Protein encapsulation of nutritional and pharmaceutical compositions

Abstract

The present disclosure relates to microencapsulated compositions comprising one or more hydrophobic substances encapsulated by an encapsulant, the encapsulant comprising one or more denatured proteins and / or peptides, the one or more denatured proteins and / or peptides being obtained from a starting protein by subjecting the starting protein to a high shear process such that the average particle size of the one or more denatured proteins and / or peptides is reduced relative to the starting protein. The present disclosure further relates to methods for protecting hydrophobic substances from oxidative degradation, improving the oxidative stability of hydrophobic substances, and reducing surface free fat in microencapsulated compositions, as well as stable emulsions and compositions comprising the hydrophobic substances.

Description

Technical Field

[0001] The present disclosure broadly relates to encapsulated compositions (hereinafter also referred to as encapsulated compositions) suitable for both nutritional and pharmaceutical uses, and means for protecting hydrophobic substances in the encapsulated compositions from oxidation and oxidative degradation.

Background Art

[0002] It is well known that various hydrophobic bioactive compounds, such as long-chain polyunsaturated fatty acids (“LCPUFAs”), carotenoids, water-insoluble vitamins, phenolic compounds, flavor components and aromatic components, provide various health benefits. Specifically, LCPUFAs are important nutritional components of the human diet, and many people are unable to consume sufficient amounts of these essential fatty acids, specifically omega-3 fatty acids such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). Numerous studies have found that omega-3 fatty acids play a significant role in the health of the heart, brain and eyes, and dietary intake is deeply related to the improvement of cardiovascular function and the suppression of various inflammation-related pathologies. For example, recent studies have suggested that EPA and DHA may have the ability to reduce heart rate and oxygen consumption during exercise, and thus contribute to the improvement of the physical and mental performance of athletes (People et al., Journal of Cardiovascular Pharmacology, 2008, 52: 540-547). Due to their essential nutritional role, compositions containing omega-3 fatty acids are important from both the perspectives of nutritional supplements and pharmaceuticals.

[0003] Therefore, there is a growing trend to incorporate omega-3 fatty acids, such as fish oil, algae oil, and certain plant seed oils, into food products to promote public health. However, these fatty acids are susceptible to oxidation or degradation upon exposure to oxygen, rising temperatures, or light, which often occur during food production and storage. Therefore, properly fortifying products with omega-3 fatty acids and maintaining their stability and activity is a challenge. Oxidation and / or degradation of omega-3 fatty acids produce undesirable oxidative degradation products that can have adverse effects on the functional or physiological properties of the formulation. Consequently, producing, transporting, and storing these functional foods is difficult.

[0004] Microencapsulation technology allows bioactive compounds to be sealed within a physically protective shell material, and this technology is well used to protect omega-3 fatty acids from oxidation and degradation. Spray drying is the most widely used technology for producing microencapsulated powders. Typically, spray-dried microencapsulated powders contain omega-3 rich oils and have an oil content of approximately 30% (w / w) and a surface free fat content of approximately 1% (w / w). Due to the excellent functionality of Maillard reaction products (MRPs), omega-3 oil-containing microencapsulated powders are produced with oil content as high as 48±2% while maintaining a surface free fat content of approximately 1% (w / w), and such products typically exhibit an induction period of just 50 hours (the number of hours before oxidation of the encapsulated oil begins). [Prior art documents] [Patent Documents]

[0005] [Patent Document 1] International Publication No. 2012 / 106777 [Patent Document 2] U.S. Patent No. 7,374,788,B2 [Non-patent literature]

[0006] [Non-Patent Document 1] People et al., Journal of Cardiovascular Pharmacology, 2008, 52: 540-547. [Overview of the Initiative] [Problems that the invention aims to solve]

[0007] There is a need to develop encapsulation and delivery systems that can improve the oxidative stability of hydrophobic compounds, particularly omega-3 oils. [Means for solving the problem]

[0008] This disclosure is based on the inventors' unexpected discovery that the use of a capsule material containing one or more denatured proteins and / or peptides (hereinafter also referred to as denatured proteins and / or peptides) (in which case, one or more denatured proteins and / or peptides are obtained from a starting protein by subjecting the starting protein to a high shear process) can result in a hydrophobic material-containing composition having both particularly high oxidative stability and especially low surface free fat content (i.e., high oil encapsulation efficiency). In certain embodiments, one or more denatured proteins and / or peptides are obtained from a starting protein by subjecting the starting protein to a high shear process such that the average particle size of the denatured proteins and / or peptides is reduced compared to the starting protein. In certain embodiments, the average particle size of the denatured proteins and / or peptides is about 70% or less of the average particle size of the starting protein, for example, about 65% or less of the average particle size of the starting protein. In some embodiments, one or more denatured proteins and / or peptides are used in the compositions or methods of this disclosure, and one or more denatured proteins are obtained from each of the one or more starting proteins.

[0009] A first aspect of the present disclosure provides a microencapsulated composition comprising one or more hydrophobic substances (hereinafter, the microencapsulated composition will also be referred to as the microencapsulated composition), wherein the capsule material comprises one or more denatured proteins and / or peptides, and the denatured proteins and / or peptides are obtained from a starting protein by subjecting the starting protein to a high-shear process such that the average particle size of the denatured proteins and / or peptides is reduced compared to the starting protein. In a preferred embodiment, the average particle size of the denatured proteins and / or peptides is about 70% or less of the average particle size of the starting protein, for example, about 65% or less of the average particle size of the starting protein.

[0010] According to some preferred embodiments, the microencapsulated composition has a surface free fat content of less than about 1.8%, for example less than about 1%, for example less than about 0.8%.

[0011] According to some embodiments, the high-shear process is carried out at an alkaline pH, for example, a pH of about 8. In some embodiments, the high-shear process includes applying a pressure of about 20 mPa to about 300 mPa to the starting protein. In some embodiments, the high-shear process is a homogenization process. In some embodiments, the high-shear process is a microfluidization process.

[0012] According to some embodiments, one or more denatured proteins and / or peptides are in the form of a protein fraction. According to some embodiments, the denatured protein is a denatured whey protein (hereinafter also referred to as denatured whey protein).

[0013] According to some embodiments, the capsule material further comprises one or more carbohydrates, such as glucose syrup and dextrose monohydrate, or a combination thereof.

[0014] According to some embodiments, one or more denatured proteins and / or peptides are present in an amount of about 3% w / w to about 25% w / w relative to the total mass of the composition.

[0015] According to some embodiments, the ratio of the denatured protein component to the carbohydrate component of the capsule material is in the range of approximately 1:10 to 1:1.

