WO2009034124A1

WO2009034124A1 - Omega-3 stabilisation towards oxidation

Info

Publication number: WO2009034124A1
Application number: PCT/EP2008/062044
Authority: WO
Inventors: Roar Hauch
Original assignee: Novozymes A/S
Priority date: 2007-09-12
Filing date: 2008-09-11
Publication date: 2009-03-19

Abstract

The present invention relates to a composition comprising a) a core comprising an oxidation labile compound; wherein the oxidation labile compound is a lipid having at least one carbon-carbon double bond that is prone to oxidation; and b) an oxidation protective coating layer comprising a lipid.

Description

Title: OMEGA-3 STABILISATION TOWARDS OXIDATION

Field of invention

The present invention relates to a product and a method for producing a product with enhanced stability towards oxidation of oxidation labile compounds.

Background of the invention

Consumption of foods rich in omega-3 polyunsaturated fatty acids (PUFAs) has been associated with decreased cardiovascular death by decreasing plasma triglycerides, blood pressure, platelet aggregation, and inflammation. While seafood is the best source of omega- 3 acids, many individuals do not like the taste of seafood, do not have ready access to seafood, or cannot afford seafood. One solution is to supplement the diet with cod liver oil or fish oil capsules, but this solution has low compliance. Another solution is to add omega-3 rich fish oils directly to foods, such as dairy products, cereal products, baked goods, and nutrition bars.

A challenge with the latter approach is to provide the benefits of omega-3 fatty acids without imparting any offending fish flavors or fish odors, which are byproducts of lipid oxidation. A need exists, therefore, for a stabilized preparation of PUFAs that can be added to low moisture or high moisture foods, such that the PUFAs are protected from oxidation.

Summary of the invention

The present invention provides compositions and methods to reduce the oxidation of an oxidation labile compound. In particular, the invention provides a capsule comprising a core of oxidation labile compound that is surrounded by a protective coating layer. It has been discovered, as demonstrated in the examples, that protection of an oxidation labile compounds, such as an omega-3 fatty acid, with a coating, such as solid palm oil fraction dramatically reduces the oxidation of the oxidation labile compound. This key discovery provides means to include omega-3 fatty acids or other oxidation labile compounds in foods without imparting offensive tastes or odours to the foods from the oxidation of the fatty acids or other oxidation labile compounds.

One aspect of the invention is a composition comprising an oxidation labile compound material and protective coating, in exemplary embodiment. Suitable oxidation labile compounds and coating materials are described below.

The present invention provides a composition comprising: i) a core comprising an oxidation labile compound ii) a protective coating layer comprising a non-polar compound

The present invention further relates to the manufacturing of said particles.

Detailed description of the invention

The present composition comprises a core comprising the oxidation labile compound and a protective coating layer that protects the oxidation labile compound against oxidation.

The core The core according to the invention comprises an oxidation labile compound, e.g., a long chain polyunsaturated fatty acid.

The core may consist of a homogenous matrix of oxidation labile compound alone or in combination with other suitable materials. The core may comprise an inert particle or mixture of compounds hereupon the oxidation labile compound is added to be adsorbed and/or absorbed .

The core may typically be formed into 10-5000 micron particles.

The core may also comprise additional materials such as antioxidants, enzymatic free radical and/or oxygen scavengers. In a particular embodiment of the present invention the core comprises an antioxidant and/or an oxygen scavenger. The core may also comprise one or more transition metal ion complexing agents, e.g., EDTA, lactoferrin and/or citric acid. In a particular embodiment of the present invention the core comprises a transition metal ion complexing agent. The matrix may also comprise one or more proteins, and one or more carbohydrates.

Oxidation labile compounds

An oxidation labile compound having utility in the present invention includes a material comprising a molecule with a carbon backbone having at least one carbon-carbon double bond that is prone to oxidation. Removal of a labile hydrogen atom from a carbon adjacent to the double bond creates a free radical that is susceptible to attack by oxygen to form a free radical peroxide, which may serve as a catalyst for further oxidation.

A variety of oxidation labile compounds are suitable for use in this invention. In general, the oxidation labile compound comprises at least one oxidation labile lipid. Oxidation labile lipids include fatty acids, fatty acid esters, fatty acid methyl esters (FAMEs), tri-, di- and/or mono- glycerides, glycolipids, phospholipids, sphingolipids, sterols, sterol esters, steroid hormones and polyisoprenoids. The oxidation labile lipids may be nautral or polar and especially include unsaturated fatty acids. In particular the oxidation labile lipid contains polyunsaturated fatty acids. Typical examples of neutral lipids are triacylglycerols, fat-soluble vitamins and waxes. A typical polar lipid class is phospholipids. According to the present invention a preferred oxidation labile compound may contain long chain polyunsaturated fatty acid, (i.e., fatty acids containing at least 2 unsaturated carbon- carbon bonds, e.g., double bonds and a carbon chain with 18 or more carbon atoms). The long chain polyunsaturated fatty acid are in a particular embodiment omega-3 and/or omega- 6 polyunsaturated fatty acids.

The polyunsaturated fatty acid may be present within triglycerides.

In still another embodiment, the oxidation labile compound may be a preparation of substantially unsaturated fats or substantially unsaturated oils. In general, fats and oils comprise monoglycerides, diglycerides, triglycerides, and free fatty acids. The glycerides of fats and oils generally comprise fatty acids that are at least 4 carbons in length, and more preferably, unsaturated fatty acids that range in length from 16 to 24 carbons. The unsaturated fatty acid may be monounsaturated or polyunsaturated.

