Microcapsules with improved shells

Description

BACKGROUND

[0001] Microcapsules are small particles of solids or liquid droplets within a thin shell of a shell material such as beeswax, starch, gelatin or polyacrylic acid. They are used, for example, to obtain liquids as free-flowing powders or compressed solids, to separate reactive materials, to reduce toxicity, to protect against oxidation and / or to control the rate of release of substances such as enzyme, flavor, nutrient, drug, etc. .

[0002] In the past, research has focused on the so-called "single-core" microcapsules. However, one of the problems with single-core microcapsules is that they are prone to breakage. Thus, others have tried to increase the wall thickness of the microcapsules to increase their strength and / or impermeability. However, this practice may lead to a reduction in the loading capacity of the microcapsules.

[0003] Another approach to improving microcapsules has been the formation of so-called "multi-core" microcapsules. For example, US Patent No. 5,780,056 discloses a "multicore" microcapsule with gelatin as the shell material. These microcapsules are formed by spray chilling an aqueous emulsion of oil or carotenoid particles such that the gelatin hardens around the "cores" of the oil or carotenoid particles. Yoshida et al. (Chemical Abstract 1990: 140735 or Japanese Patent Publication JP 01-148338) discloses a complex method of coacervation for the production of microcapsules in which an emulsion of gelatin and paraffin wax is added to a gum arabic solution and then mixed with a surfactant to form multi-core microcapsules. ". Ijichi et al. (J. Chem. Eng. Jpn. (1997) 30 (5): 793-798) microencapsulated large droplets of biphenyl using a complex coacervation method to form multi-layer microcapsules. U.S. Patents No. 4,219,439 and 4,222,891 disclose "multinucleated" oil containing microcapsules having an average diameter of 3-20 gm and an oil droplet size of 1-10 gm for use in pressure sensitive copying and heat sensitive recording papers. While some improvement in microcapsule strength can be realized using such methods, there is still a need to provide microcapsules with improved impermeability and good oxidative barrier to encapsulated material, preferably in combination with large batch volumes. Compositions and methods that meet these and other needs are described herein.

[0004] Additional relevant prior art are for example US 6,969,530, JP H02 261534, US 5,780,056 and US 6,974,592.

SUMMARY

[0005] In accordance with the purpose of the disclosed materials, compounds, compositions, compositions, articles and methods, as an embodiment and broadly, the disclosed subject matter in one aspect relates to compositions and methods of making and using such compositions. In a further aspect, the disclosed subject matter relates to microcapsules and methods for their preparation and

2 uses as well as methods of improving various properties of the microcapsules such as impermeability.

[0006] Additional advantages will be identified in part in the description below, and in part will be apparent from the description, or may be apparent from the practice of the aspects described below. The advantages described below will be realized and attained by means and combinations, especially as set out in the appended claims. It should be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not limiting.

SHORT DESCRIPTION OF THE FIGURES

[0007] The accompanying figures, which are incorporated into and form part of this description, illustrate several aspects described below.

Fig. 1 is a schematic of transglutaminase catalyzed reactions. In particular, Fig. 1a shows the cross-linking reaction between lysine and glutamine residues. Fig. Ib shows the acyl transfer reaction. Fig. Ic shows the deamidation reaction. Figure Id is a diagram of a cross-linking reaction between the two chains of gelatin particles with transglutaminase.

Fig. 2 is a pair of diagrams of two multi-core microcapsules, one where the gelatin shell secondary material is cross-linked with transglutaminase and the other where the gelatin secondary (outer) chitosan shell material is cross-linked with transglutaminase.

Fig. 3 is a group of three patterns of multicore microcapsules, one formed without the addition of wax, one formed by adding a wax emulsion before emulsifying and agglomerating the microcapsule, and one formed by adding wax particles after shell formation, where the wax particles block the pores of the secondary (outer) shell material.

Fig. 4 is a schematic of a multicore microcapsule with wax particles added after shell formation (e.g., before spray drying).

Fig. 5 is a plot of dissolved oxygen (mg / L) during the preparation of a non-fish gelatin Bloom suspension.

Fig. 6 is a group of micrographs from Example 10.1. Fig. 6A is a micrograph of agglomerated multi-core fish oil particles prior to addition of a CoQio emulsion with 100 mg CoQio / 500 mg EPA / DHA loading. Fig. 6B is a microchip of CoQio coated multi-core fish oil particles (loaded with 100 mg CoQ10 / 500 mg EPA / DHA). Fig. 6C is a micrograph of finished CoQ10 coated microcapsules (loaded with 100mg CoQ 10/500 mg EPA / DHA).

Fig. 7 is a group of micrographs from Example 10.2. Fig. 7A is a micrograph of agglomerated multi-core fish oil particles prior to addition of a CoQio emulsion with 30 mg CoQio / 500 mg EPA / DHA loading. Fig. 7B is a micrograph

-3 CoQio-coated multi-core fish oil particles (loaded with 30mg CoQio / 500mg EPA / DHA).

Fig. 8 is a micrograph of the finished CoQ10 coated microcapsules (loaded with 200 mg CoQ 10/500 mg EPA / DHA) of Example 10.3.

Fig. 9 is a graph showing the prediction of zinc levels in fish oil powder by co-spraying ZnCbz with a microcapsule suspension.

DETAILED DESCRIPTION

[0008] The invention is defined in the appended claims.

[0009] The materials, compounds, compositions and methods described herein may be more readily understood by reference to the following detailed description of specific aspects of the disclosed subject matter and the Examples and Figures incorporated herein.

[0010] Before disclosing and describing the present materials, compounds, compositions and methods, it should be understood that the aspects described below are not limited to particular synthetic methods or specific reagents as such may, of course, vary. It should also be understood that the terminology used herein is intended to describe specific aspects only and is not intended to be limiting.

General Definitions

[0011] In the description and the following claims, reference will be made to a number of terms which will be defined as having the following meanings.

Throughout the specification and claims, the word "comprise" and other forms of the word such as "comprising" and "comprises" mean including, but not limited to, and is not intended to exclude, for example, other additives, components, integers or steps.

[0012] As used in the description and the appended claims, the singular form "some", "one", "this" and the like includes plural references unless the context clearly dictates otherwise. Thus, for example, reference to "a compound" includes mixtures of two or more such compounds, reference to "omega 3 fatty acid" includes mixtures of two or more such acids, reference to "microcapsule" includes mixtures of two or more such microcapsules, and so on.

[0013] "Optional" or "optionally" means that the later described event or circumstance may or may not occur, and that the description includes instances where the event or circumstance occurs and instances in which it does not. For example, the phrase "adding a loading agent, a second polymeric component, and optionally a composition to the emulsion" includes instances where the composition is added to the emulsion and instances where it is not added to the emulsion.

[0014] A range may be expressed herein as from "about" one specific value, and / or to "about" another specific value. When expressing such a range, another aspect ranges from one specific value and / or to another specific value. Similarly, when values ​​are expressed as approximations, using the preceding "about" application,

It will be understood that a particular value creates another aspect. It will be further understood that the endpoints of each range are significant both with respect to the different endpoint and independently of the other endpoint. It is also understood that there are a number of values ​​disclosed herein and that each value is also disclosed herein as "about" that particular value in addition to the value itself. For example, if the value "10" is disclosed, "about 10" is also disclosed. It is also understood that when a value "less than or equal to" a value is disclosed, "greater than or equal to a value" and possible ranges between the values ​​are also disclosed as is appropriately understood by those skilled in the art. For example, if the value "10" is disclosed, then "less than or equal to 10" as well as "greater than or equal to 10" is also disclosed. It is also understood that throughout the application, data is provided in a variety of different formats and that the data represents endpoints and starting points and ranges for any combination of data points. For example, if a specific data point "10" and a specific data point "15" are disclosed, it is understood that greater than, greater than or equal to, less than, less than or equal to, and equal to 10 and 15 are deemed to be disclosed as also between 10 am and 3 pm. It is also understood that any entity between two specific entities is also a disclosure. For example, if 10 and 15 are disclosed, 11, 12, 13 and 14 are also disclosed.

[0015] References throughout the specification and summarizing the claims for a portion by weight of a given component in the composition define the weight ratio between the component and any other components in the composition for which the portion by weight is expressed. Accordingly, in a compound containing 2 parts by weight of component X and 5 parts by weight of component Y, X and Y are present in a 2: 5 weight ratio and are present in this ratio whether or not additional components are included in the compound.

[0016] The weight percent (wt.%) Of a component, unless specifically stated otherwise, is based on the total weight of the formulation or composition into which the component is incorporated.

[0017] "Subject" as used herein means an entity. In one aspect, the subject is a mammal, such as a primate, and in another aspect, the subject is a human. The term "subject" also includes domesticated animals (e.g., cats, dogs, etc.), farm animals (e.g., cattle, horses, pigs, sheep, goats, etc.), and laboratory animals (e.g., mice, rabbits, rats, mumps, etc.) sea ​​fly, fruit fly, etc.).

[0018] Reference will now be made in detail to specific aspects of the disclosed materials, compounds, compositions, articles, and methods, examples of which are illustrated in the accompanying Examples.

Materials and Compositions

[0019] Materials, compounds, compositions, and components disclosed herein may be used, may be used in conjunction with, may be used to prepare, or are products of the disclosed methods and compositions. These and other materials are disclosed herein, and it is understood that when combinations, subsets, interactions, groups, etc. of these materials are disclosed, although specific mention of each of the various units and aggregate combinations and permutations of these compounds may not be explicitly disclosed, each is herein

-5specifically considered and described. For example, when a compound is disclosed and a series of modifications are discussed that can be made to a series of components or moieties of the compound, without exception, any combination and permutation that is possible are specifically considered, unless specifically indicated to the contrary. Hence, if component class A, B, and C is disclosed, as well as component class D, E, and F, and an example of a combined AD composition is disclosed, then even if each is not individually listed, each is individually and collectively considered. . Accordingly, in this example, each of the combinations of AE, AF, BD, BE, BF, CD, CE, and CF is specifically contemplated and should be considered as disclosed in disclosure of A, B, and C; D, E and F; and an exemplary combination of AD. Likewise, any subset or combination of these are specifically contemplated and disclosed. Accordingly, for example, the subgroups AE, BF, and CE are specifically contemplated and should be considered as disclosed in disclosure of A, B, and C; D, E and F; and an exemplary combination of AD. This concept applies to all aspects of this disclosure, including but not limited to, steps in the methods of making and using the disclosed compositions. Accordingly, when there are a number of additional steps that can be performed, it is understood that each of these additional steps can be performed with any specific aspect or combination of aspects of the disclosed methods and that each such combination is specifically contemplated and should be construed as disclosed herein. .

Microcapsules

[0020] The shells of a plurality of microcapsules, eg, microcapsules with gelatin shells, are often "porous", which may allow oxygen in the air or dissolved water to diffuse into the charging substance cores (s). Oxidation of the loading substance can cause stability and sensor problems. To overcome these problems, microcapsules with improved shells and methods for their preparation are disclosed herein. Generally, methods for making microcapsules are disclosed that involve the use of waxes, saccharides, proteins, and small molecules such as amino acids and sugars to block the pores of the microcapsule shell and / or to increase the number of network connections in the microcapsule shell. Accordingly, the microcapsules disclosed herein generally have a combination of structural strength, impermeability, and high loading capacity.

[0021] In certain aspects, microcapsules are disclosed that contain agglomeration of primary microcapsules and a loading material, each individual primary microcapsule having a primary shell, the loading substance being encapsulated by a primary shell and the agglomeration being encapsulated by the outer shell. These microcapsules are referred to herein as "multi-core microcapsules". Also disclosed are "single-core" microcapsules which contain a core, the core containing a charging material, a primary sheath surrounding the core, and an outer sheath surrounding the primary sheath. Unless otherwise stated, the term "microcapsule" is used herein to refer to multi-core, single-core, or mixture of multi-core and single-core microcapsules. In these mi kr okap sulki (and others disclosed here) the primary sheath,

The outer coat or both the primary and the outer coat comprises the remainder of one or more compositions comprising an amino acid, protein, saccharide, wax, or a combination thereof.

[0022] The term "residue" as used herein refers to a moiety that is the obtained product of certain chemicals in a specific reaction scheme or further formulation or chemical product, whether or not the moiety is actually derived from the specified chemicals. For example, an "amino acid residue" refers to a moiety that is formed when an amino acid participates in a particular reaction (e.g. the residue may be the product of an amino acid undergoing a transglutaminase-catalyzed cross-linking reaction with another amino acid). In this case, the amino acid residue is "derived" from an amino acid. It is understood that this moiety can be obtained by reacting with a compound other than the specified amino acid, for example, by reacting a protein or peptide containing an amino acid, and the like. This concept covers other chemical compounds disclosed herein, such as protein, saccharides like chitosan, lactose, and sucrose and waxes. Accordingly, when specific compounds undergo particular reactions or treatments (e.g. acid / base reactions, cross-linking reactions with other chemicals, and functional group transformations), they are referred to herein as the rest of the corresponding chemical compound.

[0023] It is also contemplated that one or more additional shell layers may be placed on the outer shell of the microcapsules. The techniques disclosed in International Publication No. WO 2004/041251 A1 can be used to add additional shell layers to the microcapsules.

[0024] As noted, the microcapsules disclosed herein may be such that the primary shell, outer shell, or both primary and outer shells contain the remainder of one or more chitosan and glutamine containing compositions; chitosan, lysine and glutamine; chitosan, glutamine, and one or more of leucine, isoleucine, methionine, cysteine, tyrosine, tryptophan, or phenylalanine; or chitosan and one or more of leucine, isoleucine, methionine, cysteine, tyrosine, tryptophan, or phenylalanine. The component of the rest may be different from the materials that make up the primary and / or outer envelopes. For example, if the primary and / or outer shell is made of a saccharide and the primary and / or outer shell is said to contain a saccharide residue, then the microcapsules disclosed are such that the saccharide residue is different from the saccharides that are used to make the shell materials . Similarly, if the primary and / or outer envelope is made of a protein, and the primary and / or outer envelope is said to contain a protein residue, then the microcapsules disclosed are such that the protein residue is different from the protein that is used to make the envelope materials.

Induction period

[0025] In many of the examples of microcapsules disclosed herein, the microcapsules have a long induction period. The induction period is a measure of the impermeability of the microcapsule. The induction period can be measured by placing a sample of the microcapsule (about 5 g) in a container (e.g. a glass container) and then placing the sample container in a metal bomb under pressure.

- oxygen. The pressurized bomb may have an initial pressure of 5 bar (500 kPa) at 65 ° C. Changes in pressure are recorded over time. The inflection point is taken as the induction period. A commercially available tool that can be used to measure the induction period is OXIPRES ™ (Mikrolab Aarhus A / S; Hojbjerg, Denmark). Generally, a more stable powder has a longer induction period at constant temperature.

[0026] Many of the microcapsules disclosed herein may have an induction period (all induction period results are obtained from a measurement at 65 ° C, unless otherwise stated) for more than about 40, 47, 50, 75, or 100 hours. For example, microcapsules are disclosed that have an induction period greater than about 40, 41, 42, 43, 44, 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, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107 , 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119 or 120 hours, where any given value can form the upper or lower endpoint of the range.

Sheath materials

[0027] A variety of polymers can be used to form the shell layers of the disclosed single-core and multi-core microcapsules. For example, the primary shell and / or outer shell material of the disclosed microcapsules can contain a surfactant, gelatin, protein, polyphosphate, polysaccharide, or mixtures thereof. Additional examples of suitable primary shell and / or outer shell materials include chitosan and glutamine; chitosan, lysine and glutamine; chitosan, glutamine, and one or more of leucine, isoleucine, methionine, cysteine, tyrosine, tryptophan, or phenylalanine; or chitosan and one or more of leucine, isoleucine, methionine, cysteine, tyrosine, tryptophan, or phenylalanine; it is also contemplated that derivatives of these polymers can also be used. One specific type of primary shell and / or outer shell material that can be used in the disclosed microcapsules is fish gelatin or pork gelatin.

[0028] In many examples of suitable microcapsules, the primary shell and / or outer shell material may have a Bloom Number from about 0 to about 350. The Bloom Number reveals the strength of a gel formed at 10 ° C with 6.67% gelled solution for 17 ± 1 hour. . The Bloom number determination of substances can be performed by methods known in the art. It is contemplated that the primary and / or outer shell material may have a Bloom Number of about 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, 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, 60, 60,61,

62, 63, 64, 65, 66, 67, 68, 69, 70, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 80, 81, 82, 83,84,

85, 86, 87, 88, 89, 90, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 100, 101, 102, 103, 104, 105,

106, 107, 108, 109, 110, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 120, 121,122,

123, 124, 125, 126, 127, 128, 129, 130, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139,140,

140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 150, 151, 152, 153, 154, 155, 156,157,

158, 159, 160, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 170, 171, 172, 173,174,

175, 176, 177, 178, 179, 180, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 190, 191,

192, 193, 194, 195, 196, 197, 198, 199, 200, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209,

210, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 220, 221, 222, 223, 224, 225, 226,

227, 228, 229, 230, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240, 240, 241, 242, 243,

244, 245, 246, 247, 248, 249, 250, 250, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 260,

261, 262, 263, 264, 265, 266, 267, 268, 269, 270, 270, 271, 272, 273, 274, 275, 276, 277, 278,

279, 280, 280, 281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 290, 291, 292, 293, 294, 295,

296, 297, 298, 299, 300, 300, 301, 302, 303, 304, 305, 306, 307, 308, 309, 310, 310, 311, 312,

313, 314, 315, 316, 317, 318, 319, 320, 320, 321, 322, 323, 324, 325, 326, 327, 328, 329, 330,

330, 331, 332, 333, 334, 335, 336, 337, 338, 339, 340, 340, 341, 342, 343, 344, 345, 346, 347,

348, 349, or 350, wherein any given value may form the upper or any endpoint, as appropriate. In some specific examples, the primary and / or outer shell material may have a Bloom Number from about 0 to about 50, and in other examples, the primary and / or outer shell material may have a Bloom Number from about 51 to about 350. Yet other specific examples include microcapsules containing a primary and / or outer shell material having a Bloom Number of about 0, about 210, about 220, or about 240. In one example, the microcapsule does not contain "low Bloom" gelatin, which is a Bloom Number gelatin. less than 50.

[0029] The sheath material may be a two-component system made up of a mixture of different types of polymer components and where a composition has been added to the system to improve impermeability. In other examples, the shell material may be a complex coacervate between two or more polymer components (e.g., gelatin A and polyphosphate). Component A may be Type A gelatin, although other polymers like those mentioned above for the shell materials are also contemplated as Component A. Component B may be Type B gelatin, polyphosphate, acacia, alginate, chitosan, carrageenan, pectin, low methoxy pectin , carboxymethyl cellulose or a mixture thereof. Again, other polymers such as those disclosed above for the shell materials are also contemplated as component B. The molar ratio of component A: component B that is used depends on the type of components, but is typically from about 1: 5 to about 15: 1. For example, when type A gelatin and polyphosphate are used as components A and B, respectively, the molar ratio of component A: component B may be about 8: 1 to about 12: 1 when type A gelatin and type B gelatin are used respectively as components. A and B, the molar ratio of component A: component B may be about 2: 1 to about 1: 2; and when type A gelatin and alginate are used as components A and B, respectively, the molar ratio of component A: component B may be about 3: 1 to about 5: 1. In many of the disclosed microcapsules, the primary shell and / or outer shell may contain a complex coacervate. For example, the primary shell and / or outer shell may comprise a complex gelatin and polyphosphate coacervate. Other examples include the complex coacervate of gelatin and alginate, gelatin and pectin, gelatin and acacia, gelatin and xanthan, gelatin and low methoxy pectin, and gelatin and whey proteins.