[0016] According to some embodiments, the hydrophobic substance is edible oil. In some embodiments, the hydrophobic substance comprises one or more long-chain polyunsaturated fatty acids (LCPUFAs). In some embodiments, the LCPUFA comprises omega-3 fatty acids and / or omega-6 fatty acids. In some embodiments, the LCPUFA exists in triglyceride form. In some embodiments, the LCPUFA exists as one or more LCPUFA-containing oils, and in some embodiments, one or more oils comprises fish oil. In some such embodiments, the fish oil is tuna oil.

[0017] According to some embodiments, the composition further comprises at least one source of vitamin C.

[0018] In some embodiments, the composition is in the form of an oil-in-water emulsion. In some embodiments, the composition is in the form of a spray-dried powder.

[0019] According to a second aspect, the present disclosure relates to a method for protecting a hydrophobic substance from oxidative degradation, A step of producing one or more denatured proteins and / or peptides by subjecting a starting protein to a high-shear process such that the average particle size of one or more denatured proteins and / or peptides is reduced compared to the starting protein, and The present invention provides a method comprising the step of encapsulating one or more hydrophobic substances in a capsule material containing one or more denatured proteins and / or peptides.

[0020] According to a third aspect, the present disclosure is a method for improving the oxidative stability of hydrophobic substances, comprising: subjecting a starting protein to a high-shear process so that the average particle size of one or more denatured proteins and / or peptides is reduced compared to the starting protein, thereby generating one or more denatured proteins and / or peptides; and encapsulating one or more hydrophobic substances with a capsule material comprising one or more denatured proteins and / or peptides.

[0021] According to a fourth aspect, the present disclosure is a method for reducing the surface-free fat of a microencapsulated composition comprising one or more hydrophobic substances encapsulated by a capsule material, comprising: subjecting one or more starting proteins and / or peptides to a high-shear process so that the average particle size of one or more denatured proteins and / or peptides is reduced compared to the one or more starting proteins and / or peptides, thereby generating one or more denatured proteins and / or peptides; and encapsulating one or more hydrophobic substances with a capsule material comprising one or more denatured proteins and / or peptides.

[0022] In some embodiments of the method according to the second, third, or fourth aspect, the average particle size of one or more denatured proteins and / or peptides is about 70% or less, such as about 65% or less, of the average particle size of the starting protein.

[0023] In some embodiments, the high-shear process is carried out at an alkaline pH, such as a pH of about 8.

[0024] In some embodiments, the hydrophobic substance comprises one or more LCPUFAs, for example, in the form of triglycerides.

[0025] According to a fifth aspect, the disclosure provides a stable emulsion comprising a hydrophobic substance, comprising one or more denatured proteins and / or peptides, wherein the one or more denatured proteins and / or peptides are obtained from a starting protein by subjecting the starting protein to a high-shear process such that the average particle size of the one or more denatured proteins and / or peptides is reduced compared to the starting protein.

[0026] In some embodiments, the average particle size of one or more denatured proteins and / or peptides is about 70% or less of the average particle size of the starting proteins, for example, about 65% or less.

[0027] In some embodiments, the high-shear process is carried out at an alkaline pH, for example, a pH of about 8.

[0028] In some embodiments, the hydrophobic substance comprises one or more LCPUFAs, for example, in triglyceride form.

[0029] According to a sixth aspect, the disclosure provides a composition comprising a hydrophobic substance and one or more denatured proteins and / or peptides, wherein the one or more denatured proteins and / or peptides are obtained from a starting protein by subjecting the starting protein to a high-shear process such that the average particle size of the one or more denatured proteins and / or peptides is reduced compared to the starting protein.

[0030] Exemplary embodiments of the present disclosure are described herein, with respect to the following drawings, merely as non-limiting examples. [Brief explanation of the drawing]

[0031] [Figure 1] This figure shows the interfacial tension of corn oil with 1.0% w / w uWPI and mWPI (original (i.e., true) pH and pH 8 solutions). [Figure 2]This scheme illustrates the process for preparing microencapsulated powders based on undenatured whey protein isolates (uWPI powder) and microencapsulated powders based on denatured whey protein isolates (mWPI powder). [Figure 3] This graph shows the results of Oxipres analysis of omega-3 oil-containing microencapsulated uWPI and mWPI powders compared to Maillard reaction products (MRP) and encapsulated omega-3 oil-containing microcapsule powders. [Figure 4] This figure shows the overall quality of microencapsulated uWPI and mWPI powders over a 4-week rapid exposure period. [Figure 5] This figure shows the rancid odor, marine odor, and flavor of microencapsulated uWPI & mWPI powders over a rapid exposure period of 4 weeks. [Modes for carrying out the invention]

[0032] Throughout this specification, unless otherwise required by context, the word “comprise,” or variations thereof such as “comprises” or “comprising,” is understood to mean encompassing a specified process, element, or integer, or a group of processes, elements, or integers, but not to mean excluding any other process, element, or integer, or a group of elements or integers. Therefore, in this specification, the term “comprises” means “primarily encompasses, but not exclusively.”

[0033] In this specification, the term "about" is understood to mean a range of numbers that a person skilled in the art would consider to be equal to the listed values ​​under the circumstances of achieving the same function or result.

[0034] With respect to this specification, a singular noun refers to one or more (i.e., at least one) grammatical objects of that thing. For example, “element” means one element or more elements.

[0035] The term "protein" refers to a polymer composed of amino acids linked together by peptide bonds. The term "peptide" may also be used to refer to such polymers, although in some examples peptides may be shorter (i.e., composed of fewer amino acid residues) than proteins. The terms "protein" and "peptide" may be used interchangeably herein.

[0036] As used herein, the term “oxidative stability” in relation to hydrophobic substances and compounds, such as LCPUFA, means the stability of a hydrophobic substance, such as LCPUFA or LCPUFA-containing oil, in the presence of oxygen, and its resistance to oxidation or oxidative degradation. Therefore, higher oxidative stability indicates greater resistance to oxidation and oxidative degradation. Typically, references to improved oxidative stability resulting from encapsulation in this disclosure mean an improvement over the oxidative stability observed without the encapsulation material of this disclosure or in the presence of an alternative encapsulation material.

[0037] According to embodiments of this disclosure, denatured whey protein was used as a capsule material in a microencapsulated composition containing tuna oil. Denaturation of the whey protein isolate by a high-shear process resulted in a decrease in average particle size, which is thought to be due to the separation of water-soluble protein aggregates. Specifically, the average particle size of the denatured protein was approximately 51% of that of the starting protein, at both the original pH and the alkaline pH (pH 8).