In another embodiment, the oxidation labile compound may be a polyunsaturated fatty acid (PUFA), which has at least two carbon-carbon double bonds generally in the cis- configuration. The PUFA may be a long chain fatty acid having at least 18 carbons atoms. The PUFA may be an omega-3 fatty acid in which the first double bond occurs in the third carbon-carbon bond from the methyl end of the carbon chain (i.e., opposite the carboxyl acid group). Examples of omega-3 fatty acids include alpha-linolenic acid (18:3, ALA), stearidonic acid (18:4), eicosatetraenoic acid (20:4), eicosapentaenoic acid (20:5; EPA), docosatetraenoic acid (22:4), n-3 docosapentaenoic acid (22:5; n-3DPA), and docosahexaenoic acid (22:6; DHA). The PUFA may also be an omega-6 fatty acid, in which the first double bond occurs in the sixth carbon-carbon bond from the methyl end. Examples of omega-6 fatty acids include linoleic acid (18:2), gamma-linolenic acid (18:3), eicosadienoic acid (20:2), dihomo-gamma- linolenic acid (20:3), arachidonic acid (20:4), docosadienoic acid (22:2), adrenic acid (22:4), n-6 docosapentaenoic acid (22:5) and calendic acid (18:3). The fatty acid may also be an omega-9 fatty acid, such as oleic acid (18:1 ), eicosenoic acid (20:1 ), mead acid (20:3), erucic acid (22:1 ), and nervonic acid (24:1 ). In one embodiment, the oxidation labile compound may be derived from a biological source, such that it may be a crude mixture of proteins, lipids, and carbohydrates. In another embodiment, the oxidation labile compound may be a mixture of lipids that is essentially devoid of proteins and/or carbohydrates. In yet another embodiment, the oxidation labile compound may be a purified lipid.

In another embodiment, the oxidation labile compound may be a marine animal-derived oil. The marine animal-derived oil may originate from a vertebrate, in-vertebrate or micro- or unicellular-organism fish or a marine organism, e.g. such that the oil may be a fish oil or a marine microorganism oil. The long chain (2OC, 22C) omega-3 and omega-6 fatty acids are found in marine organisms. The ratio of omega-3 to omega-6 fatty acids in marine organisms ranges from about 8:1 to 20:1. Marine organisms from which oil rich in omega-3 fatty acids may be derived include, but are not limited to tuna, abalone scallops, albacore tuna, anchovies, catfish, clams, cod, gem fish, herring, lake trout, mackerel, menhaden, orange roughy, salmon, sardines, sea mullet, sea perch, shark, shrimp, squid and trout. In yet another embodiment, the oxidation labile compound may be a plant-derived oil. Plant and vegetable oils are rich in omega-6 fatty acids. Some plant-derived oils, such as flaxseed oil, are especially rich in omega-3 fatty acids. Plant or vegetable oils are generally extracted from the seeds of a plant, but may also be extracted from other parts of the plant. Plant or vegetable oils that are commonly used for cooking or flavoring include, but are not limited to, acai oil, almond oil, amaranth oil, apricot seed oil, argan oil, avocado seed oil, babassu oil, ben oil, blackcurrant seed oil, Borneo tallow nut oil, borage seed oil, buffalo gourd oil, canola oil, carob pod oil, cashew oil, castor oil, coconut oil, coriander seed oil, corn oil, cottonseed oil, evening primrose oil, false flax oil, flax seed oil, grapeseed oil, hazelnut oil, hemp seed oil, kapok seed oil, lallemantia oil, linseed oil, macadamia oil, meadowfoam seed oil, mustard seed oil, okra seed oil, olive oil, palm oil, palm kernel oil, peanut oil, pecan oil, pequi oil, perilla seed oil, pine nut oil, pistachio oil, poppy seed oil, prune kernel oil, pumpkin seed oil, quinoa oil, ramtil oil, rice bran oil, safflower oil, sesame oil, soybean oil, sunflower oil, tea oil, thistle oil, walnut oil, or wheat germ oil. The plant derived oil may also be hydrogenated or partially hydrogenated.

The oxidation labile compound ma be derived from microorganisms such as algae, fungi and bacteria. In a particular embodiment the fungi is Mortierella alpine. In still a further embodiment, the oxidation labile compound may be an algae derived oil. Commercially available algae-derived oils include those from and

Suitable algae, from which oil is extracted, include Aphanizomenon, Bacilliarophy, Botryococcus, Chlorophyceae, Crypthecodinium, Dunaliella, Euglena, Isochrysis, Nannochloropsis, Nannochloris, Neochloris, Phaeodactylum, Pleurochrysis, Prymnesiumparvum, Scenedesmus, Schizochytrium, Spirulina and Tetraselmis. In one particular embodiment the algae, from which oil is extracted, include Aphanizomenon flos-aquae, Bacilliarophy sp., Botryococcus braunii, Chlorophyceae sp., Crypthecodinium cohnii, Dunaliella tertiolecta, Euglena gracilis, Isochrysis galbana, Nannochloropsis salina, Nannochloris sp., Neochloris oleoabundans, Phaeodactylum tricornutum, Pleurochrysis carterae, Prymnesiumparvum, Scenedesmus dimorphus, Schizochytrium sp., Spirulina sp., and Tetraselmis chui.

In one embodiment embodiment the core comprising the oxidation labile compound is a lipid enriched biomass produced from a microorganism, in particular from an auxotrophic marine microorganism containing - at least in part - the antioxidants produced by said microorganism.

In a particular embodiment auxotrophic marine microorganism according to the invention is an algae, in particular a micro algae or an algae-like microorganism, preferably a member of the Stramenopiles group, more preferably a Hamatores sp, a Proteromonads sp, a Opalines sp, a Developayella sp, a Diplophrys sp, a Labrinthulids sp, a Thraustochytrids sp, a Biosecids sp, an Oomycetes sp, a Hypochytridiomycetes sp, a Commation sp, a Reticulosphaera sp, a Pelagomonas sp, a Pelagococcus sp, an Ollicola sp, an Aureococcus sp, a Parmales sp, a Diatoms sp, a Xanthophytes sp, a Phaeophytes sp (brown algae), a Eustigmatophytes sp, a Raphidophytes sp, a Synurids sp, an Axodines sp, a Chrysomeridales sp, a Sarcinochrysidales sp, a Hydrurales sp, a Hibberdiales sp, or a Chromulinales sp.

The lipid enriched biomass may be produced as known in the art, e.g., as described in WO 2005/021735.

In another embodiment, the oxidation labile compound may be a spice or fragrance oil. Suitable examples of spice or fragrant oils include angelica oil, anise oil, basil oil, bergamont oil, orange oil, black pepper oil, calamus oil, citronella oil, calendula oil, camphor oil, cardamom oil, celery oil, chamomile oil, cinnamon oil, clove oil, coriander oil, lemon grass oil, cypress oil, cumin seed oil, davana oil, dill seed oil, eucalyptus oil, fennel seed oil, garlic oil, geranium oil, ginger oil, grape seed oil, hyssop oil, jasmine oil, juniper berry oil, lavender oil, lemon oil, lime oil, myrrh oil, neroli oil, neem oil, nutmeg oil, palm Rosa oil, parsley oil, peppermint oil, rose oil, rosemary oil, rose wood oil, sage oil, sesame oil, spearmint oil, tarragon oil, tea tree oil, thyme oil, tangerine oil, turmeric root oil, vetiver oil, wormwood oil, and yara yara oil.