[0030] In the disclosed microcapsules, the outer shell may have an average diameter of from about 1 µm to about 2000 µm, from about 20 µm to about 1000 µm, or from about 30 µm to about 80 µm. In further examples, the diameter of the outer sheath may be about 1, 10, 20, 30, 40, 50, 60, 70, 80, 90, 200, 300, 400, 500, 600, 700, 800, 900, 1000, 1200, 1300. , 1400, 1500, 1600, 1700, 1800, 1900, or 2000 µm, where any given value may form an upper or lower endpoint, as appropriate.

[0031] The primary shells of the disclosed microcapsules can have an average diameter of from about 40 nm to about 10 µm, or from about 0.1 µm to about 5 µm. In further examples, the primary sheath diameter may be about 40 nm, 50 nm, 60 nm, 70 nm, 80 nm, 90 nm, 100 nm, 200 nm, 300 nm, 400 nm, 500 nm, 600 nm, 700 nm, 800 nm. , 900 nm, 1000 nm, 2 pm, 3 pm, 4 pm, 5 pm, 6 pm, 7 pm, 8 pm, 9 pm, 10 pm where any of the given values ​​can form an upper or lower end point when it is appropriate.

[0032] Particle size can be measured using any conventional equipment known in the art, for example, Coulter LS230 Particle Size Analyzer, Miami, Fla., USA.

Additional compositions

[0033] As disclosed, the microcapsules may have shell (s) (primary and / or outer) that contain additional compositions to improve the impermeability of the microcapsule. These additional compositions can be incorporated into the shell (s) at various points during the preparation of the microcapsule, as discussed more fully herein. In general, additional compositions can be associated with the shell (s) through physical, electrostatic, ionic, van der Waals, steric, or chemical interactions. For example, the additional composition may be physically trapped inside the pore present in the shell, thereby blocking the pore. In another example, the additional composition may be chemically bonded to the shell material via a covalent bond (e.g., by an enzyme catalyzed cross-linking reaction).

[0034] Some specific examples of additional compositions that may be present in the (primary and / or outer) shell (s) of the disclosed microcapsules include, but are not limited to, amino acids, peptides, proteins, saccharides (ie, mono-, di-, oligo- or polysaccharides) and waxes, including combinations thereof and residues thereof. For further illustration, the chitosan polysaccharide may be present in the shells of the disclosed microcapsules and may participate in an enzymatically catalyzed cross-linking reaction between the first and / or second polymer components that are used to form the shell material. Chitosan, with its multiple cross-linking sites, can thus be chemically bonded to other polymer components in the shell material and thus increase the impermeability of the shell. In other examples, a small molecule such as an amino acid or sugar can be physically entrapped, entangled with, or even chemically bound to the shell (s) of the microcapsule, thereby strengthening the shell and / or blocking any pores. Larger wax and protein particles can also be

Incorporated into the shell of the microcapsule to increase strength, reinforcement and / or increase impermeability by blocking any pores.

[0035] It is also contemplated that any combination of such additional compositions may be used and may be present in the shell material of the disclosed microcapsules. That is, one or more amino acids, one or more proteins and one or more polysaccharides or one or more waxes may be used. Additionally, one or more amino acids and proteins, one or more amino acids and saccharides, or one or more amino acids and waxes may be used. Still additionally, one or more proteins and saccharides or one or more proteins and waxes may be used. In addition, one or more saccharides and waxes can be used. In yet another example, one or more amino acids, proteins and saccharides, one or more amino acids, proteins and waxes, one or more proteins, saccharides and waxes, one or more amino acids, saccharides and waxes may be used.

[0036] Specific examples of amino acids, including their residues, that can be used in the disclosed shell (s) of the microcapsules include the 20 naturally occurring amino acids that make up proteins and polypeptides. Additionally, they further include less typically ingredients that also occur naturally, such as, but not limited to, formylmethionine and selenocysteine, analogs of commonly encountered amino acids, and amino acid mimetics or amino acid functionality. Polymers are also contemplated, in the case of amino acids such as polylysine. Non-limiting examples of these and other molecules are discussed herein. In many examples, the additional composition comprises lysine, leucine, isoleucine, glutamine, methionine, tyrosine, phenylalanine, tyrosine, tryptophan, cysteine, or any combination thereof. Amino acids may be present in the shell material in a ratio of from about 1: 5 to about 5: 1 (e.g., about 2: 1) as compared to the second polymer component. Further examples include microcapsules with an amino acid to second polymer component ratio of about 1: 5, 1: 4, 1: 3, 1: 2, 1: 1, 2: 1, 3: 1, 4: 1, and 5: 1, where any ratio can form the upper or lower endpoint of the ratio range.

[0037] Suitable proteins, which also include "peptides", are compounds consisting of amino acids chemically bound together. In general, amino acids are linked to each other via amide bonds ("CONH"); however, the amino acids can be linked together by any other chemical bond known in the art. For example, amino acids can be linked by amino bonds. It is also possible to use peptides and proteins linked to other molecules (e.g. conjugates). For example, carbohydrates (e.g., glycoproteins) can be linked to a protein or a peptide. Such derivatives, variants and analogs of peptides and proteins are contemplated herein with the meaning of the term protein. Some specific proteins include, but are not limited to, milk protein, gelatin, whey protein isolate, whey protein concentrate, caseinate, soy protein, BSA, and other albumin, including mixtures thereof. The proteins may be present in the envelope material at a ratio to the second polymer component of from about 1: 1 to about 40: 1 (e.g., about 28.5: 1). Further examples include microcapsules with a protein to one ratio

-11 polymer component of approximately 1: 1, 5: 1, 10: 1, 15: 1, 20: 1, 25: 1, 30: 1, 35: 1 and 40: 1, where any ratio may form the upper or lower point final scope of relations.

[0038] Also suitable are polymeric amines that are olefin-based polymers that contain one or more amine functional groups. Many such polyamines can be obtained commercially or can be obtained by methods known in the art. Suitable examples of polyamines that can be used as the first active ingredient in the disclosed target / active composites include, but are not limited to, polyvinylamine and polyalkyleneimines such as polyethyleneimine.

[0039] Saccharides, including residues thereof, also are suitable compositions that may be present in the disclosed microcapsule shells. Specific examples include an N-acetylglucosamine polymer such as chitosan and chitin. Chitosan is a naturally occurring polymer found in many fungi. However, for convenience reasons, chitosan is derived from chitin, which is the second most abundant natural polymer (after cellulose). Chitin is easily isolated from crustaceans or insect exoskeletons and is also found in molluscs and fungi. Chitin is a water-insoluble copolymer of N-acetyl-D-glucosamine and D-glucosamine, but the vast majority of the monomer units are N-acetyl-D-glucosamine residues. Chitosan is a copolymer of the same two monomer units, but most of the monomer units are D-glucosamine residues. Since the D-glucosamine residues carry the basic amino acid function, they readily form salts with acids. Many of these salts are water soluble. Treatment of chitin with concentrated caustic at elevated temperature converts N-acetyl-D-glucosamine residues to D-glucosamine residues and thereby converts chitin to chitosan. There are a number of compositions possible between pure poly-N-acetyl-D-glucosamine and pure poly-D-glucosamine. These compositions are within the skill of the art to be prepared and are all suitable for the uses described herein.

[0040] Suitable acids for the preparation of the chitosan salts for use in the methods described herein are those which form water-soluble salts with chitosan. It is not necessary that the acid itself is water-soluble; however, such water-soluble acids can facilitate handling. The inorganic acids that form the water-soluble salts of chitosan include halogen acids and nitric acid, but excluding sulfuric and phosphoric acids, as they do not form water-soluble salts with chitosan. Organic acids are particularly suitable and include, but are not limited to, lactic acid, glycolic acid, glutamic acid, formic acid, acetic acid, and mixtures thereof. Mono- or polyfunctional carboxylic acids can also be used. They can be aliphatic or aromatic as long as they form water-soluble salts with chitosan.

[0041] Other polysaccharides and their residues which are suitable saccharides for the disclosed microcapsules are maltodextrin (DE18, DE 21, DE40 etc.), modified starch (NLOK), oligofructans, cyclodextrins (alpha-, beta- and gamma-cyclodextrins), carboxymethylcellulose, hydroxypropylmethylcellulose (HPMC) (Methocel), ethylcellulose

-12 (Ethocel), hydroxypropyl cellulose (HPC), (e.g. Klucel), cellulose ether (e.g. Benecel), agar, alginate, pectin, low methoxyl pectin, gum arabic, carrageenan, cellulose gum, dilutan gum, gellan gum, carob gum , welan gum, and xanthan gum.

[0042] Other suitable saccharides, including residues thereof, are monosaccharides such as glucose, fructose, galactose, arabinose, ribose, ribulose, xylose, mannose and xylulose. Still further, suitable saccharides, including residues thereof, include disaccharides or trisaccharides, where the saccharide is in the form of pyranose or furanose (6- or 5-membered rings). Non-limiting examples of di- and tri-saccharides include sucrose, lactose, cellobiose, sorbose, cellotriose, trehalose, maltose and raffinose and the like. Particularly useful forms of saccharides that can be used are maple syrup, honey and corn syrup which are safe and can add flavor to the microcapsules. Various saccharide derivatives such as xylitol, sorbitol, isomalt and glucosamine are also suitable for use in the disclosed microcapsules.

[0043] The saccharides disclosed herein may be present in the shell material in a ratio of the total shell material (first and second polymer components) of from about 1: 0.2 to about 1: 5, or about 1: 0.02 to 1: 0. 5 ratio to the second polymer component (e.g. polyphosphate). Further examples include microcapsules with a ratio of saccharide to total polymer component of about 1: 0.2, 1: 0.5, 1: 1, 1: 1.5, 1: 2.0, 1: 2.5, 1: 3 , 0, 1: 3.5, 1: 4.0, 1: 4.5, and 1: 5.0 wherein any ratio may form the upper or lower endpoint of the ratio range. Still further examples include microcapsules with a ratio of saccharide to second polymer component of about 1: 0.02, 1: 0.05, 1: 0.1, 1: 0.15, 1: 0.2, 1: 0.25, 1: 0.3, 1: 0.35, 1: 0.4, 1: 0.45, and 1: 0.5 wherein any ratio may form the upper or lower endpoint of the ratio range.

[0044] A suitable wax that may be present in the disclosed microcapsule shells is carnauba wax, which may be present as a microemulsion. Other suitable waxes include, but are not limited to, candelilla, ceresin, (synthetic) Japanese wax, orange peel wax, rice bran wax, shellac, paraffin, montan, microcrystalline wax, polyethylene, and beeswax. Wax may be present in the shell material in a ratio of from 1: 1 to about 1:10 to the second polymer component. (e.g. 1: 6). Further examples include microcapsules with a ratio of wax to second polymer component of about 1: 1, 1: 2, 1: 3, 1: 4, 1: 5, 1: 6, 1: 7, 1: 8, 1: 9, : 10, where any ratio can form the upper or lower endpoint of the ratio range.

Charging substances

[0045] In the disclosed microcapsules, the loading substance can be any substance that one wishes to microencapsulate (e.g., a substance one wishes to deliver to the subject). In many examples, the appropriate charging agent is not fully soluble in the aqueous mixture. The charging substance can be a solid, a hydrophobic liquid, or a mixture of a solid substance and a hydrophobic liquid. In many of the examples included herein, the loading agent may include a long chain polyunsaturated fatty acid.

-13, specific examples of which are included below. Additionally, the loading agent may contain a biologically active substance, a nutrient such as a nutritional supplement, a flavoring agent, a polyunsaturated fatty acid such as an omega-3 fatty acid, a vitamin, a mineral, a carbohydrate, a steroid, a trace element and / or a protein, and the like, including mixtures and combinations thereof. In other examples, the loading agent may include microbial oil, algae oil (e.g. dinoflagellate oil such as Crypthecodinium cohnii), mushroom oil (e.g. Thraustochytrium, Schizochytrium oil or a mixture thereof) and / or vegetable oil (e.g. linseed, vegetable) including mixtures and combinations thereof. In other examples, the loading agent can be a pharmaceutical composition (e.g., drug and / or enzyme) or a flavor. The charging substance can also be a hydrophobic liquid such as a lubricant, oil, or a mixture thereof. Typical oils can be fish oils, vegetable oils (e.g. rapeseed, olive oil, corn oil), mineral oils, their derivatives or mixtures thereof. The loading agent may contain a purified or partially purified oily substance such as a fatty acid, triglyceride or a mixture thereof.

[0046] In still other examples, a suitable loading agent may include sea oil, such as natural and refined and concentrated fish oil. Examples of suitable fish oils include, but are not limited to, Atlantic fish oil, Pacific fish oil, Mediterranean fish oil, light pressed fish oil, alkali processed fish oil, heat treated fish oil, light and heavy brown fish oil. , bonito oil, sprats oil, tuna oil, sea bass oil, halibut oil, marlin oil, barracuda oil, cod oil, menhadena oil, sardine oil, anchovy oil, capelin oil, Atlantic cod oil, Atlantic herring oil, Atlantic mackerel oil, Atlantic menhadena oil, salmonid oil and shark oil, including mixtures and combinations thereof. Untreated fish oil is also a suitable charging agent. Other sea oils suitable for use include, but are not limited to, squid oil, cuttlefish oil, octopus oil, krill oil, seal oil, whale oil, and the like, including mixtures and combinations thereof. Any sea oil and combination of sea oil can be used in the disclosed delivery items and the disclosed food items and methods.

[0047] Many of the microbial, algae, fungal, vegetable, and marine oils described herein contain omega-3 fatty acids. As such, certain delivery devices disclosed herein may include a loading agent that includes an omega-3 fatty acid, an omega-3 fatty acid alkyl ester, an omega-3 fatty acid triglyceride ester, an omega-3 fatty acid phytosterol ester, and / or mixtures thereof. and combinations. Omega-3 fatty acid is an unsaturated fatty acid that contains CH3 — CH2-CH = CH — as its end. Overall, omega-3 fatty acid has the following formula:

AT 11 ₂

CH₃“CH₂CH — CH-R¹—C - OR²

-14 with R.¹ is a C3-C40 alkyl or alkenyl group containing at least one double bond and R² is H or an alkyl group. The term "alkane" or "alkyl" as used herein is a saturated hydrocarbon group (e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, s-butyl, t-butyl, n-pentyl, isopentyl, s-pentyl, neopentyl, hexyl, heptyl, octyl, nonyl, decyl, dodecyl, tetradecyl, hexadecyl, eicosyl, tetracosyl and the like). The term "alkene" or "alkenyl" as used herein is a hydrocarbon group containing at least one carbon-carbon double bond. Asymmetric structures such as (AB) C = C (CD) are intended to link both the E and Z isomers (cis and trans). In a further example, R.¹may be a C5-C38, C6-C36, C8-C34, C10-C32, C12-C30, C14-C28, C16-C26 or C18-C24 alkenyl group. In yet another example, the R 1 alkenyl group may have 2 to 6, 3 to 6, 4 to 6, or 5 to 6 double bonds. In addition, the alkenyl group R 1 may have 1, 2, 3, 4, 5 or 6 double bonds, each of the values ​​indicated may form the upper or lower endpoint, respectively.

[0048] Specific examples of omega-3 fatty acids that are suitable loading agents that can be used in the disclosed articles for delivery include, but are not limited to, alpha-linolenic acid (18: 3ω 3), octadecatetraenoic acid (18: 4ω3). ), eicosapentaenoic acid (20: 5ω3) (EPA), eicosapentaenoic acid (20: 4ω3), henicosapentaenoic acid (21: 5ω3), docosahexaenoic acid (22: 6ω3) (DHA), docosapentaenoic acid (22: 5ω3) (DPA) including derivatives and mixtures thereof. Many types of fatty acid derivatives are well known to those skilled in the art. Examples of suitable derivatives are esters such as phytosterol esters, furanoid esters, branched or unbranched C1-C30 alkyl esters, branched or unbranched C2-C30 alkenyl esters, or branched or unbranched C3-C30 cycloalkyl esters, especially C1-C6 alkyl esters and C1-C30 alkyl esters . In a further example, the loading agent may be docosahexaenoic acid and / or eicosapentaenoic acid phytosterol ester, docosahexaenoic acid and / or eicosapentaenoic acid C 1 -C 6 alkyl ester, docosahexaenoic acid and / or eicosapentaenoic acid triglyceride ester, and / or a mixture thereof.

[0049] Other examples of suitable loading materials that may be present in the disclosed delivery articles include at least 4, at least 6, at least 8, at least 10, at least 12, at least 14, at least 16, at least 18, or at least 20 carbon atoms. In some other examples, the loading agent may include about 8, 9, 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 or 45 carbon atoms, any of the values ​​given may where appropriate form the upper or lower end point. In still other examples, the loading agent may contain a mixture of fatty acids (including derivatives thereof) having the carbon range. For example, the loading agent may contain from about 8 to about 40, from about 10 to about 38, from about 12 to about 36, from about 14 to about 34, from about 16 to about 32, from about 18 to about 30, or from about about 20 to about 28 carbon atoms.

[0050] Some further examples of loaders are those that contain at least one unsaturation (i.e., a carbon-carbon double or triple bond). For example, the loading agent may contain at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, or at least 8 carbon-carbon double bonds, triple bonds, or any combination thereof. In another example, the loading agent may contain 1, 2, 3, 4, 5, 6, 7, or 8 unsaturations, any of which may be the upper or lower endpoint, respectively.

[0051] Some specific examples of loading agents that are unsaturated fatty acids are shown in the tables below. Derivatives of these fatty acids are also suitable and are therefore contemplated.

