usually mixed continuously or intermittently until the liquids form an emulsion, for example a nanoemulsion. In one example, the mixing speed is maintained to emulsify the oil and water phases. In one example, the mixer baffle is adjusted, for example by moving the baffle further down to the mixture or further up from the mixture, to control the type of mixing, e.g. to switch from downward flow to upward flow and vice versa while mixing the emulsion. In another example, the homogenizer can be adjusted to increase or decrease shear or in

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EP2 268 160 B1 to maintain shear at a certain speed. Methods for homogenizing the oil and water phases are well known and other methods can be used in the methods to homogenize the oil and water phases.

iii. Cooling [0418] Typically, the emulsion is cooled under agitation, for example by rapid cooling. In one example, the emulsion is cooled to support emulsion stability and phase emulsification, for example by preventing and minimizing oxidation, e.g., oxidation of a non-polar compound. Cooling, e.g. rapid cooling, is usually carried out using one or more cooling devices, e.g. any of the cooling devices described herein, or any known cooling device. In one example, the cooling device is a recirculated cooler. In another example, the cooling device is a water bath or oil bath. In one example, when the device is a recirculated cooler, the fluid from the vessel used for the emulsification step is recirculated through the cooler and then returns to the vessel to rapidly cool and maintain the temperature of the mixture during mixing. Typically, the forming emulsion is mixed and cooled until the phases are emulsified and the temperature reaches between 25 ° C or about 25 ° C and 43 ° C or 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 rapid cooling, the temperature is reached in less than 2 hours or about 2 hours, usually in 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.

[0419] Once the oily and aqueous phases are emulsified, thereby forming an emulsion, e.g. a liquid nanoemulsion concentrate, the emulsion can be used, for example, in the dilution methods provided, to produce a liquid diluted composition, e.g., a concentrate-containing beverage. Optionally, one or more additional steps may be performed prior to using the concentrate.

d. Additional steps [0420] Typically, one or more additional steps are performed after phase emulsification, before using the concentrate. For example, the emulsion can be evaluated (e.g., by measuring the pH and / or temperature of the concentrate). In another example, one or more additional ingredients may be added to the emulsion. In another example, the nanoemulsion concentrate is transferred to a storage vessel or packaging vessel, e.g. storage / packaging vessel, e.g. storage / packaging vessel. In another example, the nanoemulsion is purified, for example, filtered before use. In one example, the addition of additional ingredients, evaluation and / or purification can be performed in a storage / packaging vessel. Other additional steps may be carried out before use.

i. Additional ingredients [0421] In one example, additional ingredients, for example pH regulators and / or agents

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Flavoriness can be added to the emulsion after it is made. 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, usually between 2.6 or about 2.6 and 3.2 or about 3.2, e.g. 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 improve the taste and / or aroma of the concentrate and / or drinks containing the concentrate. In another example, an additional polar solvent, e.g. water, may be added to the emulsion, for example in the event of evaporation, to bring the concentrate to the appropriate volume. Other additional ingredients may also be added to the emulsion. Typically, additional ingredients are added to the emulsion containing vessel, e.g., water phase vessel, oil phase vessel, emulsion vessel, or other vessel, e.g. storage / packaging vessel. Typically, the emulsion is mixed (e.g., using any of the described agitators, usually standard agitators) during the addition of additional ingredients.

ii. Evaluation of the concentrate [0422] Usually, the concentrate is evaluated prior to use. Usually the pH and / or temperature is measured, for example using a pH and temperature meter. In one example, the pH and / or temperature is assessed after the addition of additional ingredients. In one example, further ingredients may be added to adjust parameters after evaluation.

iii. Concentrate filtration [0423] In one example, the concentrate is purified (e.g., using any of the purifiers described), e.g., using a final product filter before using the concentrate, e.g., before diluting the concentrate in an aqueous medium.

4. Laboratory scale process [0424] In one example of a method for producing liquid nanoemulsion concentrates, the method steps are performed using a laboratory scale production process that is carried out on a work table, counter, table or other surface. Typically, a laboratory-scale process is used to make emulsions having relatively smaller volumes than those produced using an enlarged scale process, e.g., volumes less than 1 or about 1 liter or less than 1 gallon or about 1 gallon, e.g. less than about 500 ml , e.g. 1000, 900, 800, 700, 600, 500, 450, 400, 350, 300, 250, 200, 150, 100, 50 or less.

[0425] For a laboratory scale process, the equipment is usually small enough to be used on the top surface of a work table or other similar surface, usually small enough to be moved, e.g., lifted, by a specialist using the method. For example, vessels, e.g., aqueous phase vessels, oil phase vessels, storage vessels, and packaging vessels are typically laboratory scale vessels made of arkorok, e.g., flasks, beakers, vials, measuring containers, bottles and / or other laboratory scale containers. In one example, the vessels in the laboratory scale process are a Pyrex® beaker. Typically, the agitators are agitators that can be used in laboratory scale vessels, e.g. standard agitators, including hand agitators, agitator rods, agitator rollers, magnetic agitators and stand agitators, e.g.

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Mechanical and / or electric tripod mixers and / or other mixers that can be used in the dishes. Examples of suitable laboratory scale mixers are standard mixers, e.g. standard mixers sold by IKA®, e.g. IKA® stand mixers, e.g. Model No. RW-14 Basic and RE-16S, which are laboratory mixers and can be used to mix ingredients , for example, the production of oil and water phases. Examples of suitable laboratory scale mixers are also homogenizers, e.g., reversible homogenizers, including The Arde Barinco reversible homogenizer, Model No. CJ-4E, which can be used for phase emulsification. Typically, heating devices are those that can be used with laboratory scale vessels, e.g. hotplates. Cooling instruments are usually instruments suitable for use with smaller vessels on a laboratory scale, e.g. ice baths and / or water baths, in which vessels can be placed, e.g. for rapid cooling. The evaluation means used in the laboratory scale process, e.g. temperature and / or pH measuring instruments, are usually suitable for placement in laboratory scale vessels.

[0426] Typically, for a laboratory scale process, the oil phase and the aqueous phase are prepared by mixing and heating in separate laboratory scale vessels, e.g., flasks, beakers, vials, measuring containers, bottles and / or other laboratory scale containers. Mixing is usually carried out using a suitable laboratory scale stirrer, e.g. a standard stirrer, such as a manual stirrer, stirring rod, stirring roller, magnetic stirrer and / or a tripod stirrer, e.g. stirrer sold by IKA®, e.g. IKA® tripod stirrers , for example model No. RW-14 Basic and RE-16S, which are laboratory mixers. Typically, heating of the oil and water phases is carried out using heating devices suitable for the laboratory scale method, e.g. a heating device on which one or more dishes, e.g. a hot plate, may be placed. To combine the oil phase and the aqueous phase, one or more phases, usually one phase, is usually transferred manually to another vessel, for example by pouring, pipetting and / or using another manual transfer means. In order to emulsify the oil and water phases, a reversible homogenizer is usually used. In order to cool the emulsion being formed, for example, rapidly cooling the emulsion, usually cooling instruments suitable for the laboratory scale method are used, e.g. cooling instruments on which or in which a vessel can be placed, e.g. a water bath or ice bath.

5. Scale-up production processes [0427] Methods for making liquid nanoemulsion concentrates can be carried out using a scale-up production process. Large scale production processes are usually used when the liquid nanoemulsion concentrate produced has a relatively larger volume than the concentrate produced using the laboratory scale method, for example volumes greater than 1L or about 1L or greater than 1 gallon or about 1 gallon, e.g. larger than about 500 ml, for example at least 0.5l, 1l, 2l or 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, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 800, 900, 1000 or more gallons.

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EP2268160B1 [0428] Generally, large scale production processes are carried out using equipment that is suitable for these larger batch volumes (batch sizes). For example, vessels used in large-scale processes are typically tanks, e.g., water-jacketed tanks that are equipped with water jackets that can be used as heating devices for heating the oil and water phase ingredients during the production of the oil and water phases. Water jackets are usually controlled using control panels. Similarly, transfer means used in an enlarged process process typically include transfer pumps and related equipment, e.g., ball valves and hoses. Examples of mixers that can be used in an enlarged process process are standard mixers (e.g. embedded mixers, e.g. LIGHTNIN® mixers, e.g. model XJC117 (permanently attached, high flow toothed agitator and model ND2). an enlarged scale is shown in figure 1 and described in this section below. The provided methods for producing concentrates can be carried out using these exemplary enlarged process processes or any variation of the enlarged process process, e.g., excluding one or more of the steps of the exemplary process, adding one or more steps according to the method and / or replacing the steps and / or equipment according to the methods described herein.

[0429] Figure 1 shows an exemplary process at an enlarged scale 100 for producing a liquid concentrate. In this example, the polar solvent is water. An example of an enlarged process process includes the following stages:

a. Water purification [0430] As mentioned herein, the polar solvent may include water (including purified water) and other polar solvents, e.g. glycerin and propylene glycol. In the example illustrated in the figure, the polar solvent is water 101 (e.g. municipal water), which is purified before the water phase is added to the vessel, by passing water through the following purifiers, in turn, in the following order: carbon filter 105, ion exchange device 106, reverse osmosis device 107, end point filter 100 micron 108i 50 micron utility point filter 109.

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

[0431] As described above, to make the aqueous phase, the polar solvent and any other components of the aqueous phase are usually weighed and / or metered, and added to the water phase vessel and mixed using a standard stirrer or other mixer such as a homogenizer or other stirrer described herein, and usually heated during stirring, where the aqueous phase is produced by mixing and heating, usually low heating (e.g. 60 ° C, 70 °, 71 ° C), according to the methods provided. In the example of the enlarged scale production process shown in Figure 1, the water phase vessel is the water phase reservoir 103, which is the water jacketed tank. In the example illustrated in Figure 1, the water phase component (s) are mixed using a standard mixer 111, e.g. LIGHTNIN® mixers (e.g. Model No. XJC117, permanently attached, high flow toothed agitator) attached to the tank, for example a top mounted tank. In the example shown in figure 1, the heating device used to heat the aqueous phase ingredients is a water jacket

112

Tank with a water jacket; the water jacket temperature is controlled using the control panel.

[0432] As described above, to produce the oil phase, the oil phase ingredients are usually weighed and / or metered and added to the oil phase vessel and mixed using a standard agitator or other mixer such as a homogenizer or other agitator described herein, and usually heats while stirring, where the oil phase is produced by stirring and heating, usually low heating (e.g. 60 ° C), according to the methods provided. In the example of an enlarged scale production process shown in Figure 1, the oil phase vessel is an oil phase oil jacket 102. In the example illustrated in figure 1, the oil phase ingredients are mixed using a standard agitator 111, e.g. a LIGHTNIN® agitator (e.g. model ND2) attached to an oil phase tank, e.g. mounted on a tank. In the example illustrated in figure 1, the heating device used to heat the oil phase ingredients is the water jacket of the water jacket oil phase tank; the water jacket temperature is controlled using the control panel.

c. Combining and emulsifying the phases [0433] As described herein, when the oil and water phases reach the desired temperature (e.g. 60 ° C, 70 ° C, 71 ° C or other temperature), after the oil phase and / or phase components have dissolved aqueous, and optionally after cooling one of the phases, e.g., cooling the aqueous phase to 60 ° C according to the provided methods, the oil and aqueous phases are combined by transfer and emulsification, usually by homogenization. In one example, the transfer is carried out slowly to prevent the forming emulsion from sticking together, such as by periodically stopping the transfer while continuing to mix the emulsion, or by slowly combining the phases with mixing. In the example of the enlarged scale production process shown in Figure 1, the phase joining is the result of transferring the oil phase to the water phase vessel, using transfer means 112, which include a transfer pump (e.g., Teel pump, model 2P377B, sold by Granger, Inc. ), plumbing fittings, transfer hose (hoses) (e.g. food grade hoses, sold by Sani-Tech West) and ball valve (s). Optionally, the aqueous phase can be transferred to the oil phase. In the example shown in Figure 1, to start the joining / emulsifying steps, a homogenizer 110 (e.g., Arde Barnico, Inc. reversible homogenizer) mounted on a water phase tank, is switched on for example at 850-1200 rpm. Then the ball valves are opened and the transfer pump is switched on, thus carrying out the transfer of the liquid oil phase to the water phase tank through the transfer hose (s). When the phases are combined, the mixture is homogenized by continuous mixing using a homogenizer 110.

[0434] In certain examples of an enlarged production process to prevent clumping, during emulsification, the pump is periodically stopped (e.g., by turning off the pump) while mixing is continued using the mixer. In one aspect of this example, this method of preventing sticking is used when the polar solvent is a non-water solvent such as propylene glycol or glycerin. During mixing, the homogenizer can be adjusted, for example by adjusting the septum further in the forming direction

113

Emulsion and / or further from the forming emulsion. In one example, the shear rate is adjusted to a speed where it can be seen that the oil phase goes to the top of the mixer. In one example, this regulation is used when the polar solvent is a solvent other than water, such as propylene glycol or glycerin.

d. Cooling [0435] As described herein, the forming emulsion is usually cooled, usually cooled quickly, during the emulsification step. In an enlarged process as shown in the example illustrated in Figure 1, rapid cooling is usually obtained by repeatedly passing the forming emulsion through recirculation cooler 115 (e.g. model No. OC-1000 RO, sold by Turmoil, West Swanzey, NH), which is attached to the water phase tank. Homogenization continues during the cooling step, for example at between 850 rpm and 1200 rpm. Cooling is continued, for example until the emulsion temperature reaches between exactly or about 25 ° C and exactly or about 43 ° C, such as between exactly or about 25 ° C and exactly or about 35 ° C, between exactly or about 35 ° C and exactly or about 43 ° C, or exactly or about 40 ° C. Typically, rapid cooling is carried out for between exactly or about 30 and exactly or about 60 minutes.

e. Additional steps [0436] As described herein, additional steps can be performed after the emulsion. In the example of the enlarged scale production process shown in Figure 1, additional steps include transferring the emulsion using transfer means 112, which includes a transfer pump (e.g., Teel pump, model 2P377B, sold by Granger, Inc.), plumbing fittings , hose (s) for transferring (e.g. food grade hoses sold by Sani-Tech West) and ball valve (s) to storage / packaging tank 104. Transfer is carried out by switching on the transfer pump and opening the ball valves. Additional ingredients may be added, e.g., pH regulators, e.g., during pH control, suitable for adjusting the nanoemulsion to a suitable pH, e.g. between about 2.6 and 3.2. Flavorings can also be added. The additional ingredients are mixed in the concentrate using a standard mixer 111. The addition and mixing of the additional ingredients and / or evaluation can be carried out in a storage / packaging tank 104. Alternatively, they can be carried out before transfer to a storage / packaging tank, e.g. in a water phase tank 103.

[0437] To prepare concentrates, variations of this exemplary process at an enlarged scale (Figure 1) can also be performed using methods, including any of the variations described herein. For example, by eliminating and / or modifying one or more steps and / or equipment in accordance with the general methods described herein.

D. METHODS OF MAKING LIQUID DILUTED COMPOSITIONS CONTAINING DILUTED CONCENTRATES [0438] Also described herein are methods for diluting liquid nanoemulsion concentrates to

114

The preparation of liquid diluted compositions, usually aqueous liquid diluted compositions containing non-polar compounds. Typically, the nanoemulsion concentrate is diluted in an aqueous medium, e.g., a beverage, e.g., carbonated drink, water, milk, juice, regeneration drink, nutritional drink, nutritional supplement, or other aqueous food or drink. The concentrate and aqueous medium can be mixed, for example by stirring and / or mixing, or using any known mixing agent. The concentrate is dispersed in an aqueous medium to produce an aqueous liquid diluted composition, for example a clear or partially clear aqueous liquid diluted composition. The aqueous liquid diluted composition can be evaluated, for example to assess the clarity, taste, smell and / or stability of the liquid.

[0439] In one example, the nanoemulsion liquid concentrate is diluted in an aqueous medium, e.g., water by heating the aqueous medium, e.g., water by heating the aqueous medium, e.g., to at least 40 ° C or at least about 40 ° C, on 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, with appropriate dilution, as described herein, to the heated aqueous medium, and mixed until it is dispersed or dissolved in water. The resulting diluted liquid composition can then be cooled, e.g. to room temperature, e.g., 25 ° C or about 25 ° C. After dilution, the aqueous liquid dilution composition may be packaged, for example, by transfer to containers, e.g. vials or beverage containers. In one example, part of the liquid diluted composition is transferred to vials for analysis, e.g., evaluation of properties such as clarity, turbidity, taste, smell, ring formation, crystal formation and / or other properties.

[0440] Examples of equipment used to dilute liquid nanoemulsion concentrates to produce liquid diluted compositions containing diluted concentrates are beakers, for example Pyrex® glass beakers, hotplates, for example a Thermolyne hob, model number 846925 or model number SP46615, stirring rods, temperature measuring instruments, e.g. temperature sensors, e.g. Cooper temperature sensors (model No. DPP400W) and scales, for example, OHUAS scale 2.0 kg (model # CS2000) and / or Sartorius Analytical Scale (model BA 110S.