[0038] This denatured protein was used as a key capsule material component to obtain spray-dried microencapsulated powder, which stabilized omega-3 oil at a high level of total oil content (>45% total oil content (w / w) (tuna oil), specifically about 49%) and a low surface free fat content of less than 1.5 ± 0.1% (1.3 ± 0.5%, and an even lower surface free fat content of 0.6 ± 0.1% at pH 8). The resulting microencapsulated powder exhibited extremely high oxidative stability over an induction period well over 100 hours, as well as acceptable primary and secondary oxidative properties (including peroxide values, p-anisidine values, overall quality, and rancid and marine odors) over a rapid exposure period of 4 weeks. The denaturation processes disclosed herein can be applied to various proteins to obtain microencapsulation systems that can be used to extend the shelf life of various sensitive hydrophobic compounds, including omega-3 oils, carotenoids, water-insoluble vitamins, phenolic compounds, flavor compounds, and aromatic compounds.

[0039] Accordingly, a particular embodiment of the present disclosure provides a microencapsulation composition comprising one or more hydrophobic materials, wherein the capsule material comprises a denatured protein and / or peptide, the denatured protein and / or peptide being obtained from a starting protein by subjecting the starting protein to a high-shear process such that the average particle size of the denatured protein and / or peptide is reduced compared to the starting protein.

[0040] Methods and compositions are also provided for encapsulating one or more hydrophobic substances using denatured proteins or peptides to protect one or more hydrophobic substances from oxidation or oxidative degradation. Protection from oxidation or oxidative degradation can be determined by any suitable means well known to those skilled in the art.

[0041] The microencapsulated compositions of this disclosure may be, for example, in the form of an emulsion or in a solid form. The emulsion may include an oil-in-water emulsion. The solid form may be a powder. The powder may be obtained, for example, by spray-drying the emulsion. In one embodiment, the composition is a highly fluid powder. The powder may have an average particle size of about 10 μm to 1000 μm, or about 50 μm to 800 μm, or about 100 μm to 300 μm. In an alternative embodiment, the composition may be in the form of granules.

[0042] The compositions of the present disclosure are produced by microencapsulation, and the capsule material comprises or consists solely of denatured proteins and / or peptides. The “denatured proteins” or “denatured peptides” are obtained from a starting protein by subjecting the starting protein or peptide to a high-shear process such that the average particle size of the denatured protein or peptide is reduced compared to the starting protein. In some embodiments, one or more denatured proteins are used in the compositions or methods of the present disclosure, and one or more denatured proteins and / or peptides are each obtained from one or more starting proteins.

[0043] In this specification, references to "protein" may refer to a starting protein, a denatured protein, or both, as can be easily understood from the context.

[0044] High shear processes can be used to alter one or more properties of a protein, such as its particle size, solubility, foaming ability, gelling ability, and / or emulsifying ability. The size of fat globules in emulsions formed from aqueous solutions of such proteins and fats can also be reduced. In particular, by subjecting denatured proteins to a high shear process, the interfacial tension with oil can be reduced when used in an aqueous solution at an alkaline pH, for example, about 8. Proteins can be denatured using any suitable high shear process, and a variety of high shear processes are well known to those skilled in the art.

[0045] In some embodiments, the high-shear process is a homogenization process. In some typical embodiments, the homogenization process may be a high-pressure homogenization process in which proteins are forced to flow at high speed through narrow gaps. In some embodiments, the homogenization process is microfluidization. Microfluidization processes use high shear rates and uniform processing pressures to advantageously obtain consistent nanoscale particle sizes and narrow particle size distributions. Exemplary microfluidization equipment is available from Microfluidics International Corporation, USA. In some typical embodiments, the homogenization process may be an ultrasonic pressure homogenization process in which ultrasonic pressure waves are generated in the medium to induce homogenization. In some embodiments, the homogenization process may be a mechanical homogenization process, for example, using a multi-stage, e.g., rotor-stator homogenizer or blade-type homogenizer. Those skilled in the art will recognize that the scope of this disclosure is not limited by reference to any particular homogenization process.

[0046] In some embodiments, a high-shear process involves multiple passes through a high-shear configuration. For example, in embodiments using a high-pressure homogenization process, the material may undergo multiple passes through narrow gaps to achieve the desired homogenization. For example, a high-shear process may involve 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more passes.

[0047] In some embodiments, the high-shear process includes applying a pressure to the starting protein from about 20 mPa to about 300 mPa, for example from about 50 mPa to about 300 mPa, for example from about 100 mPa to about 300 mPa, for example from about 100 mPa to about 250 mPa, for example from about 100 mPa to about 200 mPa, for example from about 125 mPa to about 175 mPa, for example from about 150 mPa, or from about 125 mPa to about 300 mPa, for example from about 150 mPa to about 300 mPa, for example from about 175 mPa to about 300 mPa, for example from about 200 mPa to about 300 mPa, for example from about 225 mPa to about 300 mPa, for example from about 250 mPa to about 300 mPa.

[0048] In some specific embodiments, the high shear process includes a homogenization process, which includes applying a pressure to the starting protein from about 20 mPa to about 300 mPa, for example from about 50 mPa to about 300 mPa, for example from about 100 mPa to about 300 mPa, for example from about 100 mPa to about 250 mPa, for example from about 100 mPa to about 200 mPa, for example from about 125 mPa to about 175 mPa, for example from about 150 mPa, or from about 125 mPa to about 300 mPa, for example from about 150 mPa to about 300 mPa, for example from about 175 mPa to about 300 mPa, for example from about 200 mPa to about 300 mPa, for example from about 225 mPa to about 300 mPa, for example from about 250 mPa to about 300 mPa.

[0049] In some specific embodiments, the high shear process includes a homogenization process, which includes applying pressure to the starting protein from about 20 mPa to about 300 mPa, for example from about 50 mPa to about 300 mPa, for example from about 100 mPa to about 300 mPa, for example from about 100 mPa to about 250 mPa, for example from about 100 mPa to about 200 mPa, for example from about 125 mPa to about 175 mPa, for example from about 150 mPa, or from about 125 mPa to about 300 mPa, for example from about 150 mPa to about 300 mPa, for example from about 175 mPa to about 300 mPa, for example from about 200 mPa to about 300 mPa, for example from about 225 mPa to about 300 mPa, for example from about 250 mPa to about 300 mPa.

[0050] In some specific embodiments, the high-shear process is carried out in an alkaline environment, for example, in an alkaline aqueous solution, and the high-shear process is applied to a protein in an aqueous solution with a pH of about 8.

[0051] The process of subjecting a starting protein to a high-shear process may be used to obtain a denatured protein or peptide having a reduced average particle size compared to the starting protein. Specifically, the average particle size of the denatured protein or peptide may be about 70% or less of the average particle size of the starting protein, for example, about 65% or less of the average particle size of the starting protein. In some embodiments, the average particle size of the denatured protein or peptide may be about 60% or less of the average particle size of the starting protein, for example, about 55% or less of the average particle size of the starting protein. The reduction in the average particle size of the protein can be readily determined by any suitable method readily available to those skilled in the art. One detailed method that may be used to determine the average particle size of the starting protein and the denatured protein or peptide, and consequently whether a reduction has occurred, is the use of the principle of dynamic light scattering, for example, by using a Malvern Zetasizer (Malvern Panalytical).