Table 1: Examples of monoenoic acids

<td rowspan="2">Total number of carbon atoms in the chain of fatty acids</td><td>Carbon number where the double bond begins.</td>

<td>("C" specifies a cis double bond; "t" specifies a trans double bond)</td>

<td> 10</td><td>4c</td>

<td> 12</td><td>4c</td>

<td> 14</td><td>4c and 9c</td>

<td> 16</td><td>3t, 4c, 5t, 6c, 6t, 9c (palmitoleic) and 1 Ic</td>

<td> 18</td><td>3t, 5c, 5t, 6c (petroselin), 6t, 9c (oleic), 10c, 1 Ic (cis-vaccene), 111 (vaccene) and 13c</td>

<td colspan="2"></td>

<td> 20</td><td>5c, 9c (gadolenic), lic, 13c, and 15c</td>

<td> 22</td><td>5c, 1 Ic (cetoleic), 13c (erucic) and 15c</td>

<td> 24</td><td>15c (selachyl, nerve)</td>

<td> 26</td><td>9c, and 17c (xymenic)</td>

<td> 28</td><td>9c, 19c (lumechin)</td>

<td> 30</td><td>21c</td>

[0052] Unsaturated fatty acids having at least one pair of broken methylene unsaturations are also suitable charging agents. By "methylene-interrupted unsaturation" is meant that one carbon-carbon double bond or triple bond is separated from the other double bond or

- a carbon-carbon triple with at least one methylene group (i.e., CH2). Specific examples of such loading agents include, but are not limited to, the n-1 family derived from 9, 12, 15-16: 3; the n-2 family derived from 9, 12, 15-17: 3, 15: 3, 17: 3, 17: 4, 20: 4; n-3 family coming from 9, 12, 15-18: 3, 15: 2, 15: 3, 15: 4, 16: 3, 16: 4, 18: 3 (a-linolenic), 18: 4, 18 : 5, 20: 2, 20: 3, 20: 4; 20: 5 (EPA), 21: 5, 22: 3, 22: 5 (DPA), 22: 6 (DHA), 24: 3, 24: 4, 24: 5, 24: 6, 26: 5, 26 : 6, 28: 7, 30: 5; the n-4 family derived from 9, 12-16: 2, 16: 2, 16: 3, 18: 2, 18: 3; the n-5 family derived from 9, 12-17: 2, 15: 2, 17: 2, 17: 3, 19: 2, 19: 4, 20: 3, 20: 4 21: 4, 21: 5; the n-6 family derived from 9, 12-18: 2, 15: 2, 16: 2, 18: 2 (linolenic acid), 18: 3 (γlinolenic acid); 20: 2, 20: 3, 20: 4 (arachidonic acid), 22: 2, 22: 3, 22: 4 (adrenic acid), 22: 5, 24: 2, 24: 4, 25: 2, 26: 2, 30: 4; the n-7 family coming from 9-16: 1, 15: 2, 16: 2, 17: 2, 18: 2, 19: 2; the n-8 family derived from 9-17: 1, 15: 2, 16: 2, 17: 2, 18: 2, 19: 2; the n-9 family coming from 918: 1, 17: 2, 18: 2, 20: 2, 20: 3, 22: 3, 22: 4; family n-11 19: 2 and family n-12 20: 2. In one specific example, the loading agent may include arachidonic acid.

[0053] In the paragraph above (and everywhere thereafter), the compounds are identified by reference first to the "nx family", where x is the position in the fatty acid where the first double bond begins. The numbering scheme starts at the end of the fatty acid where, e.g., the end of the CH3 group is at position 1. In this sense, the n-3 family would be an omega-3 fatty acid as described above. The next number represents the total number of carbon atoms in the fatty acid. The third number after the colon is the total number of double bonds in the fatty acid. Thus, for example, in the n-1 family, 16: 3 refers to a 16-carbon fatty acid with 3 double bonds, each separated by methylene, the first double bond starting at the 1 position, i.e. the end of the fatty acid. In another example, in the n-6 family, 18: 3 refers to an 18-carbon fatty acid with 3 double bonds separated by methylene, starting at position 6, i.e. the sixth carbon from the end of the fatty acid, and so on.

[0054] Further examples of loading agents that contain at least one pair of methylene-interrupted unsaturations are shown in Table 2.

Table 2: Examples of polyenoic acids

<td>Total number of carbon atoms in the chain of fatty acids</td><td>Carbon number at which a double bond begins. ("C" means cis double bond, "t" means trans double bond)</td>

<td> 18</td><td> 5, 9</td>

<td></td><td>5, H.</td>

<td></td><td>2t, 9, 12</td>

<td></td><td>3t, 9, 12</td>

<td></td><td>5t, 9, 12</td>

<td></td><td>5, 9, 12 5, H, 14 3t, 9, 12, 15 5, 9, 12, 15</td>

<td> 20</td><td>5, H.</td>

<td></td><td> 5, 13</td>

<td></td><td> 7, 11</td>

<td></td><td> 7, 13</td>

<td></td><td>5, H, 14</td>

<td></td><td> 7, 11, 14</td>

<td></td><td> 5, 11, 14, 17</td>

<td> 22</td><td>5, H.</td>

<td></td><td> 5, 13</td>

<td></td><td> 7, 13</td>

<td></td><td> 7, 15</td>

<td></td><td> 7, 17</td>

<td></td><td> 9, 13</td>

<td></td><td> 9, 15</td>

[0055] Specific examples of suitable loaders that contain conjugated unsaturations include, but are not limited to, those listed in Table 3. By "conjugated unsaturation" it is meant that at least one pair of carbon-carbon double bonds and / or triple bonds is bound to each other, with no methylene (CH2) group between them (e.g., -CH = CH-CH = CH-).

Table 3: Examples of Conjugated Polyacids

<td>Total number of carbon atoms in the chain of fatty acids</td><td>Carbon number at which a double bond begins. ("C" means cis double bond, "t" means trans double bond)</td>

<td> 10</td><td>2t, 4t, 6c</td>

<td></td><td>2c, 4t, 6t</td>

<td></td><td>3t, 5t, 7c</td>

<td></td><td>3c, 5t, 7t</td>

<td> 12</td><td> 3, 5, 7, 9, 11</td>

<td> 14</td><td> 3, 5, 7, 9, 11</td>

<td> 18</td><td>lOt, 12t 8c, lOt, 12c (marjoram) 8t, lOt, 12c (calendar) 8t, 10t, 12t 9t, lithium, 13c (catalp) 9c, lithium, 13c (a-eleostearic) 9c, lit, 13c (punch) 9t, lithium, 13t (β-eleostearic) 9c, lithium, 13t, 15c (a-parinars) 9t, lithium, 13t, 15t (β-paraNar)</td>

[0056] Derivatives of the disclosed loading materials may also be used in the above examples of suitable loaders. By "derivatives" is meant fatty acid esters (e.g., methyl and ethyl esters), fatty acid salts (e.g., sodium and potassium salts) and triglycerides, diglycerides and monoglycerides, sterol esters, antioxidant-oil conjugates (e.g., ascorbyl palmitate) and natural derivatives such as furanoid fatty acid derivatives.

[0057] Loaders disclosed herein can also be crude oil, semi-refined oils (also called refined alkali) or refined oils from the sources disclosed herein. Yet additionally disclosed compositions and methods can use oils containing reesterified triglycerides.

[0058] It is contemplated that one or more of the disclosed loading materials may be used. For example, the disclosed delivery articles may contain two or more different loading substances. Additionally, the loading agent may be present in an amount from about 1% to about 50% by weight of the microcapsule. In specific examples, the charging agent may be present in an amount from about 1% to about 40%, from about 1% to about 30%, from about 1% to about 20%, from about 1% to about 15%, or from about 1%. % to about 10% by weight of the microcapsule.

[0059] In one example, the loading agent is not a fatty acid conjugate. A fatty acid conjugate is a fatty acid that has been conjugated (e.g., bonded) to another chemical moiety, such as a metal (e.g., chromium) or a cofactor (CoQio). In other examples, the loading agent is not low interfacial tension (IT) oil (ie, has an interfacial tension less than about 15 dynes / cm). In other examples, the loading agent is such a fatty acid conjugate or low IT oil.

[0060] In one example, the loading agents may or may include an antioxidant. Suitable examples of antioxidants include, but are not limited to, a phenolic compound, a plant extract, or a sulfur-containing compound. In certain examples disclosed herein, the antioxidant may be ascorbic acid or a salt thereof, e.g., sodium ascorbate. In other examples, the antioxidant can be citric acid or a salt thereof. In still other examples, the antioxidant may be vitamin E, CoQio, lutein, zeaxanthane, carotene (e.g. beta-carotene), tocopherols, lipid soluble derivatives of more polar antioxidants such as ascorbyl esters of a fatty acid (e.g. ascorbyl palmitate). plant extracts (e.g. rosemary, sage and oregano oils), algae extracts and synthetic antioxidants (e.g. BHT, TBHQ, ethoxyquin, alkyl gallates, hydroquinones, tocotrienols) or mixtures thereof.

[0061] The disclosed loading substance may also be or may contain other nutrients such as vitamins, other trace elements (e.g., zinc), minerals, and the like. Additionally, the loading agents may contain other components such as preservatives, antimicrobials, antioxidants, chelating agents, thickening agents, flavoring agents, diluents, emulsifiers, dispersants, or binders, including any mixture thereof.

[0062] Additionally, the charging substance may have a low interfacial tension. For example, a suitable charging agent may have an interfacial tension of less than about 20, less than about 15, less than about 11, less than about 9, less than about 7, or less than about 5 dynes / cm. In other examples, the charging agent can have an interfacial tension from about 0.1 to about 20, from about 1 to about 15, from about 2 to about 9, from about 3 to about 9, from about 4 to about 9, from about 5 to about 9 or from about 2 to about 7 dyne / cm. In still further examples, the charging agent may have an interfacial tension of about 0.1, 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4. , 5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5, 10.0, 10.5 , 11.0, 11.5, 12.0, 12.5, 13.0, 13.5, 14.0, 14.5, 15.0, 15.5, 16.0, 16.5, 17 , 0, 17.5, 18.0, 18.5, 19.0, 19.5, or 20.0, where any of the given values ​​may form the upper or lower endpoint where appropriate. In specific examples, the loading agent may be an algae oil with an interfacial tension of about 0.5, 0.6, 0.7, 0.8, 0.9, or 1.0 dyne / cm. The loading agent may be mushroom oil with an interfacial tension of about 3.0, 3.1, 3.2, 3.3 or 3.4 dyne / cm.

[0063] The interfacial tension of the charging substance can be determined by methods known in the art. For example, the interfacial tension from a loading substance to a standard gelatin solution or from a loading substance to distilled water can be determined using the Fisher surface tensjornate. Generally, a standard solution of gelatin or distilled water can be poured into the sample vessel which is placed on the tensiometer test table. The charging substance can then be added to the sample vessel. The sample may be introduced such that the tensiometer ring is immersed in the charging material. The interfacial stress is a measure of the force acting on the ring that passes through the contact of the charging substance and the standard gelatin solution or contact

-20 charger and distilled water, whichever method is used.

[0064] The interfacial tension measurements for the loading substances presented herein refer to the values ​​determined as described using a standard gelatin solution (50 ° C) which contains 3.3% (w / w) of Kosher Fish Gelatin 240 Bloom (e.g. from LAPI, Tuscany, Italy), 0.5% (w / w) sodium ascorbate and 0.33% (w / w) polyphosphate solution dissolved in water.

[0065] Additionally, the loading charges of the disclosed microcapsules may be from about 20% to about 90%, from about 50% to about 70% by weight, or about 60% by weight of the microcapsule . In other examples, the disclosed microcapsules may contain about 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, or 90% by weight of the microcapsule, the values ​​provided may form the upper limit if desired. or the lower endpoint.

Concrete examples

[0066] Specific examples of microcapsules are disclosed, which include any shell material and any loading substance. Some specific examples include, but are not limited to, microcapsules, where the shell materials are complex coacervates, e.g., gelatin and polyphosphate coacervates. The shell material may, in some examples, contain gelatin with a Bloom Number from about 0 to about 50. Loading agents that may be used may in many cases include marine oils (e.g. fish oils and algae oils). Loaders containing omega-3 fatty acids such as EPA and DHA may be desirable. Additionally, loaders for omega-3 fatty acid derivatives such as mono-, di- and triglycerides, alkyl esters, sterol esters, antioxidant esters (e.g. ascorbyl and citrile esters) and furanoid esters may also be suitable.

[0067] Some particularly suitable microcapsules include microcapsules containing fish oils. Examples of such fish oils include, but are not limited to, sardine, anchovy, bonito, and / or tuna oil. Fish oils may also be referred to herein as the approximate ratio of EPA and DHA, or derivatives thereof, present in the oil. For example, 18:12 oils typically contain a ratio of EPA to DHA (or, for example, their triglyceride esters) of about 18:12. Likewise, 5:25 oils generally contain an EPA to DHA ratio of about 5:25. Each of these oils can be encapsulated in a complex coacervate containing either fish or pork gelatin. Such microcapsules can be generally Regarded as Safe GRAS, Kosher and / or Halal. Such microcapsules may have at least about 130 mg of DHA or at least about 150 mg of EPA and DHA per gram of powder. Additionally, antioxidants such as ascorbic acid, citric acid, and / or phosphoric acid (or their salts) may be present in such microcapsules.

[0068] Some specific examples of the disclosed foodstuffs include microcapsules containing about 130 mg DHA per gram of microcapsule (e.g., a microcapsule in which the loading substance contains 5:25 tuna-derived oil and / or bonito) and the outer shell of the microcapsules comprises pork or fish gelatin . In another

In a specific example, the disclosed foodstuff may contain a microcapsule containing about 150 mg of DHA and EPA per gram of microcapsule (e.g., a microcapsule in which the loading agent contains 18:12 of oil derived from sardines and / or anchovies) and the outer shell of the microcapsules comprises pork or fish gelatin. .

[0069] Particularly suitable microcapsules are disclosed in US Patent Nos. 6,974,592 and 6,969,530 and US Publication Nos. 2005-0019416-A1.

The method of producing microcapsules

[0070] Microcapsules made by the methods disclosed herein typically have a combination of payload and structural strength that is suitable for the comestibles, supplements, formulation carriers, and methods disclosed herein. In one example, the methods disclosed in US Patent Nos. 6,974,592 and 6,969,530, and US Publication Nos. 2005-0019416-A1 can be used to prepare microcapsules. It is also contemplated that one or more additional sheath layers may be placed on the outer sheath of the single or multicore microcapsules. In one example, the techniques described in International Publication No. WO 2004/041251 A1 can be used to add additional shell layers to single and multicore microcapsules.

[0071] In general, suitable microcapsules can be made by a method that comprises providing an emulsion containing the first polymer component and a loading agent; adding a second polymer to the emulsion; adjusting the pH, temperature, concentration, agitation rate, or combinations thereof to form an aqueous mixture containing the primary shell material, the primary shell material comprising the first and second polymeric components and surrounding the charging agent; cooling the aqueous mixture to a temperature above the gelling temperature of the primary shell material until the primary shell material agglomerates; and further cooling the aqueous mixture to form a shell around the agglomeration.

[0072] In these methods, the first polymer component and the second polymer component may be the same as any of the primary and secondary coating material described herein. That is, the first and second polymeric components can become the primary and / or outer shell materials in the disclosed microcapsule preparation methods. Additionally, any loading material described herein may be used in these methods of making microcapsules.

[0073] In the disclosed methods, an aqueous charging substance mixture, a first polymeric component of the shell material and a second polymeric component of the shell material are formed. The aqueous mixture can be a mechanical mixture, a suspension or an emulsion. When a liquid loading agent, especially a hydrophobic liquid, is used, the aqueous mixture may be an emulsion of the loading substance and polymer components. In another example, the first polymer component is provided in an aqueous solution along with processing aids such as antioxidants. The charging substance can then be dispersed into the aqueous mixture, for example using a homogenizer. If the charging substance is a hydrophobic liquid, an emulsion is formed in which a fraction of the first

The polymer component begins to settle around the individual drops of the charging substance to begin forming the original shells. If the loading agent is a solid particle, a suspension is formed in which a fraction of the first polymer component begins to settle around the individual particles to begin forming primary shells. Another aqueous solution of the second polymer component may be added to the aqueous mixture at this point.

[0074] In the disclosed methods for preparing microcapsules, an emulsion of the first polymer component is provided and the loading substance can be achieved by methods and devices known in the art, e.g., homogenization and high pressure / high shear pumps. For example, emulsification can occur by emulsifying from about 1,000 to about 15,000 rpm. The emulsification step can be monitored by removing a sample of the mixture and analyzing it by methods such as microscopy, light scattering, turbidity, etc. In general, emulsification can be performed until the average droplet size is less than about 1000, 750, 500, 100, or 10 nm. Without wishing to be bound by theory, it is believed that single-core or multi-core microcapsules can be produced by varying the rate of emulsification. For example, when lower emulsification rates are used (e.g. 1000 up to 2000 rpm), the droplets of the loading substance are large enough to form a single particle which, when encapsulated, produces a single-core microcapsule. Conversely, when high emulsification rates are used (e.g. 5,000 to 15,000 rpm), the resulting loading droplets are usually small (e.g. 1 to 10 µm). These small droplets can exhibit higher surface energy and can easily agglomerate when the pH and / or temperature are appropriately controlled, resulting in the formation of multi-core microcapsules upon encapsulation. Particle size can be measured using any conventional device known in the art, for example COULTER.TM. LS230 Particie Size Analyzer, Miami, Florida, USA,

[0075] The emulsifying step may be carried out at a temperature greater than room temperature, greater than 30, 40, 50, 60, 70 or 80 ° C, if any of the values ​​given may constitute an upper or lower endpoint as desired. Specific examples include emulsifying the mixture at a temperature from about 30 ° C to about 60 ° C or from about 40 °. to about 50 ° C.

[0076] Additionally, it is contemplated that antioxidants and / or surfactants, which are also described herein, may be added to the emulsion and / or the aqueous mixture. Such antioxidants and / or surfactants may be added before, during and / or after delivery of the emulsion. Additionally, in the entire system containing the loading agent, shell materials, antioxidants, and additional compositions, the antioxidant capacity is at some level when the amount of antioxidants is administered. Therefore, in the methods for making microcapsules disclosed herein, purging with an inert gas such as nitrogen during any or all of the emulsification, mixing, coacervation, and cooling methods can prevent air oxygen from consuming antioxidants and delay oxidation of the loading agent during storage. It can also prevent the formation of aroma disturbing compounds due to oxidation during microencapsulation.

[0077] It is also contemplated that the chelators may be added to the emulsion and / or the aqueous mixture. Lipid autooxidation is catalyzed by metal ions, especially iron and copper ions. Metal ion chelation can therefore delay oxidation and extend its "delay phase", thereby extending the shelf life of bulk oil or encapsulated oils. Like antioxidants, chelators can be added before, during and / or after delivery of the emulsion. Examples of suitable chelators include, but are not limited to, disodium ethylenediaminetetraacetic acid, which is one of the most widely used chelating agents in food processing, citric acid, phytic acid, malic acid, tartaric acid, oxalic acid, succinic acid, polyphosphoric acids, etc.

[0078] The amount of polymeric components of the shell material provided in the aqueous mixture is usually sufficient to form both the primary shells and the outer shells of the charging agglomeration of the microcapsules. The loading agent may be provided in an amount of from about 1% to about 15% by weight of the aqueous mixture, from about 3% to about 8% by weight, or about 6% by weight.

[0079] The PH, temperature, concentration, agitation rate, or a combination thereof can be adjusted to form an aqueous mixture comprising the primary coating material, the primary coating material comprising the first and second polymer components and surrounding the charging agent. If more than one type of polymer component is present, complex coacervation will occur between the elements to form a coacervate which then settles around the charging substance to form the primary envelopes of the shell material. The adjustment of the pH depends on the type of shell material to be prepared. For example, the pH can be adjusted to between 3.5 and 5.0, or between 4.0 and 5.0. If the pH of the mixture begins in the required range, little or no pH adjustment is required.

[0080] The initial temperature of the aqueous mixture may be from about 20 ° C to about 60 ° C, or about 30 ° C to about 50 ° C.

[0081] The mixing can be controlled such that there is good mixing without destroying the microcapsules as they are formed. The particular mixing parameters depend on the type of equipment used. Any of the many types of mixing devices known in the art can be used. In one example, an axial flow rotor such as a LIGHTNIN ™ A310 or A510 may be used.