1. Dilutions [0441] Usually, the concentrates provided can be diluted in an aqueous medium to produce aqueous liquid compositions diluted over a wide dilution range. In one example, the concentrate can be diluted such that the aqueous liquid dilution composition contains between 0.05 g or about 0.05 g and 10 g or about 10 g, usually between 0.05 g and 5 g of liquid concentrate per 8 ounces of liquid volume, at least 8 ounces liquid volume or less than 8 ounces liquid volume, or per serving of liquid. For example, the concentrate can be diluted such that the aqueous liquid dilution composition contains 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, I g, 2 g, 3 g, 4 g, 5 g, 6 g, 7 g, 8 g, 9 g or 10 g of concentrate per 8 ounces volume, about 8 ounces volume or at least 8 ounces volume or at least about 8 ounces volume of aqueous medium, e.g. 8, 9, 10, 11, 12 , 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 100, 200 or more ounces of water medium volume.

115

[0442] 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 liquid concentrate, for example 1 ml, 2 ml, 3 ml, 4ml, 5ml, 6ml, 7ml, 8ml, 9ml or 10ml concentrate per 8 oz volume, about 8 oz volume, at least 8 oz volume or at least about 8 oz volume, or less than 8 ounces of volume or less than about 8 ounces of volume, or per portion of the aqueous medium, e.g. 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 100, 200 or more ounces of water medium volume.

[0443] In another example, the liquid concentrate is diluted such that the aqueous liquid dilution composition contains at least 10 mg or about 10 mg, usually at least 25 mg or about 25 mg, usually at least 35 mg of a non-polar compound, e.g. a non-polar compound active ingredient, on 8 ounces volume (0.236588 liters) or about 8 ounces volume, at least 8 ounces volume or at least about 8 ounces volume (0.236588 liters) of aqueous medium, either me than 8 ounces or less than about 8 ounces, or on a portion size of an 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 non-polar compound in at least 8 ounces volume (0.236588 liter) or at least about 8 ounces volume aqueous medium.

2. Analysis of aqueous liquid diluted compositions containing diluted concentrates [0444] The properties of aqueous liquid diluted compositions containing liquid concentrates can be evaluated using a variety of different evaluation methods. For example, clarity can be assessed; indications for human consumption, for example, a pleasant taste and / or smell, no "fish" taste / smell, no "ring formation" and no crystal formation; stability, for example, no oxidation, "ring formation", precipitation and / or visible phase separation over time; and safety for human consumption. Many of these properties can be judged empirically, for example by observing the liquid at once or over time, or by sniffing and / or tasting the liquid. In one example, after evaluating the aqueous liquid diluted compositions, the concentrates are reformulated to correct one or more parameters. In another example, the dilution factor can be corrected.

a. Clarity / turbidity [0445] The clarity of the aqueous liquid dilution compositions can be assessed using one or more approaches, for example empirical observation, particle size measurement and / or turbidity value measurement. The measurement can be qualitative or quantitative. In one example, a specific qualitative or quantitative value of clarity is determined. In another example, the clarity of a liquid can be expressed in terms of the clarity of another liquid, e.g., an aqueous dilute liquid composition prepared according to the provided methods, or a beverage, e.g., a beverage that does not contain a liquid concentrate. In this example, the liquid may be as clear as, less clear or more clear than other liquid. For example, an aqueous liquid dilution composition containing a liquid concentrate diluted in a drink may be like this

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EP2 268 160 B1 clear or almost as clear as the same beverage that does not contain the concentrate. Any type of evaluation can be performed qualitatively, e.g. by empirical evaluation, or quantitatively, e.g. by measuring particle size or turbidity.

i. Empirical evaluation [0446] In one example, the clarity / turbidity of the aqueous liquid dilution composition is qualitatively assessed, for example by observation. In one example, a liquid is considered clear if it does not appear cloudy and / or contains no particles or has few particles that can be seen with the naked eye. In another example, the liquid may be considered relatively clear or relatively cloudy based on comparison with other liquids, e.g., water, fruit juice, carbonated drink and / or milk and / or other aqueous liquid composition (s) diluted, prepared according to methods provided. For example, the aqueous liquid dilution composition may be as clear or almost as clear as water or another liquid, for example a drink. For example, a liquid containing a liquid concentrate diluted in a drink may be as clear or nearly as clear as a drink that does not contain a liquid concentrate. In a related example, the liquid may be clear or partially clear when there is no substantial difference, e.g., no observable difference, between the aqueous diluted liquid 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 very little) visible particles or turbidity may be clear. In another example, the lack of crystal formation or "ring formation" may indicate liquid clarity.

ii. Particle size [0447] In another example, clarity / turbidity is assessed by quantifying the particle size and / or number of particles in an aqueous liquid dilution composition. In this example, the clarity can be expressed as a representation of the particle size represented by the number, or as a comparison with the particle size of another liquid.

[0448] Methods of measuring liquid particle size are well known. Any method can be used to measure particle size as long as it is sensitive to the size of the particles in the expected and / or appropriate ranges of the provided aqueous liquid diluted compositions. For example, granulometric 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, e.g., using Delta Analytical Instruments, Inc., using a light scattering analyzer, e.g., dynamic light scattering analyzer, e.g., Horiba® LB-550, which can measure particle sizes in the 0.001 micron to 6 micron range and uses the Fourier transform / iterative deconvolution technique for the data obtained and can measure sample concentrations from ppm to 40% solids; Horiba® LA-920, which is a laser light scattering device having a He-Nu laser and tungsten lamp which can determine particle sizes from 0.02 microns to 2000 microns using Mie theory; or other analyzers available from Delta Analytical Instruments, Inc.

[0449] Optionally, particle size can be measured by observing the liquid under a microscope under magnification, for example 640X magnification. Particle size can be measured by comparison

117

EP2 268 160 B1 with a measuring standard, e.g. a ruler, which is also observed under magnification. In one example, particles of about 25 nm or larger than about 25 nm are visible, while particles smaller than 25 nm are not visible, for example at 640X magnification.

iii. Turbidity measurement [0450] In another example, the clarity / turbidity of liquids is assessed and / or expressed using a turbidity measurement, for example Nephelometric Turbidity Units (NTU). In this example, turbidity is measured optically to obtain a value indicating the turbidity or haze of the liquid that is correlated with the number and size of particles suspended in the liquid. The clearer the liquid, the lower its turbidity value. Turbidity can be measured optically, for example using a nephelometer, an instrument with a light source and a detector. The nephelometer measures turbidity by detecting the scattered light obtained by irradiating an aqueous liquid composition diluted with incident light. The amount of scattered light is correlated with the amount and size of the solid particles in the liquid, and thus the clarity. For example, a light beam passes through a low-turbidity sample with little distortion, giving very poorly diffused light, giving a low turbidity (NTU) reading. Other turbidity measurement methods may be used, including commercial turbidity measurement devices, e.g., devices available from ACZ Laboratories, Inc., Steamboat Springs, CO.

[0451] The following examples are for illustrative purposes only and are not intended to limit the scope of the invention.

E. EXAMPLES

Example 1: General procedure used to prepare liquid nanoemulsion concentrates in Examples 2-4.

[0452] The following tables 2A (i) to 4A show the ingredients that were used to make the many liquid nanoemulsion concentrates described in more 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 as a polar solvent contained water and a tocopherol polyethylene glycol succinate (TPGS) surfactant (TPGS surfactant sold under the name Vitamin E TPGS® by Eastman Chemical Company).

[0453] Tables 2A (i) to 4A each show milligrams (mg) per serving (serving size is indicated) of each ingredient in the concentrate, percentage, weight (total concentrate), for each ingredient, and amount (g) of each ingredient per batch of the indicated batch size (g). In each table, in the "phase" column, it is also indicated whether each component was added to the water phase ("water"), oil phase ("oil") or was added later to the emulsion that was formed by emulsifying the oil phase and water ("emulsion").

[0454] Each of the liquid nanoemulsion concentrates shown in Examples 2-4 were prepared using a laboratory scale process according to the provided methods. Each concentrate can be prepared alternatively by extending the process on a laboratory scale to

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The preparation of concentrates using the manufacturing process provided methods at an enlarged scale, for example to produce larger batch sizes in the examples below.

[0455] To prepare the concentrates in Examples 2-4, the laboratory scale process was carried out using the following general steps (details are indicated in the specific examples).

[0456] To make each of the liquid nanoemulsion concentrates shown in Examples 2-4 below, the indicated amounts of each component were weighed using Toledo Scale (model GD13x / USA), Sartorius Basic Analytical Scale (model BA110S) or OHAUS Scale (Model CS2000) . The choice of weight (weights) depended on the weight of the specific ingredient (s).

[0457] To form the water phase, the water phase ingredients (indicated by "water" in each table in the "phase" column) were added, in the indicated amounts (g / batch) to the water phase vessel (beaker Pyrex®) and mixed using standard stirrer (IKA® model No. RE-16 1S, which is a stand stirrer (laboratory stirrer) compatible with the process on a laboratory scale). While stirring, the aqueous phase ingredients were heated using a heating device. The heating device was a hot plate (Thermolyne hot plate model # SP46615, Barnstead International, Dubuque, Iowa). In addition, where indicated, when the aqueous phase contained water and an emulsion stabilizer, these components were added sequentially, in the following order: 1) water; 2) emulsion stabilizer. The aqueous phase ingredients were heated using a hot plate until the temperature reached 60 ° C. The aqueous phase was then kept at 60 ° C until the oil and water phases were combined and emulsified. To determine (measure) the temperature of the aqueous phase, a temperature measuring instrument (temperature sensor (model # DPP400W, Cooper-Atkins)) was used.

[0458] The oil phase ingredients (indicated by "oil" in each table in the "phase" column) were added to the oil phase vessel (Pyrex® beaker) and mixed using a standard stirrer (IKA® model No. RE-16 1S, which was the stirrer stand (laboratory stirrer) compatible with the process on a laboratory scale). In general, unless otherwise indicated, when the oil phase contained two or more of a surfactant, a preservative, a non-polar solvent and an emulsion stabilizer, these components were added sequentially, in the following order: 1) surfactant; 2) preservative; 3) non-polar solvent; 4) surfactant; 5) non-polar compound and 6) emulsion stabilizer. For example, when the oil phase contained a surfactant, preservative and non-polar compound, these ingredients were added in the following order (unless otherwise indicated): 1) surfactant, 2) preservative; 3) non-polar relationship.

[0459] When the oil phase ingredients were mixed, they were heated using a heating apparatus. The heating device was a hot plate (Thermolyne hot plate model # SP46615, Bamstead International, Dubuque, Iowa). The oil phase ingredients were heated until the mixture reached 60 ° C. The oily phase was stirred at this temperature until all ingredients were dissolved and kept at 60 ° C until mixed with the aqueous phase. To determine (measure) the oil phase temperature, a temperature measuring instrument (temperature sensor (model # DPP400W, Cooper-Atkins)) was used.

[0460] After both phases reached 60 ° and the oil phase components dissolved, the phases were combined and

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EP2 268 160 B1 was emulsified using the following steps. A reversible homogenizer (Arde Barinco, Inc .; Model CJ-4E) was placed in the aqueous phase vessel and turned on at 850-1200 rpm. The oil phase was then transferred to the water phase vessel by pouring the oil phase from the oil phase vessel to the water phase vessel. Mixing in the homogenizer was continued by adjusting the septum in the homogenizer to obtain and preserve the emulsion, for example by moving the septum further towards the forming emulsion and / or from the forming emulsion.

[0461] Homogenization of the forming emulsion was continued between 850 and 1200 rpm, with rapid cooling. Rapid cooling was achieved by placing an oil phase vessel (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 (specified temperatures indicated: typically lasting between about 30 minutes and about 60 minutes).

[0462] After emulsification and rapid cooling, additional ingredients were added as indicated in the individual examples / tables. For example, any ingredient that has been added after the emulsification step (instead of to one of the individual phases) is indicated by the word "emulsion" in the "phase" column. During the addition of any additional ingredients, the emulsion was mixed using a standard stirrer (IKA® model No. RE-16 1S). Exemplary additional ingredients that were added in the examples below are flavors (D-limonene and lemon oil) and pH regulators (e.g. citric acid). In several examples (where indicated), the pH of the emulsion was measured using a pH and temperature measuring instrument (Hanna Instruments, model HI 8314). When needed, the pH was adjusted using an appropriate amount of pH adjuster (the amount is indicated in the tables), e.g. citric acid or phosphoric acid, until the emulsion reached a pH between 2.6 and 3.2. Each of the concentrates prepared in the examples below had a pH between about 2.6 and 3.2.

[0463] As a final step, the concentrates were filtered using a 100 micron end product filter before further evaluation, dilution and / or use.

Example 2: Liquid nanoemulsion concentrates with non-polar PUFA containing compounds [0464] Examples 2A-2D show details of liquid nanoemulsion concentrates containing non-polar compounds (non-polar active ingredients), which include polyunsaturated fatty acids (PUFA) (e.g., non-polar active ingredients containing fatty acids omega-3, omega-6 fatty acids, conjugated fatty acids and other fatty acids). These concentrates were prepared using the general procedures described in Example 1 above.

Example 2A: Liquid nanoemulsion concentrates with non-polar compounds containing omega-3 [0465] Examples 2A (i) -2A (xii) provide details of liquid nanoemulsion concentrates containing non-polar compounds that include omega-3 fatty acids (e.g., DHA, EPA, ALA). These concentrates were prepared using the general procedures described in Example 1 above.

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EP 2 268 160 B1

Example 2A (i): Liquid nanoemulsion concentrate with 5% non-polar compound (fish oil) containing DHA and 18% TPGS surfactant [0466] Table 2A (i) below shows the ingredients used to make 500 g of a batch of liquid nanoemulsion concentrate containing DHA (portion size 2 ml), which was prepared according to the procedure set out in Example 1 above, with the following details and modifications:

[0467] To form the aqueous phase using the method described in Example 1 above, the following components of the aqueous phase were sequentially added in the following order to the aqueous phase vessel: 1) water, 2) emulsion stabilizer. The water was purified municipal water, which, prior to adding the aqueous phase to the vessel, was purified according to the methods provided by passing through using the following purifiers, in order: carbon filter, ion exchange purifier, reverse osmosis purifier and endpoint filter, e.g. endpoint filter 100 microns. The emulsion stabilizer was the SALADIZER® emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

[0468] To form the oil phase using the method described in Example 1 above, the following oil phase ingredients were added in succession, in the following order, to the oil phase vessel: 1) surfactant, 2) preservative; 3) surfactant; 4) non-polar active ingredient and 5) emulsion stabilizer.

[0469] The non-polar active ingredient was fish oil containing about 70% (74% DHA and about 10% (9.3%) EPA (Omega-3 Fish Oil EE, manufactured by O3C Nutraceuticals, supplied by

Jedwards International Inc., Quincy, MA). The non-polar active ingredient was added in such an amount that the active ingredient should constitute 5% by weight of the final concentrate. The co-surfactant was a phosphatidylcholine co-surfactant, sold under the trade name S-100 by Lipoid, LLC, Newark, NJ. Phosphatidylcholine was obtained from soybean extract and contained more than 95% phosphatidylcholine. The preservative was a natural (GRAS certified) preservative, benzyl alcohol. The emulsion stabilizer was the SALADIZER® emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

[0470] Flavors (lemon oil sold by Mission Flavors, Foothill Ranch, CA; and GRAS 99% D-Limonene certified, sold by Florida Chemical, Winter Haven, FL) were added after emulsification and the oil and water phases were cooled quickly . After forming the emulsion, an appropriate amount of citric acid (shown 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.

Table 2A (i): Liquid nanoemulsion concentrate with 5% non-polar compound (fish oil) containing DHA and 18% surfactant TPGS

<td>Ingredient</td><td>Portion mg / 2 ml</td><td>Phase</td><td>Percentage (by weight) of concentrate</td><td>g / batch</td>

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

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

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<td>Ingredient</td><td>Portion mg / 2 ml</td><td>Phase</td><td>Percentage (by weight) of concentrate</td><td>g / batch</td>

<td>Mark emulsion stabilizer SALADIZER® trademark (a mixture of xanthan gum, guar gum and sodium alginate)</td><td> 6,8</td><td>Water</td><td> 0,34</td><td> 1,7</td>

<td>Tocopherol polyethylene glycol succinate (surfactant)</td><td> 360</td><td>Oil</td><td> 18</td><td> 90</td>

<td>Mark emulsion stabilizer SALADIZER® trademark (a mixture of xanthan gum, guar gum and sodium alginate)</td><td> 1,27</td><td>Oil</td><td> 0,0635</td><td> 0,3175</td>

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

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

<td>Lemon oil (flavoring agent</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>Oil</td><td> 0,669</td><td> 3,345</td>

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

<td>In total</td><td> 2000,000</td><td></td><td> 100,000</td><td> 500</td>

Example 2A (ii): Liquid nanoemulsion concentrate with 5% non-polar compound (algae oil) containing DHA and 18% TPGS surfactant [0471] The following Table 2A (ii) shows the ingredients used to make 200 g of a lot of liquid nanoemulsion concentrate, containing DHA (portion size 2 ml), which was prepared according to the procedure set out in Example 1 above, with the following details and modifications:

[0472] To form the oil phase using the method described in Example 1 above, the following oil phase ingredients were sequentially added in the following order to the oil phase vessel: 1) surfactant, 2) preservative; 3) non-polar active ingredient and

4) emulsion stabilizer.

[0473] The non-polar active ingredient was algae oil containing 35% DHA. The non-polar active ingredient was added in such an amount that the ingredient should constitute 5% by weight (w / w) of the final concentrate. The preservative was a natural (GRAS certified) preservative, benzyl alcohol. The emulsion stabilizer was a trademark emulsion stabilizer

SALADIZER®, obtained from TIC Gums, Inc., Belcamp, MD.