[0052] The scope of this disclosure should not be limited by reference to any particular protein. Any suitable starting protein and denatured protein or peptide may be used in the compositions and methods of this disclosure. The protein may be, for example, in the form of a protein fraction obtained from a natural source, such as a cell or tissue source. The cell or tissue source may be obtained from any suitable source, such as an animal or plant source. In a typical embodiment, the protein is a whey protein isolate, the starting protein is an undenatured whey protein isolate, and the denatured protein is a denatured whey protein isolate. In another typical embodiment, the protein is a whey protein concentrate, the starting protein is an undenatured whey protein concentrate, and the denatured protein is a denatured whey protein concentrate. In other embodiments, the protein may be derived from a plant source, such as a pea or soybean protein, or a pea protein isolate or soybean protein isolate.

[0053] Any suitable protein of any molecular weight can be used in accordance with this disclosure. For example, the starting protein and / or denatured protein or peptide may have a molecular weight in the range of about 500 Da to about 150 kDa. For example, proteins can have molecular weights of approximately 500 kDa, 1 kDa, 2 kDa, 3 kDa, 4 kDa, 5 kDa, 10 kDa, 15 kDa, 20 kDa, 25 kDa, 30 kDa, 35 kDa, 40 kDa, 45 kDa, 50 kDa, 55 kDa, 60 kDa, 65 kDa, 70 kDa, 75 kDa, 80 kDa, 85 kDa, 90 kDa, 95 kDa, 100 kDa, 105 kDa, 110 kDa, 115 kDa, 120 kDa, 125 kDa, 130 kDa, 135 kDa, 140 kDa, 145 kDa, or up to approximately 150 kDa.

[0054] Any suitable protein of any molecular size can be used in accordance with this disclosure. For example, the starting protein and / or denatured protein or peptide may have a particle radius in the range of about 0.5 nm to about 5 nm. For example, proteins may have particle radii of approximately 0.5 nm, 0.6 nm, 0.7 nm, 0.8 nm, 0.9 nm, 1.0 nm, 1.1 nm, 1.2 nm, 1.3 nm, 1.4 nm, 1.5 nm, 1.6 nm, 1.7 nm, 1.8 nm, 1.9 nm, 2.0 nm, 2.1 nm, 2.2 nm, 2.3 nm, 2.4 nm, 2.5 nm, 2.6 nm, 2.7 nm, 2.8 nm, 2.9 nm, 3.0 nm, 3.1 nm, 3.2 nm, 3.3 nm, 3.4 nm, 3.5 nm, 3.6 nm, 3.7 nm, 3.8 nm, 3.9 nm, 4.0 nm, 4.1 nm, 4.2 nm, 4.3 nm, 4.4 nm, 4.5 nm, 4.6 nm, 4.7 nm, 4.8 nm, 4.9 nm, or approximately 5.0 nm.

[0055] Denatured proteins and / or peptides may be introduced into the emulsion or composition at any stage in the preparation of the emulsion or composition so as to form a homogeneous aqueous dispersion or slurry. Those skilled in the art can optimize the amount and molecular weight of the introduced proteins and / or peptides without excessive burden or experimentation. In some preferred embodiments, the molecular weight of the proteins and / or peptides may be low enough to facilitate microencapsulation, while the amount of said proteins and / or peptides may be sufficient to produce effective protection as a capsule material. In the case of oil-in-water emulsions, viscosity can also be controlled. If the viscosity is too high, spray drying may be hindered. Determining the appropriate protein content and appropriate viscosity is well within the capabilities of those skilled in the art.

[0056] In a typical embodiment, denatured proteins and / or peptides may be present in amounts of about 3% (w / w) to about 30% (w / w) of the total mass of the composition, or about 3% (w / w) to about 25% (w / w) of the total mass of the composition. In the case of an oil-in-water emulsion, this means about 3% (w / w) to about 30% (w / w) of the total mass of the aqueous and oil phases, or about 3% (w / w) to about 25% (w / w). For example, proteins and / or peptides may be present in amounts of about 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%, or 30% w / w of the total mass of the composition.

[0057] In certain embodiments described herein, the capsule material comprises compounds, substances, or parts in addition to denatured proteins and / or peptides. For example, the capsule material may comprise a combination of denatured proteins and one or more polysaccharide or carbohydrate components. For example, carbohydrates having reducing sugar functional groups may react with proteins, dextrose (including dextrose monohydrate), glucose, lactose, sucrose, oligosaccharides, and dry glucose syrup. In further embodiments, polysaccharides, high-methoxyl pectin, or carrageenan may be added to the protein-carbohydrate mixture in some formulations. Care must be taken when reacting proteins and carbohydrates to ensure that this state does not result in large-scale gelation or coagulation of the protein, as this would prevent the protein from forming a good film.

[0058] In a typical embodiment, the compositions of the present disclosure may be prepared by solubilizing a polysaccharide or carbohydrate component of the capsule material in an aqueous phase containing a denatured protein, optionally using a high-shear mixer. The mixture may then be heated to a temperature of about 50°C to 80°C, after which one or more antioxidants may be added as needed. The hydrophobic material may be added inline to the aqueous mixture and passed through a high-shear mixer to form a crude emulsion. The crude emulsion may then be subjected to homogenization. If it is desirable to prepare a powdered product, the emulsion may be pressurized and spray-dried at an inlet temperature of about 180°C and an outlet temperature of 80°C.

[0059] For example, suitable polysaccharide and carbohydrate components may include maltodextrin, dextrose (including dextrose monohydrate), glucose, lactose, sucrose, oligosaccharides, and dry glucose syrup, or a combination of one or more thereof. In further embodiments, polysaccharides, high-methoxyl pectin, or carrageenan may be added to the protein-carbohydrate mixture in some formulations. Care must be taken when reacting the protein and carbohydrate to ensure that this state does not result in large-scale gelation or coagulation of the protein, as this would prevent the protein from forming a good film. The ratio (by mass) of denatured protein to polysaccharide or carbohydrate component of the capsule material may be, for example, about 3:1, 2.5:1, 2:1, 1.5:1, 1:1, 1:1.5, 1:2, 1:2.5, 1:3, 1:3.5, 1:4, 1:4.5, 1:5, 1:5.5, 1:6, 1:6.5, 1:7, 1:7.5, 1:8, 1:8.5, 1:9, 1:9.5, or 1:10.