[0082] In many examples disclosed herein, the primary shell and outer shell of the disclosed microcapsules may contain a complex coacervate. The complex coacervate may be formed from the first and second polymer components. For example, the primary coating and the outer coating may contain a complex coacervate between gelatin and polyphosphate. All combinations of the first and second polymer components are contemplated herein for complex coacervate and primary and outer shells.

[0083] The aqueous mixture may then be cooled under a controlled cooling rate and mixing parameters to allow agglomeration of the primary shells to form encapsulated primary shell agglomerations. Not wanting to be bound by theory,

The encapsulated agglomerations are self-contained molecules. It is preferable to control the formation of the encapsulated agglomerates at a temperature above the gelling temperature of the shell material and allow excess shell material to form a thicker outer shell. It is also possible to add more polymer at this stage where the polymer is the same or different than the shell material to thicken the outer shell and / or to produce microcapsules having primary and outer shells of different composition. The outer shell encapsulates the agglomeration of the primary shells to form a rigid encapsulated agglomeration of the microcapsules.

[0084] Cooling of the aqueous mixture can be carried out by methods known in the art (e.g. using a refrigerator). The cooling rate may be about 1 ° C for about 1 to about 100 minutes. For example, the cooling rate may be about 1 ° C for about 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90 , 95, or 100 minutes, if any of the values ​​provided can create an upper or lower endpoint as needed. In specific examples, the cooling rate may be about 1 ° C / 5 minutes. Cooling may take place until the mixture is at a temperature of from about 5 ° C to about 10 ° C, for example about 5 ° C.

[0085] Processing aids may be included in the shell material (e.g., primary and / or outer shells). Processing aids can be used for a variety of reasons. For example, they can be used to stimulate agglomeration of primary microcapsules, stabilize the emulsion system, improve outer shell properties, control microcapsule size, and / or act as an antioxidant. In one aspect, the processing aid may be an emulsifier, fatty acid, lipid, wax, microbe (e.g. a yeast cell line), clay, or an inorganic compound (e.g. calcium carbonate). Without wishing to be bound by theory, these processing aids can improve the barrier properties of the microcapsules. In one aspect, one or more antioxidants may be added to the shell material. Antioxidant properties are useful both during the process (e.g. during coacervation and / or spray-drying) and in the microcapsules after their formation (i.e. to extend shelf life, etc.). Advantageously, a small number of processing aids that perform a large number of functions may be used. In one aspect, the antioxidant can be a phenolic compound, a plant extract, or a sulfur-containing amino acid. In one aspect, ascorbic acid or citric acid (or a salt thereof, such as sodium or potassium ascorbate, sodium or potassium citrate) may be used to promote agglomeration of primary microcapsules to control microcapsule size and act as an antioxidant. The antioxidant may be used in an amount of about 100 ppm to about 12,000 ppm, or from about 1,000 ppm to about 5,000 ppm. Other processing aids, such as, for example, metal chelators, can also be used. For example, ethylenediaminetetraacetic acid can be used to bind metal ions, which can reduce catalytic oxidation of the loading agent.

[0086] In the disclosed microcapsules, the shell material may also be cross-linked. Thus, the disclosed methods may further comprise adding a crosslinking agent. You can

Add a cross-linker to further increase the stiffness of the microcapsules by cross-linking the shell material in the outer and primary shells and to make the shells insoluble in both aqueous and oily media. In one example, the crosslinking agent is added after the outer shell of the microcapsule is formed. Any suitable crosslinker can be used, and the selection of the crosslinker may vary with the selection of the first and second polymer components. In another example, the crosslinkers can be enzymatic crosslinkers (e.g., transglutaminase), aldehydes (e.g., formaldehyde or glutaraldehyde), tannic acid, alum, or a mixture thereof. In another aspect, the cross-linking agent can be a plant extract or a phenolic compound. It is also believed that one or more loading agents (e.g., antioxidants) can be used with the crosslinker. When the microcapsules are to be used in a formulation that is to be delivered to the body, the cross-linking agents are preferably non-toxic or have a sufficiently low toxicity. The amount of crosslinker used depends on the components selected and can be adjusted to provide more or less structural stiffness as needed. In one aspect, the amount of the crosslinker may be from about 0.1% to about 5.0%, from about 0.5% to about 5.0%, from about 1.0% to about 5.0%, from about 2.0% to about 4.0%, or about 2.5% by weight of the first polymer component. In general, one skilled in the art can routinely determine the desired amount in any case by simple experimentation. The crosslinking sodium can be added at any stage of the process; it can usually be added after the cooling step.

[0087] Additionally, in some applications, the use of transglutaminase to cross-link microcapsules may not be desirable (eg, temperature, pH are too low, and / or transglutaminase is expensive). Accordingly, it is contemplated herein that the disclosed methods may employ glutaraldehyde to cross-link the disclosed microcapsules. In certain instances, the use of one or more amino acid or protein containing compositions may react with residual glutaraldehyde that was completely or partially unreacted from the crosslinking reaction. This means that unreacted and semi-reacted glutaraldehyde (i.e. with one aldehyde group still reactive) can be neutralized by a protein with the ε-amino group of lysine or a different amino group, making the end product safer. In this sense, compositions containing amino acids and / or proteins can improve the shell of the microcapsule by filling any pores and neutralizing glutaraldehyde from the cross-linking reaction. This approach can also eliminate the need to wash the microcapsule after cross-linking as the microcapsule will be substantially free of glutaraldehyde. Cross-linking can also be achieved by genipin (e.g. with genipin and carboxymethyl chitosan).

[0088] Furthermore, the disclosed microcapsules can be washed with water and / or dried to provide a free flowing powder. Thus, the disclosed methods for making microcapsules may include a drying step for the microcapsules. Drying may be achieved by a number of methods known in the art, such as, for example, freeze drying, ethanol drying, or spray drying. In one aspect, spray drying can be used to dry the microcapsules. Spray drying techniques are disclosed

-26 in "Spray Drying Handbook", K. Masters, Wyd. 5., Longman Scientific Technical UK, 1991, the disclosure of which is incorporated herein by reference at least in relation to its description of spray drying methods.

Addition of saccharides before coacervation

[0089] In some instances, saccharides such as the chitosan polysaccharide, chitin and others disclosed herein may be added prior to emulsification and coacervation to provide microcapsules with enhanced impermeability. Without wishing to be bound by theory, adding saccharides to the polymer component solution (e.g. gelatin) increases the viscosity of the base and therefore can help stabilize the oil droplets after emulsification, to illustrate, the chitosan polysaccharide, consisting of D-glucosamine units, contains many amino groups as shown below.

OH H

[0090] At some pH the cationic molecule will participate in electrostatic interactions during coacervation. The chitosan then forms a "composite" shell material together with the first and second polymeric materials (eg, gelatin polyphosphate coacervates).

[0091] Furthermore, transglutaminases (TGases) can cross-link proteins (ie gelatin) (Fig. 1), including gelatin incorporated into chitosan. While not all of the amine groups on the lysine and glutamine residue on gelatin are cross-linked by TGases, including saccharides such as chitosan to the shell material, they may form additional cross-links to form bridging between gelatin molecules. Therefore, the shell strength would be greater and the pore size could be reduced (better oxygen barrier) (Fig. 2).

Addition of saccharides and / or amino acids after shell formation and cross-linking

[0092] In another example, amino acids such as lysine and / or glutamine may be added to the microcapsules after their formation, but before or after cross-linking with the transglutaminase. As discussed above, in order to form bonds between the amino groups of lysine and glutamine, these two amino residues must be in the correct spatial position such that TGase can catalyze the reaction. It can be assumed that not all amine groups can form cross-links. Therefore, after shell formation and cross-linking, amino groups are available on the gelatin shell materials. When lysine and glutamine are added, TGase will be able to attach them to the glutamine and lysine residues on the gelatin molecules. This can therefore form amino acid attachments inside the pores of the shell and can improve the barrier properties of the microcapsules.

[0093] A combination of polysaccharides such as chitosan and amino acids can also be used. For example, when chitosan is added after shell formation and shell cross-linking, it may attach to a lysine and glutamine residue or form a bridge between gelatin molecules or domains with an available NH2 moiety and / or an available glutamine NH2 moiety.

[0094] When chitosan is added to lysine and glutamine, the effect may be better under certain circumstances as they can match pores of different sizes.

[0095] In some circumstances, the use of lysine and glutamine may promote moisture sorption, which may not be desirable. Thus, the use of amino acids such as cysteine, leucine, isoleucine, phenylalanine, tyrosine, tryptophan and tyrosine in combination or combination with glutamine and / or chitosan is disclosed. Such a microcapsule powder may have a better moisture barrier as these amino acids are more hydrophobic than lysine. Therefore, powder agglomeration can be slowed down.

Adding wax

[0096] Hydrophobic materials such as waxes can have good moisture barrier properties, especially when compared to proteins and carbohydrates. Thus, disclosed herein are microcapsules wherein the void volume within the multi-core agglomeration comprises wax particles. The addition of the wax particles can fill the space in the agglomeration as well as the pores of the shell (Fig. 3). Wax can be added at various points during the preparation of the microcapsule. For example, wax (e.g. in a microemulsion of wax particles) can be added to the emulsion and / or the aqueous mixture prior to coacervation. Alternatively, or additionally, the wax may be added after shell formation and cross-linking (e.g., before spray drying). In this way, the wax can form a protective layer, thus improving the moisture and oxygen barrier of the microcapsules (Fig. 4).

Co-spray drying of the protective saccharides and / or proteins after shell formation and cross-linking

[0097] After the shell is formed and cured by cross-linking, the microcapsules can be used directly in suitable slurry applications or made into a dry powder during dehydration such as spray drying. Co-spraying of the disclosed microcapsules with protective materials can further improve stabilization of the loading substance. Protective compositions include, but are not limited to, lipids and waxes, carbohydrates, saccharides, amino acids, peptides, and proteins as described herein. By filling the shell and / or coating the surface of the shell, the protective materials can provide additional barriers to moisture and oxygen after co-spray drying. One or more of these protective compositions may be added to the microcapsule suspension either in dry form or as a solution (e.g., dissolved in water). The protective agents can be applied just prior to spray drying the slurry, which allows dissolving and mixing.

[0098] Carbohydrates have higher glass transition temperatures (i.e., more stable to molecular mobility) than proteins and lipids. Carbohydrates are also a better barrier to oxygen than proteins and lipids (when dry). Spray-drying microcapsules together with carbohydrates can form a more stable matrix that can provide better protection against oxygen attack on the encapsulated loading substances. Co-spray dried polysaccharides with microcapsules can provide increased impermeability mainly by forming a protective matrix as a coating layer on the surface of the microcapsule shell. When the coating materials contain amphiphilic moieties, these film-forming materials exhibit improved properties for both moisture and oxygen barriers due to their hydrophobic moieties. Examples of such protective materials are disclosed herein and include gum arabic and a modified starch such as sodium octenyl succinate. In addition to the matrix coating on the shell surface, molecules of medium size carbohydrates or small sugars also disperse in the porous network of shell polymers and block the pathway of oxygen and / or volatile compounds, such as those in malodorous and malodorous ones.

[0099] The inclusion of proteins in the microcapsule suspension prior to spray-drying can produce an inert and stable powder which improves the drying efficiency. Thermally denatured proteins can undergo irreversible thermal gelation which forms a stable coating on the surface of the microcapsules. Heating the mixture before drying can also reduce the amount of aromatics. Protein co-spray drying compositions may also contain plasticizers such as glycerol, sorbitol, mono-, di- or oligosaccharides (e.g. lactose). Small molecules such as oligopeptides and hydrophobic amino acids can also fill the porous molecular network of the shell materials in addition to film formation on the surface of the coating microcapsules.

Inclusion of drying / anti-caking agents to improve fluidity

[0100] Drying agents or anti-caking agents may also be used to prepare free flowable powders. Typically desiccants have a high porosity, which can help adsorb surface oil and aromatics due to raw materials or lipid oxidation. Examples of suitable drying and / or anti-caking agents include, but are not limited to, HUBERSORB ™ and ΖΕΟΤΗΙΧ ™ (JM Huber Corp; Harve de Grace, Md.) And CAPSUL ™ (from National Starch &amp; Chemical Co.) and VITACEL ™ (J. Rettenmair USA; Schoolcraft, Mich.).

Incorporating antioxidants into the powder

[0101] Other examples disclose methods of incorporating antioxidants into and / or onto the primary shell, outer shell, or both primary and outer shell materials. The disclosed methods include providing a microcapsule, providing an emulsion containing a polymer component and an antioxidant; combining the emulsion and the microcapsule, producing a microcapsule with an antioxidant-containing shell material. Suitable antioxidants include, but are not limited to,

-29CoQίο, lutein, zeaxanthan, carotene, and combinations thereof. They can be used alone or in addition to the amino acids, proteins, saccharides, or waxes disclosed herein.

[0102] The microcapsule can be any microcapsule, but particularly suitable microcapsules are those disclosed. Such microcapsules can be formed, for example, by providing an emulsion comprising a first polymer component, a loading agent, a second polymer component; by adjusting the pH, temperature, concentration, agitation rate or a combination thereof to agglomerate the primary microcapsules, each primary microcapsule having a primary shell, the loading agent being encapsulated by the primary shell, the agglomeration being encapsulated by the exterior shell, with the primary shell and the outer one include first and second polymer components. The resulting agglomeration may then be combined with an antioxidant emulsion of the third polymer component, which may be the same or different from the first or second polymer components. The resulting suspension may then be cooled and the coated microcapsules can be dried. In many suitable examples, the microcapsules can be contained in a suspension that includes antioxidants, and the suspension can be spray-dried.

Incorporation of zinc into the powder

[0103] Other examples disclose methods for incorporating zinc into and / or on the primary shell, outer shell, or both primary and outer shells. The disclosed methods include microcapsule delivery, providing an emulsion containing a polymer component and zinc; combining the emulsion and the microcapsule to provide the microcapsule with the zinc-containing shell material. Zinc may be used alone or in addition to the amino acids, proteins, saccharides, or waxes disclosed herein.

[0104] The microcapsule can be any microcapsule, but particularly suitable microcapsules are those disclosed. Such microcapsules can be formed, for example, by providing an emulsion comprising a first polymer component, a loading agent, a second polymer component; by adjusting the pH, temperature, concentration, agitation rate, or a combination thereof to agglomerate the primary microcapsules, each primary microcapsule having a primary shell, the loading agent being encapsulated by the primary shell, the agglomeration being encapsulated by the outer shell, and the shell the primary and outer components contain first and second polymer components. The resulting agglomeration can then be combined with an antioxidant emulsion and a third polymer component, which may be the same or different from the first or second polymer components. The resulting suspension may then be cooled and the coated microcapsules can be dried. In many suitable examples, the microcapsules can be contained in a suspension that includes zinc and the suspension can be spray dried.

Concrete examples

[0105] In a specific example, methods for preparing a microcapsule are disclosed herein, comprising providing an emulsion comprising a first polymeric component and a composition.

Containing a saccharide, a wax, or a combination thereof; adding a loading agent, a second polymer component and, optionally, a composition, to the emulsion; adjusting the pH, temperature, concentration, agitation rate, or combinations thereof to form an aqueous mixture containing the primary shell material, the primary shell material comprising the first and second polymeric components and surrounding the charging agent; cooling the aqueous mixture to a temperature above the gelling temperature of the primary shell material until the primary shell material agglomerates; and further cooling the aqueous mixture to form an outer shell around the agglomeration, wherein the primary shell, outer shell or both material comprises a saccharide, a wax, or a combination thereof.

[0106] In another specific example, methods for preparing a microcapsule are disclosed, comprising providing an emulsion comprising a first polymer component, a loading agent and a second polymer component; adjusting the pH, temperature, concentration, agitation rate, or a combination thereof to form an aqueous mixture containing the primary shell material, the primary shell material comprising the first and second polymer components and surrounding the loading agent; cooling the aqueous mixture to a temperature above the gelling temperature of the primary shell material until the primary shell material agglomerates; further adding the saccharide-containing composition to the aqueous mixture; and further cooling the aqueous mixture to form an outer shell around the agglomeration, wherein the primary shell material, the outer shell or both contain saccharide.

[0107] In yet another specific example, methods for preparing a microcapsule are disclosed, comprising providing a suspension of one or more microcapsules, wherein the microcapsule comprises a shell material and a loading agent; adding a composition containing one or more of an amino acid, protein, saccharide, wax, antioxidant, zinc, or combinations thereof to the suspension; and then drying the slurry.

[0108] In yet another specific example, disclosed herein are methods for preparing a microcapsule, comprising providing an emulsion comprising a first polymer component, a loading agent, a second polymer component, and an emulsion chelator; adjusting the pH, temperature, concentration, agitation rate, or a combination thereof to form an aqueous mixture containing the primary shell material, the primary shell material comprising the first and second polymer components and surrounding the loading agent; cooling the aqueous mixture to a temperature above the gelling temperature of the primary shell material until the primary shell material agglomerates; and further cooling the aqueous mixture to form an outer shell around the agglomeration.

Formula carriers

[0109] Also disclosed herein are formulation vehicles containing the disclosed microcapsules. Any of the microcapsules described herein can be incorporated into a formulation carrier. Examples of formulation carriers are provided herein and include, but are not limited to, foodstuffs, beverages, nutritional preparations, pharmaceutical preparations, lotions, creams or sprays.

In some other specific examples, the disclosed emulsions and / or microcapsules can be incorporated into gels, gelcaps, or tablets. Other carriers include powders or polymer coated powders. Such carriers may be administered orally or, for example, in a powder, sprinkled on food or beverages.

Supplements

[0110] Nutritional supplements are also disclosed herein, which include the microcapsules as disclosed herein. A nutritional supplement is any compound or composition that can be administered to a subject or that can ingest, provide or enhance nutrient (s) (e.g., vitamins, minerals, essential trace element, amino acid, peptide, nucleic acid, oligonucleotide) , lipid, cholesterol, steroid, carbohydrate and the like). For example, a dietary supplement may contain a composition containing one or more of the substances disclosed herein.

[0111] A dietary supplement may include any number of microcapsules disclosed in this specification, but typically includes an amount determined to provide the subject with the required dose of loading agent (e.g., EPA and / or DHA). The exact amount of microcapsules required in nutritional supplements will vary from subject to subject, depending on the species, age, weight and general condition of the patient, the severity of any dietary deficiency, the particular mode of administration, and the like. Therefore, it is not possible to define the exact amount for each nutritional supplement. However, the appropriate amount can be determined by one of ordinary skill in the art using only routine experimentation, taking into account the knowledge contained herein.

[0112] The nutritional supplement may contain other nutrients such as vitamins, other trace elements, minerals, and the like. In addition, the nutritional supplement may contain other ingredients such as preservatives, antimicrobials, antioxidants, chelating agents, thickening agents, flavoring agents, diluents, emulsifiers, dispersants, or binders.

[0113] Nutritional supplements are generally taken orally and may be in any form suitable for oral administration. For example, a nutritional supplement may typically be in a tablet, gel, capsule, liquid, sachet, or syrup form.

[0114] The nutritional supplements may be intended for humans or animals, based on the recommended dietary intake for an individual. Such considerations are based generally on various factors such as species, age, and sex as described above that are known or can be determined by those skilled in the art. In one example, the disclosed supplements can be used as an ingredient in feed for animals such as, but not limited to, livestock (e.g. pigs, chickens, cows, goats, horses etc.) and domestic animals (e.g. cats, dogs, birds and the like).