[0474] To form 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 vessel: 1) water, 2) emulsion stabilizer. The water was purified municipal water, which, prior to adding the aqueous phase to the vessel, was purified according to the methods provided by passing through using the following purifiers, in order: carbon filter, ion exchange purifier, reverse osmosis purifier and endpoint filter, e.g. endpoint filter 100 microns. The emulsion stabilizer was the SALADIZER® emulsion stabilizer obtained from TIC

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Gums, Inc., Belcamp, MD.

[0475] After forming the emulsion, an appropriate amount of citric acid (shown 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): Liquid nanoemulsion concentrate with 5% non-polar compound (algae oil) containing DHA and 18% TPGS surfactant

<td>Ingredient</td><td>Portion mg / 2 ml</td><td>Phase</td><td>Percentage (by weight) of concentrate</td><td>g / batch</td>

<td>Algae oil (35% DHA)</td><td> 100</td><td>Oil</td><td> 5</td><td> 10</td>

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

<td>Stabilizer of the SALADIZER® trademark (a mixture of xanthan gum, guar gum and sodium alginate)</td><td> 6,8</td><td>Water</td><td> 0,34</td><td> 0,68</td>

<td>Tocopherol polyethylene glycol succinate (surfactant)</td><td> 360</td><td>Oil</td><td> 18</td><td> 36</td>

<td>Stabilizer of the SALADIZER® trademark (a mixture of xanthan gum, guar gum and sodium alginate)</td><td> 1,27</td><td>Oil</td><td> 0,0635</td><td> 0,127</td>

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

<td>Citric acid (pH regulator)</td><td> 5,6</td><td>Emulsion</td><td> 0,28</td><td> 0,56</td>

<td>In total</td><td> 2000,000</td><td></td><td> 100,000</td><td> 200</td>

Example 2A (iii): Liquid nanoemulsion concentrate with 5% non-polar compound (algae oil) containing DHA and 25.2% TPGS surfactant [0476] The following Table 2A (iii) shows the ingredients used to make 150 g batches of liquid nanoemulsion concentrate , containing DHA (portion size 2 ml), which was prepared according to the procedure set out in Example 1 above, with the following details and modifications:

[0477] To form the oil phase using the method described in Example 1 above, the following oil phase ingredients were sequentially added 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 algae oil containing 35% DHA. The non-polar active ingredient was added in such an amount that the ingredient should constitute 5% by weight (w / w) of the final concentrate. The preservative was a natural (GRAS certified) preservative, benzyl alcohol. The emulsion stabilizer was the SALADIZER® emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

[0478] To form the aqueous phase using the methods described in Example 1 above, the following components of the aqueous phase were sequentially added in the following order to the aqueous phase vessel: 1) water, 2) emulsion stabilizer. The water was purified municipal water, which, prior to adding the aqueous phase to the vessel, was purified according to the methods provided by passing through, using the following purifiers, in order: carbon filter, purifier

123

Ion exchange, reverse osmosis purifier and endpoint filter, e.g. 100 micron endpoint filter. The emulsion stabilizer was the SALADIZER® emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

[0479] After forming the emulsion, an appropriate amount of citric acid (shown 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): Liquid nanoemulsion concentrate with 5% non-polar compound (algae oil) containing DHA and 25.2% TPGS surfactant

<td>Ingredient</td><td>Portion mg / 2 ml</td><td>Phase</td><td>Percentage (by weight) of concentrate</td><td>g / batch</td>

<td>Algae oil (35% DHA)</td><td> 100</td><td>Oil</td><td> 5</td><td> 7,5</td>

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

<td>Stabilizer of the SALADIZER® trademark (a mixture of xanthan gum, guar gum and sodium alginate)</td><td> 3,4</td><td>Water</td><td> 0,17</td><td> 0,255</td>

<td>Tocopherol polyethylene glycol succinate (surfactant)</td><td> 504</td><td>Oil</td><td> 25,2</td><td> 37,8</td>

<td>Stabilizer of the SALADIZER® trademark (a mixture of xanthan gum, guar gum and sodium alginate)</td><td> 1,27</td><td>Oil</td><td> 0,0635</td><td> 0,09525</td>

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

<td>Citric acid (pH regulator)</td><td> 5,6</td><td>Emulsion</td><td> 0,28</td><td> 0,42</td>

<td>In total</td><td> 2000,000</td><td></td><td> 100,000</td><td> 150</td>

Example 2A (iv): Liquid nanoemulsion concentrate with 5% non-polar compound (linseed oil) containing ALAi 17.75% TPGS surfactant [0480] The following Table 2A (iv) shows the ingredients used to make 500 g of a batch of liquid nanoemulsion concentrate containing acid alpha-linolenic (ALA) (2 ml serving size), which was prepared according to the procedure set out in Example 1 above, with the following details and modifications:

[0481] To form the oil phase using the method described in Example 1 above, the following oil phase ingredients were sequentially added in the following order to the oil phase vessel: 1) surfactant, 2) preservative; 3) non-polar solvent; 4) surfactant; 5) non-polar active ingredient and 6) emulsion stabilizer.

[0482] 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), which contained not less than (NLT) 50% alpha linolenic acid C18: 3. The non-polar active ingredient was added in such an amount that the ingredient should constitute 5% by weight (w / w) of the final concentrate.

[0483] The co-surfactant was a phosphatidylcholine co-agent

124

EP 2 268 160 B1 surfactant, sold under the trade name S-100, by Lipoid, LLC, Newark, NJ. This phosphatidylcholine surfactant was obtained from soy extract and contained more than 95% phosphatidylcholine. The non-polar solvent was vitamin E oil, sold by ADM Natural Health and Nutrition, Decatur, IL, under the name Novatol ™ 5-67 Vitamin E (D-alpha5 Tocopherol; product code ADM 410217). This oil contained at least 67.2% tocopherol and about 32.8% soybean oil. The preservative was a natural (GRAS certified) preservative, benzyl alcohol. The emulsion stabilizer was the SALADIZER® emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

[0484] To form the aqueous phase using the methods described in Example 1 above, the following components of the aqueous phase were sequentially added in the following order to the aqueous phase vessel: 1) water, 2) emulsion stabilizer.

[0485] The water was purified municipal water, which, prior to adding the aqueous phase to the vessel, was purified according to the provided methods by passing through, using the following purifiers in turn: a carbon filter, an ion exchange purifier, a reverse osmosis purifier and an endpoint filter, for example 100 micron endpoint filter. The emulsion stabilizer was the SALADIZER® emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

[0486] After forming the emulsion, an appropriate amount of citric acid (shown 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): Liquid nanoemulsion concentrate with 5% non-polar compound (linseed oil) containing ALA and 17.75% TPGS surfactant

<td>Ingredient</td><td>Portion mg / 2 ml</td><td>Phase</td><td>Percentage (by weight) of concentrate</td><td>g / batch</td>

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

<td>Stabilizer of the SALADIZER® trademark (a mixture of xanthan gum, guar gum and sodium alginate)</td><td> 1,270</td><td>Oil</td><td> 0,06</td><td> 0,3</td>

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

<td>Stabilizer of the SALADIZER® trademark (a mixture of xanthan gum, guar gum and sodium alginate)</td><td> 6,800</td><td>Water</td><td> 0,34</td><td> 1,7</td>

<td>Tocopherol polyethylene glycol succinate (surfactant)</td><td> 355,0</td><td>Oil</td><td> 17,75</td><td> 88,75</td>

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

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

<td>Linseed oil 50% omega acids</td><td> 100,0</td><td>Oil</td><td> 5,0000</td><td> 25,000</td>

<td>Citric acid (pH regulator)</td><td> 3,800</td><td>Emulsion</td><td> 0,1900</td><td> 0,9500</td>

<td>In total</td><td> 2000,00</td><td></td><td> 100,000</td><td> 500</td>

125

EP 2 268 160 B1

Example 2A (v): Liquid nanoemulsion concentrate with 10% non-polar compound (algae oil) containing DHA and 20.2% TPGS surfactant [0487] The following Table 2A (v) shows the ingredients used to make 500 g batches of liquid nanoemulsion concentrate containing DHA (portion size 2 ml), which was prepared according to the procedure set out in Example 1 above, with the following details and modifications:

[0488] To form the oil phase using the method described in Example 1 above, the following oil phase ingredients were sequentially added in the following order to the oil phase vessel: 1) surfactant, 2) preservative; 3) non-polar active ingredient and 4) emulsion stabilizer.

[0489] The non-polar active ingredient was algae oil containing 35% DHA. The non-polar active ingredient was added in such an amount that the ingredient should constitute 10% by weight (w / w) of the final concentrate. The preservative was a natural (GRAS certified) preservative, benzyl alcohol. The emulsion stabilizer was the SALADIZER® emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

[0490] To form the aqueous phase using the methods described in Example 1 above, the following components of the aqueous phase were sequentially added in the following order to the aqueous phase vessel: 1) water, 2) emulsion stabilizer.

[0491] The water was purified municipal water, which, prior to adding the aqueous phase to the vessel, was purified according to the provided methods by passing through using the following purifiers in turn: a carbon filter, an ion exchange purifier, a reverse osmosis purifier and an endpoint filter, for example 100 micron endpoint filter. The emulsion stabilizer was the SALADIZER® emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

[0492] After forming the emulsion, an appropriate amount of citric acid (shown 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 above.

Table 2A (v): Liquid nanoemulsion concentrate with 10% non-polar compound (algae oil) containing DHA and 25.2% TPGS surfactant

<td>Ingredient</td><td>Portion mg / 2 ml</td><td>Phase</td><td>Percentage (by weight) of concentrate</td><td>g / batch</td>

<td>Algae oil (35% DHA)</td><td> 200</td><td>Oil</td><td> 10</td><td> 50</td>

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

<td>Stabilizer of the SALADIZER® trademark (a mixture of xanthan gum, guar gum and sodium alginate)</td><td> 1,2</td><td>Water</td><td> 0,06</td><td> 0,3</td>

<td>Tocopherol polyethylene glycol succinate (surfactant)</td><td> 504</td><td>Oil</td><td> 25,2</td><td> 126</td>

<td>Stabilizer of the SALADIZER® trademark (a mixture of xanthan gum, guar gum and sodium alginate)</td><td> 0,44</td><td>Oil</td><td> 0,022</td><td> 0,11</td>

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

126

EP 2 268 160 B1

<td>Ingredient</td><td>Portion mg / 2 ml</td><td>Phase</td><td>Percentage (by weight) of concentrate</td><td>g / batch</td>

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

<td>In total</td><td> 2000,000</td><td></td><td> 100,0000</td><td> 500</td>

Example 2A (vi): Liquid nanoemulsion concentrate with 5% non-polar compound (fish oil) containing DHA and 25.2% TPGS surfactant [0493] Table 2A (vi) below shows the ingredients used to make 250 g of a batch of liquid nanoemulsion concentrate containing DHA (portion size 2 ml), which was prepared according to the procedure outlined in example 1 above, with the following details and modifications:

[0494] To form the oil phase using the method described in Example 1 above, the following oil phase ingredients were sequentially added in the following order to the oil phase vessel: 1) surfactant, 2) preservative; 3) non-polar active ingredient and 4) emulsion stabilizer.

[0495] The non-polar active ingredient was Denomega ™ 100, a fish oil that contained about 13% DHA and about 13 EPA. The non-polar active ingredient was added in such an amount that the ingredient should constitute 5% by weight of the final concentrate. The preservative was a natural (GRAS certified) preservative, benzyl alcohol. The emulsion stabilizer was the SALADIZER® emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD. To form the aqueous phase using the methods described in Example 1 above, the following aqueous phase ingredients were added sequentially to water phase vessels: 1) water; 2) emulsion stabilizer.

[0496] The water added to the aqueous phase was purified municipal water, which, prior to the addition of the aqueous phase to the vessel, was purified according to the provided methods by passing through, using the following purifiers in turn: a carbon filter, ion exchange purifier, reverse osmosis purifier and endpoint filter, for example, a 100 micron endpoint filter.

[0497] The emulsion stabilizer was the SALADIZER® emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

[0498] After forming the emulsion, an appropriate amount of citric acid (shown 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): Liquid nanoemulsion concentrate with 5% non-polar compound (fish oil) containing DHA and 25.2% TPGS surfactant

<td>Ingredient</td><td>Portion mg / 2 ml</td><td>Phase</td><td>Percentage (by weight) of concentrate</td><td>g / batch</td>

<td>Fish oil (about 13% EPA; 13% DHA)</td><td> 100</td><td>Oil</td><td> 5</td><td> 12,5</td>

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

<td>Stabilizer of the SALADIZER® trademark (a mixture of xanthan gum, guar gum and sodium alginate)</td><td> 3,4</td><td>Water</td><td> 0,17</td><td> 0,425</td>

127

EP 2 268 160 B1

<td>Ingredient</td><td>Portion mg / 2 ml</td><td>Phase</td><td>Percentage (by weight) of concentrate</td><td>g / batch</td>

<td>Tocopherol polyethylene glycol succinate (surfactant)</td><td> 504</td><td>Oil</td><td> 25,2</td><td> 63</td>

<td>Stabilizer of the SALADIZER® trademark (a mixture of xanthan gum, guar gum and sodium alginate)</td><td> 1,27</td><td>Oil</td><td> 0,0635</td><td> 0,15875</td>

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

<td>Citric acid (pH regulator)</td><td> 5,6</td><td>Emulsion</td><td> 0,28</td><td> 0,7</td>

<td>In total</td><td> 2000,000</td><td></td><td> 100,0000</td><td> 250</td>

Example 2A (vii): Liquid nanoemulsion concentrate with 5% non-polar compound (fish oil) containing DHA and 25.2% TPGS surfactant [0499] The following Table 2A (vii) shows the ingredients used to make 250 g of a batch of liquid nanoemulsion concentrate containing DHA (portion size 2 ml), which was prepared according to the procedure set out in Example 1 above, with the following details and modifications:

[0500] To form the oil phase using the method described in Example 1 above, the following oil phase ingredients were sequentially added in the following order to the oil phase vessel: 1) surfactant, 2) preservative; 3) non-polar active ingredient and

4) emulsion stabilizer.

[0501] The non-polar active ingredient was fish oil, containing about 70% (74%) DHA and about 10% (9.3%) EPA (Omega-3 Fish Oil EE, manufactured by O3C Nutraceuticals, supplied by Jedwards International Inc., Quincy, MA). The non-polar active ingredient was added in such an amount that the ingredient should constitute 5% by weight of the final concentrate. The preservative was a natural (GRAS certified) preservative, benzyl alcohol. The emulsion stabilizer was the SALADIZER® emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

[0502] To form the aqueous phase using the methods described in Example 1 above, the following components of the aqueous phase were sequentially added in the following order to the aqueous phase vessel: 1) water; 2) emulsion stabilizer.

[0503] The water was purified municipal water, which, prior to adding the aqueous phase to the vessel, was purified according to the provided methods by passing through using the following purifiers in turn: a carbon filter, an ion exchange purifier, a reverse osmosis purifier and an endpoint filter, for example 100 micron endpoint filter. The emulsion stabilizer was the SALADIZER® emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD. [0504] After forming the emulsion, an appropriate amount of citric acid (shown 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.

128

EP 2 268 160 B1

Table 2A (vii): Liquid nanoemulsion concentrate with 5% non-polar compound (fish oil) containing DHA and 25.2% TPGS surfactant

<td>Ingredient</td><td>Portion mg / 2 ml</td><td>Phase</td><td>Percentage (by weight) of concentrate</td><td>g / batch</td>

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

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

<td>Stabilizer of the SALADIZER® trademark (a mixture of xanthan gum, guar gum and sodium alginate)</td><td> 3,4</td><td>Water</td><td> 0,17</td><td> 0,425</td>

<td>Tocopherol polyethylene glycol succinate (surfactant)</td><td> 504</td><td>Oil</td><td> 25,2</td><td> 63</td>

<td>Stabilizer of the SALADIZER® trademark (a mixture of xanthan gum, guar gum and sodium alginate)</td><td> 1,27</td><td>Oil</td><td> 0,0635</td><td> 0,15875</td>

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

<td>Citric acid (pH regulator)</td><td> 5,6</td><td>Emulsion</td><td> 0,28</td><td> 0,7</td>

<td>In total</td><td> 2000,000</td><td></td><td> 100,0000</td><td> 250</td>

Example 2A (viii): Liquid nanoemulsion concentrate with 10% non-polar compound (fish oil) containing DHA and 20.2% TPGS surfactant [0505] The following Table 2A (viii) shows the ingredients used to make 250 g of a batch of liquid nanoemulsion concentrate containing DHA (portion size 2 ml), which was prepared according to the procedure set out in Example 1 above, with the following details and modifications:

[0506] To form the oil phase using the method described in Example 1 above, the following oil phase ingredients were sequentially added in the following order to the oil phase vessel: 1) surfactant, 2) preservative; 3) non-polar active ingredient and 4) emulsion stabilizer.

[0507] The non-polar active ingredient was Denomega ™ 100, a fish oil that contained about 13% DHA and about 13% EPA. The non-polar active ingredient was added in such an amount that the ingredient should constitute 10% by weight of the final concentrate. The preservative was a natural (GRAS certified) preservative, benzyl alcohol. The emulsion stabilizer was the SALADIZER® emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD. [0508] To form the aqueous phase using the methods described in Example 1 above, the following components of the aqueous phase were sequentially added, in the following order, to the aqueous phase vessel: 1) water; 2) emulsion stabilizer.