[0060] In certain embodiments, the ratio (by mass) of the protein component to the carbohydrate component of the capsule material may range from about 1:10 to about 1:1.5. For example, the ratio of protein component to carbohydrate component may be about 1:10, 1:9.5, 1:9, 1:8.5, 1:8, 1:7.5, 1:7, 1:6.5, 1:6, 1:5.5, 1:5, 1:4.5, 1:4, 1:3.5, 1:3, 1:2.5, 1:2, or 1:1.5. The ratio of protein to carbohydrate components may range from approximately 1:5 to approximately 1:1.5, for example, approximately 1:4, 1:3, 1:2.9, 1:2.8, 1:2.7, 1:2.6, 1:2.5, 1:2.4, 1:2.3, 1:2.2, 1:2.1, 1:2, 1:1.9, 1:1.8, 1:1.7, 1:1.6, or 1:1.5. The ratio of protein to carbohydrate components may range from approximately 1:2 to approximately 1:1.9, for example, approximately 1:2, 1:1.99, 1:1.98, 1:1.97, 1:1.96, 1:1.95, 1:1.94, 1:1.93, 1:1.92, 1:1.91, or 1:1.9. In a typical embodiment, the ratio of protein to carbohydrate components is approximately 1:1.99.

[0061] Polysaccharide or carbohydrate components may have DE values ​​of approximately 0 to 100, approximately 10 to 70, approximately 20 to 60, or approximately 20 to 40. DE values ​​can be approximately 0, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100.

[0062] Those skilled in the art will recognize that alternative carbohydrate sources may also be used in capsule materials in combination with one or more denatured proteins. For example, the carbohydrate sources may include octenyl succinic anhydride-denatured starch and one or more reducing sugar sources having dextrose equivalent values ​​of about 0 to 80, as previously described in WO2012 / 106777, the disclosure of which is incorporated herein by reference. In short, the starch may include primary and / or secondary denaturation and may be ester or half-ester. Suitable octenyl succinic anhydride-denatured starch includes, for example, waxy corn-based starch and those sold by Ingredion ANZ Pty Ltd, Seven Hills, NSW, Australia under the trade names PURITY GUM®, CAPSUL® IMF and HI CAP® IMF. Octenyl succinic anhydride-modified starch may be present in amounts of approximately 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6.5%, 6%, 5.5%, 5%, 4.5%, 4%, 3.5%, 3%, 2.5%, less than 2%, or less than 1% of the total mass of the composition.

[0063] Reducing sugar sources are well known to those skilled in the art and include monosaccharides and disaccharides, such as glucose, fructose, maltose, galactose, glyceraldehyde, and lactose. Preferred reducing sugar sources also include oligosaccharides, such as glucose polymers, dextrin and maltodextrin, as well as glucose syrup solids. Reducing sugars can also be derived from glucose syrup, which typically contains 20% or more reducing sugars by mass.

[0064] The surface free fat content of the microencapsulated compositions according to this disclosure may be about 10% or less, about 9% or less, about 8% or less, about 7% or less, about 6% or less, about 5% or less, about 4% or less, about 3% or less, about 2.5% or less, about 2.4% or less, about 2.3% or less, about 2.2% or less, about 2.1% or less, about 2% or less, about 1.9% or less, about 1.8% or less, about 1.7% or less, about 1.6% or less, about 1.5% or less, about 1.4% or less, about 1.3% or less, about 1.2% or less, about 1.1% or less, about 1% or less, or less than about 0.8%. In certain preferred embodiments, the surface free fat content is less than about 1.8%, for example less than about 1.5%, for example less than about 1.4%, for example less than about 1%. For example, in some embodiments in which a protein is subjected to a high-shear process at an alkaline pH, for example, a pH of about 8, the surface free fat content may be less than about 1%, for example less than about 0.8%. In certain embodiments, this surface free fat content is determined by the powder derived from or produced from the emulsion.

[0065] The oxidative stability of the microencapsulated compositions according to this disclosure can be measured in terms of the induction period, for example, as described in Example 7 below, using ML Oxipres (Mikrolab Aarhus) (where "Oxipres" is an indirect measurement of potential oxidative stability). In certain preferred embodiments, the induction period of the microencapsulated compositions according to this disclosure is at least about 50 hours, e.g., at least about 60 hours, e.g., at least about 70 hours, e.g., at least about 80 hours, e.g., at least about 90 hours, when measured at 70°C and a pressure of 5 bar. In some embodiments, the induction period is at least about 100 hours. In some embodiments, the induction period is at least about 120 hours, e.g., at least about 130 hours.

[0066] The compositions and emulsions of this disclosure comprise one or more hydrophobic substances. The term “hydrophobic substance” includes pure hydrophobic compounds, hydrophobic mixtures, and hydrophobic compositions. A hydrophobic substance may be any hydrophobic compound or composition that is desirable to be microencapsulated in accordance with this disclosure. Examples of hydrophobic substances that may be used in accordance with this disclosure include bioactive substances, e.g., LCPUFAs and oils containing LCPUFAs, carotenoids, water-insoluble vitamins, e.g., vitamins A, D, E, and K, phenolic compounds, flavor and aromatic compounds, and edible oils. A hydrophobic substance may provide one or more health benefits when administered to a subject. In certain embodiments, a hydrophobic substance may be one or more LCPUFAs, or oils containing one or more LCPUFAs. Such oils may be naturally occurring, derived from nature, or synthesized from genetically modified or non-genetically modified sources. In relation to this disclosure, the terms “naturally occurring” and “derived from nature” include oils and lipid compositions that may be extracted from natural sources, for example, the organisms listed herein, or that may be derived from, or modified from, oils or one or more lipids found in such natural sources. Those skilled in the art will recognize that the scope of this disclosure is not limited by reference to hydrophobic substances, or one or more LCPUFAs, or oil entities or sources containing one or more LCPUFAs.

[0067] Exemplary oils containing or rich in LCPUFA, or that can be modified to contain LCPUFA, or that can be used without modification to the LCPUFA content, include oils from marine organisms, such as crustaceans, such as krill; mollusks, such as oysters; and fish, such as tuna, salmon, trout, sardines, mackerel, sea bass, menheden, herring, pilchard, kipper, eel, or sardines. The oil may be from the eggs of one or more marine organisms, such as those listed herein. In typical embodiments, the oil is or includes lipid extracts from tuna oil, krill oil, or fish eggs. In certain embodiments, the hydrophobic substance is tuna oil.

[0068] Other typical oils that contain or are rich in LCPUFA, or can be modified to contain LCPUFA, or can be used without modification to their LCPUFA content, include plant and microbial sources. Plant sources include, but are not limited to, linseed, walnut, sunflower seed, canola, safflower, soybean, wheat germ, corn, and leafy green plants such as kale, spinach, and parsley. Microbial sources include algae and fungi.