In yet another embodiment, the oxidation labile compound may be a pharmaceutical formulation comprising an oxidatively unstable pharmaceutical, such as arachadonic/arachidonic acid or a prostaglandin. The formulation may also comprise an unstable oil as a carrier. Suitable examples of pharmaceutical grade carrier oils include cod liver oil, corn oil, cottonseed oil, eucalyptus oil, lavender oil, olive oil, peanut oil, peppermint oil, safflower oil, sesame oil, and soybean oil. The oxidation labile compound may also be a formulation comprising a fat-soluble vitamin, such as vitamin A, D, K, or E. In another embodiment, the oxidation labile compound may be preparation of fish materials or fish meal, which is the solid material that remains after most of the water and oil have been removed from the starting fish material. Non-limiting examples of fish or marine organism that may be used for the preparation of fish meal include anchovy, blue whiting, capelin, crab, herring, mackerel, menhaden, pollack, salmon, shrimp, squid, tuna, and whitefish.

In still another embodiment, the oxidation labile compound may be an animal-derived fat. Non-limiting examples of suitable animal-derived fats include poultry fat, beef tallow, mutton tallow, butter, pork lard, whale blubber, and yellow grease (which may be a mixture of vegetable and animal fats).

In one embodiment the oxidation labile compound is an oil from marine organisms comprising omega-3 and omega-6 fatty acids. In another particular embodiment, the oxidation labile compound is an omega-3 fish oil. In yet another particular embodiment, the oxidation labile compound is an omega-3 fatty acid. In a particular embodiment of the invention the oxidation labile compound content of the core is in amounts of more thaniO % w/w, in particular in amounts of 10-20% w/w. In one embodiment the oxidation labile compound is present in the core in amounts of more than 5%. In another embodiment the oxidation labile compound is present in the core in amounts of more than 10%. In a further embodiment the oxidation labile compound is present in the core in amounts of more than 15%. In a particular embodiment the oxidation labile compound is present in the core in amounts of more than 20%. In a further particular embodiment the oxidation labile compound is present in the core in amounts of more than 25%, more than 30%, more than 35%, more than 40%, more than 50%, more than 60% or even more than 75%.

Antioxidants

Antioxidants may be present in the core together with the oxidation labile compound. The antioxidant may be natural or synthetic. Suitable antioxidants include, but are not limited to, ascorbic acid and its salts, ascorbyl palmitate, ascorbyl stearate, anoxomer, N- acetylcysteine, benzyl isothiocyanate, o-, m-or p-amino benzoic acid (o is anthranilic acid, p is PABA), butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), caffeic acid, canthaxantin, alpha-carotene, beta-carotene, beta-caraotene, beta-apo-carotenoic acid, camosol, carvacrol, cetyl gallate, chlorogenic acid, citric acid and its salts, clove extract, coffee bean extract, p-coumaric acid, 3,4-dihydroxybenzoic acid, N,N'-diphenyl-p- phenylenediamine (DPPD), dilauryl thiodipropionate, distearyl thiodipropionate, 2,6-di-tert- butylphenol, dodecyl gallate, edetic acid, ellagic acid, erythorbic acid, sodium erythorbate, esculetin, esculin, 6-ethoxy-1 ,2-dihydro-2,2,4-trimethylquinoline, ethyl gallate, ethyl maltol, ethylenediaminetetraacetic acid (EDTA), eucalyptus extract, eugenol, ferulic acid, flavonoids (e.g., catechin, epicatechin, epicatechin gallate, epigallocatechin (EGC), epigallocatechin gallate (EGCG), polyphenol epigallocatechin-3-gallate), flavones (e.g., apigenin, chrysin, luteolin), flavonols (e.g., datiscetin, myricetin, daemfero), flavanones, fraxetin, fumaric acid, gallic acid, gentian extract, gluconic acid, glycine, gum guaiacum, hesperetin, alpha- hydroxybenzyl phosphinic acid, hydroxycinammic acid, hydroxyglutaric acid, hydroquinone, N-hydroxysuccinic acid, hydroxytryrosol, hydroxyurea, lactic acid and its salts, lecithin, lecithin citrate; R-alpha-lipoic acid, lutein, lycopene, malic acid, maltol, 5-methoxy tryptamine, methyl gallate, monoglyceride citrate; monoisopropyl citrate; morin, beta-naphthoflavone, nordihydroguaiaretic acid (NDGA), octyl gallate, oxalic acid, palmityl citrate, phenothiazine, phosphatidylcholine, phosphoric acid, phosphates, phytic acid, phytylubichromel, pimento extract, propyl gallate, polyphosphates, quercetin, trans-resveratrol and/or combinations thereof. Resveratrol is a polyphenols phytoalexin. It is a stilbenoid, a derivative of stilbene. It exists as two structural isomers: cis-resveratrol and trans-resveratrol. Resveratrol is produced by plants. It is found in the skins of certain red grapes, in peanuts, blueberries, some pines, the roots and stalks of Japanese knotweed and giant knotweed.

Further antioxidants are rice bran extract, rosemary extract, rosmarinic acid, sage extract, sesamol, silymarin, sinapic acid, succinic acid, stearyl citrate, syringic acid, tartaric acid, thymol, tocopherols, or vitamin E, is a fat-soluble vitamin. Natural vitamin E exists in eight different forms or isomers four tocopherols (i.e., alpha-, beta-, gamma-and delta-tocopherol), and four tocotrienols (i.e., alpha-, beta-, gamma-and delta-tocotrienols). Further antioxidants include tyrosol, vanilic acid, 2,6-di-tert-butyl-4-hydroxymethylphenol (i.e., lonox 100), 2,4- (tris-3',5'-bi-tert-butyl-4'-hydroxybenzyl)-mesitylene (i.e., lonox 330), 2,4,5- trihydroxybutyrophenone, ubiquinone, tertiary butyl hydroquinone (TBHQ), thiodipropionic acid, trihydroxy butyrophenone, tryptamine, tyramine, uric acid, vitamin K and derivates, vitamin Q10, wheat germ oil, zeaxanthin, and/or combinations thereof. Preferred antioxidants include tocopherols, ascorbyl palmitate, propyl gallate and rosemary extracts. The concentration of the antioxidant or combination of antioxidants may range from about 0.001% to about 5% by weight. In a particular embodiment the concentration of the antioxidant or combination of antioxidants may range from about 0.01-1%. In a most particular embodiment the concentration of the antioxidant or combination of antioxidants may range from about 0.03-0.3% by weight. In a further embodiment the antioxidant is selected from the group consisting of tocopherol, resveratrol and/or derivatives thereof, Propyl gallat, rosemary extracts, ascorbic acid, ascorbyl palmitate and combinations thereof.