[0509] The water was purified municipal water, which, prior to adding the aqueous phase to the vessel, was purified according to the provided methods by passing through, using the following purifiers in turn: a carbon filter, an ion exchange purifier, a reverse osmosis purifier and an endpoint filter, for example 100 micron endpoint filter. The emulsion stabilizer was the SALADIZER® emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

129

[0510] After forming the emulsion, an appropriate amount of citric acid (shown 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): Liquid nanoemulsion concentrate with 10% non-polar compound (fish oil) containing DHA and 20.2% TPGS surfactant

<td>Ingredient</td><td>Portion mg / 2 ml</td><td>Phase</td><td>Percentage (by weight) of concentrate</td><td>g / batch</td>

<td>Fish oil (about 13% EPA; 13% DHA)</td><td> 200</td><td>Oil</td><td> 10</td><td> 25</td>

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

<td>Stabilizer of the SALADIZER® trademark (a mixture of xanthan gum, guar gum and sodium alginate)</td><td> 3,4</td><td>Water</td><td> 0,17</td><td> 0,425</td>

<td>Tocopherol polyethylene glycol succinate (surfactant)</td><td> 404</td><td>Oil</td><td> 20,2</td><td> 50,5</td>

<td>Stabilizer of the SALADIZER® trademark (a mixture of xanthan gum, guar gum and sodium alginate)</td><td> 1,27</td><td>Oil</td><td> 0,0635</td><td> 0,15875</td>

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

<td>Citric acid (pH regulator)</td><td> 5,6</td><td>Emulsion</td><td> 0,28</td><td> 0,7</td>

<td>In total</td><td> 2000,000</td><td></td><td> 100,0000</td><td> 250</td>

Example 2A (ix): Liquid nanoemulsion concentrate with 10% non-polar compound (fish oil) containing DHA and 20.2% TPGS surfactant [0511] The following Table 2A (ix) shows the ingredients used to make 250 g of a batch of liquid nanoemulsion concentrate containing DHA (portion size 2 ml), which was prepared according to the procedure set out in Example 1 above, with the following details and modifications:

[0512] To form the oil phase using the method described in Example 1 above, the following oil phase ingredients were sequentially added in the following order to the oil phase vessel: 1) surfactant, 2) preservative; 3) non-polar active ingredient and 4) emulsion stabilizer.

[0513] The non-polar active ingredient was fish oil containing about 70% (74%) DHA and about 10% (9.3%) EPA (Omega-3 Fish Oil EE, manufactured by O3C Nutraceuticals, supplied by Jedwards International Inc., Quincy, MA). The non-polar active ingredient was added in such an amount that the ingredient should constitute 10% by weight of the final concentrate. The preservative was a natural (GRAS certified) preservative, benzyl alcohol. The emulsion stabilizer was the SALADIZER® emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

[0514] To form the aqueous phase using the methods described in example 1 above, the following components of the aqueous phase were sequentially added in the following order to the phase vessel

130

EP 1 268 160 B1: 1) water; 2) emulsion stabilizer.

[0515] The water was purified municipal water, which, prior to adding the aqueous phase to the vessel, was purified according to the provided methods by passing through, using the following purifiers in turn: a carbon filter, an ion exchange purifier, a reverse osmosis purifier and an endpoint filter, for example 100 micron endpoint filter. The emulsion stabilizer was the SALADIZER® emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD. [0516] After forming the emulsion, an appropriate amount of citric acid (shown 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): Liquid nanoemulsion concentrate with 10% non-polar compound (fish oil) containing DHA and 20.2% surfactant TPGS

<td>Ingredient</td><td>Portion mg / 2 ml</td><td>Phase</td><td>Percentage (by weight) of concentrate</td><td>g / batch</td>

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

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

<td>Stabilizer of the SALADIZER® trademark (a mixture of xanthan gum, guar gum and sodium alginate)</td><td> 3,4</td><td>Water</td><td> 0,17</td><td> 0,425</td>

<td>Tocopherol polyethylene glycol succinate (surfactant)</td><td> 404</td><td>Oil</td><td> 20,2</td><td> 50,5</td>

<td>Stabilizer of the SALADIZER® trademark (a mixture of xanthan gum, guar gum and sodium alginate)</td><td> 1,27</td><td>Oil</td><td> 0,0635</td><td> 0,15875</td>

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

<td>Citric acid (pH regulator)</td><td> 5,6</td><td>Emulsion</td><td> 0,28</td><td> 0,7</td>

<td>In total</td><td> 2000,000</td><td></td><td> 100,0000</td><td> 250</td>

Example 2A (x): Liquid nanoemulsion concentrate with 5% non-polar compound (linseed oil) containing ALAi 25.2% TPGS surfactant [0517] The following table 2A (x) shows the ingredients used to make 250 g of the batch liquid nanoemulsion concentrate containing acid alpha-linolenic (ALA) (2 ml serving size), which was prepared according to the procedure set out in Example 1 above, with the following details and modifications:

[0518] To form the oil phase using the method described in Example 1 above, the following oil phase ingredients were sequentially added in the following order to the oil phase vessel: 1) surfactant, 2) preservative; 3) non-polar active ingredient and 4) emulsion stabilizer.

[0519] 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), which contained not less than (NLT) 50% alpha linolenic acid C18: 3. Non-polar active ingredient

131

This amount was added in such an amount that the component should constitute 5% by weight (w / w) of the final concentrate. The surfactant was also added to the oil phase. The preservative was a natural (GRAS certified) preservative, benzyl alcohol. The emulsion stabilizer was the SALADIZER® emulsion stabilizer obtained from TIC

Gums, Inc., Belcamp, MD.

[0520] To form the aqueous phase using the methods described in Example 1 above, the following components of the aqueous phase were sequentially added, in the following order, to the aqueous phase vessel: 1) water; 2) emulsion stabilizer. The water was purified municipal water, which, prior to adding the aqueous phase to the vessel, was purified according to the methods provided by passing through using the following purifiers, in order: carbon filter, ion exchange purifier, reverse osmosis purifier and endpoint filter, e.g. endpoint filter 100 microns. The emulsion stabilizer was the SALADIZER® emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

[0521] After forming the emulsion, the appropriate amount of citric acid (shown in the table) was added

2A (x)) to adjust the pH of the emulsion to between 2.6 and 3.4 as described in Example 1 above.

Table 2A (x): Liquid nanoemulsion concentrate with 5% non-polar compound (linseed oil) containing ALAi 25.2% TPGS surfactant

<td>Ingredient</td><td>Portion mg / 2 ml</td><td>Phase</td><td>Percentage (by weight) of concentrate</td><td>g / batch</td>

<td>Linseed oil 50% omega acids</td><td> 100</td><td>Oil</td><td> 5</td><td> 12,5</td>

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

<td>Stabilizer of the SALADIZER® trademark (a mixture of xanthan gum, guar gum and sodium alginate)</td><td> 3,4</td><td>Water</td><td> 0,17</td><td> 0,425</td>

<td>Tocopherol polyethylene glycol succinate (surfactant)</td><td> 504</td><td>Oil</td><td> 25,2</td><td> 63</td>

<td>Stabilizer of the SALADIZER® trademark (a mixture of xanthan gum, guar gum and sodium alginate)</td><td> 1,27</td><td>Oil</td><td> 0,0635</td><td> 0,15875</td>

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

<td>Citric acid (pH regulator)</td><td> 5,6</td><td>Emulsion</td><td> 0,28</td><td> 0,7</td>

<td>In total</td><td> 2000,000</td><td></td><td> 100,0000</td><td> 250</td>

Example 2A (xi): Liquid nanoemulsion concentrate with 10% non-polar compound (linseed oil) containing ALAi 20.2% TPGS surfactant [0522] The following table 2A (xi) shows the ingredients used to make 250 g batch of liquid nanoemulsion concentrate containing alpha-linolenic acid (ALA) (2 ml serving size), which was prepared according to the procedure set out in Example 1 above, with the following details and modifications:

132

[0523] To form the oil phase using the method described in Example 1 above, the following oil phase ingredients were sequentially added in the following order to the oil phase vessel: 1) surfactant, 2) preservative; 3) non-polar active ingredient and 4) emulsion stabilizer.

[0524] 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), which contained not less than (NLT) 50% alpha linolenic acid C18: 3. The non-polar active ingredient was added in such an amount that the ingredient should constitute 10% by weight (w / w) of the final concentrate. The preservative was a natural (GRAS certified) preservative, benzyl alcohol. The emulsion stabilizer was the SALADIZER® emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

[0525] To form the aqueous phase using the methods described in Example 1 above, the following components of the aqueous phase were sequentially added in the following order to the aqueous phase vessel: 1) water; 2) emulsion stabilizer. The water was purified municipal water, which, prior to adding the aqueous phase to the vessel, was purified according to the methods provided by passing through using the following purifiers, in order: carbon filter, ion exchange purifier, reverse osmosis purifier and endpoint filter, e.g. endpoint filter 100 microns. The emulsion stabilizer was the SALADIZER® emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

[0526] After forming the emulsion, the appropriate amount of citric acid (shown in the table) was added

2A (xi)) 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 (linseed oil) containing ALAi 20.2% TPGS surfactant

<td>Ingredient</td><td>Portion mg / 2 ml</td><td>Phase</td><td>Percentage (by weight) of concentrate</td><td>g / batch</td>

<td>Linseed oil 50% omega acids</td><td> 200</td><td>Oil</td><td> 10</td><td> 25</td>

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

<td>Stabilizer of the SALADIZER® trademark (a mixture of xanthan gum, guar gum and sodium alginate)</td><td> 3,4</td><td>Water</td><td> 0,17</td><td> 0,425</td>

<td>Tocopherol polyethylene glycol succinate (surfactant)</td><td> 404</td><td>Oil</td><td> 20,2</td><td> 50,5</td>

<td>Stabilizer of the SALADIZER® trademark (a mixture of xanthan gum, guar gum and sodium alginate)</td><td> 1,27</td><td>Oil</td><td> 0,0635</td><td> 0,15875</td>

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

<td>Citric acid (pH regulator)</td><td> 5,6</td><td>Emulsion</td><td> 0,28</td><td> 0,7</td>

<td>In total</td><td> 2000,000</td><td></td><td> 100,0000</td><td> 250</td>

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Example 2A (xii): Liquid nanoemulsion concentrate with 5% non-polar compound (fish oil) containing DHAi 25.2% TPGS surfactant [0527] The following Table 2A (xii) shows the ingredients used to produce 250 g batch of liquid DHA containing nanoemulsion concentrate (portion size 2 ml), which was prepared according to the procedure set out in Example 1 above, with the following details and modifications:

[0528] To form the oil phase using the method described in Example 1 above, the following oil phase ingredients were sequentially added in the following order to the oil phase vessel: 1) surfactant, 2) preservative; 3) non-polar active ingredient and 4) emulsion stabilizer.

[0529] The non-polar active ingredient was fish oil, containing about 20% DHA and about 40% EPA (manufactured by O3C Nutraceuticals, supplied by Jedwards International Inc., Quincy, MA). The non-polar active ingredient was added in such an amount that the active ingredient should constitute 5% by weight of the final concentrate. The preservative was a natural (GRAS certified) preservative, benzyl alcohol. The emulsion stabilizer was the SALADIZER® emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

[0530] To form the aqueous phase using the methods described in Example 1 above, the following components of the aqueous phase were sequentially added, in the following order, to the aqueous phase vessel: 1) water; 2) emulsion stabilizer.

[0531] The water was purified municipal water which, prior to adding the aqueous phase to the vessel, was purified according to the provided methods by passing through using the following purifiers, in order: a carbon filter, an ion exchange purifier, a reverse osmosis purifier and an endpoint filter, for example 100 micron endpoint filter. The emulsion stabilizer was the SALADIZER® emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD. [0532] After forming the emulsion, the appropriate amount of citric acid (shown in the table) was added

2A (xii)) to adjust the pH of the emulsion to between 2.6 and 3.4, as described in Example 1 above.

Table 2A (xii): Liquid nanoemulsion concentrate with 5% non-polar compound (fish oil) containing DHAi 25.2% TPGS surfactant

<td>Ingredient</td><td>Portion mg / 2 ml</td><td>Phase</td><td>Percentage (by weight) of concentrate</td><td>g / batch</td>

<td>Fish oil (40% EPA; 20% DHA)</td><td> 100</td><td>Oil</td><td> 5</td><td> 12,5</td>

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

<td>Stabilizer of the SALADIZER® trademark (a mixture of xanthan gum, guar gum and sodium alginate)</td><td> 3,4</td><td>Water</td><td> 0,17</td><td> 0,425</td>

<td>Tocopherol polyethylene glycol succinate (surfactant)</td><td> 504</td><td>Oil</td><td> 25,2</td><td> 63</td>

<td>Stabilizer of the SALADIZER® trademark (a mixture of xanthan gum, guar gum and sodium alginate)</td><td> 1,27</td><td>Oil</td><td> 0,0635</td><td> 0,15875</td>

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EP2 268 160 B1

<td>Ingredient</td><td>Portion mg / 2 ml</td><td>Phase</td><td>Percentage (by weight) of concentrate</td><td>g / batch</td>

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

<td>Citric acid (pH regulator)</td><td> 5,6</td><td>Emulsion</td><td> 0,28</td><td> 0,7</td>

<td>In total</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): Liquid nanoemulsion concentrate with 5% non-polar compound (borage oil) containing GLA and 17.75% TPGS surfactant [0533] The following Table 2B (i) shows the ingredients used to make 500 g batches of liquid concentrate nano-emulsion containing GLA (gamma-linoleic acid) (portion size 2 ml), which was prepared according to the procedure set out in Example 1 above, with the following details and modifications:

[0534] To form the oil phase using the method described in Example 1 above, the following oil phase ingredients were sequentially added in the following order to the oil phase vessel: 1) surfactant, 2) preservative; 3) non-polar solvent; 4) surfactant; 5) non-polar active ingredient and 6) emulsion stabilizer.

[0535] The non-polar active ingredient was borage oil compound, obtained from Sanmark LLC, Greensboro, NC (Sanmark Limited, Dalian, Liaoning Province, China), which was obtained by extrusion and isolation from seeds of Borago officinalis L. This oil contained no less than (NLT) 22% gamma-linolenic acid (GLA) C18: 3. The non-polar active ingredient was added in such an amount that the ingredient should constitute 5% by weight of the final concentrate. The co-surfactant was a phosphatidylcholine co-surfactant, sold under the trade name S-100, by Lipoid, LLC, Newark, NJ. This phosphatidylcholine co-surfactant was obtained from soy extract and contained more than 95% phosphatidylcholine. The nonpolar solvent was 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). This oil contained at least 67.2% tocopherol and about 32.8% soybean oil. The preservative was a natural (GRAS certified) preservative, benzyl alcohol, and the emulsion stabilizer under the trademark SALADIZER®, obtained from TIC Gums, Inc., Belcamp, MD.

[0536] To form the aqueous phase using the methods described in Example 1 above, the following components of the aqueous phase were sequentially added, in the following order, to the aqueous phase vessel: 1) water; 2) emulsion stabilizer.

[0537] The water was purified municipal water, which, prior to adding the aqueous phase to the vessel, was purified according to the provided methods by passing through using the following purifiers, in order: a carbon filter, an ion exchange purifier, a reverse osmosis purifier and an endpoint filter, for example 100 micron endpoint filter. The emulsion stabilizer was the SALADIZER® emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD. [0538] After forming the emulsion, the appropriate amount of citric acid (shown in the table) was added

135

EP 2 268 160 B1

2B (i)) to adjust the pH of the emulsion to between 2.6 and 3.4 as described in Example 1 above.

Table 2B (i): Liquid nanoemulsion concentrate with 5% non-polar compound (borage oil) containing GLA and 17.75% TPGS surfactant

<td>Ingredient</td><td>Portion mg / 2 ml</td><td>Phase</td><td>Percentage (by weight) of concentrate</td><td>g / batch</td>

<td>Vitamin E (5-67) oil (nonpolar solvent)</td><td> 75,00</td><td>Oil</td><td> 3,750</td><td> 18,75</td>

<td>Stabilizer of the SALADIZER® trademark (a mixture of xanthan gum, guar gum and sodium alginate)</td><td> 1,270</td><td>Oil</td><td> 0,06</td><td> 0,3</td>

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

<td>Stabilizer of the SALADIZER® trademark (a mixture of xanthan gum, guar gum and sodium alginate)</td><td> 6,800</td><td>Water</td><td> 0,34</td><td> 1,7</td>

<td>Tocopherol polyethylene glycol succinate (surfactant)</td><td> 355,0</td><td>Oil</td><td> 17,75</td><td> 88,75</td>

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

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

<td>Borage oil 22% GLA</td><td> 100,0</td><td>Oil</td><td> 5,0000</td><td> 25,000</td>

<td>Citric acid (pH regulator)</td><td> 3,800</td><td>Emulsion</td><td> 0,1900</td><td> 0,9500</td>

<td>In total</td><td> 2000,00</td><td></td><td> 100,000</td><td> 500</td>

Example 2B (ii): Liquid nanoemulsion concentrate with 5% non-polar compound (borage oil) containing GLA and 25.2% TPGS surfactant [0539] The following Table 2B (ii) shows the ingredients used to make 250 g batches of liquid concentrate nano-emulsion containing GLA (gamma-linoleic acid) (portion size 2 ml), which was prepared according to the procedure set out in Example 1 above, with the following details and modifications:

[0540] To form the oil phase using the method described in Example 1 above, the following oil phase ingredients were sequentially added in the following order to the oil phase vessel: 1) surfactant, 2) preservative; 3); non-polar active ingredient and 4) emulsion stabilizer.