Pharmaceutical formulation

[0115] Also disclosed is a pharmaceutical formulation containing the disclosed microcapsules. A suitable pharmaceutical formulation may contain any of the compositions disclosed

With a pharmaceutically acceptable carrier. For example, a pharmaceutical formulation may contain one or more of the disclosed emulsions and / or microcapsules and a pharmaceutically acceptable carrier. The disclosed pharmaceutical formulations can be used therapeutically or prophylactically.

[0116] By "pharmaceutically acceptable" is meant a material that is not biologically or otherwise undesirable, ie, the material can be administered to a subject without causing any undesirable biological effects or adversely affecting any of the other ingredients of the pharmaceutical formulation in which it is contained. . The carrier will of course be chosen to minimize any degradation of the active ingredient and to minimize any adverse side effects on an individual as would be well known to those skilled in the art.

[0117] Pharmaceutical carriers are well known to those skilled in the art. These would most often be the standard vehicle for administering drugs to humans, including solutions such as sterile water, saline, and buffered solutions at physiological pH. Suitable carriers and their formulations are described in Remington: The Science and Practice of Pharmacy, Ed. 21 .., Lippincott Williams &amp; Wilkins, Philadelphia, Pa., 2005. Typically, an appropriate amount of a pharmaceutically acceptable salt is used in the formulation to render the formulation isotonic. Examples of a pharmaceutically acceptable vehicle include, but are not limited to, saline, Ringer's solution, and dextrose solution. The pH of the solution may be from about 5 to about 8 (e.g., from about 7 to about 7.5). Further carriers include sustained release preparations, such as semi-permeable matrices of solid hydrophobic polymers containing the disclosed compounds, which matrices are in the form of shaped articles, e.g., films, liposomes, microparticles, or microcapsules. It will be apparent to those skilled in the art that certain carriers may be more advantageous depending, for example, on the route of administration and the concentration of the composition. The other compounds can be administered according to standard procedures used by those of skill in the art.

[0118] The pharmaceutical preparations can contain additional carriers as well as thickeners, diluents, buffers, preservatives, surfactants and the like in addition to the compounds disclosed herein. The pharmaceutical preparations can also contain one or more additional active ingredients such as antibacterial agents, anti-inflammatory agents, anesthetics, and the like.

[0119] The pharmaceutical formulation may be administered in a variety of ways depending on whether local or systemic treatment is desired and to the area to be treated. Administration may be topical (including ophthalmologically, vaginally, rectally, nasally), orally, by inhalation, or parenterally, such as by intravenous drip, subcutaneous, intraperitoneal, or intramuscular injection. The disclosed compounds can be administered intravenously, intraperitoneally, intramuscularly, subcutaneously, intravenously or transdermally.

[0120] Preparations for parenteral administration include sterile aqueous or non-aqueous solutions, suspensions, and emulsions. Examples of non-aqueous solvents are propylene glycol, polyethylene glycol, vegetable oils such as olive oil, sea oils

Injectable organic esters such as ethyl oleate. Aqueous carriers include water, alcoholic / aqueous solutions, and emulsions or suspensions including saline and buffered media. Parenteral vehicles include sodium chloride solution, Ringer's dextrose, dextrose and sodium chloride, lactated Ringer's solutions, and hydrogenated oils. Intravenous vehicles include fluid and nutritional supplements, electrolyte fillers (such as based on Ringer's dextrose), and the like. Preservatives and other additives may also be present, such as, for example, antimicrobials, antioxidants, chelating agents, and inert gases, and the like.

[0121] Pharmaceutical formulations for topical administration may include ointments, lotions, creams, gels, drops, suppositories, sprays, liquids, and powders. Conventional pharmaceutical carriers, aqueous, powdered or oily bases, thickening agents and the like may be desired.

[0122] Pharmaceutical formulations for oral administration include, but are not limited to, powders or granules, suspensions or solutions in water or non-aqueous media, capsules, sachets or tablets. Thickening agents, flavoring agents, diluents, emulsifiers, dispersion aids or binders may be desirable.

[0123] Some of the formulations can potentially be administered as a pharmaceutically acceptable acid or base addition salt formed by reaction with inorganic acids such as hydrochloric acid, hydrobromic acid, perchloric acid, nitric acid, thiocyanic acid, sulfuric acid, and phosphoric acid and acids organic acid, such as formic acid, acetic acid, propionic acid, glycolic acid, lactic acid, pyruvic acid, oxalic acid, malonic acid, succinic acid, maleic acid and fumaric acid, or by reaction with an inorganic base such as sodium hydroxide, ammonium hydroxide, potassium hydroxide, and organic bases such as mono-, di-, trialkyl, and arylamines and substituted ethanolamines.

Groceries

[0124] Also disclosed herein are comestibles that contain any of the disclosed microcapsules. By "foodstuff" is meant any item that can be consumed (eg, eaten, drunk, or assimilated) by the subject. In one example, the disclosed compositions can be used as nutritional supplements to be added to a foodstuff. For example, the disclosed microcapsules can be added to food or beverages. In this sense, the disclosed compositions may be prepared, for example, in powdered form and contained in articles such as sachets or shakes which may be used to pour or spray the disclosed compositions onto and into foods and drinks.

[0125] In some examples, the foodstuff is a baked product, a pasta product, a meat product, a frozen dairy product, a dairy product, a cheese product, an egg product, a seasoning, a soup mix, a snack, a nut product, a vegetable protein product, a hard candy, a soft candy. candy, poultry product, processed fruit juice, granulated sugar (e.g. white or brown), sauce, syrup, nutritional bar, drink

-34 dry powdered drink, jam or jelly, fish product, or accompanying food. In other examples, the food item is bread, tortillas, cereals, sausage, chicken, ice cream, yogurt, milk, salad dressing, rice bran, fruit juice, dry drink powder, liquid drink, rolls, cookies, crackers, fruit pie or pies.

Emulsions

[0126] Also disclosed are compositions comprising a spray dried emulsion comprising a first polymer component and a loading agent and the remainder of one or more compositions comprising an amino acid, protein, saccharide, wax, or a combination thereof. The first polymer component may be any of the first polymer components disclosed herein. Likewise, the loading material may be any of the substances disclosed herein. Still further, mention may be made of amino acid, protein, saccharide, wax, and combinations thereof.

Application methods

[0127] The disclosed microcapsules have wide application as well. For example, disclosed are methods of delivering a loading substance to a subject by administering a microcapsule to the subject in accordance with the disclosure. Also disclosed is the use of a microcapsule of the disclosure for the preparation of a medicament for delivering a loading substance to a subject.

[0128] The use of microcapsules can protect certain compositions from oxidation and degradation while keeping the loading substance fresh. In addition, because microcapsules can hide an unpleasant odor or taste from certain compositions, the methods disclosed herein may be particularly useful for delivering and replenishing unpleasant compositions. Still further, the use of microcapsules can allow various charging substances to be added to foodstuffs that cannot otherwise be refilled. For example, omega-3 fatty acid can degrade or oxidize in air and can be sensitive to food manufacturing techniques (e.g. baking). By using microencapsulated omega-3 fatty acids, the compositions can be added to foods without significant degradation during food preparation.

[0129] Particularly suitable microcapsules include those that are resistant to breakage during the preparation of the food product (including the packaging, transport and storage of the foodstuffs). In some examples, the microcapsules can be of a size and texture that does not reduce the texture and constitution of the foodstuff.

[0130] In a specific example, the disclosed microcapsules (including dietary supplements, pharmaceutical formulations, delivery products, and foodstuffs containing the disclosed microcapsules) can be used as a source of fatty acids (eg, omega-3 fatty acids), lowering triglycerides and affecting the biochemistry of diabetes. In another specific example, methods of replenishing omega-3 fatty acids in a subject are disclosed herein by administering an effective amount of a microcapsule disclosed herein, wherein the loading agent comprises an omega-3 fatty acid. In another

As an example, disclosed herein are methods of lowering cholesterol levels, triglyceride levels or combinations thereof in a subject by administering an effective amount of the disclosed emulsion and / or microcapsule.

[0131] Omega-3 fatty acids are essential for everyday life and function. For example, the beneficial effects of omega-3 fatty acids such as cis5,8,11,14,17-eicosapentaenoic acid (EPA) and cis-4,7,10,13,16,19-docosahexaenoic acid (DHA) in reducing the concentration of Serum triglycerides are well known. These compounds are also known to have other cardioprotective benefits such as preventing heart disorders, stabilizing atherosclerotic plaques, reducing platelet aggregation and reducing blood pressure. See, e.g., Dyrberg et al., In Omega-3 Fatty Acids: Prevention and Treatment of Vascular Disease. Kristensen et al., Eds., Bi &amp; Gi Publ., Verona-Springer-Verlag, London, pp. 217-26,1995; O'Keefe and Harris, Am. J. Cardiology 2000, 85: 1239-41; Radack et al., "The effects of Iow doses of omega-3 fatty acid supplementation on blood pressure in hypertensive subjects: a randomized controlled trial." Arch. Intern. Med. 1991,151: 1173-80; Harris, "Extending the cardiovascular benefits of omega-3 fatty acids." Curr Atheroscler Rep 2005, 7: 375-80; Hołub, "Clinical nutrition: 4 omega-3 fatty acids in cardiovascular care." CMAJ 2002,166 (5): 608-15. In fact, the American Heart Association also reported that omega-3 fatty acids can reduce the risk of cardiovascular disease and heart disease. Other benefits of omega-3 fatty acids include activities related to the prevention and / or treatment of inflammation and neurodegenerative conditions, and the improvement of cognitive development. See, e.g., Sugano and Michihiro, "Balanced intake of polyunsaturated fatty acids for health benefits." J. Oleo Sci. 2001, 50 (5): 305-11.

[0132] The fatty acids EPA and DHA can be synthesized in the human body from alinolenic acid (18: 3); however, the conversion rate from this precursor molecule is limited (Muskiet et al., "Is docosahexaenoic acid (DHA) essential? Lessons from DHA status regulation, our ancient diet, epidemiology and randomized controlled trials." J. Nutr. 2004, 134 (1 ): 183-6). Accordingly, EPA and DHA in the body are mainly derived from dietary sources (e.g. fatty fish). Diets high in fish oils are known to have many beneficial effects on heart disease, cancer, arthritis, allergies, and other chronic diseases. Epidemiological studies have found that increasing omega-3 fatty acid intake, in the form of fish or fish oil supplements, may reduce various risk factors associated with cardiovascular disease. See, e.g., The American Heart Association, Scientific Statement, "Fish Consumption, Fish Oil, Omega-3 Fatty Acids and Cardiovascular Disease," November 2002; Appel et al., "Does supplementation of diet with 'fishy oil' reduce blood pressure? A meta-analysis of controlled clinical trials. " Arch. Intern. Med. 1993, 153 (12): 1429-1438; GISSI-Prevenzione Investigators. "Dietary supplementation with omega-3 polyunsaturated fatty acids and vitamin E after myocardial infarction: results of the GISSIPrevenzione trial." Lancet 1999, 354: 447-55.

[0133] Despite strong evidence for the benefit of omega-3 fatty acids such as EPA and DHA in preventing cardiovascular disease, the average daily intake of these fatty acids is

-36 fatty acids by North American Americans are estimated at 0.1 to 0.2 grams, compared to a suggested daily intake of 0.65 grams to confer benefit (Webb, "Alternative sources of omega-3 fatty acids." Natural Foods Merchandiser 2005 , XXVI (8): 40-4). Since changing the diet of the population is difficult and many people do not like fish, supplementing with EPA and DHA is an important approach to solving this problem. Unfortunately, many omega-3 fatty acid supplements are sensitive to oxidation and can have an unpleasant odor and taste. In addition, adherence to dietary supplementation regimes requires discipline that is often lacking. In view of the health benefits of omega-3 fatty acids, the disclosed microcapsules can be used to deliver omega-3 fatty acids to individuals.

[0134] In the disclosed methods of use, the emulsions and / or microcapsules that are administered can be any of the compositions disclosed herein. For example, the disclosed microcapsules can be used in the disclosed methods in the form of any of the disclosed nutritional supplements. In another example, the disclosed microcapsules can be used in the disclosed methods in the form of any of the disclosed pharmaceutical formulations. In yet another example, the disclosed microcapsules can be incorporated into any of the disclosed articles or incorporated into any disclosed foodstuff and used in the disclosed methods.

[0135] It is believed that the disclosed methods can be accomplished by administering various forms of the disclosed microcapsules. For example, any of the pharmaceutical formulations may be administered with any of the disclosed foodstuffs. In another example, a tablet or capsule may be administered with any of the nutritional supplements disclosed. In yet another example, any of the pharmaceutical formulations may be administered with any of the disclosed delivery products and nutritional supplements, and the like.

Dosage

[0136] When used in the above-described methods or other methods of treatment or in dietary supplements, pharmaceutical formulations, products for administration, or foodstuffs disclosed herein, an "effective amount" of one of the disclosed microcapsules may be used in pure form or where such forms exist. , in a pharmaceutically acceptable salt form with or without a pharmaceutically acceptable excipient, carrier, or other additive.

[0137] The specific level of the effective dose for any particular subject will depend on a number of factors, including the disorder being treated and the severity of the disorder; the identity and specific activity of the composition used; the age, body weight, general health, sex and diet of the patient; time of administration, route of administration; the rate of excretion of the specific composition used; duration of treatment; drugs used in combination or concomitantly with the specific composition to be used; and the like, well known in medical techniques. For example, it is within the skill of the art to initiate dosages of the compositions at levels lower than those required for

The desired therapeutic effect is achieved and the dosage is gradually increased until the desired effect is achieved. If desired, the effective daily dose can be divided into multiple doses for administration. Consequently, single dose compositions may contain such amounts or submultiples thereof to make up a daily dose.

[0138] The dosage may be adjusted by the individual physician or the person in the event of any contraindications. The dosage may vary and may be given in one or more doses per day for one or more days. Guidance can be found in the literature on the appropriate dosages for each class of pharmaceutical product.

[0139] Further disclosed are methods of delivering the disclosed composition to a subject by administering to the subject any of the disclosed dietary supplements, pharmaceutical formulations, delivery products, and / or foodstuffs. The disclosed compositions (including dietary supplements, delivery products, and pharmaceutical formulations) can typically be administered orally.

EXAMPLES

[0140] The following examples are presented to illustrate the methods and results of the disclosed subject matter. All the examples, except examples 2.2.4, 2.2.5 and 9, are outside the scope of the claims. Efforts have been made to ensure accuracy with respect to numbers (e.g., amounts, temperature, pH, etc.) but some errors and deviations should be considered. Unless otherwise stated, parts are parts by weight, the temperature is at or is at ambient temperature, and the pressure is at or about atmospheric pressure. There are many variations and combinations of conditions, e.g., ingredient concentrations, temperatures, pressures, and other reaction ranges and conditions, that can be used to optimize the purity and yield of the product obtained from the described process. Only reasonable and routine experimentation will be required to optimize such process conditions.

[0141] Certain materials, compounds, compositions, and ingredients disclosed herein may be obtained commercially or readily synthesized using techniques well known to those skilled in the art. For example, the starting materials and reagents used in preparing the disclosed compositions are either available from commercial suppliers such as Ocean Nutrition Canada, Ltd. (Dartmouth, Canada), Aldrich Chemical Co., (Milwaukee, Wis.), Acros Organics (Morris Plains, NJ), Fisher Scientific (Pittsburgh, Pa.), Or Sigma (St. Louis, Mo.), or are prepared by methods known to those skilled in the art by following the procedures outlined in sources such as Fieser and Fieser's Reagents for Organic Synthesis, Tomy 1-17 (John Wiley and Sons, 1991); Rodd's Chemistry of Carbon Compounds, Tomy 1-5 and Supplements (Elsevier Science Publishers, 1989); Organic Reactions, Volumes 1-40 (John Wiley and Sons, 1991); March's Advanced Organic Chemistry, (John Wiley and Sons, 4th Edition); and Larock's Comprehensive Organic Transformat! ons (VCH Publishers Inc., 1989).

Control Example A: Preparation of omega-3 microcapsules using 275 Bloom gelatin

Pigskin 275 Bloom Gelatin (44 g) was dissolved in water (482 g) and the solution was heated to 50 ° C. The initial pH of the gelatin solution was 4.638. Sodium ascorbate (7.3 g) was then added to the gelatin solution and the pH was 5.271.

[0143] High DHA fish oil (72.0 g; XODHA from Ocean Nutrition Canada Ltd.) was added to the gelatin solution and emulsified with a POLYTRON ™ homogenizer at 7,500 rpm for 4 minutes. The emulsion was examined under a microscope after emulsification to verify that the oil droplets were small and uniform (diameter about 1-5 µm).

[0144] Distilled water (890 g) was added to the 2 L reactor and the temperature was maintained at 50 ° C. The emulsion was then added to distilled water in the reactor and the pH was found to be 5.058. Sodium polyphosphate (4.4 g) dissolved in distilled water (84 g) was added to the diluted emulsion in the reactor and the resulting mixture had a pH of 5.821.

[0145] The pH was then lowered with 10% phosphoric acid to agglomerate the primary microcapsules. Upon further lowering the pH to 4.686, the secondary microcapsules formed 30-50 µm agglomerations. The mixture was cooled with an average cooling rate of 1 ° C / 5 minutes from 50 ° C to 4 ° C.

[0146] After adjusting the pH to 6.0 by adding 10% NaOH, a 1% w / w formulation was added. transglutaminase (Ajinomoto USA Inc., Fort Lee, NJ) and the temperature was kept at room temperature (~ 25 ° C) for 16 hours.

[0147] The slurry was then ready for use in the food industry. It was also spray dried to produce a free flowing powder. This powder had an induction period of 44.7 hours, determined at 65 ° C with an initial oxygen pressure of about 550 kPa using Oxipres (Mikrolab Aarhus A / S, Hojbjerg, DNK).

Control Example B: Preparation of omega-3 microcapsules using 240 Bloom Gelatin

[0148] Fish gelatin 240 Bloom (44 g) was dissolved in water (320 g) and the solution was heated to 40 ° C. The initial pH of the gelatin solution was 5.807. Sodium ascorbate (7.3 g) was then added to the gelatin solution and the pH was 5.902.

[0149] High DHA fish oil (72.0 g; XODHA from Ocean Nutrition Canada Ltd.) was added to the gelatin solution and emulsified with a POLYTRON ™ homogenizer at 7,500 rpm for 4 minutes. The emulsion was examined under a microscope after emulsification to verify that the oil droplets were small and uniform (diameter approximately 15 µm).

[0150] Distilled water (1051 g) was added to the 2 L reactor and the temperature was maintained at 40 ° C. The emulsion was then added to distilled water in the reactor and the pH was found to be 5.812. To the diluted emulsion in the reactor was then added sodium polyphosphate (4.4 g) dissolved in distilled water (84 g) and the resulting mixture had a pH of 6.512.

[0151] The pH was then lowered with 10% phosphoric acid to form agglomeration of the primary microcapsules. Upon further lowering the pH to 4.773, the secondary microcapsules formed 30-50 µm agglomerations. The mixture was cooled with an average cooling rate of 1 ° C / 5 minutes from 40 ° C to 5 ° C.