[0069] Hydrophobic substances may be present in amounts of about 0.1% to 80% of the total mass of the composition, or in amounts of about 1% to 80%, or about 1% to 75%, or about 5% to 80%, or about 5% to 75%, or about 5% to 70%. In a typical embodiment where the oil is tuna oil, the oil may be present in an amount of about 2%, 4%, 6%, 8%, 10%, 12%, 14%, 16%, 18%, 20%, 22%, 24%, 26%, 28%, 30%, 32%, 34%, 36%, 38%, 40%, 42%, 44%, 46%, 48%, 49%, 50%, 52%, 54%, 56%, 58%, 60%, 62%, 64%, 66%, 68%, 70%, 72%, 74%, 76%, 78%, or 80% of the total mass of the composition.

[0070] Hydrophobic substances containing LCPUFA typically include one or more omega-3 fatty acids and / or one or more omega-6 fatty acids, or mixtures thereof. The fatty acids may include DHA, AA, EPA, DPA, and / or stearidonic acid (SDA), or mixtures thereof. In one embodiment, the fatty acids include DHA and EPA.

[0071] The compositions envisioned by this disclosure may further include additional components, such as antioxidants, anti-caking agents, flavoring agents, coloring agents, vitamins, minerals, amino acids, and chelating agents.

[0072] Suitable antioxidants are well known to those skilled in the art and may be water-soluble or oil-soluble. Suitable water-soluble antioxidants include, for example, sodium ascorbate, calcium ascorbate, potassium ascorbate, ascorbic acid, glutathione, lipoic acid, and uric acid. In some embodiments, the water-soluble antioxidant may be present in the composition in the range of about 0 to 10% wt / wt of the total composition. Suitable oil-soluble antioxidants include, for example, tocopherol, ascorbyl palmitate, tocotrienol, phenol, and polyphenol. In some embodiments, the oil-soluble antioxidant is present in the oil phase in the range of about 0 to 10% wt / wt of the total composition.

[0073] Anticaking agents compatible with the compositions of this disclosure are well known to those skilled in the art and include calcium phosphate, such as tricalcium phosphate and tricalcium carbonate, for example, calcium carbonate and magnesium carbonate, and silicon dioxide.

[0074] The composition may further contain one or more low molecular weight emulsifiers. Suitable low molecular weight emulsifiers include, for example, mono and diglycerides, lecithin, and sorbitan esters. Other suitable low molecular weight emulsifiers are well known to those skilled in the art. The low molecular weight emulsifier may be present in an amount of about 0.1% to 3% of the total mass of the composition, or in an amount of about 0.1% to 2%, or about 0.1% to 0.5%, or about 0.1% to 0.3% of the total mass of the composition. For example, the low molecular weight emulsifier may be present in an amount of about 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, or 2% of the total mass of the composition.

[0075] The compositions discussed herein may be formulated for administration to a subject via any preferred route, typically by oral administration. The compositions may be in liquid or solid form and may be consumed as is (e.g., in the form of a syrup or other preferred liquid, or as a capsule or other preferred solid). Alternatively, the compositions may be incorporated into food or beverage products.

[0076] Those skilled in the art will recognize that numerous modifications and / or changes to the present invention can be made without departing from the spirit or scope of the invention as broadly described. This embodiment should therefore be considered illustrative in all respects and not restrictive.

[0077] Any reference in this specification to prior art (or information derived therefrom) or any known substance shall not be, and should not be, construed as an acknowledgment, understanding, or suggestion in any form that the prior art (or information derived therefrom) or the known substance constitutes part of the common general knowledge in the art relating to this specification.

[0078] The present invention is described in more detail below by reference to certain subsequent embodiments, which should not be construed as limiting the scope of the invention. [Examples]

[0079] (Example 1 - Denaturation of whey protein isolate) Whey protein isolate (WPI) and whey protein concentrate (WPC) were dissolved in water at a 10% (w / w) concentration, respectively. The pH of several WPI solutions was adjusted to 8. The solutions (WPI, WPC, and WPI (pH 8)) were stirred at 50°C for 30 minutes under gentle shear. Subsequently, the mixtures were homogenized six times at 1500 bar (150 mPa) to induce denaturation of the whey protein. Ice packs were used to maintain the temperature of the WPI and WPC below 60°C during homogenization.

[0080] (Example 2 - Particle size analysis of protein aqueous solution) The protein particle sizes of 1.0% w / w solutions of the undenatured whey protein isolate (uWPI) and denatured whey protein isolate (mWPI) obtained in Example 1, as well as the undenatured whey protein concentrate (uWPC) and denatured whey protein concentrate (mWPC), were measured using a Malvern Zetasizer based on the principle of dynamic light scattering. Backscattering (BS) examines a wide range of particle sizes, while forward scattering (FS) captures the larger particle size range. The results are shown in Table 1 below.

[0081] [Table 1]

[0082] All samples are polydispersible, and therefore, as shown in Table 1, the average particle size (Z-Av) can be significantly affected by the presence of very large particles. Denaturation of whey protein isolate (mWPI) results in a significant decrease in protein particle size, regardless of pH, in contrast to the undenatured sample (uWPI). This is thought to be because pressure denaturation separates most of the soluble protein aggregates. uWPC has a similar average particle size to uWPI, but has substantially smaller soluble protein aggregates, as seen in the FS results. The degree of pressure denaturation in relation to particle size reduction is similar to that observed when applied to whey protein concentrate (WPC). For both WPI and WPC, pressure denaturation at 1500 bar / 6 cycles resulted in approximately a 50% reduction in average protein particle size.

[0083] (Example 3 - Surface charge analysis of aqueous protein solution) In Example 1, the surface charge of 1.0% w / w aqueous solutions / aqueous dispersions of uWPI and mWPI obtained at both the original pH (i.e., no pH adjustment) and pH 8 was measured using a Zetasizer to measure the interfacial dynamic potential. The results are shown in Table 2 below.

[0084] [Table 2]

[0085] Slight differences in the surface charge (zeta potential; ZP) of mWPI solutions may be due to a decrease in soluble protein aggregates. We improved the hydration of WPI using an alkaline pH of 8 and further observed a negative surface charge.

[0086] (Example 4 - Interfacial tension between protein aqueous solution and corn oil) The interfacial tensions of the 1.0% w / w aqueous solutions of uWPI and mWPI obtained in Example 1 are shown in Figure 1.

[0087] The interfacial tension between corn oil and water is approximately 27 mN / m. A 1.0% w / w WPI solution efficiently reduced the interfacial tension between corn oil and water, demonstrating good surface activity and adsorption behavior at the interface. In contrast to the uWPI solution, the mWPI solution, which has a smaller average particle size, hydrates well (pH 8), and therefore disperses quickly at the interface, further reduced the interfacial tension value.