[0541] The non-polar active ingredient was borage oil compound, obtained from

Sanmark LLC, Greensboro, NC (Sanmark Limited, Dalian, Liaoning Province, China), which was obtained by extrusion and isolation from seeds of Borago officinalis L. This oil contained not less than (NLT) 22% of gamma-linolenic acid (GLA) C18: 3. The non-polar active ingredient was added in such an amount that the ingredient should constitute 5% by weight of the final concentrate. The preservative was a natural (GRAS certified) preservative, benzyl alcohol. Emulsion stabilizer

136

EP2 268 160 B1 was the SALADIZER® emulsion stabilizer obtained from TIC Gums, Inc., Belcamp, MD.

[0542] To form the aqueous phase using the methods described in Example 1 above, the following components of the aqueous phase were sequentially added, in the following order, to the aqueous phase vessel: 1) water; 2) emulsion stabilizer.

[0543] The water was purified municipal water, which prior to the addition of the aqueous phase to the vessel, was purified according to the provided methods by passing through using the following purifiers in turn: a carbon filter, an ion exchange purifier, a reverse osmosis purifier and an endpoint filter, for example 100 micron endpoint filter. The emulsion stabilizer was the SALADIZER® emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD. [0544] After forming the emulsion, an appropriate amount of citric acid (shown 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): Liquid nanoemulsion concentrate with 5% non-polar compound (borage oil) containing GLA and 25.2% TPGS surfactant

<td>Ingredient</td><td>Portion mg / 2 ml</td><td>Phase</td><td>Percentage (by weight) of concentrate</td><td>g / batch</td>

<td>Borage oil (22% GLA)</td><td> 100</td><td>Oil</td><td> 5</td><td> 12,5</td>

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

<td>Stabilizer of the SALADIZER® trademark (a mixture of xanthan gum, guar gum and sodium alginate)</td><td> 3,4</td><td>Water</td><td> 0,17</td><td> 0,425</td>

<td>Tocopherol polyethylene glycol succinate (surfactant)</td><td> 504</td><td>Oil</td><td> 25,2</td><td> 63</td>

<td>Stabilizer of the SALADIZER® emulsion (a mixture of xanthan gum, guar gum and sodium alginate)</td><td> 1,27</td><td>Oil</td><td> 0,0635</td><td> 0,15875</td>

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

<td>Citric acid (pH regulator)</td><td> 5,6</td><td>Emulsion</td><td> 0,28</td><td> 0,7</td>

<td>In total</td><td> 2000,000</td><td></td><td> 100,0000</td><td> 250</td>

Example 2B (iii): Liquid nanoemulsion concentrate with 10% non-polar compound (borage oil) containing GLA and 20.2% TPGS surfactant [0545] Table 2B (iii) below shows the ingredients used to make 250 g batches of liquid concentrate nano-emulsion containing GLA (gamma-linoleic acid) (portion size 2 ml), which was prepared according to the procedure set out in Example 1 above, with the following details and modifications:

[0546] To prepare the oil phase using the method described in Example 1 above, the following oil phase ingredients were sequentially added in the following order to the phase vessel

137

Oily: 1) a surfactant, 2) a preservative; 3); non-polar active ingredient and 4) emulsion stabilizer.

[0547] The non-polar active ingredient was borage oil compound, obtained from Sanmark LLC, Greensboro, NC (Sanmark Limited, Dalian, Liaoning Province, China), which was obtained by extrusion and isolation from seeds of Borago officinalis L. This oil contained no less than (NLT) 22% gamma-linolenic acid (GLA) C18: 3. The non-polar active ingredient was added in such an amount that the ingredient should constitute 10% by weight of the final concentrate. The preservative was a natural (GRAS certified) preservative, benzyl alcohol. The emulsion stabilizer was the SALADIZER® emulsion stabilizer, obtained from TIC Gums, Inc.,

Belcamp, MD.

[0548] To form the aqueous phase using the methods described in Example 1 above, the following components of the aqueous phase were sequentially added in the following order to the aqueous phase vessel: 1) water; 2) emulsion stabilizer.

[0549] The water was purified municipal water, which, prior to adding the aqueous phase to the vessel, was purified according to the provided methods by passing through using the following purifiers, in order: a carbon filter, an ion exchange purifier, a reverse osmosis purifier and an endpoint filter, for example 100 micron endpoint filter. The emulsion stabilizer was the SALADIZER® emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD. [0550] After forming the emulsion, the appropriate amount of citric acid (shown in the table) was added

2B (ii)) to adjust the pH of the emulsion to between 2.6 and 3.4 as described in Example 1 above.

Table 2B (iii): Liquid nanoemulsion concentrate with 10% non-polar compound (borage oil) containing GLA and 20.2% surfactant TPGS

<td>Ingredient</td><td>Portion mg / 2 ml</td><td>Phase</td><td>Percentage (by weight) of concentrate</td><td>g / batch</td>

<td>Borage oil (22% GLA)</td><td> 200</td><td>Oil</td><td> 10</td><td> 25</td>

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

<td>Stabilizer of the SALADIZER® trademark (a mixture of xanthan gum, guar gum and sodium alginate)</td><td> 3,4</td><td>Water</td><td> 0,17</td><td> 0,425</td>

<td>Tocopherol polyethylene glycol succinate (surfactant)</td><td> 404</td><td>Oil</td><td> 20,2</td><td> 50,5</td>

<td>Stabilizer of the SALADIZER® trademark (a mixture of xanthan gum, guar gum and sodium alginate)</td><td> 1,27</td><td>Oil</td><td> 0,0635</td><td> 0,15875</td>

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

<td>Citric acid (pH regulator)</td><td> 5,6</td><td>Emulsion</td><td> 0,28</td><td> 0,7</td>

<td>In total</td><td> 2000,000</td><td></td><td> 100,0000</td><td> 250</td>

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EP2 268 160 B1

Example 2C: Liquid nanoemulsion concentrates with non-polar compounds containing conjugated fatty acid

Example 2C (i): Liquid nanoemulsion concentrate with 5% non-polar compound containing CLA and 17.75% surfactant TPGS [0551] The following table 2C (i) shows the ingredients used to make 500 g of lot of liquid nanoemulsion concentrate containing CLA (portion size 2 ml), which was prepared according to the procedure in Example 1 above, with the following details and modifications:

[0552] To prepare the oil phase using the method described in Example 1 above, the following oil phase ingredients were sequentially added in the following order to the oil phase vessel: 1) surfactant, 2) preservative; 3) non-polar solvent; 4) surfactant; 5) non-polar active ingredient and 6) emulsion stabilizer.

[0553] The non-polar active ingredient was conjugated linolenic acid (CLA), obtained from Sanmark, LTD (Dalian, Liaoning Province, China; product code 01057-A80) containing 80% CLA. The non-polar active ingredient was added in such an amount that the ingredient should constitute 5% by weight (w / w) of the final concentrate. The co-surfactant was a phosphatidylcholine co-surfactant, sold under the trade name S-100, by Lipoid, LLC, Newark, NJ. This phosphatidylcholine co-surfactant was obtained from soy extract and contained more than 95% phosphatidylcholine. The non-polar solvent was 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). This oil contained at least 67.2% tocopherol and about 32.8% soybean oil. The preservative was a natural (GRAS certified) preservative, benzyl alcohol. The emulsion stabilizer was the SALADIZER® emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

[0554] To form the aqueous phase using the methods described in Example 1 above, the following components of the aqueous phase were sequentially added, in the following order, to the aqueous phase vessel: 1) water; 2) emulsion stabilizer.

[0555] The water was purified municipal water, which prior to the addition of the aqueous phase to the vessel, was purified according to the provided methods by passing through using the following purifiers in turn: a carbon filter, ion exchange purifier, reverse osmosis purifier and endpoint filter, for example 100 micron endpoint filter. The emulsion stabilizer was the SALADIZER® emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD. [0556] After forming the emulsion, an appropriate amount of citric acid (shown in Table 2C (i)) was added to adjust the pH of the emulsion to between 2.6 and 3.4, as described in Example 1 above.

139

EP 2 268 160 B1

Table 2C (i): Liquid nanoemulsion concentrate with 5% non-polar compound containing CLA and 17.75% surfactant TPGS

<td>Ingredient</td><td>Portion mg / 2 ml</td><td>Phase</td><td>Percentage (by weight) of concentrate</td><td>g / batch</td>

<td>Vitamin E (5-67) oil (nonpolar solvent)</td><td> 75,00</td><td>Oil</td><td> 3,750</td><td> 18,75</td>

<td>Stabilizer of the SALADIZER® trademark (a mixture of xanthan gum, guar gum and sodium alginate)</td><td> 1,270</td><td>Oil</td><td> 0,06</td><td> 0,3</td>

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

<td>Stabilizer of the SALADIZER® trademark (a mixture of xanthan gum, guar gum and sodium alginate)</td><td> 6,800</td><td>Water</td><td> 0,34</td><td> 1,7</td>

<td>Tocopherol polyethylene glycol succinate (surfactant)</td><td> 355,0</td><td>Oil</td><td> 17,75</td><td> 88,75</td>

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

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

<td>Oil containing 80% conjugated linoleic acid</td><td> 100,0</td><td>Oil</td><td> 5,0000</td><td> 25,000</td>

<td>Citric acid (pH regulator)</td><td> 3,800</td><td>Emulsion</td><td> 0,1900</td><td> 0,9500</td>

<td>In total</td><td> 2000,00</td><td></td><td> 100,000</td><td> 500</td>

Example 2C (ii): Liquid nanoemulsion concentrate with a 5% non-polar compound containing CLA and

25.2% TPGS surfactant [0557] The following table 2C (ii) shows the ingredients used to make 250 g batch of liquid CLA-containing nanoemulsion concentrate (2 ml batch size) which was prepared according to the procedure set out in Example 1 above, with the following details and modifications:

[0558] To form the oil phase using the method described in Example 1 above, the following oil phase ingredients were sequentially added in the following order to the oil phase vessel: 1) surfactant, 2) preservative; 3) non-polar active ingredient and 4) emulsion stabilizer.

[0559] The non-polar active ingredient was conjugated linolenic acid (CLA), obtained from Sanmark, LTD (Dalian, Liaoning Province, China; product code 01057-A80) containing

80% CLA. The non-polar active ingredient was added in such an amount that the ingredient should constitute 5% by weight (w / w) of the final concentrate. The preservative was a natural (GRAS certified) preservative, benzyl alcohol, and the emulsion stabilizer under the trademark SALADIZER®, obtained from TIC Gums, Inc., Belcamp, MD.

[0560] To form the aqueous phase using the methods described in Example 1 above, the following components of the aqueous phase were sequentially added in the following order to the phase vessel

140

EP 2 268 160 B1: 1) water; 2) emulsion stabilizer.

[0561] The water was purified municipal water, which, prior to adding the aqueous phase to the vessel, was purified according to the provided methods by passing through, using the following purifiers in turn: a carbon filter, an ion exchange purifier, a reverse osmosis purifier and an endpoint filter, for example 100 micron endpoint filter. The emulsion stabilizer was the SALADIZER® emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD. [0562] After forming the emulsion, an appropriate amount of citric acid (shown 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): Liquid nanoemulsion concentrate with 5% non-polar compound containing CLA and

25.2% TPGS surfactant

<td>Ingredient</td><td>Portion mg / 2 ml</td><td>Phase</td><td>Percentage (by weight) of concentrate</td><td>g / batch</td>

<td>Oil containing 80% conjugated linoleic acid</td><td> 100</td><td>Oil</td><td> 5</td><td> 12,5</td>

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

<td>Stabilizer of the SALADIZER® trademark (a mixture of xanthan gum, guar gum and sodium alginate)</td><td> 3,4</td><td>Water</td><td> 0,17</td><td> 0,425</td>

<td>Tocopherol polyethylene glycol succinate (surfactant)</td><td> 504</td><td>Oil</td><td> 25,2</td><td> 63</td>

<td>Stabilizer of the SALADIZER® trademark (a mixture of xanthan gum, guar gum and sodium alginate)</td><td> 1,27</td><td>Oil</td><td> 0,0635</td><td> 0,15875</td>

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

<td>Citric acid (pH regulator)</td><td> 5,6</td><td>Emulsion</td><td> 0,28</td><td> 0,7</td>

<td>In total</td><td> 2000,000</td><td></td><td> 100,0000</td><td> 250</td>

Example 2C (iii): Liquid nanoemulsion concentrate with 10% CLA-containing non-polar compound and 20.2% TPGS surfactant [0563] Table 2C (iii) below shows the ingredients used to make 250 g of a lot of CLA-containing liquid nanoemulsion concentrate (size 2 ml), which was prepared according to the procedure in Example 1 above, with the following details and modifications:

[0564] To form the oil phase using the method described in Example 1 above, the following oil phase ingredients were sequentially added in the following order to the oil phase vessel: 1) surfactant, 2) preservative; 3) non-polar active ingredient and

4) emulsion stabilizer.

[0565] The non-polar active ingredient was conjugated linolenic acid (CLA), obtained from Sanmark, LTD (Dalian, Liaoning Province, China; product code 01057-A80) containing 80% CLA. The non-polar active ingredient was added in such an amount that the ingredient should constitute 10%

141

% By weight (w / w) of the final concentrate. The preservative was a natural (GRAS certified) preservative, benzyl alcohol. The emulsion stabilizer was the SALADIZER® emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

[0566] To form the aqueous phase using the methods described in Example 1 above, the following components of the aqueous phase were sequentially added in the following order to the aqueous phase vessel: 1) water; 2) emulsion stabilizer.

[0567] The water was purified municipal water, which, prior to adding the aqueous phase to the vessel, was purified according to the provided methods by passing through using the following purifiers in turn: a carbon filter, an ion exchange purifier, a reverse osmosis purifier and an endpoint filter, for example 100 micron endpoint filter. The emulsion stabilizer was the SALADIZER® emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD. [0568] After forming the emulsion, an appropriate amount of citric acid (shown 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): Liquid nanoemulsion concentrate with 10% non-polar compound containing CLA and

20.2% TPGS surfactant

<td>Ingredient</td><td>Portion mg / 2 ml</td><td>Phase</td><td>Percentage (by weight) of concentrate</td><td>g / batch</td>

<td>Oil containing 80% conjugated linoleic acid</td><td> 200</td><td>Oil</td><td> 10</td><td> 25</td>

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

<td>Stabilizer of the SALADIZER® trademark (a mixture of xanthan gum, guar gum and sodium alginate)</td><td> 3,4</td><td>Water</td><td> 0,17</td><td> 0,425</td>

<td>Tocopherol polyethylene glycol succinate (surfactant)</td><td> 404</td><td>Oil</td><td> 20,2</td><td> 50,5</td>

<td>Stabilizer of the SALADIZER® trademark (a mixture of xanthan gum, guar gum and sodium alginate)</td><td> 1,27</td><td>Oil</td><td> 0,0635</td><td> 0,15875</td>

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

<td>Citric acid (pH regulator)</td><td> 5,6</td><td>Emulsion</td><td> 0,28</td><td> 0,7</td>

<td>In total</td><td> 2000,000</td><td></td><td> 100,0000</td><td> 250</td>

Example 2D: Liquid nanoemulsion concentrates with non-polar compounds that are sabal palm extract

Example 2D (i): Liquid nanoemulsion concentrate with a 5% non-polar sabal palm extract and 17.75% TPGS surfactant [0569] The following table 2D (i) shows the ingredients used to make a 250 g batch of liquid nanoemulsion concentrate Sabal palm extract (2 ml serving size), which was prepared according to the procedure set out in Example 1 above, with the following details and

142

EP 2 268 160 B1 modifications:

[0570] To form the oil phase using the method described in Example 1 above, the following oil phase ingredients were sequentially added in the following order to the oil phase vessel: 1) surfactant, 2) preservative; 3) non-polar solvent; 4) surfactant; 5) non-polar active ingredient and 6) emulsion stabilizer.

[0571] The non-polar active ingredient was a sabal palm extract, a sabal palm lipophilic extract commercially available from Natural Medicinals, Inc., Felda, FL, which contained about 90% of the total amount of 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% palmitic 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% moisture. The non-polar active ingredient was added in such an amount that the ingredient should constitute 5% by weight (w / w) of the final concentrate.

[0572] The co-surfactant was a phosphatidylcholine co-surfactant, sold under the trade name S-100, by Lipoid, LLC, Newark, NJ. This phosphatidylcholine co-surfactant was obtained from soy extract and contained more than 95% phosphatidylcholine.

[0573] The non-polar solvent was 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). This oil contained at least 67.2% tocopherol and about 32.8% soybean oil. The preservative was a natural (GRAS certified) preservative, benzyl alcohol. The emulsion stabilizer was the SALADIZER® emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

[0574] To form the aqueous phase using the methods described in Example 1 above, the following components of the aqueous phase were sequentially added, in the following order, to the aqueous phase vessel: 1) water; 2) emulsion stabilizer.

[0575] The water was purified municipal water, which, prior to adding the aqueous phase to the vessel, was purified according to the provided methods by passing through, using the following purifiers in turn: a carbon filter, an ion exchange purifier, a reverse osmosis purifier and an endpoint filter, for example 100 micron endpoint filter. The emulsion stabilizer was the SALADIZER® emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD. [0576] After forming the emulsion, an appropriate amount of citric acid (shown 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.