[0152] After adjusting the pH to 6.0 by adding 10% NaOH, a 1% w / w formulation was added. transglutaminases (Ajinomoto USA Inc., Fort Lee, NJ) for cross-linking and curing of the microcapsule shell at 5 ° C for 1 hour, 15 ° C for 8 hours, 20 ° C for 9 hours.

[0153] The slurry was then ready for use in the food industry. It was also spray dried to produce a free flowing powder. This powder had an induction period of 43.5 hours, determined at 65 ° C, with an initial oxygen pressure of about 550 kPa using Oxipres (Mikrolab Aarhus A / S, Hojbjerg, DNK).

Control example C: Preparation of omega-3 microcapsules using 0 Bloom Gelatin

[0154] Fish gelatin 0 Bloom (44 g; Kenny &amp; Ross Ltd., Shelburne, NS) was dissolved in water (323 g) and the solution was heated to 35.6 ° C. The initial pH of the gelatin solution was 5.807. Sodium ascorbate (7.3 g) was then added to the gelatin solution and the pH was 6.042. Sodium polyphosphate (4.4 g) dissolved in distilled water (84 g) was then added to the gelatin solution. The mixture had a pH of 6.306 at 34.1 ° C and it was adjusted to 4.9 using 10% phosphoric acid.

[0155] High DHA fish oil (72.6 g; XODHA from Ocean Nutrition Canada Ltd.) was mixed with a gelatin solution and emulsified with a POLYTRON ™ homogenizer at 7,500 rpm for 4 minutes. The emulsion was examined under a microscope after emulsification to verify that the oil droplets were small and uniform (diameter approximately 15 µm).

[0156] Distilled water (1060 g) was added to the 2 L reactor and the temperature was maintained at 35 ° C. The emulsion was then added to the distilled water in the reactor and the pH was found to be 4.9412. While stirring the mixture, the pH was lowered with 10% phosphoric acid to agglomerate the primary microcapsules. After lowering the pH to 4.751, the secondary microcapsules had a diameter of about 40 µm. The mixture was cooled with an average cooling rate of 1 ° C / 5 minutes from 35 ° C to 5 ° C.

[0157] After adjusting the pH to 6.0 by adding 10% NaOH, a 1% w / w formulation was added. transglutaminase (Ajinomoto USA Inc., Fort Lee, NJ) to cross-link the microcapsule shell at 5 ° C for 5 hours, followed by enzymatic curing at 20 ° C for 10 hours.

[0158] The final microcapsule suspension was then ready for food applications. It was also spray dried to produce a free flowing powder. This powder had an induction period of 36.9 hours determined at temperature

-4065 ° C, with an initial oxygen pressure of about 550 kPa using Oxipres (Mikrolab Aarhus A / S, Hojbjerg, DNK).

Example 1: Preparation of omega-3 microcapsule including chitosan before coacervation

Example 1.1: Preparation of omega-3 microcapsules with 240 Bloom fish gelatin and chitosan (added before emulsification and coacervation)

[0159] Fish gelatin 240 Bloom (44 g; from Lapi Gelatine SpA, Empoli, Italy) was dissolved in water (256 g) with sodium ascorbate (7.3 g) and heated to 41 ° C. A 1% solution of chitosan in 1% acetic acid (44 g) was added to the gelatin solution, taking into account the amount of additional water to make a total weight of 320 g. Phosphoric acid (10% solution, 17.6 ml) was added to the gelatin solution to obtain the pH about 4.5. High DHA fish oil (72.0 g; XODHA from Ocean Nutrition Canada Ltd.) was then added to the gelatin-chitosan solution and emulsified with a POLYTRON ™ homogenizer at 7,500 rpm for 4 minutes.

[0160] Distilled water (752 g) was added to the 2 L reactor and the temperature was kept at 41 ° C. Then, the emulsion was added to the distilled water in the reactor, and the mixture was stirred at 41 ° C. Sodium polyphosphate (4.4 g) dissolved in distilled water (300 g) was added to the diluted emulsion in the reactor in 50 ml portions. (The ratio of sodium polyphosphate to chitosan can range from 50: 1 to 5: 1; however, a ratio of 10: 1 was used in this particular example.) After the addition of all the sodium polyphosphate solution, the reaction mixture had a pH of about 4.7.

[0161] While stirring the mixture, the pH was adjusted to 4.301 using 10% phosphoric acid, creating 30-70 µm agglomerations of primary microcapsules. The mixture was then cooled with an average cooling rate of 1 ° C / 5 minutes from 41 ° C to 3 ° C.

[0162] After adjusting the pH to 6.0 by adding 10% NaOH, a 1 wt% transglutaminase preparation (Ajinomoto USA Inc., Fort Lee, NJ) was added. The slurry was then held at 3 C for 1 hour to cross-link and then enzymatically cured at 15 C for 8 hours and 20 C for 10 hours.

[0163] The final microcapsule suspension was then ready for food applications. It was also spray dried to produce a free flowing powder. This powder had an induction period of 61 hours, determined at 65 ° C, with an initial oxygen pressure of about 550 kPa using Oxipres (Mikrolab Aarhus A / S, Hojbjerg, DNK). The induction period improved at 17.4 hours, better than that of Control B.

Example 1.2: Preparation of Omega-3 Microcapsules with 240 Bloom Fish Gelatin and Chitosan (Using a Two Step Process)

[0164] Fish Gelatin 240 Bloom (44 g; from Lapi Gelatine SpA, Empoli, Italy) was dissolved in water (289 g) with sodium ascorbate (7.3 g) and heated to 41 ° C. Phosphoric acid (10% solution) was added to the gelatin solution to produce a pH of about 4.5.

A 1% solution of chitosan in 1% acetic acid (31.4 g) was then added to the gelatin solution. High DHA fish oil (72.0 g; XODHA from Ocean Nutrition Canada Ltd.) was then added to the gelatin-chitosan solution and emulsified with a POLYTRON ™ homogenizer at 7,500 rpm for 4 minutes.

[0165] Distilled water (752 g) and sodium polyphosphate (3.14 g) were added to the 2 L reactor and the temperature was kept at 41 ° C. The emulsion was then added to the distilled water in the reactor, and the mixture was stirred at 41 ° C.

[0166] Sodium polyphosphate (1.26 g) dissolved in distilled water (192 g) was added to a 1% acetic acid solution (192 g) containing 0.13 g of chitosan and mixed. (In this particular example, the ratio of sodium polyphosphate to chitosan was 10: 1.) A mixture of chitosan and polyphosphate was then added to the dilute emulsion in the reactor to obtain agglomerated particles. The mixture was then cooled with an average cooling rate of 1 ° C / 5 minutes from 41 ° C to 3 ° C.

[0167] After adjusting the pH to 6.0 by adding 10% NaOH, a 1 wt% transglutaminase preparation (Ajinomoto USA Inc., Fort Lee, NJ) was added. The slurry was then held at 3 ° C for 1 hour to cross-link and then enzymatically cured at 15 ° C for 8 hours and 20 ° C for 10 hours.

[0168] The final microcapsule suspension was then ready for food applications. It was also spray dried to produce a free flowing powder. This powder had an induction period of 49.7 hours, determined at 65 ° C, with an initial oxygen pressure of about 550 kPa using Oxipres (Mikrolab Aarhus A / S, Hojbjerg, DNK). The induction period was 6.2 hours longer than the Control B samples.

Examples 2: Preparation of omega-3 microcapsules including chitosan, lysine and / or glutamine after coacervation and envelope formation

Example 2.1: Preparation of omega-3 microcapsules using 240 Bloom fish gelatin and adding chitosan after agglomeration but before shell formation

[0169] Fish Gelatin 240 Bloom (44 g; from Lapi Gelatine SpA, Empoli, Italy) was dissolved in water (320 g) and heated to 40 ° C. Sodium ascorbate (7.3 g) was added to the gelatin solution. High DHA fish oil (72.0 g; XODHA from Ocean Nutrition Canada Ltd.) was then added to the gelatin solution and emulsified with a POLYTRON ™ homogenizer at 7,500 rpm for 4 minutes.

[0170] Distilled water (944 g) and sodium polyphosphate (4.4 g) were added to the 2 L reactor and the temperature was kept at 40 ° C. The emulsion was then added to the reactor. While the mixture was stirred, the pH was adjusted to 4.3 using 10% phosphoric acid, creating about 30-60 µm agglomeration of the primary microcapsules.

[0171] The mixture was then cooled with an average cooling rate of 1 ° C / 5 minutes from 40 ° C to 3 ° C. When the temperature reached 23 ° C, chitosan (192 g of 1% acetic acid solution containing 0.44 g of chitosan) was added to the reactor. Cooling was continued without interruption.

[0172] After adjusting the pH to 6.0 by adding 10% NaOH, a 1 wt% transglutaminase preparation (Ajinomoto USA Inc., Fort Lee, NJ) was added. The slurry was then held at 3 ° C for 1 hour to cross-link and then enzymatically cured at 15 ° C for 8 hours and 20 ° C for 10 hours.

[0173] The final microcapsule suspension was then ready for food applications. It was also spray dried to produce a free flowing powder. This powder had an induction period of 49.7 hours, determined at 65 ° C, with an initial oxygen pressure of about 550 kPa using Oxipres (Mikrolab Aarhus A / S, Hojbjerg, DNK). This induction period was 6.2 hours longer than that of the corresponding Control B sample.

Example 2.2: Preparation of omega-3 microcapsules using 0 Bloom fish gelatin combined with chitosan, lysine and glutamine

[0174] Fish Bloom gelatin (88 g; Kenny &amp; Ross Ltd., Shelburne, NS) was dissolved in water (640 g) and the solution was heated to 35 ° C. Sodium ascorbate (14.6 g) was added to the gelatin solution. High DHA fish oil (144.0 g; XODHA from Ocean Nutrition Canada Ltd.) was mixed with a gelatin solution and emulsified with a POLYTRON ™ homogenizer at 7,500 rpm for 4 minutes. The emulsion was examined under a microscope after emulsification to verify that the oil droplets were small and uniform (diameter about 1-5 µm).

[0175] Distilled water (2000 g) was added to the 3 L reactor and the temperature was maintained at 35 ° C. The emulsion was then added to distilled water in the reactor and the pH was found to be 5.98. Sodium polyphosphate (6.0 g) dissolved in distilled water (160 g) was then added to the diluted emulsion in the reactor. The resulting mixture in the reactor had a pH of 6.50.

[0176] While stirring, the pH was adjusted to 4.78 with 10% phosphoric acid to agglomerate the original microcapsules with a diameter of approximately 50 µm. The mixture was then cooled from 35 ° C to 4 ° C with an average cooling rate of 1 ° C / 5 minutes.

[0177] After adjusting the pH to 6.0 by adding 10% NaOH, a 1 wt% transglutaminase preparation (Ajinomoto USA Inc., Fort Lee, NJ) was added. The slurry was held at 4 ° C for 5 hours and then at 8 ° C for 6 hours for cross-linking. The solution was then heated to 20 ° C.

[0178] Two identical batches of this stock slurry were prepared and mixed together for further processing.

Example 2,2,1: Control

[0179] The stock suspension of Example 2.2 (1000 g) was further cross-linked at room temperature (~ 25 ° C) for 6 hours. The control suspension was then spray dried.

Example 2,2,2: Treatment with high molecular weight chitosan

[0180] The stock slurry of Example 2.2 (1000 g) was treated with high molecular weight chitosan (131.3 kDa) by first transferring the slurry to a 1.5 L reactor. A solution (250 g) of 1.0% w / w was obtained. chitosan in 1.0% w / w. acetic acid and diluted to 0.5% w / w. with distilled water. The 0.5% chitosan solution was then added slowly to the stock suspension in the 1.5 L reactor. The pH was adjusted to 6.0 and the mixture was stirred at room temperature (~ 25 ° C) for 5 hours.

Example 2,2.3. Treatment with low molecular weight chitosan

[0181] The stock slurry of Example 2.2 (1000 g) was treated with a low molecular weight (5.3 kDa) chitosan slurry by first transferring the slurry to a 1.5 L reactor. A solution (200 g) of 1.0% w / w was obtained. chitosan in 1.0% w / w. acetic acid and diluted to 0.4% w / w. with distilled water. This 0.4% chitosan solution was then slowly added to the slurry in the 1.5 L reactor. The pH was adjusted to 5.6 and the mixture was stirred at room temperature (~ 25 ° C) for 5 hours.

Example 2,2,4 Treatment of lysine and glutamine

[0182] Stock slurry from Example 2.2 (1000 g) v lysine and glutamine by first transfering the slurry to a 1.5 L reactor. Lysine (5.0 g) in distilled water (40 g) was slowly added to the slurry in the 1.5 L reactor. The pH was adjusted to 6.0. After 2 hours, glutamine (2.0 g) in distilled water (60.0 g) was also slowly added to the slurry. The mixture was stirred at room temperature (~ 25 ° C) for 3 hours.

Example 2.2.5: Treatment with high molecular weight chitosan and glutamine

[0183] The stock slurry of Example 2.2 (1000 g) was treated with high molecular weight chitosan and glutamine by first transferring the slurry to a 1.5 L reactor. A solution (250 g) of 1.0% w / w was obtained. chitosan in 1.0% w / w. acetic acid and diluted to 0.5% w / w. with distilled water. This 0.5% chitosan solution was then slowly added to the slurry in the 1.5 L reactor. The pH was adjusted to 6.0. After 2 hours, glutamine (2.0 g) in distilled water (60.0 g) was slowly added to the suspension. The mixture was stirred at room temperature (~ 25 ° C) for 3 hours.

[0184] Samples of the finished microcapsule suspension of Examples 2.2.1 to 2.2.5 were then spray dried to produce free flowing powder products. All of these samples had a better induction period compared to Control 2.2.1 and Control C (Table 1).

Table 4: Results of chitosan, lysine and glutamine treatments

<td>Example #</td><td>Free Oil (%)</td><td>Induction period (h)</td>

<td> 2.2.1</td><td> 0,032</td><td> 44,4</td>

<td> 2.2.2</td><td> 0,027</td><td> 55,9</td>

<td> 2.2.3</td><td> 0,081</td><td> 68,0</td>

<td></td><td></td><td></td>

<td> 2.2.4</td><td> 0,035</td><td> 80,2</td>

<td> 2.2.5</td><td> 0,016</td><td> 83,0</td>

Example 3 Preparation of omega-3 microcapsules using 0 Bloom gelatin with inclusion of wax before aggloremation and shell formation

[0185] Fish Gelatin 0 Bloom (44.1 g) was dissolved in water (323.8 g) and heated to 35 ° C. Sodium ascorbate (7.32 g) and a kamauba wax microemulsion (7.90 g; ME28230 from Michelman Inc., Cincinnati, Ohio) were added to the gelatin solution. High DHA fish oil (73.54 g; XODHA from Ocean Nutrition Canada Ltd.) was added to the gelatin solution and emulsified with a POLYTRON ™ homogenizer at 7,500 rpm for 4 minutes.

[0186] The emulsion was transferred to a 2 L reactor containing distilled water (1061.4 g), held at 35 ° C. The emulsion had a pH of 5.88 at 35 ° C. A 5% solution of sodium polyphosphate (88.0 g) was added to the mixture and the pH was adjusted to 35 ° C to 6.59. While the mixture was stirred, the pH was adjusted to 4.68 at 35 ° C with 10% phosphoric acid to form a 3060 µm agglomeration of primary microcapsules.

[0187] The obtained mixture of multicore microcapsules was then cooled with an average cooling rate of 1 ° C / 5 minutes from 35 ° C to 4 ° C. After adjusting the pH to 6.0 by adding 10% NaOH, a 1% w / w formulation was added. transglutaminases (Ajinomoto USA Inc., Fort Lee, NJ). The slurry was then held at 5 ° C for 5 hours to cross-link and then enzymatically cured at 20 ° C for 10 hours.

[0188] The final microcapsule suspension was then ready for food applications. It was also spray dried to produce a free flowing powder. This powder had an induction period of 70.5 hours compared to 36.9 hours for the no-wax control (e.g., Control Example C).

Example 4: Preparation of omega-3 microcapsules using 275 Bloom gelatin with inclusion of wax after shell formation

[0189] Fish gelatin 275 Bloom (40.92 g) was dissolved in water (452 ​​g) and heated to 50 ° C. Sodium ascorbate (6.82 g) was added to the gelatin solution. High DHA fish oil (68.25 g; XODHA from Ocean Nutrition Canada Ltd.) was added to the gelatin solution and emulsified with a POLYTRON ™ homogenizer at 6,400 rpm for 11 minutes.

[0190] The emulsion was transferred to a 2 L reactor containing distilled water (833.3 g), held at 50 ° C. The emulsion had a pH of 5.23 at 51.8 ° C. A 5% solution of sodium polyphosphate (82.5 g) was added to the mixture, and the pH was adjusted to 50.4 ° C to 5.66. While mixing

The pH of the mixture was adjusted to 4.80 at 50.4 ° C with 10% phosphoric acid to form about 30-60 µm agglomeration of primary microcapsules.

[0191] The mixture of multicore microcapsules was then cooled with an average cooling rate of 1 ° C / 5 minutes from 50 ° C to 4 ° C. After adjusting the pH to 6.0 by adding 10% NaOH, a 1% w / w formulation was added. transglutaminases (Ajinomoto USA Inc., Fort Lee, NJ). The slurry was kept at room temperature (about 25 ° C) for 16 hours for cross-linking and curing.

[0192] The pH was adjusted to 9.3 and a carnauba wax microemulsion (187 g; ME62125Am, Michelman Inc.) was added. The mixture had a pH of 8.69 and contained 46.7 g of total weight carnauba wax.

[0193] The final microcapsule suspension was then ready for food applications. It was also spray dried to produce a free flowing powder. The powder had an induction period of 80.0 hours compared to 44.7 hours for the no-wax control (eg, Control Example A).

Example 5: Preparation of omega-3 microcapsules using 240 Bloom gelatin with carbohydrates and proteins included after coating

Example 5.1 Preparation of Fish Oil Microcapsule Stock Suspension Using 240 Bloom Fish Gelatin

[0194] Bloom Fish Gelatin 240 (325.8 g) was dissolved in water (3599 g) in a 10,000 g reactor and heated to 40 ° C with stirring. Sodium ascorbate (49.4 g) and a 20% phosphoric acid solution (60 ml) were added to the gelatin solution. Fish oil with high DHA content (565 g; XODHA from Ocean Nutrition Canada Ltd.) was added to the gelatin solution and emulsified using a high shear pump until the droplets were 1-5 µm in diameter. Distilled water (5453.4 g) was added to the reactor and kept at 40 ° C.

[0195] Sodium polyphosphate (32.6 g) dissolved in distilled water (100 g) was then added to the diluted emulsion in the reactor. The pH was adjusted to 4.57 using 20% ​​phosphoric acid (about 100 ml) to form about 30 µm agglomeration of the primary microcapsules.

[0196] The mixture was then cooled from 40 ° C to 6 ° C with an average cooling rate of 1 ° C / 5 minutes. After adjusting the pH to 6.0 by adding 10% NaOH, a 1% w / w formulation was added. transglutaminases (Ajinomoto USA Inc., Fort Lee, NJ). The slurry was then cross-linked at 15 ° C for 9 hours and 20 ° C for 8 hours.

[0197] The final microcapsule suspension was then ready for coating. The slurry could also be spray dried to produce a free flowing powder.