Enzymatic oxygen scavenger

Enzymatic oxygen scavengers may be present in the core.

The enzymatic oxygen scavenger may be a mixture of a glucoseoxidase and a catalase. In a particular embodiment of the invention the antioxidant content of the core is in an amount of less than 0.2% w/w (glucoseoxidase and catalase).

Transition metal ion complexing agents. Transition metal ion complexing aents may be present in the core. A useful transition metal ion complexing agent is selected from the group consisting of EDTA and citric acid. The transition metal ion complexing agent would normally be added in amounts of 25-100 ppm, in particular 50 ppm of EDTA is preferred.

Carbohydrates

The core may also comprise one or more carbohydrates. In a particular embodiment the carbohydrate is one having four or more carbon atoms; preferably from 4 to 500 carbon atoms; in particular from 6 to 120 carbon atoms.

Among the mono-saccharides glucose, fructose, mannose and galactose may be suitable. Among the di-saccharides sucrose is preferred.

Among the oligosaccharides, dextrins, limit dextrins, cyclodextrins, amylose and amylopectins are preferred.

In a particular embodiment of the invention a mixture of modified waxy maize starch, more specifically an octenyl succinate starch compound, native starch and dextrins are preferred. In a particular embodiment of the invention the carbohydrate content of the core is in amounts of 5-60% w/w, more preferably in amounts of 10-50% w/w.

Protein

The core may further comprise at least one protein. In a particular embodiment of the invention the protein content of the core is at an amount of 1- 60 w/w . In a more particular embodiment the protein content of the core is 10-40% w/w. In a most particular embodiment the protein content of the core is in an amount of 2-30%. The protein may be a vegetable protein, an animal protein, a fungal protein, a microbial protein, or a mixture thereof. Non-limiting examples of an animal protein suitable for use in this invention include casein, dairy whey protein, gelatin, or a mixture thereof. Non-limiting examples of a vegetable protein include soy protein, corn protein, wheat protein, rice protein, canola protein, pea protein, or a mixture thereof. Examples of wheat protein being gluten (wheat protein), wheat protein derivates, e.g. soluble wheat protein and deamidated soluble wheat protein. The corn protein may be corn gluten meal, or more preferably, zein. A preferred vegetable protein is deamidated soluble wheat protein.

Potato protein, The soy protein may be provided by a preparation of soy flour, soy protein concentrate, or soy protein isolate. These preparations of soy protein are typically formed from a soybean starting material, which may be soybeans or a soybean derivative. Preferably, the soybean starting material may be soybean cake, soybean chips, soybean meal, soybean flakes, or a mixture of these materials. The soybean cake, chips, meal, or flakes may be formed from soybeans according to conventional procedures in the art. That is, soybean cake and soybean chips are generally formed by extraction of part of the oil from soybeans by pressure or solvents; soybean flakes are generally formed by cracking, heating, and flaking soybeans and reducing the oil content of the soybeans by solvent extraction; and soybean meal is generally formed by grinding soybean cake, chips, or flakes.

The protein may be modified using procedures known in the art to improve the utility or characteristics of the protein. The modifications include, but are not limited to deamidation, denaturation or hydrolysis of the protein. The denaturation or hydrolysis may be chemically mediated or it may be enzymatic.

A non-polar coating layer

According to the invention the matrix described above will have a coating layer comprising a non-polar coating.

This non-polar coating is a protective coating that keeps oxygen away from the oxygen labile compound.