[0088] (Example 5 - Protein / Oil Emulsion) An oil-in-water emulsion (protein-to-oil mass ratio 1:3) was prepared using 1% w / w aqueous solutions of uWPI and mWPI obtained in Example 1, and refined tuna oil containing mixed natural tocopherols. The WPI solution and tuna oil mixture was roughly homogenized using an UltraTurrax at 10,000 RPM for 10 minutes. The median diameter d (0.5) and average size D [4.3] (in micrometers) of the oil droplets were measured using a particle size analyzer (Malvern Instruments, Mastersizer MS3000) based on the laser diffraction principle, and the results are shown in Table 3 below. In Table 3, "EAI" refers to the emulsion activity index, and "ESI" refers to the emulsion stability index. EAI reflects the protein adsorption capacity at the oil-water interface, and ESI reflects the emulsion's resistance to instability, such as soft aggregation and creaming.

[0089] [Table 3]

[0090] The median diameter d(0.5) of the oil droplets and the average size D[4.3] of the oil droplets were smaller with mWPI (pH 8). All parameters increased over time regardless of denaturation or pH treatment, but the scale of change was minimal with mWPI (pH 8) after 1 week of storage time. This is consistent with the excellent surface activity of the mWPI (pH 8) solution shown in Example 4 and the adsorption behavior that gives good emulsion stability over time.

[0091] (Example 6 - Encapsulation of Hydrophobic Substances Using Unmodified and Modified Protein Capsule Materials) Using the unmodified whey protein isolate (uWPI) and modified whey protein isolate (mWPI) obtained in Example 1, microencapsulated compositions were prepared. Purified tuna oil containing mixed natural tocopherols was used as the hydrophobic core material. The respective formulations of the microencapsulated compositions are shown in Table 4 below. The preparation method for each composition is shown in Figure 2 and will be discussed in detail below.

[0092]

Table 4

[0093] <Preparation of Microencapsulated Powdered Composition Using uWPI and Carbohydrate Capsule Material (「uWPI-Microencapsulated Powder」)> WPI (15.00% (w / w)), dextrose monohydrate (14.50% (w / w)), dried glucose syrup (DE value 30) (15.10% (w / w)), and sodium ascorbate (5.35% (w / w)) were dissolved in water. This aqueous phase was stirred at 50 °C for 35 minutes under gentle shear. An antioxidant containing tuna oil (50.05% (w / w)) was then added, and thereafter, an emulsion was prepared as follows: High shear mixing was used at 10,000 rpm for 10 - 15 minutes to produce a coarse emulsion, followed by two - stage homogenization three times at 400 / 200 bar (total 600 bar) to produce a fine emulsion. The final oil - in - water emulsion was spray - dried using a bench - top spray dryer with inlet and outlet temperatures of 170 and 90 - 100 °C, respectively. The produced powder was filled into aluminum sachets under N2 as a protective gas. The uWPI powder was stored at 25 °C and then used. The total oil filling amount in the uWPI powder was 50% (w / w).

[0094] <Preparation of microencapsulated powdered composition using mWPI and carbohydrate capsule materials (「mWPI - microencapsulated powder」)> WPI was modified as described in Example 1. To an aqueous phase of mWPI dextrose monohydrate (14.50% (w / w)), dried glucose syrup (DE value 30) (15.10% (w / w)) and sodium ascorbate (5.35% (w / w)) were added at 50 °C. Purified tuna oil containing mixed natural tocopherols was added (50.05% (w / w)), and thereafter, an emulsion was prepared as follows: High shear mixing was used at 10,000 rpm for 10 - 15 minutes to produce a coarse emulsion, followed by two - stage homogenization three times at 400 / 200 bar (40 / 20 mPa) (total 600 bar (60 mPa)) to produce a fine emulsion. The final oil - in - water emulsion was spray - dried using a bench - top spray dryer with inlet and outlet temperature ranges of 170 and 90 - 100 °C, respectively. The produced powder was filled into aluminum sachets under N2 as a protective gas. The total oil filling amount in the mWPI powder was 50% (w / w).

[0095] <Preparation of microencapsulated powdered composition using mWPI (pH 8 solution) and carbohydrate capsule material (``mWPI - microencapsulated powder (pH 8)'')> The WPI solution was adjusted to pH 8 and denatured as described in Example 1. Dried glucose syrup (DE value 30) (15.10% (w / w)) and sodium ascorbate (5.35% (w / w)) were added to the aqueous phase of mWPI dextrose monohydrate (14.50% (w / w)) at 50 °C. Purified tuna oil containing mixed natural tocopherols was added (50.05% (w / w)), and then an emulsion was prepared as follows: High - shear mixing was used at 10,000 rpm for 10 - 15 minutes to produce a coarse emulsion, followed by two - stage homogenization at 400 / 200 bar (40 / 20 mPa) (total 600 bar (60 mPa)) three times to produce a fine emulsion. The final oil - in - water emulsion was spray - dried using a bench - top spray dryer with inlet and outlet temperatures in the ranges of 170 and 90 - 100 °C, respectively. The resulting powder was filled into aluminum sachets under N2 as a protective gas. The total oil filling amount in the mWPI powder was 50% (w / w).

[0096] (Example 7 - Evaluation of surface - free fat and oxidative stability of tuna oil in microcapsule powder) The oxidative stability of the microencapsulated powder was analyzed using Oxipres as a rapid and reliable measurement method. A microencapsulated powder with a total of 4 g of oil was sealed in a container and heated at 70 °C under oxygen at 5 bar (0.5 mPa). The time when the oxygen pressure in the container began to decrease was recorded as the induction period (IP), which indicated the occurrence of oxidation. Figure 3 shows the Oxipres analysis of omega - 3 - containing microencapsulated powder compared with the Maillard reaction product (MRP) described in US7374788B2 and the microencapsulated powder containing encapsulated omega - 3 oil.

[0097] The percentage of surface free fat was measured by exposing the powder to diesel fuel for a short time (15 minutes) to extract the surface free fat; the wall material / encapsulating oil was removed using filter paper, and the solvent containing the "washed" fat was then evaporated to obtain the percentage of surface free fat by dividing the residual mass, i.e., the oil, by the mass of powder used (in grams) and multiplying by 100%. The results are shown in Table 5 below.

[0098] [Table 5]

[0099] As shown in Figure 3, the induction period was significantly extended by using denatured protein capsule material. As shown in Table 5, mWPI microencapsulated powders exhibited significantly lower surface free fat (SFF) values ​​than uWPI. mWPI (pH 8) had an SFF of less than 1%, which met the requirements of a typical powder, while also possessing a high (approximately 50%) oil content and good oxidative stability. All samples showed good oxidative stability in IP for over 100 hours with Oxipres (70°C, 5 bar).