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EP 2 268 160 B1

Table 2D (i): Liquid nanoemulsion concentrate with a 5% non-polar sabal palm extract and 17.75% TPGS surfactant

<td>Ingredient</td><td>Portion mg / 2 ml</td><td>Phase</td><td>Percentage (by weight) of concentrate</td><td>g / batch</td>

<td>Vitamin E oil (5-67)</td><td> 75,00</td><td>Oil</td><td> 3,750</td><td> 9,375</td>

<td>Stabilizer of the SALADIZER® trademark (a mixture of xanthan gum, guar gum and sodium alginate)</td><td> 1,270</td><td>Oil</td><td> 0,06</td><td> 0,2</td>

<td>Water</td><td> 1435</td><td>Water</td><td> 71,74</td><td> 179,3</td>

<td>Stabilizer of the SALADIZER® trademark (a mixture of xanthan gum, guar gum and sodium alginate)</td><td> 6,800</td><td>Water</td><td> 0,34</td><td> 0,9</td>

<td>Tocopherol polyethylene glycol succinate (surfactant)</td><td> 355,0</td><td>Oil</td><td> 17,75</td><td> 44,38</td>

<td>Phosphatidylcholine S-100 (co-surfactant)</td><td> 13,38</td><td>Oil</td><td> 0,6690</td><td> 1,673</td>

<td>Benzyl alcohol (preservative)</td><td> 10,00</td><td>Oil</td><td> 0,5000</td><td> 1,2500</td>

<td>Sabal palm oil (90% fatty acids)</td><td> 100,0</td><td>Oil</td><td> 5,0000</td><td> 25,500</td>

<td>Citric acid (pH regulator)</td><td> 3,800</td><td>Emulsion</td><td> 0,1900</td><td> 0,4750</td>

<td>In total</td><td> 2000,00</td><td></td><td> 100,000</td><td> 250</td>

Example 2D (ii): Liquid nanoemulsion concentrate with 5% non-polar sabal palm extract and 25.2% TPGS surfactant [0577] The following table 2D (ii) shows the ingredients used to make 250 g of a batch of liquid nanoemulsion concentrate containing the extract from Sabal palm (2 ml serving size), which was prepared according to the procedure set out in Example 1 above, with the following details and modifications:

[0578] To form the oil phase using the method described in Example 1 above, the following oil phase ingredients were sequentially added in the following order to the oil phase vessel: 1) surfactant, 2) preservative; 3) non-polar active ingredient and 4) emulsion stabilizer.

[0579] The non-polar active ingredient was sabal palm extract, sabal palm lipophilic extract commercially available from Natural Medicinals, Inc., Felda, FL, which contained about 85-90% of 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% palmitic acid, 2% stearic acid, 26.7 oleic acid, 4.9% linoleic acid, 0.7% linolenic acid, 0.42%; 0.42% phytosterols, incl

0.42% beta-sitosterol, 0.09% campesterol, 0.03% Stigmasterol and 0.2% moisture. The non-polar active ingredient was added in such an amount that the ingredient should constitute 5% by weight (w / w) of the concentrate

144

EP2 268 160 B1 final. The preservative was a natural (GRAS certified) preservative, benzyl alcohol. The emulsion stabilizer was the SALADIZER® emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

[0580] To form the aqueous phase using the methods described in Example 1 above, the following components of the aqueous phase were sequentially added, in the following order, to the aqueous phase vessel: 1) water; 2) emulsion stabilizer.

[0581] The water was purified municipal water, which, prior to adding the aqueous phase to the vessel, was purified according to the provided methods by passing through using the following purifiers, in order: a carbon filter, an ion exchange purifier, a reverse osmosis purifier and an endpoint filter, for example 100 micron endpoint filter. The emulsion stabilizer was the SALADIZER® emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD. [0582] After forming the emulsion, an appropriate amount of citric acid (shown 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): Liquid nanoemulsion concentrate with a 5% non-polar sabal palm extract and 25.2% TPGS surfactant

<td>Ingredient</td><td>Portion mg / 2 ml</td><td>Phase</td><td>Percentage (by weight) of concentrate</td><td>g / batch</td>

<td>Sabal palm oil containing 85-90% fatty acids</td><td> 100</td><td>Oil</td><td> 5</td><td> 12,5</td>

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

<td>Stabilizer of the SALADIZER® trademark (a mixture of xanthan gum, guar gum and sodium alginate)</td><td> 3,4</td><td>Water</td><td> 0,17</td><td> 0,425</td>

<td>Tocopherol polyethylene glycol succinate (surfactant)</td><td> 504</td><td>Oil</td><td> 25,2</td><td> 63</td>

<td>Stabilizer of the SALADIZER® trademark (a mixture of xanthan gum, guar gum and sodium alginate)</td><td> 1,27</td><td>Oil</td><td> 0,0635</td><td> 0,15875</td>

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

<td>Citric acid (pH regulator)</td><td> 5,6</td><td>Emulsion</td><td> 0,28</td><td> 0,7</td>

<td>In total</td><td> 2000,000</td><td></td><td> 100,0000</td><td> 250</td>

Example 2D (iii): Liquid nanoemulsion concentrate with a 10% non-polar sabal palm extract and 20.2% TPGS surfactant [0583] The following table 2D (ii) shows the ingredients used to make 250 g of a batch of liquid nanoemulsion concentrate containing the extract from Sabal palm (2 ml serving size), which was prepared according to the procedure set out in Example 1 above, with the following details and modifications:

145

[0584] To form the oil phase using the method described in Example 1 above, the following oil phase ingredients were sequentially added in the following order to the oil phase vessel: 1) surfactant, 2) preservative; 3) non-polar active ingredient and 4) emulsion stabilizer.

[0585] The non-polar active ingredient was sabal palm extract, a sabal palm lipophilic extract commercially available from Natural Medicinals, Inc., Felda, FL, which contained about 85-90% of the total amount of 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% palmitic 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% moisture. The non-polar active ingredient was added in such an amount that the ingredient should constitute 10% by weight (w / w) of the final concentrate. The preservative was a natural (GRAS certified) preservative, benzyl alcohol. The emulsion stabilizer was the SALADIZER® emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

[0586] To form the aqueous phase using the methods described in Example 1 above, the following components of the aqueous phase were sequentially added in the following order to the aqueous phase vessel: 1) water; 2) emulsion stabilizer.

[0587] The water was purified municipal water, which, prior to adding the aqueous phase to the vessel, was purified according to the methods provided by passing through using the following purifiers, in order: a carbon filter, an ion exchange purifier, a reverse osmosis purifier and an endpoint filter, for example 100 micron endpoint filter. The emulsion stabilizer was the SALADIZER® emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD. [0588] After forming the emulsion, the appropriate amount of citric acid (shown in the table) was added

2D (iii)) to adjust the pH of the emulsion to between 2.6 and 3.4 as described in Example 1 above.

Table 2D (iii): Liquid nanoemulsion concentrate with 10% non-polar sabal palm extract and 20.2% TPGS surfactant

<td>Ingredient</td><td>Portion mg / 2 ml</td><td>Phase</td><td>Percentage (by weight) of concentrate</td><td>g / batch</td>

<td>Sabal palm oil containing 85-90% fatty acids</td><td> 200</td><td>Oil</td><td> 10</td><td> 25</td>

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

<td>Stabilizer of the SALADIZER® trademark (a mixture of xanthan gum, guar gum and sodium alginate)</td><td> 3,4</td><td>Water</td><td> 0,17</td><td> 0,425</td>

<td>Tocopherol polyethylene glycol succinate (surfactant)</td><td> 404</td><td>Oil</td><td> 20,2</td><td> 50,5</td>

146

EP2 268 160 B1

<td>Ingredient</td><td>Portion mg / 2 ml</td><td>Phase</td><td>Percentage (by weight) of concentrate</td><td>g / batch</td>

<td>Stabilizer of the SALADIZER® trademark (a mixture of xanthan gum, guar gum and sodium alginate)</td><td> 1,27</td><td>Oil</td><td> 0,0635</td><td> 0,15875</td>

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

<td>Citric acid (pH regulator)</td><td> 5,6</td><td>Emulsion</td><td> 0,28</td><td> 0,7</td>

<td>In total</td><td> 2000,000</td><td></td><td> 100,0000</td><td> 250</td>

Example 3: Liquid nanoemulsion concentrates with non-polar compounds containing coenzyme Q

Example 3A Liquid nanoemulsion concentrate with 5% non-polar CoQ10 compound and 17.75% TPGS surfactant [0589] The following Table 3A shows the ingredients used to make 650 g batch of liquid nanoemulsion concentrate containing CoQ10 (portion size 2 ml), which was prepared according to the procedure in Example 1 above, with the following details and modifications:

[0590] To form the oil phase using the method described in Example 1 above, the following oil phase ingredients were sequentially added in the following order to the oil phase vessel: 1) surfactant, 2) preservative; 3) non-polar solvent; 4) surfactant; 5) non-polar active ingredient and 6) emulsion stabilizer.

[0591] The non-polar active ingredient was coenzyme Q 10 (CoQ10 compound, sold under the name Kaneka Q10 ™ (USP Ubidecarenone) by Kaneka Nutrients, LP, Pasadena, TX, which contains more than 98% ubidecarenone (ubiquinone). The non-polar active ingredient was added in such that the component should constitute 5% by weight (w / w) of the final concentrate. The co-surfactant was a phosphatidylcholine co-surfactant, sold under the trade name S-100, by Lipoid, LLC, Newark, NJ. This phosphatidylcholine co-surfactant was obtained from soy extract and contained more than 95% phosphatidylcholine. The nonpolar solvent was vitamin E oil, sold by ADM Natural Health and Nutrition, Decatur, IL, under the name Novatol ™ 5-67 Vitamin E (D-alphaTocopherol; product code ADM 410217). This oil contained at least 67.2% tocopherol and about 32.8% soybean oil. The preservative was a natural (GRAS certified) preservative, benzyl alcohol, and the emulsion stabilizer under the trademark SALADIZER®, obtained from TIC Gums, Inc., Belcamp, MD.

[0592] To form the aqueous phase using the methods described in Example 1 above, the following components of the aqueous phase were sequentially added, in the following order, to the aqueous phase vessel: 1) water; 2) emulsion stabilizer.

[0593] The water was purified municipal water, which, prior to adding the aqueous phase to the vessel, was purified according to the provided methods by passing through using the following purifiers, in order: a carbon filter, an ion exchange purifier, a reverse osmosis purifier and an endpoint filter, for example 100 micron endpoint filter. The emulsion stabilizer was

147

EP 2 268 160 B1 emulsion stabilizer under the trademark SALADIZER®, obtained from TIC Gums, Inc., Belcamp, MD. [0594] After forming the emulsion, an appropriate amount of citric acid (shown 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 Liquid nanoemulsion concentrate with 5% non-polar compound containing coenzyme Q 5 (CoQ10) and 17.75% TPGS surfactant

<td>Ingredient</td><td>Portion mg / 2 ml</td><td>Phase</td><td>Percentage (by weight) of concentrate</td><td>g / batch</td>

<td>Vitamin E (5-67) oil (nonpolar solvent)</td><td> 75,00</td><td>Oil</td><td> 3,750</td><td> 24,375</td>

<td>Stabilizer of the SALADIZER® trademark (a mixture of xanthan gum, guar gum and sodium alginate)</td><td> 1,270</td><td>Oil</td><td> 0,06</td><td> 0,4</td>

<td>Water</td><td> 1435</td><td>Water</td><td> 71,74</td><td> 466,3</td>

<td>Stabilizer of the SALADIZER® trademark (a mixture of xanthan gum, guar gum and sodium alginate)</td><td> 6,800</td><td>Water</td><td> 0,34</td><td> 2,2</td>

<td>Tocopherol polyethylene glycol succinate (surfactant)</td><td> 355,0</td><td>Oil</td><td> 17,75</td><td> 115,38</td>

<td>Phosphatidylcholine S-100 (co-surfactant)</td><td> 13,38</td><td>Oil</td><td> 0,6690</td><td> 4,349</td>

<td>Benzyl alcohol (preservative)</td><td> 10,00</td><td>Oil</td><td> 0,5000</td><td> 3,2500</td>

<td>CoQ10</td><td> 100,0</td><td>Oil</td><td> 5,0000</td><td> 32,500</td>

<td>Citric acid (pH regulator)</td><td> 3,800</td><td>Emulsion</td><td> 0,1900</td><td> 1,2350</td>

<td>In total</td><td> 2000,00</td><td></td><td> 100,000</td><td> 650</td>

Example 4: Liquid nanoemulsion concentrates with non-polar compounds containing phytosterols

Example 4A: Liquid nanoemulsion concentrate with 5.25% non-polar phytosterol compound and 20% TPGS surfactant [0595] Table 4A below shows the ingredients used to make 250 g of a batch of liquid nanoemulsion concentrate containing phytosterols (portion size 2 ml), which was prepared according to with the procedure outlined in example 1 above, with the following details and modifications:

[0596] To form the oil phase using the method described in Example 1 above, the following oil phase ingredients were sequentially added in the following order to the oil phase vessel: 1) surfactant, 2) preservative; 3) non-polar solvent; 4) surfactant; 5) non-polar active ingredient and 6) emulsion stabilizer.

[0597] The non-polar active ingredient was a phytosterol compound, sold under the name CardioAid ™, distributed by B&amp;D Nutrition and manufactured by ADM Natural Health and

Nutrition, Decatur, IL. This compound contained kosher, not containing meat substances and

148

Dairy products and plant-based sterols, which are produced according to current GMP, and contained at least 95% plant sterols. The non-polar active ingredient was added in such an amount that the ingredient should constitute 5.25% by weight (w / w) of the final concentrate. The co-surfactant was a phosphatidylcholine co-surfactant, sold under the trade name S-100, by Lipoid, LLC, Newark, NJ. This phosphatidylcholine co-surfactant was obtained from soy extract and contained more than 95% phosphatidylcholine. The nonpolar solvent was linseed oil, obtained from Sanmark LLC, Greensboro, NC (Sanmark Limited, Dalian, Liaoning Province, China), which contains not less than (NLT) 50% C18: 3 alpha linolenic acid. The preservative was a natural (GRAS certified) preservative, benzyl alcohol. The emulsion stabilizer was the SALADIZER® emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD. [0598] To form the aqueous phase using the methods described in Example 1 above, the following components of the aqueous phase were sequentially added in the following order to the aqueous phase vessel: 1) water; 2) emulsion stabilizer and 3) pH regulator.

[0599] The water was purified municipal water, which, prior to adding the aqueous phase to the vessel, was purified according to the provided methods by passing through using the following purifiers in turn: a carbon filter, an ion exchange purifier, a reverse osmosis purifier and an endpoint filter, for example 100 micron endpoint filter. The emulsion stabilizer was the SALADIZER® emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

The emulsion stabilizer was phosphoric acid, which was added to the concentrate indicated to ensure that the concentrate pH was between 2.6 and 3.4.

[0600] After formation of the emulsion, the pH of the concentrate was measured using a pH meter as described above to make sure it was between 2.6 and 3.4.

Table 4A: Liquid nanoemulsion concentrate with 5.25% non-polar phytosterol compound and 20% TPGS surfactant

<td>Ingredient</td><td>Portion mg / 2 ml</td><td>Phase</td><td>Percentage (by weight) of concentrate</td><td>g / batch</td>

<td>Linseed oil (50% ALA)</td><td> 105,00</td><td>Oil</td><td> 5,25</td><td> 13,13</td>

<td>Water</td><td> 1365,70</td><td>Water</td><td> 68,29</td><td> 170,71</td>

<td>Stabilizer of the SALADIZER® trademark (a mixture of xanthan gum, guar gum and sodium alginate)</td><td> 3,30</td><td>Water</td><td> 0,17</td><td> 0,41</td>

<td>Tocopherol polyethylene glycol succinate (surfactant)</td><td> 400,00</td><td>Oil</td><td> 20,00</td><td> 50,00</td>

<td>Phosphatidylcholine (co-surfactant)</td><td> 3,00</td><td>Oil</td><td> 0,15</td><td> 0,38</td>

<td>Benzyl alcohol (preservative)</td><td> 10,00</td><td>Oil</td><td> 0,50</td><td> 1,25</td>

<td>phytosterols</td><td> 105,00</td><td>Oil</td><td> 5,25</td><td> 13,13</td>

<td>Phosphoric acid (pH regulator)</td><td> 8,00</td><td>Water</td><td> 0,40</td><td> 1,00</td>

<td>In total</td><td> 2000,00</td><td></td><td> 100,000</td><td> 250</td>

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EP 2 268 160 B1

Example 5: Liquid nanoemulsion concentrates with various non-polar active ingredients [0601] Examples 5A-B show details of liquid nanoemulsion concentrates containing two or more non-polar compounds (non-polar active ingredients) selected from polyunsaturated fatty acids (PUFAs) (e.g., non-polar active ingredients containing omega-3 fatty acids and omega-6 fatty acids), coenzyme Q10 and vitamins. These concentrates were prepared using the general procedure described in Example 1 above.

Example 5A: Liquid nanoemulsion concentrate formulated for women [0602] Table 5A below shows the ingredients used to make 250g batches (2ml serving size) of a liquid nanoemulsion concentrate containing non-polar compounds that include omega-3 fatty acids (e.g. DHA, GLA, ALA), omega-6 fatty acids, coenzyme Q10 and vitamins, which were prepared according to the procedure set out in Example 1 above, with the following details and modifications:

[0603] To form the oil phase using the method described in Example 1 above, the following oil phase ingredients were sequentially added in the following order to the oil phase vessel: 1) surfactant, 2) preservative; 3) non-polar active ingredients and 4) emulsion stabilizer.