The non-polar coating may be a pure lipid or a mixture of lipids. The non-polar coating may also be prepared from a mixture of lipid(s), structuring agents and/or plasticizers. The structuring agents and/or plasticizers may be chosen from this non-limiting list (structuring agents); Cellulose, Microcrystalline cellulose, Methyl cellulose, Ethyl cellulose, Hydroxypropyl cellulose, Hydroxypropylmethyl cellulose, Methylethyl cellulose, Carboxymethyl cellulose, Kaolin, titanium dioxide, clay, CaCo₃, Diatomic earth, (plasticizers) Dicapryl adipate, Di-(2- ethylhexyl adipate, Di(n-heptyl, n-nonyl) adipate, Diisobutyl adipate, Diisodecyl adipate, Dinomyl adipate, Di-(tridecyl), adipate, Di-(2-ethylheyl azelate), Diisodecyl azelate, Diisoctyl azelate, Dimethyl azelate, Di-n-hexyl azelate, ENZOIC ACID DERIVATIVES, Diethylene glycol dibenzoate, Dipropylene glycol dibenzoate, Polyethylene glycol 200 dibenzo, Acetyl tri- n-butyl citrate, Acetyl triethyl citrate, Tri-n-butyl citrate, Triethyl citrate, Bis-(2-hydroxyethyl dimerate), Epoxidized, linseed oil, Epoxidized soy bean oil, 2-Ethylhexyl epoxytallate, n-Octyl epoxystearate, Dibutyl fumarate, Glycerol triacetate, 2,2,4-Trimethyl-1 ,3-pentanediol, Diisobutyrate, Di-(2-ethylhexyl) isophthalate, Dimethyl isophthalate, Diphenyl isophthalate, Methyl laurate, Methyl linoleate, Di-(2-ethylhexyl) maleate, Di-n-butyl maleate, Tricapryl trimellitate, Tri-(2-ethylhexyl) trimellitate, Triisodecyl trimellitate, Tri-(n-octyl,n-decyl) trimellitate, lsopropyl myristate, Butyl oleate, Glycerol monooleate, Glycerol trioleate, Methyl oleate, n-Propyl oleate, Tetrahydrofurfuryl oleate, lsopropyl palmitate, Methyl palmitate, Chloroparaffin, 41% Cl, Chloroparaffin, 50% Cl, Chloroparaffin, 60% Cl, Chloroparaffin, 70% Cl, 2-Ethylhexyl diphenyl phosphate, lsodecyl diphenyl phosphate, t-Butylphenyl diphenyl phosphate, Tri-butoxyethyl phosphate, Tributyl phosphate, Tricresyl phosphate, Triphenyl phosphate, Butyl benzyl phthalate, Butyl octyl phthalate, Dicapryl phthalate, Dicyclohexyl phthalate, Di-(2-ethylhexyl) phthalate, Diethyl phthalate, Dihexyl phthalate, Diisobutyl phthalate, Diisodecyl phthalate, Diisononyl phthalate, Diisooctyl phthalate, Dimethyl phthalate, Ditridecyl phthalate Diundecyl phthalate, Butyl ricinoleate, Glyceryl tri(acetyl) ricinoleate), Methyl acetyl ricinoleate , Methyl ricinoleate, n-Butyl acetyl ricinoleate, Propylene glycol ricinoleate, Dibutyl sebacate, Di-(2-ethylhexyl) sebacate, Dimethyl sebacate, Ethylene glycol monostearate, Glycerol monostearate, lsopropyl isostearate, Methyl stearate, n-Butyl stearate, Propylene glycol monostearate, Diethyl succinate, N-Ethyl o,p-toluenesulfonamide, o,p-toluenesulfonanamide, Polyesters, adipic acid polyester, Paraplex G-40, adipic acid polyester, Santicizer 334F, azelaic acid polyester, Plastolein 9720, azelaic acid polyester, Plastolein 9750, sebacic acid polyester, Paraplex G-25, Sucrose derivatives, sucrose acetate-isobutyrate (SAIB), Tartaric acid derivative, dibutyl tartrate, Terephthalic acid derivative, bis(2-ethylhexyl) terephthalate, Trimellitic acid derivatives, tris(2-ethylhexyl) trimellitate, heptyl nonyl trimellitate, heptyl nonyl undecyl trimellitate, triisodecyl trimellitate, Glycol derivatives, diethylene glycol dipolargonaic, triethylene glycol di-2, ethylbutyrate, poly(ethylene glycol) (200) di 2-ethylhexanoate, Glycolates, methyl phthalyl ethyl glycolnic, butyl phthalyl butyl glycolnic, Hydrocarbons, hydrogenated terphenyls HR-40, poly(alkyl naphthalene)s, Panaflex, aliphatic aromatics Leromoll, chlorinated paraffin (52 wt % Cl), Cereclor S-52, Camphor, Hydrogenated methyl ester or rosin, Chlorinated Polyphosphanate, Polydimethyl siloxane, Polyco-dimethyl/propylamine siloxanes with various amount of propylamine content, Polydiphenyl siloxanes, Polyco-dimethylphenyl siloxanes, Silanol terminated polysiloxanes, Amino terminated polysiloxanes, Epoxy terminated polysiloxanes, Carbirol terminated polysiloxanes and Polysilanes. An example of such a mixture of lipids, structuring agents and/or plasticizers is SepiFilm^tm (by Seppic).

In a particular embodiment of the present invention the lipid containing coating material is selected from neutral lipids such as oil, fat or wax. In a particular embodiment the coating material is a triglyceride dominated neutral lipid. In one embodiment the oil is a plant oil. Preferably the oil or fat consists of tri-, di- or mono- acylglycerols, free fatty acids or fatty acids derivates (e.g. fatty acid esters). Examples of coating oils or fats that may be suitable include native, partially hydrogenated or fully hydrogenated Aconite Oil, Ajwain Oil/Ajwoin Oil, Alfalfa Oil, Allspice Oil, Almond Oil, Aloe Vera Oil, Angelica Oil, Anise Oil, Apricot Oil, Arnica Oil, Artichoke Oil, Avocado Oil, Basil Oil, Bayleaf Oil, Benzoin Oil, Bergamot Oil, Birch Oil, Blackberry Oil, Black Pepper Oil, Blessed Thistle Oil, Boldo Oil, Borage Oil, Buchu Oil, Buckthorn Oil, Cajuput Oil, Calendula Oil, Camphorwood Oil, Canola Oil, Caraway Oil, Cardamom Oil, Capsicum Oleoresin, Carrot Oil, Cassia Oil, Castor Oil, Catmint Oil, Cayenne Pepper Oil, Cedar Oil, Celery Oil, Centella Oil, Chamomile Oil, Chaste Tree Oil, Chickweed Oil, Chives Oil, Cinnamon Oil, Citronella Oil, Citrus Oil, Clary-sage Oil, Clove Oil, Coconut Oil, Comfrey Oil, Coriander Oil, Corn Oil, Copra Oil, Cotton Seed Oil, Cranberry Oil, Cypress Oil, Cubeb Oil, Cumin Oil, Dandelion Oil, Davana Oil, Devil's Claw Oil, Dill Oil, Echinacea Oil, Elder Flower Oil, Eleuthero Extracts , Ephedra Oil, Eucalyptus Oil, Evening Primrose Oil, Fennel Oil, Fenugreek Oleoresin, Flax Oil, Frankincense Oil, Gardenia Perfume Oil, Garlic Oil, Geranium Oil, Ginger Oil, Ginkgo Biloba Oil, Ginseng Oil, Gold of Pleasure (False Flax) Oil, Golden Rod Oil, Goldenseal Oil, Gotu Kola Oil, Grapefruit Oil, Green Tea Oil, Hawthorn Oil, Hazelnut Oil, Hemp Seed Oil, Heena Perfume Oil, Hibiscus Oil, Hop Oil, Horse Chestnut Oil, Horseradish Oil, Horsetail Oil, Hyssop Oil, Iceland Moss Oil, Immortelle/Everlasting Oil, Ivy Oil, Jasmine Oil, Jatropha Oil, Jojoba Oil, Juniper Oil, Kava Kava Oil, Kewra Perfume Oil, Kola Nut Oil, Kukui Nut Oil, Lady's Mantle Oil, Lavender Oil, Lavandin Oil, Lemon Oil, Lemongrass Oil, Lemon Verbena Oil, Licorice Root Oil, Lily (White Lily) Oil, Lime Oil, Linden Flowers Oil, Linseed/Flax Seed Oil, Lotus Oil, Macadamia Nut Oil, Manuka Oil, Marjoram Oil, Marshmallow Oil, Meadowsweet Oil, Melaleuca Oil, Melissa Oil/Lemon Balm Oil, Menthol Oil, Milk Thistle Oil, Macodonia Nut Oil, Mint Oil, Mullein Oil, Mustard Oil, Myrrh Oil, Neem Oil, Neroli Oil, Nettle Oil, Niaouli Oil, Nutmeg Oil, Oak Oil, Oat Oil, Olive Oil, Onion Oleoresin, Orange Oil, Oregano Oil, Palmarosa Oil, Paprika Oleoresin, Parsley Oil, Passionflower Oil, Patchouli Oil, Palm Oil, palm stearin, palm kernel oil, palm olein, Peanut/Groundnut Oil, Peppermint Oil, Petitgrain Oil, Pine Oil, Plantain Oil, Poplar Extract, Primrose Oil, Psyllium seed husk (Blonde) Oil, Pumpkin Seed Oil, Rapeseed oil, Radish Oil, Rice Bran Oil, Rosehip Oil, Rosemary Oil, Rosewood Oil, Common-sage Oil, Saffron Oil/Zafri Perfume Oil, Saw Palmetto Oil, Sandalwood Oil, Seaweed Oil, Sesame Oil, Spearmint Oil, St. John's Wort Oil, Stinging Nettle Oil, Soybean Oil, Sunflower Oil, Tagetes Oil, Tangerine Oil, Tea Oil, Thyme Oil, Turmeric Oleoresin, Valerian Oil, Vetiver Oil, Walnut Oil, Wheat Germ Oil, Wintergreen Oil, Witch Hazel Oil, Yarrow Oil, Ylang-Ylang Oil and mixtures thereof. In one embodiment the coating materials are selected from the group consisting of palm oil, palm stearin, palm kernel oil, palm olein, coconut oil, soybean oil, sunflower oil, rice bran oil, olive oil, corn oil, castor oil, canola oil, rapeseed oil, jojoba oil and mixtures thereof. Non-limiting examples of oils are native, partially hydrogenated or fully hydrogenated variants of; palm stearin, palm kernel oil, palm olein, coconut oil, cocoa butter, soybean oil, sunflower oil, rice bran oil, olive oil, corn oil, castor oil, canola oil, rapeseed oil, jojoba oil and mixtures thereof.