[0100] The good oxidative stability, in terms of primary and secondary oxidative properties, is further shown in Table 6 below, and the microencapsulated powders were exposed to a temperature of 40°C for 4 weeks. Up to the end of the 4 weeks, all samples were well within specifications with respect to peroxide values ​​(POV; 5 meq O2 / Kg fat) and p-anisidine values ​​(p-AV; 20).

[0101] [Table 6]

[0102] The perceptual properties of microencapsulated uWPI & mWPI were also evaluated over a 4-week rapid exposure period. The results are shown in Figures 4 and 5. The 15 highest scores indicate excellent quality / characteristics. Overall quality (Figure 4) was rated as good across all samples by the end of the storage period, with no perceptible rancid or marine odor, nor any noticeable flavor (Figure 5).

Claims

1. A method for producing a microencapsulated composition comprising one or more hydrophobic substances encapsulated by a capsule material, wherein the capsule material comprises one or more denatured proteins and / or peptides, A step of obtaining one or more denatured proteins and / or peptides from a starting protein by subjecting the starting protein to a high-shear process such that the average particle size of one or more denatured proteins and / or peptides is 70% or less of the average particle size of the starting protein, wherein the high-shear process is carried out at an alkaline pH, and thereafter, A method for producing a microencapsulated composition, comprising the step of encapsulating one or more hydrophobic substances in a capsule material containing one or more denatured proteins and / or peptides.

2. The method according to claim 1, wherein the average particle size of one or more of the denatured proteins is 65% or less of the average particle size of the starting protein.

3. The method according to claim 1 or 2, wherein the microencapsulated composition has a surface free fat content of less than 1.8%.

4. The method according to claim 3, wherein the microencapsulated composition has a surface free fat content of less than 1%.

5. The method according to claim 4, wherein the microencapsulated composition has a surface free fat content of less than 0.8%.

6. The method according to any one of claims 1 to 5, wherein the high shear process is carried out at a pH of 8.

7. The method according to any one of claims 1 to 6, wherein the high shear process includes a step of applying a pressure of 20 MPa to 300 MPa to the starting protein.

8. The method according to any one of claims 1 to 7, wherein the high shear process is a homogenization process.

9. The method according to any one of claims 1 to 8, wherein the high shear process is a microfluidization process.

10. The method according to any one of claims 1 to 9, wherein the one or more denatured proteins and / or peptides are in the form of a protein fraction.

11. The method according to any one of claims 1 to 10, wherein the denatured protein is denatured whey protein.

12. The method according to any one of claims 1 to 11, wherein the capsule material further comprises one or more carbohydrates.

13. The method according to claim 12, wherein one or more of the carbohydrates are selected from glucose syrup and dextrose monohydrate, or a combination thereof.

14. The method according to any one of claims 1 to 13, wherein the one or more denatured proteins and / or peptides are present in an amount of 3% w / w to 25% w / w of the total mass of the composition.

15. The method according to any one of claims 1 to 14, wherein the ratio of the denatured protein component of the capsule material to the carbohydrate component of the capsule material is in the range of 1:10 to 1:

1.

16. The method according to any one of claims 1 to 15, wherein the hydrophobic substance is edible oil.

17. The method according to any one of claims 1 to 16, wherein the hydrophobic substance comprises one or more long-chain polyunsaturated fatty acids (LCPUFAs).

18. The method according to claim 17, wherein the LCPUFA comprises omega-3 fatty acids and / or omega-6 fatty acids.

19. The method according to claim 17 or 18, wherein the LCPUFA exists in triglyceride form.

20. The method according to any one of claims 17 to 19, wherein the LCPUFA is present as one or more LCPUFA-containing oils.

21. The method according to claim 20, wherein one or more of the oils include fish oil.

22. The method according to claim 21, wherein the fish oil is tuna oil.

23. The method according to any one of claims 1 to 22, wherein the composition further comprises at least one source of vitamin C.

24. The method according to any one of claims 1 to 23, wherein the composition is in the form of an oil-in-water emulsion.

25. The method according to any one of claims 1 to 24, wherein the composition is in the form of a spray-dried powder.

26. A method for protecting hydrophobic substances from oxidative decomposition, A step comprising subjecting a starting protein to a high-shear process to produce one or more denatured proteins and / or peptides, wherein the average particle size of the one or more denatured proteins and / or peptides is 70% or less of the average particle size of the starting protein, wherein the high-shear process is carried out at an alkaline pH; and thereafter, A method comprising the step of encapsulating one or more hydrophobic substances in a capsule material containing the one or more denatured proteins and / or peptides.

27. A method for improving the oxidative stability of hydrophobic substances, A step of subjecting a starting protein to a high-shear process to produce one or more denatured proteins and / or peptides, wherein the average particle size of the one or more denatured proteins and / or peptides is 70% or less of the average particle size of the starting protein, wherein the high-shear process is carried out at an alkaline pH; and thereafter A method comprising the step of encapsulating one or more hydrophobic substances in a capsule material containing the one or more denatured proteins and / or peptides.

28. A method for reducing surface free fat of a microencapsulated composition comprising one or more hydrophobic substances encapsulated by a capsule material, A step comprising subjecting one or more starting proteins and / or peptides to a high-shear process to produce one or more denatured proteins and / or peptides, wherein the average particle size of the one or more denatured proteins and / or peptides is 70% or less of the average particle size of the one or more starting proteins and / or peptides, wherein the high-shear process is carried out at an alkaline pH, and thereafter A method comprising the step of encapsulating one or more hydrophobic substances in a capsule material containing the one or more denatured proteins and / or peptides.

29. The method according to any one of claims 26 to 28, wherein the average particle size of one or more denatured proteins and / or peptides is 65% or less of the average particle size of the starting protein.

30. The method according to any one of claims 26 to 29, wherein the high shear process is carried out at a pH of 8.

31. The method according to any one of claims 26 to 30, wherein the hydrophobic substance comprises one or more LCPUFAs (long-chain polyunsaturated fatty acids).

32. The method according to claim 31, wherein one or more of the LCPUFAs are in the form of triglycerides.

33. A method for producing a stable emulsion containing a hydrophobic substance, wherein the emulsion further comprises one or more denatured proteins and / or peptides, the one or more denatured proteins and / or peptides being obtained from a starting protein by subjecting the starting protein to a high-shear process before emulsification, wherein the average particle size of the one or more denatured proteins and / or peptides is 70% or less of the average particle size of the starting protein, and the high-shear process is performed at an alkaline pH.

34. The method according to claim 33, wherein the average particle size of one or more denatured proteins and / or peptides is 65% or less of the average particle size of the starting protein.

35. The method according to claim 33 or 34, wherein the high shear process is carried out at a pH of 8.

36. The method according to any one of claims 33 to 35, wherein the hydrophobic substance comprises one or more LCPUFAs (long-chain polyunsaturated fatty acids).

37. The method according to claim 36, wherein the LCPUFA is in the form of a triglyceride.

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