Example 5.2: Incorporation of modified starch into microcapsules

[0198] Modified starch (40 g; N-LOK from National Starch &amp; Chemical Co., Bridgewater, NJ) was dissolved in water (60 g) with agitation.

The stock suspension obtained in Example 5.1 (600 g) was transferred to a 1000 ml beaker and the suspension was stirred by the magnetic bar on a hot plate. The modified starch solution was then added to the slurry and mixing was continued for 30 minutes. The slurry was spray dried to produce a free flowing powder.

Example 5.3 Incorporation of modified starch and lactose into microcapsules

[0199] Modified starch (20 g; N-LOK from National Starch &amp; Chemical Co., Bridgewater, NJ) was dissolved in water (30 g) with agitation to make a 40% slurry. Lactose (25 g) was dissolved in water (25 g) with stirring to form a 50% solution. The stock suspension obtained in Example 5.1 (600 g) was transferred to a 1000 ml beaker and the suspension was stirred by a magnetic bar on a hot plate. The starch and lactose solutions were mixed thoroughly and added to the stock slurry which was mixed for 30 minutes. The slurry was spray dried to produce a free flowing powder.

Example 5.4 Incorporation of lactose in microcapsules

[0200] Lactose (50 g) was dissolved in water (50 g) with heating and stirring. Tween 80 (5 g) was then added to the lactose solution. The stock suspension obtained in Example 5.1 (600 g) was transferred to a 1000 ml beaker and the suspension was stirred by the magnetic bar on a hot plate. The remaining lactose-Tween 80 solution was added to the slurry and stirring continued for 30 minutes. The slurry was spray dried to produce a free flowing powder.

Example 5.5 Incorporation of maple syrup into microcapsules

[0201] The stock suspension obtained in Example 5.1 (600 g) was transferred to a 1000 ml beaker and the suspension was stirred by the magnetic bar on a hot plate. Maple syrup (100g; from supermarket) was added to the slurry and stirring continued for 30 minutes. The slurry was spray dried to produce a free flowing powder.

Example 5.6 Incorporation of sucrose in microcapsules

[0202] Sucrose (50 g) was dissolved in water (50 g) with heating and stirring. Tween 80 (5 g) was then added to the sucrose solution. The stock suspension obtained in Example 5.1 (600 g) was transferred to a 1000 ml beaker and the suspension was stirred by a magnetic bar on a hot plate. Sucrose solution Tween 80 was added to the suspension and stirring continued for 30 minutes. The slurry was spray dried to produce a free flowing powder.

Example 5.7 Incorporation of methylcellulose in microcapsules

[0203] Hydroxypropyl methylcellulose (HPMC) (5 g; Methocel E3, from DOW Chemical Co., Midland, Mich.) Was suspended in water (95 g) with heating and stirring. The stock suspension obtained in Example 5.1 (600 g) was transferred to a 1000 ml beaker and the suspension was stirred by a magnetic bar on a hot plate. For the suspension

HPMC solution was added and stirring continued for 30 minutes. The slurry was spray dried to produce a free flowing powder.

Example 5.8 Incorporation of milk protein in microcapsules

[0204] High calcium milk protein (50 g; Alaco 9090 from NZMP (North America) Inc., Santa Rosa, California) was suspended in water (50 g) with heating and stirring. The stock suspension obtained in Example 5.1 (600 g) was transferred to a 1000 ml beaker and the suspension was stirred by the magnetic bar on a hot plate. The milk protein solution was then added to the slurry and mixing was continued for 30 minutes. The slurry was spray dried to produce a free flowing powder.

Example 5.9 Incorporation of whey protein and glycerin in microcapsules

[0205] Whey protein (50 g; Alacen 841 from NZMP (North America) Inc., Santa Rosa, California) was dissolved in water (50 g) with heating and stirring. Glycerin (5 g) was also added. The stock suspension obtained in Example 5.1 (600 g) was transferred to a 1000 ml beaker and the suspension was stirred by a magnetic bar on a hot plate. The whey protein-glycerin solution was added to the slurry and mixing was continued for 30 minutes. The slurry was spray dried to produce a free flowing powder.

Table 5: Effect of various carbohydrates and proteins on the stability of fish oil microcapsules

<td>Example #</td><td>Induction period (h)</td>

<td> 5.1</td><td> 36,0</td>

<td> 5.2</td><td> 91,0</td>

<td> 5.3</td><td> 91,0</td>

<td> 5.4</td><td> 116,0</td>

<td> 5.5</td><td> 116,0</td>

<td> 5.6</td><td> &gt;116</td>

<td> 5.7</td><td> 36</td>

<td> 5.8</td><td> 63,0</td>

<td></td><td></td>

<td> 5.9</td><td> 62,0</td>

-48 Example 6: Preparation of omega-3 microcapsules using 0 Bloom fish gelatin with improved sensory properties by nitrogen purification

[0206] An amount of 720 g of 0 Bloom Fish Gelatin solution (12% w / w, 35 ° C) was obtained. Sodium ascorbate (3.6 g) was then added to the gelatin solution. High DHA fish oil (140 g; XODHA from Ocean Nutrition Canada Ltd.) was also added and the solution was emulsified with a POLYTRON ™ homogenizer at 7,500 rpm for 4 minutes and purged with nitrogen.

[0207] Distilled water (1050 g) was added to each of the two 2 L reactors, and the temperature was maintained at 35 ° C. Sodium ascorbate (5.7 g) was also added to the water in each reactor. Half of the emulsion was transferred to each reactor (approximately 430 g). One reactor was used as a control reactor (Example 6.1, under atmospheric conditions), while the other (Example 6.2) was continuously purged with nitrogen to exclude oxygen from the air and minimize oxidative deterioration of the fish oil. The mixture in each reactor was stirred continuously and had a temperature of 36.0 ° C and a pH of 6.086.

[0208] A 5% sodium polyphosphate solution (89.4 g) was added to each reactor, and its pH increased to 6.607. After adjusting the pH to 4.888 with 5% phosphoric acid, secondary microcapsules were formed and the agglomerations were about 50 µm in diameter in each reactor. The samples were then cooled from 35 ° C to 5 ° C at an average rate of 1 ° C / 5 minutes.

[0209] After adjusting the pH to 6.0 with 10% NaOH, a 1% w / w transglutaminase preparation (Ajinomoto USA Inc., Fort Lee, NJ) was added. The slurry was then held at 5 ° C for 5 hours to cross-link and then enzymatically cured at 20 ° C for 10 hours.

[0210] The final microcapsule suspension was spray dried to form a free flowing powder. The powder samples had an induction period of 50.8 and 50.3 hours, respectively. It was found, as shown in Table 3, that nitrogen purging contributed to the sensory improvement of the final product.

Table 6: Effect of exposure of the slurry to air or nitrogen on the sensory properties of fish oil microcapsules.

<td>Example</td><td>Treatment</td><td>Smell</td><td>Aroma</td><td>IP (h)</td>

<td> 6.1</td><td>No N2</td><td>Very sour, fishy</td><td>Sour, milky, slightly salty, slightly fishy</td><td> 50,8</td>

<td> 6.2</td><td>cleansing n₂</td><td>Sour milky, very slightly green</td><td>Sour milky, salty, musty (i.e. not fishy)</td><td> 50,3</td>

Example 7: Preparation of omega-3 microcapsules using 275 Bloom gelatin with 200 mg / L Na2EDTA in suspension incorporated

[0211] Sodium ethylenediaminetheacetate (Na2EDTA) (0.2919 g) was dissolved in water (464 g); The pH of the solution was 4.63. Great Lakes pork gelatin (42 g) was then added to the solution (pH 4.73). Then sodium ascorbate (7.0 g) was added and the pH was 5.23.

[0212] High DHA fish oil (73.54 g; XODHA from Ocean Nutrition Canada Ltd.) was added to the gelatin solution and emulsified with a POLYTRON ™ homogenizer at 7,500 rpm for 4 minutes. The emulsion was examined under a microscope after emulsification to verify that the oil droplets were small and uniform (diameter approximately 15 µm).

[0213] Distilled water (855 g) was added to the reactor 2 L and the temperature was maintained at 53 ° C. The emulsion was then added to the distilled water in the reactor, and the pH was 5.25. Sodium polyphosphate (4.25 g) dissolved in distilled water (80 g) was then added to the diluted emulsion in the reactor. The mixture in the reactor then had a pH of 5.92. The oil droplets were 1-5 µm in diameter and appeared similar to a normal fish oil-in-gelatin emulsion.

[0214] The pH was then lowered with 10% phosphoric acid to form agglomeration of the primary microcapsules. A conventional method of microencapsulating pork oil would typically need to be carried out at about pH 4.5-5. In this case, when the pH was lowered to 4.67, the oil droplets had a diameter of 20-40 µm.

[0215] The slurry was then cooled to 5 ° C with an average cooling rate of 1 ° C / 5 minutes. When the temperature reached 4 ° C, a 1% w / w formulation was added to the slurry. transglutaminases (Ajinomoto USA Inc., Fort Lee, NJ). The pH was then adjusted to 6.0 with 10% NaOH. The suspension of microcap sulkom was allowed to cross-link and cure at room temperature (~ 25 ° C) for 16 hours.

[0216] The slurry was spray dried and tested for various quality and stability parameters. The powder was free flowing and had an induction period of 56.4 hours. Although the induction period was similar to the control without Na2EDTA, the level of lipid oxidation product measured by the peroxide value (PV) was different. The microcapsule powders with and without the addition of Na2EDTA had a PV of 1.18 and 2.35 meq / kg, respectively.

Example 8: Preparation of omega-3 microcapsule including anti-caking agents with improved free-flow properties

[0217] Fish oil microcapsule suspensions were prepared following Control Example A and tested for final product flowability. Desiccants tested included Hubersorb 600 (JM Huber Corp., Harve de Grace, Md.), Zeothix 265 (JM Huber Corp.), Capsul modified starch (National Starch &amp; Chemical Co.) and Vitacel cellulose (J. Rettenmaier USA LP, Schoolcraft, Mich.). Examples and the resulting flowability of the powder product are shown in Table 4. All desiccants were found to improve the free-flowing property of the microcapsules.

Table 7: Comparison of powder free flow properties

<td>Example #</td><td>Treatment</td><td>Powder appearance</td>

<td> 8.1</td><td>Control (no measure anti-caking agent)</td><td>Fluffy coarse powder with some very large lumps.</td>

<td> 8.2</td><td>Hubersorb 600 (1 g / l)</td><td>Fine, flowing freely with small lumps</td>

<td> 8.3</td><td>Zeothix 265 (1 g / l)</td><td>Fine, flowing freely</td>

<td> 8.4</td><td>Capsul (lg / 1)</td><td>Fine, flowing freely</td>

<td> 8.5</td><td>Yitacel (lg / 1)</td><td>Mostly fine flowing freely</td>

Example 9: Preparation of an omega-3 microcapsule cross-linked with glutaraldehyde and additional amino acids

[0218] A microcapsule suspension may be prepared as disclosed herein. The slurry may be treated with about 2.5% glutaraldehyde based on the weight of the gelatin to cross-link the microcapsules. Since the MW of glutaraldehyde and lysine are 100 g / mol and 146.2 g / mol, respectively, three times as much lysine is needed to neutralize the residual aldehyde. Then, at least 480 mg of lysine / kg of suspension (about 0.05% by weight of the suspension) is needed. The addition of 0.25% lysine (or leucine, isoleucine and other amino acids) was found to extend the induction period from the pretest. Hydrophobic amino acids also improved powder agglomeration when tested in an open dish at 30 ° C / 75% RH (RH). It is then a five-fold reactive amino acid group in excess. As much as 0.5% of amino acids such as lysine can be used. Amino acids or proteins may be added 1-2 hours before cross-linking is complete.

Example 10: CoQio encapsulation and co-delivery with omega-3 without mixing with oil

[0219] To demonstrate that CoQio can be provided in a microcapsule without mixing with fish oil prior to microencapsulation, the following examples were made.

[0220] As described, the fish oil was emulsified into the gelatin solution and the resulting oil droplets were agglomerated by means of a complex coacervation with polyphosphate. After agglomeration, a CoQio emulsion in gelatin solution at the level of 30-200 mg CoQio was added to each 500 mg of EPA + DHA supplied. On cooling, the CoQio droplets became part of the shell and deposited on the surface of the agglomerates. Cross-linking then took place after cooling, hardening the gelatin-based shell.

[0221] Below are three examples of 30, 100 and 200 mg CoQio loading levels per serving. The powder samples from these experiments had a free oil content of less than 0.1% and an induction period of 13.5-14.5 hours, tested at 80 ° C.

-51 Example 10.1: Microencapsulation of DHA oil using a loading ratio of 100 mg CoQio / 500 mg EPA / DHA in a porcine gelatin shell

[0222] 39.1 g of pig gelatin was dissolved in 464.0 g of distilled water at 50 ° C. The reactor was connected to a circulation pump and the temperature was set to 50 ° C. Then 690.0 g of distilled water was added to the reactor and the temperature was kept at 50 ° C. An amount of 72.0 g of fish oil was mixed with the gelatin solution and emulsified at 7500 rpm for 4 minutes. An emulsion is formed and contains oil droplets having a diameter of about 1-5 µm. The emulsion was added to the reactor containing water at 50 ° C. The mixture had a pH of 5.045. Then 6.4 g of sodium ascorbate was added to the mixture. A further 85.2 g of 5% w / w was added to the reactor. sodium polyphosphate solution at room temperature. The pH was adjusted to 4.488 and the agglomeration was allowed to grow to about 40 µm as examined with an optical microscope. The multi-core fish oil particles formed in this step are shown in Fig. 6A.

[0223] 16.0 g of pork gelatin was mixed with 184.0 g of distilled water. Gelatin dissolved after being dispersed in water and heated to and held at 57 ° C. Then 24.0 g of CoQio powder was added to the gelatin solution and emulsified at 6000 rpm for 2 minutes and 7500 rpm for 1 minute. A CoQio emulsion was formed and contained droplets approximately 1-5 µm in diameter. Then 41.0 g of CoQio emulsion was mixed with the agglomerated slurry in the reactor at 50 ° C. The coating of the CoQio droplets around the multi-core fish oil particles was visible as shown in Fig. 6B.

[0224] The above agglomerations containing the slurry were then cooled to 4 ° C over 2.5 hours. The transglutaminase enzyme preparation was added at 0.2% w / w. and the temperature was adjusted to 20 ° C for enzymatic curing for at least 12 hours. The finished microcapsule suspension as shown in Fig. 6C was spray dried. The powder of the microcapsules was free flowing and the free surface oil was less than 0.1% w / w.

Example 10.2: Microencapsulation of DHA oil with a loading ratio of 30 mg Co-Q 10/500 mg EPA / DHA in a porcine gelatin shell

[0225] 41.1 g of pork gelatin was dissolved in 464.0 g of distilled water at 50 ° C. The reactor was connected to a circulation pump and the temperature was set to 50 ° C. 717.0 g of distilled water was added to the reactor and the temperature was maintained at 50 ° C. 72.0 g of fish oil was mixed with this freshly prepared gelatin solution and emulsified at 7500 rpm for 4 minutes. An emulsion is formed and contains oil droplets having a diameter of about 1-5 µm. The emulsion was added to the reactor containing water at 50 ° C. The mixture had a pH of 5.045. Then 6.4 g of sodium ascorbate was added to the mixture. A further 85.2 g of 5% w / w was added to the reactor. sodium polyphosphate solution at room temperature. The pH was adjusted to 4.488 and the agglomeration was allowed to grow to about 40 µm as examined with an optical microscope. The multi-core fish oil particles formed in this step are shown in Fig. 7A.

[0226] 16.0 g of pork gelatin was mixed with 184.0 g of distilled water. Gelatin dissolved after dispersion in distilled water and heating to and holding at 57 ° C. 24.0 g of CoQio powder was added to the gelatin solution and emulsified at 6000 rpm for 2 minutes and 7500 rpm for 1 minute. A CoQio emulsion was formed and contained droplets approximately 1-5 µm in diameter. Then 12.2 g of the CoQio emulsion was mixed with the agglomerated slurry in the reactor at 48.3 ° C. CoQio coated microcapsules are shown in Fig. 7B.

[0227] The above microcapsule agglomerations containing the suspension were then cooled to 4 ° C over 2.5 hours. The transglutaminase enzyme preparation was added at 0.2% w / w. and the temperature was adjusted to 20 ° C for enzymatic curing for at least 12 hours. The finished microcapsule suspension was spray dried. The powder of the microcapsules was free flowing and the free surface oil was less than 0.1% w / w.

Example 10.3: Microencapsulation of DHA oil using pork gelatin with a loading ratio of 200 mg Co-Q10 / 500 mg EPA / DHA

[0228] 36.1 g of pork gelatin was mixed with 396.7 g of distilled water. Gelatin dissolved upon dispersion in water and heating to and holding at 50 ° C. The reactor was connected to a circulation pump and the temperature was set to 50 ° C. 728.0 g of distilled water was added to the reactor and the temperature was maintained at 50 ° C. To this freshly prepared gelatin solution was added 72.0 g of fish oil and emulsified at 7500 rpm for 4 minutes. An emulsion is formed and contains oil droplets having a diameter of about 1-5 µm. The emulsion was added to the reactor containing water at 50 ° C. The mixture had a pH of 5.045. Then 6.4 g of sodium ascorbate was added to the mixture. A further 85.2 g of 5% w / w was added to the reactor. sodium polyphosphate solution at room temperature. The pH was adjusted to 4.488 which allowed the agglomeration to grow to about 40 µm as judged by an optical microscope.

[0229] 16.0 g of pork gelatin was mixed with 184.0 g of distilled water. Gelatin dissolved after being dispersed in water and heated to and held at 57 ° C. Then 24.0 g of CoQio powder was added to the gelatin solution and emulsified at 6000 rpm for 2 minutes and 7500 rpm for 1 minute. A CoQio emulsion was formed and contained droplets approximately 1-5 µm in diameter. Then 82.0 g of CoQio emulsion was mixed with the agglomerated slurry in the reactor at 48.3 ° C. The above suspension, containing the microcapsule agglomerations, was then cooled to 4 ° C over 2.5 hours. The transglutaminase enzyme preparation was added at 0.2% w / w. and the temperature was adjusted to 20 ° C for enzymatic curing for at least 12 hours. The finished microcapsule suspension as shown in Fig. 8 was spray dried. The powder of the microcapsules was free flowing and the surface oil free was less than 0.1% w / w.

Example 11 Co-delivery of zinc and fish oil in microcapsule powder

[0230] The omega-3 microcapsule powder used had an average of 180.5 mg / g DHA + EPA powder and 210.9 mg / g total omega-3 powder. For zinc supply at 2, 5, 10, 50

- 53 and 100 mg per 500 mg EPA + DHA powder, ZnCl 2 was added to the finished slurry prior to spray drying. The formulations used are described in (Table 8).