The fatty acids of the lipids may have varying degree of unsaturation and chain length and thus varying melting points. The desired degree of fatty acid unsaturation (e.g. measured by iodine value, AOCS Official Method Tg 1-64) and fatty acids chain length of the oils/fats can be achieve be using single or multiple oils/fats or oils/fats that are partial or fully hydrogenated.

It is important that the melting point of the coating and/or coating material is above 30⁰C, to prevent the coating from melting during storage.

In a particular embodiment of the present invention the melting point of the coating and/or coating material may be between 30-80⁰C. In a more particular embodiment, the melting point is between 35 to 70⁰C. In an even more particular embodiment the melting point of the coating and/or coating material is 37 to 60°C. In a most particular embodiment the coating and/or coating material has a melting point of 37 to 50⁰C. In another particular embodiment the melting point of the coating and/or coating material is 42 to 60°C, such as 42 to 55°C. In a further embodiment the melting point of the coating and/or coating material is 42 to 60⁰C. In a most particular embodiment the coating and/or coating material has a melting point of 50 to 60°C.

Furthermore if the particles are for human or animal digestion, it is important that the coating material does not have a too high melting point.

In one embodiment of the present invention the coating material has a melting point of below 45°C, such as below 40°C, such as below 38°C or even below 35°C. The coating layer is in a particular embodiment of the present invention below 50% w/w of the finished particles. In another embodiment the coating layer constitute 2-50% w/w of the finished particles. In a further embodiment the coating layer constitute 5-30% w/w of the finished particles. In one embodiment the coating layer constitute 8-25% w/w or 10-20% w/w of the finished particles.

In an embodiment of the present invention the coating layer material comprises at least 10% w/w of non-polar material. In a particular embodiment the coating layer material comprises at least 20% of non-polar material. In a more particular embodiment the coating layer comprises at least 30% w/w of non-polar material. In a most particular embodiment the coating layer comprises at least 45%, such at least 55% or even at least 75% w/w of non- polar material. In a further embodiment the coating layer comprises at least 85% or 95% w/w of non-polar material.

The non-polar coat may also comprise a wax.

A "wax" in the context of the present invention is to be understood as a single wax or a composition of waxes. A wax is a polymeric material having a melting point between 25 -150 ⁰C, particularly 30 to 100⁰C more particularly 35 to 85°C most particularly 40 to 75°C. The wax is preferably in a solid state at room temperature, 25°C. The lower limit is preferred to set a reasonable distance between the temperature at which the wax starts to melt to the temperature at which the particles or compositions comprising the particles are usually stored, 20 to 30⁰C.

The wax composition of the invention may comprise any wax, which is chemically synthesized. It may also equally well comprise waxes isolated from a natural source or a derivative thereof. Accordingly, the wax composition of the invention may comprise waxes selected from the following non limiting list of waxes: Waxes isolated from a natural source, such as Candelilla wax (melting point between 68- 70⁰C). Other waxes include castor wax, esparto wax (from esparto gras/Macrochloa tenacissima), Ouricury wax (from the Brazilian Feather palm, Syagrus coronata) or synsthetic waxes e.g. PEG/PEO (Polyethylene glycol and polyethylene oxide (PEO) or derivatives thereof. The waxes may be derived from animals or plants, e.g. of marine origin. Permeability (of oxygen, active ingredients e.i. PUFA) through the non-polar coat can be varied by choosing coat materials with varying degree of polarity (generally polarity of waxes<tri-glycerids<di-glycerids<mono-glycerids) Also permeability can be controlled by varying the coating layer thickness.

A Pullulan coating layer

The core described above may further have a coating layer comprising pullulan.

Pullulan is an extracellular polysaccharide excreted by the fungus Aureobasidium pullulans.

It is a linear polysaccharide made up of alpha-1 ,6-linked maltotriose residues. Pullulan is available from numerous manufacturers. The pullulan used according to the invention is preferably having a purity higher than 25% by weight, preferably a purity higher than 50% by weight, and in particular a purity higher than

75% by weight. The amount of pullulan coated onto the core particles is preferably within 1-10% by weight of the core particles, in particular within 1-5% by weight of the core particles.

Preparation of the particles The particles of the invention may be produced by any suitable methods known in the art. Methods for preparing the particles of the invention include known particle formulation technologies e.g. spray drying, fluid bed, fluidized spray drying, spray fluidizing, spray cooling, spray chilling, mixer granulation and extrusion. Relevant particles may be granulated products, layered products, absorbed products, pelletized products, prilled products. The cores and particles may optionally be dried after granulation.