[0604] Non-polar active ingredients containing omega-3 fatty acid and omega-6 fatty acid included:

[0605] Borage oil compound, obtained from Sanmark LLC, Greensboro, NC (Sanmark Limited, Dalian, Liaoning Province, China), was obtained by extruding and isolating from seeds of Borago officinalis L. This borage oil non-polar active ingredient contained not less than (NLT) 22% C18: 3 gamma-linolenic acid (GLA), and it was added in an amount of 4.55% by weight (w / w) of the final concentrate, whereby the concentrate contained 1.0% GLA;

[0606] The linseed oil compound Fresh Flax Oil, obtained from Barleans Organic Oils, LLC, Ferndale, WA, contained not less than (NLT) 55% C18: 3 alpha-linolenic acid, and was added in an amount of 2.4 % w / w (w / w) of the final concentrate, whereby the concentrate contained 1.2% ALA; and [0607] Fish oil containing about 30% DHA / EPA (sold under the name Omega 30 TG Food Grade (Non-GMO) MEG-3 ™ Fish Oil by Ocean Nutrition Canada Limited, Nova Scotia, MA). The non-polar active ingredient fish oil was added in an amount of 0.2% by weight of the final concentrate, whereby the concentrate contained 0.06% EPA + DHA.

[0608] The non-polar active ingredient containing coenzyme Q was coenzyme Q10 (CoQ10), sold under the name Kaneka Q10 ™ (USP Ubidecarenone) by Kaneka Nutrients, LP, Pasadena, TX, which contains more than 98% ubidecarenone (ubiquinone). The non-polar active ingredient was added in such an amount that the ingredient should constitute 0.5% by weight (w / w)

150

EP2 268 160 B1 final concentrate.

[0609] Vitamin non-polar active ingredients included vitamin E and vitamin D3. Vitamins were added in amounts that corresponded to the recommended dietary reference intake (DRI) for women. Vitamin D3 was obtained from DSM Nutritional Products, Parsippany, NJ.

Vitamin E, in the form of vitamin E oil, was sold 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 about 32.8% soybean oil.

[0610] The preservative was a natural (GRAS certified) preservative, benzyl alcohol. The emulsion stabilizer was the SALADIZER® emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

[0611] To form the aqueous phase using the methods described in Example 1 above, the following components of the aqueous phase were sequentially added in the following order to the aqueous phase vessel: 1) water and 2) emulsion stabilizer.

[0612] The water was purified municipal water which, prior to adding the aqueous phase to the vessel, was purified according to the provided methods by passing through using the following purifiers, in order: a carbon filter, an ion exchange purifier, a reverse osmosis purifier and an endpoint filter, for example 100 micron endpoint filter. The emulsion stabilizer was the SALADIZER® emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD. [0613] After forming the emulsion, the appropriate amount of citric acid (shown in the table) was added

5A to adjust the pH of the emulsion to between 2.6 and 3.4, as described in Example 1 above.

Example 5: Liquid nanoemulsion concentrates with various non-polar active ingredients

<td>Ingredient</td><td>Portion mg / 2 ml</td><td>Phase</td><td>Percentage (by weight) of concentrate</td><td>g / batch</td>

<td>Borage oil (22% GLA) (non-polar active ingredient)</td><td> 91,00</td><td>Oil</td><td> 4,55</td><td> 11,38</td>

<td>Linseed oil 50% omega acids (non-polar active ingredient)</td><td> 48,00</td><td>Oil</td><td> 2,40</td><td> 6,0</td>

<td>Omega 30 TG Food Grade (Non-GM) MEG-3 ™ Fish Oil (non-polar active ingredient)</td><td> 4,00</td><td>Oil</td><td> 0,20</td><td> 0,5</td>

<td>Kaneka Q10 ™ (non-polar active ingredient)</td><td> 10,000</td><td>Oil</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>Oil</td><td> 0,75</td><td> 1,875</td>

<td>Vitamin D3 (non-polar active ingredient)</td><td> 0,010</td><td>Oil</td><td>5.0 x 10 ⁶</td><td> 0,00125</td>

<td>Water</td><td> 1310,75</td><td>Water</td><td> 65,538</td><td> 163,844</td>

<td>Stabilizer of the SALADIZER® trademark (a mixture of xanthan gum, guar gum and sodium alginate)</td><td> 1,200</td><td>Water</td><td> 0,06</td><td> 0,15</td>

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<td>Ingredient</td><td>Portion mg / 2 ml</td><td>Phase</td><td>Percentage (by weight) of concentrate</td><td>g / batch</td>

<td>Tocopherol polyethylene glycol succinate (surfactant)</td><td> 504,0</td><td>Oil</td><td> 25,20</td><td> 63,00</td>

<td>Stabilizer of the SALADIZER® trademark (a mixture of xanthan gum, guar gum and sodium alginate)</td><td> 0,44</td><td>Oil</td><td> 0,0220</td><td> 0,06</td>

<td>Benzyl alcohol (preservative)</td><td> 10,000</td><td>Oil</td><td> 0,50</td><td> 1,25</td>

<td>Citric acid (pH regulator)</td><td> 5,60</td><td>Emulsion</td><td> 0,2800</td><td> 0,700</td>

<td>In total</td><td> 2000,000</td><td></td><td> 100,0000</td><td> 250</td>

Example 5B: Liquid nanoemulsion concentrate formulated for children [0614] Table 5B below shows the ingredients used to make 250g batch (2ml serving size) of a liquid nanoemulsion concentrate containing non-polar compounds that include omega-3 fatty acids (e.g., DHA, ALA) and vitamins, which were prepared according to the procedure set out in Example 1 above, with the following details and modifications:

[0615] To form the oil phase using the method described in Example 1 above, the following oil phase ingredients were sequentially added in the following order to the oil phase vessel: 1) surfactant, 2) preservative; 3) non-polar active ingredients and

4) emulsion stabilizer.

[0616] Non-polar active ingredients containing omega-3 fatty acid included:

[0617] The linseed oil compound Fresh Flax Oil, obtained from Barleans Organic Oils, LLC, Ferndale, WA, contained not less than (NLT) 55% C18: 3 alpha-linolenic acid, and was added in an amount of 4.2 % w / w (w / w) of the final concentrate, resulting in a concentrate containing 2.1%

ALA; and [0618] Fish oil containing about 30% DHA / EPA (sold under the name Omega 30 TG Food Grade (Non-GMO) MEG-3 ™ Fish Oil by Ocean Nutrition Canada Limited, Nova Scotia, MA). The non-polar active ingredient fish oil was added in an amount of 0.2% by weight of the final concentrate, whereby the concentrate contained 0.06% EPA + DHA.

[0619] Vitamin non-polar active ingredients included vitamin E, vitamin D3, and vitamin A palmitate. Vitamins were added in amounts that corresponded to the recommended daily allowance (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 sold 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 32.8% soybean oil. [0620] The preservative was a natural (GRAS certified) preservative, benzyl alcohol. The emulsion stabilizer was the SALADIZER® emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD.

[0621] To form the aqueous phase using the methods described in example 1 above,

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The following ingredients of the aqueous phase were sequentially added in the following order to the aqueous phase vessel: 1) water and 2) emulsion stabilizer.

[0622] The water was purified municipal water, which, prior to adding the aqueous phase to the vessel, was purified according to the provided methods by passing through using the following purifiers, in order: a carbon filter, an ion exchange purifier, a reverse osmosis purifier and an endpoint filter, for example 100 micron endpoint filter. The emulsion stabilizer was the SALADIZER® emulsion stabilizer, obtained from TIC Gums, Inc., Belcamp, MD. [0623] After forming the emulsion, an appropriate amount of citric acid (shown 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>Portion mg / 2 ml</td><td>Phase</td><td>Percentage (by weight) of concentrate</td><td>g / batch</td>

<td>Linseed oil 50% omega acids (non-polar active ingredient)</td><td> 84,00</td><td>Oil</td><td> 4,20</td><td> 10,50</td>

<td>Omega 30 TG Food Grade (Non-GM) MEG-3 ™ Fish Oil (non-polar active ingredient)</td><td> 4,00</td><td>Oil</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>Oil</td><td> 1,50</td><td> 3,75</td>

<td>Vitamin D3 (non-polar active ingredient)</td><td> 0,800</td><td>Oil</td><td> 0,04</td><td> 0,1</td>

<td>Vitamin A palmitate (non-polar active ingredient)</td><td> 2,800</td><td>Oil</td><td> 0,14</td><td> 0,35</td>

<td>Water</td><td> 1357,16</td><td>Water</td><td> 67,858</td><td> 169,645</td>

<td>Stabilizer of the SALADIZER® trademark (a mixture of xanthan gum, guar gum and sodium alginate)</td><td> 1,200</td><td>Water</td><td> 0,06</td><td> 0,15</td>

<td>Tocopherol polyethylene glycol succinate (surfactant)</td><td> 504,0</td><td>Oil</td><td> 25,20</td><td> 63,00</td>

<td>Stabilizer of the SALADIZER® trademark (a mixture of xanthan gum, guar gum and sodium alginate)</td><td> 0,44</td><td>Oil</td><td> 0,0220</td><td> 0,06</td>

<td>Benzyl alcohol (preservative)</td><td> 10,000</td><td>Oil</td><td> 0,50</td><td> 1,25</td>

<td>Citric acid (pH regulator)</td><td> 5,60</td><td>Emulsion</td><td> 0,2800</td><td> 0,70</td>

<td>In total</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 diluted compositions

To evaluate the various properties, the selected nanoemulsion liquid concentrates prepared in the above examples were diluted, according to the provided examples, in an aqueous medium to produce aqueous liquid diluted compositions. The results are detailed in Examples 6A-B below.

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Example 6A: Dilution and Clarity Assessment of Diluted Compositions: Turbidity Analysis [0624] The concentrates prepared in Examples 2A (vi), 2A (xii), 2A (iii), 2A (v) and 3A were diluted in an aqueous medium according to the provided methods dilution of concentrates. The resulting aqueous liquid dilution compositions were then evaluated for clarity by measuring turbidity using a nephelometer. Dilution parameters and evaluation results are shown in Table 6A below. For each sample listed in Table 6A, the example in which the concentrate was made was indicated.

[0625] Each of the liquid nanoemulsion concentrates listed in Table 6A was diluted by adding the amount of concentrate indicated in Table 6A to the amount of water (purified according to the provided methods) indicated in Table 6A. The approximate dilution factors are also listed. The concentrates were diluted in an aqueous medium in accordance with the concentrate dilution methods provided, using the following steps:

[0626] The indicated amount of water was heated in a Pyrex® beaker by placing the beaker on a hot plate

Thermolyne (model # 846925) until the water reached 49.8 ° C. Then the indicated amount of nanoemulsion liquid concentrate (about 1 g) was added to the heated water and mixed using a stir bar until it dispersed. The resulting aqueous liquid dilution composition containing the non-polar active ingredient was cooled to room temperature (about 25 ° C). For evaluation, the cooled liquid dilution composition was added to a brown glass vial (Alcon). Each of the diluted liquid compositions containing DHA was prepared from the concentrates of Examples 2A (iii) and 2A (v) containing 17.5 mg DHA (in 250 ml and 500 ml water, respectively). The diluted liquid composition containing DHA, prepared from the concentrate of Example 2A (xii), contained 10 mg DHA and 20 mg EPA in 250 ml water. The diluted liquid composition containing DHA, prepared from the concentrate of Example 2A (vi), contained 6.5 mg DHA and 6.5 mg EPA in 250 ml water.

[0627] Vials containing liquid diluted compositions were sent to ACZ Laboratories, Inc., Steamboat Springs, CO for turbidity analysis using a nephelometer. The results are presented in the form of Nephelometric Turbidity Units (NTU) and are indicated in Table 6A below. As shown in Table 6A, each of the liquid aqueous compositions containing the diluted concentrates had an NTU value of less than about 300. Several compositions had a

NTU between about 10 and about 12.

Table 6A: Turbidity (NTU) of liquid aqueous dilute compositions containing liquid nanoemulsion concentrates

<td>Concentrate:</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>Containing DHA (algae oil)</td><td> 1,0085</td><td> 250</td><td> 1:250</td><td> 12,1</td>

<td>Example 2A (v)</td><td>Containing DHA (algae oil)</td><td> 1,0153</td><td> 500</td><td> 1:500</td><td> 159</td>

<td>Example 2A (vi)</td><td>Containing DHA (fish oil)</td><td> 1,0215</td><td> 250</td><td> 1:250</td><td> 10,7</td>

<td>Example 2A (xii)</td><td>Containing DHA (fish oil)</td><td> 1,0013</td><td> 250</td><td> 1:250</td><td> 10,4</td>

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<td>Concentrate:</td><td>Non-polar active ingredient</td><td>Concentrate (grams)</td><td>Water (grams)</td><td>dilution</td><td>NTU</td>

<td>Example 3A</td><td>CoQ10</td><td> 1,0246</td><td> 250</td><td> 1:250</td><td> 239</td>

Examples concentrate, followed by analysis using a Horiba® LB-550 light scattering analyzer. The liquid nanoemulsion concentrate of Example 3A was mixed well and three drops of concentrate were added to 25 mL of water. The sample was then placed in a pan which was used to measure the average particle size on a Horiba® LB-550 light scattering analyzer. The results include measuring the average particle size in the diluted composition, which was measured three times, in separate series. The measurement for each series and the average of three measurements are indicated in Table 6B below. As indicated in Table 6B, the particle size of the liquid diluted composition was less than 150 nm.

Table 6B: Particle size of the aqueous liquid dilution composition containing the liquid coenzyme Q emulsion concentrate

<td></td><td>Average particle size (nm)</td>

<td>Series 1</td><td> 147,5</td>

<td>Series 2</td><td> 143,8</td>

<td>Series 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 [0629] The following tables 8A (i) to 9C show the ingredients that were used to prepare the example liquid nanoemulsion 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 further contained a natural, GRAS certified preservative (benzyl alcohol) and was prepared according to the present general method, in 1000 gram (g) or 500 g batches (batch sizes are indicated in the tables).

[0630] Tables 8A (i) to 9C each show milligrams (mg) of each ingredient per 2 ml serving in an exemplary concentrate, percentage, weight (total concentrate), for each ingredient, and amount in grams (g) of each ingredient per 1000 g a lot. In each table, the "phase" column also indicates whether each component has been added to the aqueous phase ("water"), oil phase ("oil") or added later to the emulsion formed after combining the water and oil phases in the emulsification step (" emulsion").

[0631] Each of the liquid nanoemulsion concentrates shown in Tables 8 and 9 were prepared

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Using the laboratory scale process provided methods. To produce larger batch sizes, the laboratory-scale process can be enlarged, to produce any of these exemplary concentrates of Examples 8-9, using the large-scale manufacturing process provided methods as described herein.

[0632] To prepare the concentrates in Examples 8 and 9, the laboratory scale process was carried out using the following general steps (further details are provided in the individual examples):

[0633] To make concentrates, the indicated amount of each component was weighed using Toledo Scale (model GD13x / USA), Sartorius Basic Analytical Scale (model BA110S) or OHAUS Scale (model CS2000). The choice of weight (weights) depends on the weight of the specific ingredient (s). To form the water phase, the water phase ingredients (indicated by "water" in each table in the "phase" column) were added, in the indicated amount (g / batch), to the water phase vessel (beaker Pyrex®) 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 heating plate as a heating device (Thermolyne model plate # SP46615, Bamstead International, Dubuque, Iowa). The water phase temperature and stirring rate were maintained until the water and oil phases were combined and emulsified. To assess (measure) the temperature of the aqueous phase, a temperature measuring instrument (temperature sensor (model # DPP400W, Cooper-Atkins)) was used. Water phase ingredients, including polar solvent (water, glycerol or propylene glycol) and additional water phase ingredients are indicated.

[0634] The oil phase ingredients (indicated by "oil" in each table in the "phase" column) were added to the oil phase vessel (Pyrex® beaker) and mixed using a standard stirrer (IKA® model No. RE-16 1 S, which is tripod stirrer (laboratory stirrer) compatible with the process on a laboratory scale). The oil phase ingredients included the non-polar active ingredient and other oil ingredients as indicated in the examples.

[0635] Once the oil phase ingredients were mixed, they were heated using a hot plate as a heating device (Thermolyne model # SP46615 hot plate, Bamstead International, Dubuque, Iowa) to the desired temperature of 60 ° C and usually stirred at this temperature until the ingredients dissolved. and kept at this temperature to mix with the aqueous phase. To assess (measure) the oil phase temperature, a temperature measuring instrument (temperature sensor (model # DPP400W, Cooper-Atkins)) was used.

[0636] After both phases had reached their respective temperatures and the oil phase ingredients had dissolved, the phases were combined and emulsified. Emulsification was carried out using a reversible homogenizer (Arde Barinco, Inc .; model CJ-4E). The reversible homogenizer, which was used to mix the components of the aqueous phase, was kept at 30 rpm for mixing during the emulsification step. While mixing using a homogenizer at this speed, the oil phase was transferred to the water phase vessel by pouring it from the oil phase vessel into the water phase vessel. Mixing using a homogenizer was continued, with the septum on the homogenizer being adjusted to obtain and maintain an emulsion, e.g.

156

By moving the septum further in the direction of the forming emulsion and / or from the forming emulsion. During emulsification, the forming emulsion was quickly cooled by placing the water phase vessel (beaker) in a water bath until the liquid temperature reached the desired temperature, as indicated in the examples, between 35 ° C and 43 ° C (usually lasting between about 30a and about 60 minutes ). [0637] In some examples, after emulsification and rapid cooling, additional components 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 the "emulsion" in the phase column) while mixing in a reversible homogenizer (Arde Barinco, Inc .; model CJ4E).