Table 8: Designed microcapsules with different levels of zinc

<td>mg Zn / 500 mg (DHA + EPA)</td><td>mg Zn / g powder</td><td>mg ZnCL / g powder</td><td>Formulated ZnCL mg / g powder</td><td>ZnCl₂ in 100g of suspension</td>

<td></td><td></td><td></td><td></td><td></td>

<td> 2</td><td> 0,72</td><td> 1,50</td><td> 1,91</td><td> 0,017</td>

<td> 5</td><td> 1,81</td><td> 3,76</td><td> 4,77</td><td> 0,042</td>

<td> 10</td><td> 3,61</td><td> 7,52</td><td> 9,53</td><td> 0,085</td>

<td> 50</td><td> 18,05</td><td> 37,62</td><td> 47,67</td><td> 0,424</td>

<td> 100</td><td> 36,10</td><td> 75,24</td><td> 95,33</td><td> 0,848</td>

<td colspan="5">Total solids content of the suspension (%): 8.90</td>

Example 11.1 Preparation of Fish Oil Microcapsule Stock Suspension Using 240 Bloom Fish Gelatin

[0231] Omega-3 fish oil microcapsule was prepared by dissolving 44g of 240 Bloom Fish Gelatin in 320g of water. The solution was then heated to 40 ° C. 7.3 g of sodium ascorbate were added to the gelatin solution. The pH of the solution increased from 5.385 to 5.650. Then 72.0 g of high DHA fish oil (OXDHA, Ocean Nutrition Canada Ltd., Dartmouth, NS) was added to the gelatin solution and then emulsified at 7500 rpm for 4 minutes using a Polytron high speed homogenizer. The emulsion was examined microscopically after emulsification and it was verified that the oil droplets were small and uniform (diameter about 1-5 µm). 1051 g of distilled water was added to the 2 L reactor and held at 40 ° C. The emulsion was added to distilled water in the reactor and the pH of the mixture was 5.662 at 39.6 ° C. Then, 4.4 g of sodium polyphosphate was dissolved in 84 g of distilled water and added to the dilute emulsion in the reactor. The mixture in the reactor had a pH of 6.401. The pH was then lowered with 10% phosphoric acid to agglomerate the primary microcapsules. After lowering the pH to 4.459, the secondary agglomeration of the microcapsules had a diameter of 30-70 µm. The slurry was then cooled from 40 ° C to 5 ° C at an average rate of 1 ° C / 5 min. After adjusting the pH to 6.0, 1% transglutaminase was added to the slurry to cross-link and cure the shell at 5 ° C for 1 hour, 15 ° C. for 8 hours, 20 ° C for 9 hours.

[0232] The previous steps were performed with four identical samples of the slurry. After cross-linking, the suspensions were mixed. One liter of the mixed suspension was then spray dried to form a free flowing powder. This sample only had omega oil

-543 and contained no supply zinc. Lipid analysis showed the powder had 129 mg DHA / g, 31 mg EPA / g, and a total omega-3 content of 176 mg / g powder.

Example 11.2: Preparation of omega-3 microcapsules using 240 Bloom gelatin with zinc incorporated into the suspension

[0233] Zinc-omega-3 microcapsules were prepared using 240 Bloom gelatin as described above for Example 11.1. One liter of the mixed suspension was taken and stirred with a magnetic stirrer. 0.15 g ZnCU was dissolved in the suspension. After stirring for 30 minutes, the slurry was spray dried to give a free flowing powder containing omega-3 oil as well as zinc for delivery. Various amounts of ZnCU (0.38, 0.76, 3.81 and 7.63 g, respectively) were incorporated into 1 L of the mixed slurry, resulting in different levels of zinc to be delivered. These are listed as Examples 11.2.1 to 11.2.5. The zinc results and analysis are shown in Table 9.

Table 9: Comparison of zinc levels from powder samples

<td>Example #</td><td>Zinc (mg / g)</td><td>Zinc (mg / 500 mg EPA + DHA)</td>

<td> 11.1</td><td> 0,006</td><td> 0,02</td>

<td> 11.2.1</td><td> 1,1</td><td> 3,4</td>

<td> 11.2.2</td><td> 2,3</td><td> 7,3</td>

<td> 11.2.3</td><td> 3,7</td><td>H, 6</td>

<td> 11.2.4</td><td> 18,5</td><td> 57,8</td>

<td> 11.2.5</td><td> 32,7</td><td> 102,2</td>

[0234] The amount of zinc in the microcapsule powder was well predicted by the amount added to the slurry prior to spray drying (Fig. 9).

Specific embodiments

[0235] Disclosed herein is a microcapsule comprising agglomeration of primary microcapsules and a loading material, each individual primary microcapsule having a primary shell, the loading agent being encapsulated by a primary shell, the agglomeration being encapsulated by an outer shell and the primary shell, outer shell or both contain the remainder of one or more compositions containing amino acid, protein, saccharide, wax, or a combination thereof. Also disclosed herein is a single core microcapsule comprising a core, the core comprising a charging substance, a primary sheath surrounding the core, and an outer sheath surrounding the primary sheath, the primary sheath, outer sheath, or both containing the remainder of one or more compositions comprising amino acid, protein, saccharide, wax. or a combination of them.

[0236] Also disclosed is a method for preparing a microcapsule comprising providing an emulsion comprising a first polymer component, a loading agent, a second polymer component and a composition comprising one or more of an amino acid, protein, saccharide, wax, or a combination thereof; adjusting the pH, temperature, concentration, agitation rate, or a combination thereof to form an aqueous mixture comprising a primary shell material, the primary shell material comprising the first and second polymer components and surrounding the loading agent; cooling the aqueous mixture to a temperature above the gelling temperature of the primary shell material until the primary shell material agglomerates; and further cooling the aqueous mixture to form an outer shell around the agglomeration, wherein the primary shell material, the outer shell or both comprises a saccharide, a wax, or a combination thereof.

[0237] Still further, disclosed is a method for preparing a microcapsule comprising providing a suspension of one or more microcapsules, the microcapsule comprising a shell material and a loading agent; adding a composition containing one or more of an amino acid, protein, saccharide, wax, antioxidant, or zinc, or combinations thereof, to the suspension; and then drying the slurry.

[0238] Further disclosed is also a method for preparing a microcapsule comprising providing an emulsion comprising a first polymer component, a loading agent and a second polymer component; adjusting the pH, temperature, concentration, agitation rate, or a combination thereof to form an aqueous mixture containing the primary shell material, the primary shell material comprising the first and second polymer components and surrounding the loading agent; cooling the aqueous mixture to a temperature above the gelling temperature of the primary shell material until the primary shell material agglomerates; adding a composition containing one or more of an amino acid, protein, saccharide, or wax to the aqueous mixture; and further cooling the aqueous mixture to form an outer shell around the agglomeration, wherein the primary shell material, the outer shell or both contain saccharide.

[0239] Also disclosed is a method of preparing a microcapsule comprising providing an emulsion comprising a first polymer component, a loading agent, a second polymer component, and an emulsion chelator; adjusting the pH, temperature, concentration, agitation rate, or a combination thereof to form an aqueous mixture comprising a primary shell material, the primary shell material comprising the first and second polymer components and surrounding the loading agent; cooling the aqueous mixture to a temperature above the gelling temperature of the primary shell material until the primary shell material agglomerates; and further cooling the aqueous mixture to form an outer shell around the agglomeration.

[0240] Also disclosed is a composition comprising a spray dried emulsion comprising a first polymer component and a loading agent and the remainder of the one or more compositions comprising an amino acid, protein, saccharide, wax, or a combination thereof.

[0241] Additionally, disclosed is a formulation carrier containing any of the microcapsules disclosed herein. The formulation carrier may be a foodstuff, beverage, nutritional formulation or pharmaceutical formulation. Also disclosed is a sachet containing any of the microcapsules disclosed herein.

[0242] Still further disclosed is a method of delivering a loading substance to a subject, comprising administering to the subject any of the microcapsules or any of the formulation carriers disclosed herein. The subject may be a mammal. The subject may be a human. The loading agent may include an omega-3 fatty acid, an omega-3 fatty acid alkyl ester, an omega-3 fatty acid triglyceride ester, an omega-3 fatty acid phytosterol ester, and / or a mixture thereof. Also disclosed is the use of any of the microcapsules disclosed herein for the preparation of a medicament to deliver a loading substance to a subject.

[0243] A microcapsule may be prepared by a method that comprises providing an emulsion comprising a first polymer component, a loading agent and a second polymer component; adjusting the pH, temperature, concentration, agitation rate, or a combination thereof to form an aqueous mixture comprising a primary shell material, the primary shell material comprising the first and second polymer components and surrounding the loading agent; cooling the aqueous mixture to a temperature above the gelling temperature of the primary shell material until the primary shell material agglomerates; and further cooling the aqueous mixture to form an outer shell around the agglomeration.

[0244] The disclosed microcapsules can have an induction period greater than about 40 hours, greater than about 50 hours, greater than about 75 hours, or greater than about 100 hours.

[0245] The composition may include an amino acid and the ratio of amino acid to the second polymer component may be about 1: 5 to about 5: 1. One or more compositions may contain the amino acid leucine, isoleucine, methionine, cysteine, tyrosine, tryptophan, phenylalanine, or a mixture thereof. One or more compositions may include the amino acid lysine. One or more compositions may include the amino acid glutamine. One or more of the compositions may contain the amino acids leucine, isoleucine, methionine, cysteine, tyrosine, tryptophan, phenylalanine or a mixture thereof, and glutamine. One or more of the compositions may include milk protein. One or more compositions may include whey protein, whey protein isolate, or whey protein concentrate; Whey protein can be combined with glycerin.

[0246] The composition may include a protein, and the ratio of protein to the second polymer component may be about 1: 1 to about 40: 1. The protein may be milk protein, gelatin, whey protein isolate, whey protein concentrate, caseinate, soy protein, BSA, or a mixture thereof. The composition may contain whey protein, whey protein isolate or whey protein concentrate. Whey protein can be combined with glycerin.

[0247] The composition may include a saccharide and the ratio of saccharide to the second polymer component may be from about 1: 0.02 to about 1: 0.5. Composition may contain

The saccharide and the ratio of saccharide to total shell material can be from about 1: 0.2 to about 1: 5.

[0248] One or more compositions may contain a saccharide having a molecular weight greater than about 100,000 Daltons or less than about 100,000 Daltons. One or more compositions may include a chitosan saccharide. One or more compositions may include chitosan and glutamine, chitosan, lysine, and glutamine, chitosan, glutamine, and one or more of leucine, isoleucine, methionine, cysteine, tyrosine, tryptophan or phenylalanine, or chitosan and one or more of leucine, isoleucine, isoleucine , cysteine, tyrosine, tryptophan or phenylalanine. One or more compositions may include a starch saccharide; the starch may be a modified starch. One or more compositions may contain a lactose saccharide. One or more compositions may contain starch and lactose saccharides. One or more compositions may contain the saccharide in the form of maple syrup, honey, corn syrup, or a mixture thereof. One or more compositions may contain a saccharide sucrose. One or more compositions may contain a hydroxypropyl methylcellulose saccharide. One or more of the compositions may contain maltodextrin saccharide, oligofructans, cyclodextrins, carboxymethyl cellulose, ethyl cellulose, hydroxypropyl cellulose, cellulose ether, agar, alginate, pectin, low methoxyl pectin, gum arabic, carrageenan, gummo gum, dilutosan gum, gummo gum, dilutosan gum. , xanthan gum, or a mixture thereof. One or more of the compositions may contain the saccharide glucose, fructose, galactose, arabinose, ribose, ribulose, xylose, xylulose, cellobiose, mannose, xylose, ribose, sorbose, celotriose, trehalose, maltose, isaltomine, sorbitol, xylitol, xylitol, or theirs, xylulose. mixtures. The saccharide may be added after cooling but before cooling the mixture further to form an outer shell around the agglomeration.

[0249] One or more of the compositions may include kamauba wax. The composition may contain carnauba wax in the form of a microemulsion. One or more of the compositions may include candelilla wax, cersines wax, Japanese wax, orange peel wax, rice bran wax, shellac, paraffin, montan, microcrystalline wax, polyethylene, beeswax, or mixtures thereof. One or more compositions may additionally contain a surfactant. The composition may include wax and the ratio of wax to the second polymer component may be about 1: 1 to about 1:10.

[0250] One or more of the compositions may include an antioxidant. The antioxidant may include coenzyme Q10, lutein, zeaxanthan, carotene (e.g., beta-carotene), or mixtures thereof.

[0251] The disclosed capsules may additionally contain a chelator. The chelator may be disodium ethylenediaminetetraacetic acid. The chelator may contain one or more of acetic acid, phytic acid, malic acid, tartaric acid, oxalic acid, succinic acid, polyphosphoric acids, or mixtures thereof. The chelator may be added to the emulsion and / or the water mixture.

[0252] The disclosed microcapsules may additionally contain an anti-caking compound. The anti-caking compound may be added to the microcapsule before, during, or after drying.

[0253] The antioxidant may be added to the emulsion and / or the aqueous mixture. The antioxidant may include a phenolic compound, a plant extract or a sulfur-containing compound. The antioxidant may include ascorbic acid or a salt thereof.

[0254] The composition may additionally include a surfactant.

[0255] The primary shell or outer shell, or both the primary shell and the outer shell, may contain a surfactant, gelatin, polyphosphate, saccharide, or a mixture thereof. The primary coat or outer coat, or both the primary coat and the outer coat, may contain Type B gelatin, polyphosphate, gum arabic, alginate, chitosan, carrageenan, pectin, low methoxy pectin, starch, modified starch, alphalactalbumin, beta-lactoglobumin, ovalbumin, polysorbitrin, polysorbitrin, polyphosphate , cyclodextrin, cellulose, methylcellulose, ethylcellulose, hydroxypropylmethylcellulose, carboxymethylcellulose, milk protein, whey protein, soy protein, rapeseed protein, albumin, kosher gelatin, non-kosher gelatin, halal gelatin, non-halal gelatin, or a mixture thereof. The primary coat or outer coat or both the primary and the outer coat may contain an antioxidant. The primary or outer coat, or both the primary and the outer coat, may contain zinc.

[0256] The primary coat or outer coat, or both the primary and the outer coat, may contain Type A gelatin. The primary coat or outer coat, or both the primary and the outer coat may contain fish gelatin. The primary coat or outer coat or both the primary and the outer coat may contain porcine gelatin. The primary coat or outer coat, or both the primary and the outer coat, may contain gelatin with a Bloom number from about 0 to about 300. The primary coat or outer coat or both primary and outer coat may contain gelatin with a Bloom number from about 0 to about 50. or the outer sheath or both the primary and the outer sheath may contain gelatin with a Bloom Number from about 51 to about 300. The primary coat or outer coat, or both the primary and the outer coat, may contain gelatin with a Bloom Number of about 0, about 210, about 220, or about 240. The primary coat or outer coat, or both primary and outer coat, may contain complex coacervate. The primary coat or outer coat or both the primary and the outer coat may contain a complex gelatin and polyphosphate coacervate. The primary shell material and the outer shell may contain a complex coacervate between gelatin and polyphosphate. The primary and outer shell material may contain a complex coacervate between gelatin and alginate, gelatin and pectin, gelatin and acacia, gelatin and xanthan, gelatin and low methoxy pectin or gelatin and whey protein.

[0257] The first polymer component may contain a surfactant, gelatin, polyphosphate, saccharide, or a mixture thereof. The first polymer component may contain gelatin of the type

-59B, polyphosphate, acacia, alginate, chitosan, carrageenan, pectin, low methoxyl pectin, starch, modified starch, alpha-lactalbumin, beta-lactoglobumin, ovalbumin, polysorbitone, maltodextrin, cyclodextrin, ethylcellulose, methylcellulose, methylcellulose, methylcellulose, methylcellulose, milk protein, whey protein, soy protein, rapeseed protein, albumin, kosher gelatin, low cost gelatin, halal gelatin, non-halal gelatin, or a mixture thereof. The first polymer component may contain type A gelatin. The first polymer component may contain fish gelatin. The first polymer component may contain pork gelatin. The first polymer component may have a Bloom Number from about 0 to about 300. The first polymer component may have a Bloom Number from about 0 to about 50. The first polymer component may have a Bloom Number from about 51 to about 300. The first polymer component may have a Bloom Number of about 0, about 210, about 220, or about 240.

[0258] The second polymer component may contain a surfactant, gelatin, polyphosphate, saccharide, or a mixture thereof. The second polymer component may contain gelatin type A, gelatin type B, polyphosphate, gum arabic, alginate, chitosan, carrageenan, pectin, low-methoxyl pectin, starch, modified starch, alpha-lactalbumin, beta-lactoglobumin, ovalbumin, polysorbitrin, cycltodrbitrin, cellulose, methyl cellulose, ethyl cellulose, hydroxypropyl methyl cellulose, carboxymethyl cellulose, milk protein, whey protein, soy protein, rapeseed protein, albumin, kosher gelatin, non-kosher gelatin, halal gelatin, non-halal gelatin, or a mixture thereof. The second polymer component may contain a polyphosphate.

[0259] The loading agent may contain a bioactive substance, nutritional supplement, microbial oil, sea oil, algae oil, dinoflagellate oil, Crypthecodinium cohnii oil, mushroom oil, Thraustochytrium oil, Schizochytrium or a mixture thereof, or a vegetable oil.

[0260] The loading agent may include fish oil, such as Atlantic fish oil, Pacific fish oil, Mediterranean fish oil, light pressed fish oil, alkaline processed fish oil, heat treated fish oil, light and heavy fish oil. brown fish oil, bonito oil, sprats oil, tuna oil, sea bass oil, halibut oil, marlin oil, barracuda oil, cod oil, menhaden oil, sardine oil, anchovy oil, capelin, Atlantic cod oil, Atlantic herring oil, Atlantic mackerel oil, Atlantic menhaden oil, salmon oil, or shark oil. The loading subsidy may contain untreated fish oil. The loading agent may include arachidonic acid. The loading agent may include an omega-3 fatty acid, an omega-3 fatty acid alkyl ester, an omega-3 fatty acid triglyceride ester, an omega-3 fatty acid phytosterol ester, and / or a mixture thereof. The loading agent may contain docosahexaenoic acid and / or eicosapentaenoic acid, their C1-C6 alkyl ester, their triglyceride ester, their phytosterol ester and / or a mixture thereof.

[0261] In the disclosed microcapsules, the outer shell may have an average diameter of from about 1 µm to about 2000 µm, from about 20 µm to about 1000 µm, or from about 30 µm to about 80 µm. The primary shell may have an average diameter of from about 40 nm to about 10 µm, or from about 0.1 µm to about 5 µm. The loading agent may be from about 20% to about 90% or from about 50% to about 70% by weight of the microcapsule.

[0262] In the disclosed methods, any or all steps may be performed under a nitrogen atmosphere.

[0263] The disclosed methods may further comprise adding a transglutaminase. The disclosed methods may additionally include the addition of glutaraldehyde.

[0264] The disclosed methods may further comprise drying the microcapsules. The microcapsules may be spray dried. The microcapsules may be spray-dried in the presence of a carbohydrate.

[0265] In the disclosed methods, an emulsion can be made by emulsification at about 1,000 to about 15,000 rpm. The emulsion may additionally contain a composition containing a saccharide, a wax, or a combination thereof.

[0266] In the disclosed methods, cooling may be at a rate of 1 ° C, for about 1 to about 100 minutes, or at a rate of 1 ° C / 5 minutes. The mixture may be cooled until it reaches a temperature of about 5 ° C to about 10 ° C or about 5 ° C.

[0267] A microcapsule obtained according to the disclosed methods is also disclosed herein.