Spray dried products, wherein a liquid solution comprising the oxidation labile compound is atomised in a spray drying tower to form small droplets which during their way down the drying tower dry to form oxidative labile compound-containing particles. Very small particles can be produced this way. Layered products, wherein the oxidation labile compound is coated as a layer around a preformed inert core particle or a preformed core made by drying a solution comprising the oxidation labile compound, wherein an oxidation labile-containing solution is atomised, typically in a fluid bed apparatus wherein the pre-formed core particles are fluidised, and the oxidation labile-containing solution adheres to the core particles and dries up to leave a layer of dry oxidation labile compound on the surface of the core particle. Particles of a desired size can be obtained this way if a useful core particle of the desired size can be found. Core particles wherein the oxidation labile compound is absorbed onto and/or into the surface of the core. Extrusion or pelletized products, wherein oxidation labile compound-containing paste is pressed to pellets or under pressure is extruded through a small opening and cut into particles which are subsequently dried. Such particles usually have a considerable size because of the material in which the extrusion opening is made (usually a plate with bore holes) sets a limit on the allowable pressure drop over the extrusion opening. Prilled products, wherein the product obtained is one wherein the oxidation labile compound is uniformly distributed throughout an inert material instead of being concentrated on its surface. Mixer granulation products, wherein a oxidation labile compound is added to additional granulating components. The composition is mixed forming granulates comprising the oxidative labile compound. In a particular product of this process a high-shear mixer is used as granulator.

The granule according to the invention may be produced by a process comprising the following steps: a) mixing an oxidation labile compound, with additional components selected from the group consisting of antioxidants, oxygen scavengers, transition metal ion complexing agents, proteins, carbohydrates, lipids and/or a combination thereof. b) adjusting the liquid content in the mixture c) homogenising the mixture after adjustment of the liquid content; and d) forming core particles from the homogenised mixture. e) coating the core particles with the non-polar and coating.

Optionally the particles are coated with an additional coating before or after applying the non- polar and coating. The additional coating may comprise pullulan. Finally the coated particles may be packed under inert gas, e.g., nitrogen atmosphere.

In one embodiment of the present invention the amount of PUFA in the finished particle is preferably above 10% by weight, such as above 15% by weight, such as above 20% by weight, such as above 25% by weight or even such as above 30% by weight. In a particular embodiment of the present invention the amount of PUFA is at least 40% w/w of the finished particle. In a more particular embodiment the amount of PUFA constitute at least 50% of the finished particle. In a most particular embodiment the PUFA constitute at least 60% w/w of the finished particle.

Food and feed products

A further aspect of the present invention is the provision of a food product comprising an edible material and a particle of the invention. The particle comprises a core and a protective coating that encapsulates the core. The core comprises oxidation labile compound. The nature of the coating will vary depending upon the type of food that the particle of the invention is to be incorporated.

In one embodiment, the food product may be a liquid beverage. Non-limiting examples of a liquid beverages include milk, flavored milk drinks, goat milk, liquid yogurt, soy milk, rice milk, fruit drinks, fruit-flavored drinks, vegetable drinks, nutritional drinks, energy drinks, sports drinks, infant formula, teas, and coffee drinks. In another embodiment, the food product may also be a dairy or an egg product. Examples of dairy products include, but are not limited to, cheese, ice cream, ice cream products, yogurt, whipping cream, sour cream, cottage cheese, buttermilk, egg whites, and egg substitutes. In an alternate embodiment, the food product may be a cereal-based product. Non-limiting examples of food products derived from cereals include breakfast cereals, pasta, breads, baked products (i.e., cakes, pies, rolls, cookies, crackers), tortillas, granola bars, nutrition bars, and energy bars. The food product may be a nutritional supplement. In still another embodiment, the food product may be a vegetable-derived product. Examples of vegetable-derived food products include textured vegetable proteins, tofu, corn chips, potato chips, vegetable chips, popcorn, and chocolate products.

In yet another embodiment, the food product may be a meat product or a meat analog. Examples of meat products include, but are not limited to, processed meats, comminuted meats, and whole muscle meat product. The meat may be animal meat or seafood meat. The meat analog may be a textured vegetable or dairy protein that mimics animal or seafood meat in texture. The meat analog may be part or all of the meat in a food product. The food product may also be a canned food product to which the microcapsule is added to prevent oxidation during the heating process.

In yet another embodiment, the food product may be a product for animals. The animal may be a companion animal, an agricultural animal, or an aquatic organism. Non-limiting examples of animal food products include canned pet foods, dried pet foods, agricultural animal feeds, and agricultural animal feed supplements. The feeds may be pelleted, extruded, or formed by other methods. The feeds or feed supplements may be liquid. Examples include a nursery diets for monogastric animals, calf milk replacer, or fish and other oils used to supplement animal feeds.

Another aspect of the invention provides for food products treated with the composition of the invention. The composition may be sprayed on or applied to a food product. Non-limiting examples of suitable food products include food bars, nutrition bars, snacks, nuts, oats, cookies, crackers, dried fish or seafood products, and pet foods or pet snacks. The composition may be added directly to oxidation sensitive foods. Examples include, but are not limited to, cooking oils, frying oils, spray-on oils, salad dressings, margarines, nut oils, herb or spice oils, cream liquors, shelf-stable cream products, fish oils, fish sauce, nutritional supplements containing fat soluble vitamins and oils, and pharmaceutical preparations containing oxidation labile lipids or oils.

In a particular embodiment the PUFA compound is for human consumption with the purpose of supplying additional PUFA to the diet. PUFA's are essential to humans because of inadequate synthesis in the human metabolism and the overall goal of supplying additional PUFA to the diet is to reach the Recommended Daily Intake for PUFA. Several types of delivery routes exist, such as a dietary supplement as capsules, as fortification of diary products, such as infant formula which is fortified products for infants ( 0-12 months of age) and growing up milk also fortified milk for children, or as fortified foods in general, examples being bread, cakes, chocolate, candy, canned food, cereals, spreads and drinks. In another particular embodiment of the present invention the particles of the invention are used in food products, as a nutritional supplement, as a dietary supplement and/or in feed products. The particles of the invention may even be incorporated into pharmaceuticals. Nonfood Products

A further aspect of the invention provides nonfood products comprising lipid-coated stabilized products comprising oxidative labile compounds. The nonfood product may be a cosmetic, a body moisturizer, or an anti-aging cream for humans, or it may be a product to prevent pet coat oil oxidation or prevent pet odor. The nonfood product may be a fragrance product or an air freshener product. The nonfood product may be a paint or varnish. The nonfood product may be a mineral oil, a synthetic oil, or a biodiesel.