[0638] As a final step, before further evaluation, dilution and / or use, the concentrates were filtered using a 100 micron end product filter.

Example 8: Liquid nanoemulsion concentrates with non-polar PUFA containing compounds. [0639] Examples 8A-8C show details of liquid nanoemulsion concentrates containing non-polar compounds (non-polar active ingredients) containing polyunsaturated fatty acids (PUFAs). Non-polar active ingredients containing PUFA in the exemplary compositions were omega-3 fatty acids, omega-6 fatty acids and conjugated fatty acids, including: [0640] Linseed oil compound, which was Fresh Flax 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 an amount of 5% w / w (w / w) of the final concentrate, resulting in the concentrate containing 2.5% ALA;

[0641] Borage oil compound, obtained from Sanmark LLC, Greensboro, NC (Sanmark Limited, Dalian, Liaoning Province, China), which was obtained by extruding and isolating from seeds of Borago officinalis L. This non-polar active ingredient constituting borage oil medicament contained not less than (NLT) 22% gamma-linolenic acid (GLA) C18: 3, and was added in an amount of 5% by weight (w / w) concentrate, whereby the concentrate contained 1.1% GLA;

[0642] The conjugated linolenic acid (CLA) compound, sold under the trade name Tonalin®, by Cognis Corporation, Cincinnati, OH, which contained 1.7% w / w (w / w) palmitic acid C16: 0, 2.6% by weight (w / w) stearic acid C: 18, 13.00% C18: 1 C9 oleic acid, 0.20% by weight (w / w) C18 linoleic acid: 2 C9 C12 and 81.00% by weight (w / w) conjugated linoleic acid (CLA) which contained 39.70% of the conjugated C9, T11 isomer and 39.50% of the conjugated T10, C12 isomer. This non-polar active ingredient containing CLA was added in an amount of 5% by weight (w / w) final concentrate; and [0643] Fish oil containing about 30% DHA / EPA (sold under the name Omega 30 TG Food Grade (Non-GMO) MEG-3 ™ Fish Oil by Ocean Nutrition Canada Limited, Nova Scotia, MA). Fish oil constituting the non-polar active ingredient was added in an amount of 5% by weight of the final concentrate, whereby the concentrate contained 1.5% EPA + DHA.

[0644] Each of the concentrates containing these non-polar active ingredients was prepared using

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EP2 268 160 B1 of the general procedure set out in Example 7 above.

Example 8A: Liquid nanoemulsion concentrates with PUFA-containing nonpolar compounds, TPGS surfactant and glycerin [0645] Tables 8A (i) -8A (iv) show the ingredients and other details of liquid nanoemulsion concentrates, each containing one of the PUFA-containing nonpolar compounds described above glycerin and TPGS surfactant (TPGS surfactant sold under the name Vitamin E TPGS® by Eastman Chemical Company). The specific non-polar active ingredient containing PUFAs is indicated in each table. [0646] These concentrates were prepared according to the general method shown in Example 7.

Glycerin was obtained from Pan Century Oleochemicals SDN, BHD, Johor, Malaysia. For each concentrate, the general method of Example 7 was used to prepare the oil phase, with the following details: The surfactant and preservative were added, and mixed and heated (60 ° C) until the surfactant melted and dissolved. The non-polar active ingredient was then added, continuing to stir and heat to 60 ° C to combine with the aqueous phase.

[0647] During the emulsification of the water and oil phases as described in Example 7, the emulsion was quickly cooled, as described in the general method, to 35-43 ° C.

Table 8A (i): Liquid nanoemulsion concentrate with 5% non-polar compound containing ALA, 25.2% surfactant TPGS and glycerin

<td>Ingredient</td><td>Portion mg / 2 ml</td><td>Phase</td><td>Percentage (by weight) of concentrate</td><td>g / batch</td>

<td>Linseed oil (55% omega-3) (non-polar active ingredient)</td><td> 100</td><td>Oil</td><td> 5</td><td> 50</td>

<td>Glycerin (polar solvent)</td><td> 1386</td><td>Water</td><td> 69,3</td><td> 693</td>

<td>Tocopherol polyethylene glycol succinate (surfactant)</td><td> 504</td><td>Oil</td><td> 25,2</td><td> 252</td>

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

<td>In total</td><td> 2000,000</td><td></td><td> 100</td><td> 1000</td>

Table 8A (ii): Liquid nanoemulsion concentrate with 5% non-polar compound (borage oil) containing GLA, 25.2% TPGS surfactant and glycerin

<td>Ingredient</td><td>Portion mg / 2 ml</td><td>Phase</td><td>Percentage (by weight) of concentrate</td><td>g / batch</td>

<td>Borage oil (22% GLA) (non-polar active ingredient)</td><td> 100</td><td>Oil</td><td> 5</td><td> 50</td>

<td>Glycerin (polar solvent)</td><td> 1386</td><td>Water</td><td> 69,3</td><td> 693</td>

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<td>Ingredient</td><td>Portion mg / 2 ml</td><td>Phase</td><td>Percentage (by weight) of concentrate</td><td>g / batch</td>

<td>Tocopherol polyethylene glycol succinate (surfactant)</td><td> 504</td><td>Oil</td><td> 25,2</td><td> 252</td>

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

<td>In total</td><td> 2000,000</td><td></td><td> 100</td><td> 1000</td>

Table 8A (iii): Liquid nanoemulsion concentrate with 5% non-polar compound containing CLA, 25.2% surfactant TPGS and glycerin

<td>Ingredient</td><td>Portion mg / 2 ml</td><td>Phase</td><td>Percentage (by weight) of concentrate</td><td>g / batch</td>

<td>Tonalin® CLA Oil (non-polar active ingredient)</td><td> 100</td><td>Oil</td><td> 5</td><td> 50</td>

<td>Glycerin (polar solvent)</td><td> 1386</td><td>Water</td><td> 69,3</td><td> 693</td>

<td>Tocopherol polyethylene glycol succinate (surfactant)</td><td> 504</td><td>Oil</td><td> 25,2</td><td> 252</td>

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

<td>In total</td><td> 2000,000</td><td></td><td> 100</td><td> 1000</td>

Table 8A (iv): Liquid nanoemulsion concentrate with 5% non-polar compound (fish oil) containing DHA, 25.2% TPGS surfactant and glycerin

<td>Ingredient</td><td>Portion mg / 2 ml</td><td>Phase</td><td>Percentage (by weight) of concentrate</td><td>g / batch</td>

<td>Omega 30 TG Food Grade NonGM) Me-3 "'Fish Oil (non-polar active ingredient)</td><td> 100</td><td>Oil</td><td> 5</td><td> 25,00</td>

<td>Glycerin (polar solvent)</td><td> 1386</td><td>Water</td><td> 69,3</td><td> 346,50</td>

<td>Tocopherol polyethylene glycol succinate (surfactant)</td><td> 504</td><td>Oil</td><td> 25,2</td><td> 126,00</td>

<td>Benzyl alcohol (preservative)</td><td> 10</td><td>Oil</td><td> 0,5</td><td> 2,50</td>

<td>In total</td><td> 2000,000</td><td></td><td> 100</td><td> 500</td>

Example 8B: Liquid nanoemulsion concentrates with PUFA-containing non-polar compounds, TPGS surfactant and propylene glycol [0648] Tables 8B (i) -8B (iv) show the ingredients and other details of liquid nanoemulsion concentrates, each containing one of the PUFA-containing nonpolar compounds described above, a polar solvent of propylene glycol and a TPGS surfactant (TPGS surfactant sold under the name Vitamin E TPGS® by Eastman Chemical Company). The concentrates were prepared according to the general method described in Example 7. Propylene glycol was prepared by Shell Chemicals, Alberta, Canada, and

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EP 2 268 160 B1 obtained from the distributor Mitsubishi International Food Ingredients, Inc., Dublin, OH. Specific non-polar active ingredients containing PUFA are indicated in the tables.

[0649] For each concentrate, the general method of Example 7 was used to produce the oil phase, with the following details: The surfactant and preservative were added, and mixed and heated (60 ° C) until the surfactant melted and did not dissolve. The non-polar active ingredient was then added, continuing to stir and heat to 60 ° C to combine with the aqueous phase.

[0650] During the emulsification of the oil and water phases, according to the general method of Example 7, the emulsion was quickly cooled, as described in Example 7, to a temperature of 35-43 ° C.

Table 8B (i): Nanoemulsion concentrate with 5% non-polar ALA-containing compound, 25.2% TPGS surfactant and propylene glycol

<td>Ingredient</td><td>Portion mg / 2 ml</td><td>Phase</td><td>Percentage (by weight) of concentrate</td><td>g / batch</td>

<td>Linseed oil (55% omega-3) (non-polar active ingredient)</td><td> 100</td><td>Oil</td><td> 5</td><td> 50</td>

<td>Propylene glycol (polar solvent)</td><td> 1386</td><td>Water</td><td> 69,3</td><td> 693</td>

<td>Tocopherol polyethylene glycol succinate (surfactant)</td><td> 504</td><td>Oil</td><td> 25,2</td><td> 252</td>

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

<td>In total</td><td> 2000,000</td><td></td><td> 100</td><td> 1000</td>

Table 8B (ii): Liquid nanoemulsion concentrate with 5% non-polar compound (borage oil) containing GLA, 25.2% surfactant TPGS and propylene glycol

<td>Ingredient</td><td>Portion mg / 2 ml</td><td>Phase</td><td>Percentage (by weight) of concentrate</td><td>g / batch</td>

<td>Borage oil (22% GLA) (non-polar component active)</td><td> 100</td><td>Oil</td><td> 5</td><td> 50</td>

<td>Propylene glycol (polar solvent)</td><td> 1386</td><td>Water</td><td> 69,3</td><td> 693</td>

<td>Tocopherol polyethylene glycol succinate (surfactant)</td><td> 504</td><td>Oil</td><td> 25,2</td><td> 252</td>

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

<td>In total</td><td> 2000,000</td><td></td><td> 100</td><td> 1000</td>

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Table 8B (iii): Liquid nanoemulsion concentrate with 5% non-polar CLA containing compound,

25.2% surfactant TPGS and propylene glycol

<td>Ingredient</td><td>Portion mg / 2 ml</td><td>Phase</td><td>Percentage (by weight) of concentrate</td><td>g / batch</td>

<td>Tonalin® CLA Oil (non-polar active ingredient)</td><td> 100</td><td>Oil</td><td> 5</td><td> 50</td>

<td>Propylene glycol (polar solvent)</td><td> 1386</td><td>Water</td><td> 69,3</td><td> 693</td>

<td>Tocopherol polyethylene glycol succinate (surfactant)</td><td> 504</td><td>Oil</td><td> 25,2</td><td> 252</td>

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

<td>In total</td><td> 2000,000</td><td></td><td> 100</td><td> 1000</td>

Table 8B (iv): Liquid nanoemulsion concentrate with 5% non-polar compound (fish oil) containing DHA, 25.2% surfactant TPGS and propylene glycol

<td>Ingredient</td><td>Portion mg / 2 ml</td><td>Phase</td><td>Percentage (by weight) of concentrate</td><td>g / batch</td>

<td>Omega30 TG Food Grade NonGM) Me-3^TM Fish Oil (non-polar active ingredient)</td><td> 100</td><td>Oil</td><td> 5</td><td> 25,00</td>

<td>Propylene glycol (polar solvent)</td><td> 1386,00</td><td>Water</td><td> 69,3</td><td> 346,50</td>

<td>Tocopherol polyethylene glycol succinate (surfactant)</td><td> 504</td><td>Oil</td><td> 25,2</td><td> 126,00</td>

<td>Benzyl alcohol (preservative)</td><td> 10</td><td>Oil</td><td> 0,5</td><td> 2,50</td>

<td>In total</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 [0651] Examples 9A-9C show details of liquid nanoemulsion concentrates each prepared from a non-polar compound (non-polar active ingredient) containing coenzyme Q, using the general procedure set forth in Example 7 above. The non-polar active ingredient in each of these concentrates was coenzyme Q10 (CoQ10), sold under the name Kaneka Q10 ™ (USP Ubidecarenone) by Kaneka Nutrients, LP, Pasadena, TX, containing more than 98% ubidecarenone (ubiquinone), and was added in an amount 5.25% w / w (w / w) of the final concentrate. In addition to the non-polar compound, surfactant, polar solvent and preservative (as described in Example 7), the concentrates in Examples 9A-9C further contained a non-polar compound (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)), containing at least 67.2% tocopherol and about 32.8% soybean oil)); and a co-surfactant (phosphatidylcholine co-surfactant, sold under the trade name S-100, by Lipoid, LLC, Newark, NJ, obtained from soy extract and containing

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EP2 268 160 B1 more than 95% phosphatidylcholine).

Example 9A: Liquid nanoemulsion concentrate with 5% non-polar compound containing coenzyme Q, 17.75% surfactant and glycerin [0652] Table 9A below lists ingredients and other details of the liquid nanoemulsion concentrate containing CoQ10 non-polar active ingredient, the glycerin polar solvent and agent TPGS surfactant (TPGS surfactant sold under the name Vitamin E TPGS® by Eastman Chemical Company). This concentrate was prepared according to the general method of Example 7. Glycerin was obtained from Pan Century Oleochemicals SDN, BHD, Johor, Malaysia).

[0653] The oily phase was prepared using the general methods described in Example 7, with the following details: The following oily phase ingredients were added to the oily phase vessel in the following order: 1) non-polar solvent; 2) a preservative and 3) a co-surfactant, and mixed using a standard stirrer and heated to 60 ° C until the co-surfactant dissolved. The TPGS surfactant was then added and dissolved at 60 ° C. CoQ10 non-polar active ingredient 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 quickly cooled to 35-43 ° C.

Table 9A: Liquid nanoemulsion concentrate with 5% non-polar compound containing coenzyme Q, 17.75% TPGS surfactant and glycerin

<td>Ingredient</td><td>Portion mg / 2 ml</td><td>Phase</td><td>Percentage (by weight) of concentrate</td><td>g / batch</td>

<td>Vitamin E oil (5-67) (non-polar solvent)</td><td> 75</td><td>Oil</td><td> 3,75</td><td> 37,5</td>

<td>Glycerin (polar solvent)</td><td> 1442</td><td>Water</td><td> 72,08</td><td> 720,8</td>

<td>Tocopherol polyethylene glycol succinate (surfactant)</td><td> 355</td><td>Oil</td><td> 17,75</td><td> 177,5</td>

<td>Phosphatidylcholine (Alcolec PC95) (co-surfactant)</td><td> 13,38</td><td>Oil</td><td> 0,669</td><td> 6,69</td>

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

<td>Kaneka Q10 ™ (non-polar active ingredient)</td><td> 105</td><td>Oil</td><td> 5,25</td><td> 52,5</td>

<td>In total</td><td> 2000,000</td><td></td><td> 100,0000</td><td> 1000</td>

Example 9B: Liquid nanoemulsion concentrate with 5% non-polar compound containing coenzyme Q, 17.75% TPGS surfactant and propylene glycol [0654] Table 9B below lists the ingredients and other details of the liquid nanoemulsion concentrate containing CoQ10 non-polar active ingredient, the polar solvent glycol

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EP 2 268 160 B1 propylene and TPGS surfactant (TPGS surfactant sold under the name Vitamin E TPGS® by Eastman Chemical Company). This concentrate was prepared according to the general method of example 7. Propylene glycol was made by Shell Chemicals, Alberta, Canada and obtained from a distributor of Mitsubishi International Food

Ingredients, Inc., Dublin, OH.

[0655] The oily phase was prepared using the general methods described in Example 7, with the following details: The following oily phase ingredients were added to the oily phase vessel in the following order: 1) non-polar solvent; 2) a preservative and 3) a co-surfactant, and mixed using a standard stirrer and heated to 60 ° C until the co-surfactant dissolved. The TPGS surfactant was then added and dissolved at 60 ° C. CoQ10 non-polar active ingredient was then added and dissolved at 60 ° C. This temperature was maintained until mixed with the aqueous phase to obtain an emulsion.

[0656] During the emulsification of the oil and water phases as described in Example 7, the mixture was quickly cooled to 35-43 ° C.

Table 9B: Liquid nanoemulsion concentrate with 5% non-polar compound containing coenzyme Q, 17.75% surfactant TPGS and propylene glycol

<td>Ingredient</td><td>Portion mg / 2 ml</td><td>Phase</td><td>Percentage (by weight) of concentrate</td><td>g / batch</td>

<td>Vitamin E oil (5-67) (non-polar solvent)</td><td> 75</td><td>Oil</td><td> 3,75</td><td> 37,5</td>

<td>Propylene glycol (polar solvent)</td><td> 1442</td><td>Water</td><td> 72,08</td><td> 720,8</td>

<td>Tocopherol polyethylene glycol succinate (surfactant)</td><td> 355</td><td>Oil</td><td> 17,75</td><td> 177,5</td>

<td>Phosphatidylcholine (Alcolec PC95) (co-surfactant)</td><td> 13,38</td><td>Oil</td><td> 0,669</td><td> 6,69</td>

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

<td>Kaneka Q10 ™ (non-polar active ingredient)</td><td> 105</td><td>Oil</td><td> 5,25</td><td> 52,5</td>

<td>In total</td><td> 2000,000</td><td></td><td> 100,0000</td><td> 1000</td>

[0657] Since the modifications will be apparent to those skilled in the art, it is believed that the invention is limited only by the scope of the appended claims.

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EP 2 268 160 B1