In a particular embodiment the particles may be used in non food products such as cosmetics, body moisterizers, anti aging creams, fragrances, air freshener, paint and or varnish

EXAMPLES

Example 1 The efficacy of Palm oil as an oxygen barrier thus preventing oxidation of oxidative labile poly unsaturated fatty acids (PUFA) was investigated by making a PUFA containing core and coating this core with combinations of two coating materials (se table 1 ).

Preparation of PUFA containing cores ("A" in table 1 ). An inert core (precipitated silica) was heated in a high-shear mixer (Lodige 10 liter horizontal mixer) to a temperature of 85°C. The molten PUFA product was added to the silica cores while distribution of the PUFA was facilitated by mixing with the horizontal mixer blades. The amount of PUFA containing product in the final cores equaled 67% (w/w).

Coating of the PUFA containing product was performed as a fluid bed (Aeromatic Strea) operation. Pullulan (Hayashibara Co., LTD. Japan) was sprayed onto "A" in a 3% solution (w/w in water) and dried at an inlet temperature of 70-80⁰C. Fully hydrogenated palm oil at a temperature of 100-120⁰C was sprayed onto "A" while maintaining the inlet temperature at 55°C.

The oxidative stability was evaluated by an Oxipres apparatus (Mikrolab Aarhus, Denmark) Results are shown in table 1. A sample of 5 grams of lipid was placed in a sealed 125 ml. steel vessel under pure O₂ atmosphere at a pressure of 5 bar. Temperature was elevated to 50⁰C and total pressure was monitored over time. The Induction Period (IP) was measured in hours and corresponded to the time period during which the product is resistant to oxidation. After the IP the oxidation is rapid and the oxygen pressure drops rapidly.

The palm oil shows to be very effective in prolonging the IP. a, b, c are duplicates. Table 1:

Claims

1. A composition comprising a) a core comprising an oxidation labile compound; wherein the oxidation labile compound is a lipid having at least one carbon-carbon double bond that is prone to oxidation; and b) an oxidation protective coating layer comprising a lipid.

2. The composition of claim 1 , wherein the core further comprises an antioxidant.

3. The composition according to claim 1 or 2, wherein the core further comprises an oxygen scavenger.

4. The composition according to any preceding claims, wherein the core further comprises a transition metal ion complexing agent.

5. The composition according to any preceding claims, wherein the oxidation labile compound is a long chain polyunsaturated fatty acid.

6. The composition according to claim 5, wherein the polyunsaturated fatty acid is eicosapentaenoic acid and/or docosahexaenoic acid.

7. The composition according to claim 5, wherein the polyunsaturated fatty acid is present within triglycerides.

8. The composition according to any preceding claims, wherein the core comprising a labile compound comprises a lipid enriched biomass produced from a microorganism.

9. The composition according to claim 8, wherein the lipid enriched biomass contains an antioxidants produced by the microorganism.

10. The composition according to claim 8, wherein the lipid enriched biomass of the microorganism has been pasteurised.

11. The composition according to claim 8, wherein the microorganism is an auxotrophic marine microorganism.

12. The composition according to claim 8, wherein the microorganism is selected from the group consisting of Aphanizomenon flos-aquae, Bacilliarophy sp., Botryococcus braunii, Chlorophyceae sp., Crypthecodinium cohnii, Dunaliella tertiolecta, Euglena gracilis, lsochrysis galbana, Nannochloropsis salina, Nannochloris sp., Neochloris oleoabundans, Phaeodactylum tricornutum, Pleurochrysis carterae, Prymnesiumparvum, Scenedesmus dimorphus, Schizochytrium sp., Spirulina sp., Tetraselmis chui, Hamatores sp, Proteromonads sp, Opalines sp, Developayella sp, Diplophrys sp, Labrinthulids sp, Thraustochytrids sp, Biosecids sp, Oomycetes sp, Hypochytridiomycetes sp, Commation sp, Reticulosphaera sp, Pelagomonas sp, Pelagococcus sp, Ollicola sp, Aureococcus sp, Parmales sp, Diatoms sp, Xanthophytes sp, Phaeophytes sp (brown algae), Eustigmatophytes sp, Raphidophytes sp, Synurids sp, Axodines sp, Chrysomeridales sp, Sarcinochrysidales sp, Hydrurales sp, Hibberdiales sp and Chromulinales sp.

13. The composition according to any preceding claims, wherein the core comprises one or more proteins and one or more carbohydrates.

14. The composition according to claim 13, wherein the protein is selected from the group consisting of plant proteins and/or milk derived proteins.

15. The composition according to claim 13, wherein the carbohydrate is selected from the group consisting of starch, glucose, sucrose and dextrins.

16. The composition according to any preceding claims, wherein core comprises an oxygen scavenger which is selected from the group consisting of an antioxidant and an enzymatic oxygen scavenger.

17. The composition according to claim 16, wherein the antioxidant is selected from the group consisting of cocoa, tocopherol, resveratrol, rosemary extracts, ascorbic acid and ascorbyl palmitate.

18. The composition according to claim 16, wherein the enzymatic oxygen scavenger is a mixture of glucoseoxidase and catalase.

19. The composition according to any preceding claims, wherein the core comprises a transition metal ion complexing agent which is selected from the group consisting of EDTA, and citric acid.

20. The composition according to any preceding claims, wherein the coating is selected from the group consisting of glycerid oils, fats and waxes.

21. A process for producing a composition according to any preceding claims, wherein the core formation comprises a) forming core particles comprising an oxidation labile compound, b) coating the particles of a) with a coating material.

22. The process according to claim 21 , wherein the particles are formed by spray drying.

23. The process according to claim 21 , wherein the particles of a) are formed in a fluidised bed.

24. The process according to claim 21 , wherein the particle formation comprises extrudation.

25. The process according to any of claims 21-24 wherein the process is operated in an atmosphere of an inert gas.

26. The process according to claim 21 , wherein the oxidation labile compound is mixed with one or more proteins, one or more carbohydrates, one or more oxygen scavengers and one or more transition metal ion complexing agents.