AU2007241642A1 - Process for separating lipid materials - Google Patents
Process for separating lipid materials Download PDFInfo
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- AU2007241642A1 AU2007241642A1 AU2007241642A AU2007241642A AU2007241642A1 AU 2007241642 A1 AU2007241642 A1 AU 2007241642A1 AU 2007241642 A AU2007241642 A AU 2007241642A AU 2007241642 A AU2007241642 A AU 2007241642A AU 2007241642 A1 AU2007241642 A1 AU 2007241642A1
- Authority
- AU
- Australia
- Prior art keywords
- feed material
- comprises greater
- product
- solvent
- material comprises
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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Links
- 239000000463 material Substances 0.000 title claims description 196
- 238000000034 method Methods 0.000 title claims description 113
- 230000008569 process Effects 0.000 title claims description 107
- 150000002632 lipids Chemical class 0.000 title claims description 92
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 257
- ZWZWYGMENQVNFU-UHFFFAOYSA-N Glycerophosphorylserin Natural products OC(=O)C(N)COP(O)(=O)OCC(O)CO ZWZWYGMENQVNFU-UHFFFAOYSA-N 0.000 claims description 80
- TZCPCKNHXULUIY-RGULYWFUSA-N 1,2-distearoyl-sn-glycero-3-phosphoserine Chemical compound CCCCCCCCCCCCCCCCCC(=O)OC[C@H](COP(O)(=O)OC[C@H](N)C(O)=O)OC(=O)CCCCCCCCCCCCCCCCC TZCPCKNHXULUIY-RGULYWFUSA-N 0.000 claims description 64
- 239000002904 solvent Substances 0.000 claims description 57
- 239000000203 mixture Substances 0.000 claims description 51
- 239000006184 cosolvent Substances 0.000 claims description 47
- ZGSPNIOCEDOHGS-UHFFFAOYSA-L disodium [3-[2,3-di(octadeca-9,12-dienoyloxy)propoxy-oxidophosphoryl]oxy-2-hydroxypropyl] 2,3-di(octadeca-9,12-dienoyloxy)propyl phosphate Chemical compound [Na+].[Na+].CCCCCC=CCC=CCCCCCCCC(=O)OCC(OC(=O)CCCCCCCC=CCC=CCCCCC)COP([O-])(=O)OCC(O)COP([O-])(=O)OCC(OC(=O)CCCCCCCC=CCC=CCCCCC)COC(=O)CCCCCCCC=CCC=CCCCCC ZGSPNIOCEDOHGS-UHFFFAOYSA-L 0.000 claims description 43
- 150000002270 gangliosides Chemical class 0.000 claims description 41
- 235000013365 dairy product Nutrition 0.000 claims description 30
- QQVDJLLNRSOCEL-UHFFFAOYSA-N (2-aminoethyl)phosphonic acid Chemical compound [NH3+]CCP(O)([O-])=O QQVDJLLNRSOCEL-UHFFFAOYSA-N 0.000 claims description 27
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 24
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 15
- 241001465754 Metazoa Species 0.000 claims description 11
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 claims description 8
- 210000003022 colostrum Anatomy 0.000 claims description 8
- 235000021277 colostrum Nutrition 0.000 claims description 8
- 210000002966 serum Anatomy 0.000 claims description 8
- 241000283690 Bos taurus Species 0.000 claims description 7
- 235000013336 milk Nutrition 0.000 claims description 7
- 239000008267 milk Substances 0.000 claims description 7
- 210000004080 milk Anatomy 0.000 claims description 7
- 210000004369 blood Anatomy 0.000 claims description 6
- 239000008280 blood Substances 0.000 claims description 6
- 235000013601 eggs Nutrition 0.000 claims description 6
- 239000012528 membrane Substances 0.000 claims description 6
- 239000007787 solid Substances 0.000 claims description 6
- 108010046377 Whey Proteins Proteins 0.000 claims description 5
- 102000007544 Whey Proteins Human genes 0.000 claims description 5
- 210000000056 organ Anatomy 0.000 claims description 5
- BDERNNFJNOPAEC-UHFFFAOYSA-N propan-1-ol Chemical compound CCCO BDERNNFJNOPAEC-UHFFFAOYSA-N 0.000 claims description 5
- 239000005862 Whey Substances 0.000 claims description 4
- 150000001298 alcohols Chemical class 0.000 claims description 4
- 235000014121 butter Nutrition 0.000 claims description 4
- 235000015155 buttermilk Nutrition 0.000 claims description 4
- 241000282414 Homo sapiens Species 0.000 claims description 3
- 108010071421 milk fat globule Proteins 0.000 claims description 3
- 244000303258 Annona diversifolia Species 0.000 claims description 2
- 235000002198 Annona diversifolia Nutrition 0.000 claims description 2
- 241000894006 Bacteria Species 0.000 claims description 2
- 241001416153 Bos grunniens Species 0.000 claims description 2
- 241000030939 Bubalus bubalis Species 0.000 claims description 2
- 241000282836 Camelus dromedarius Species 0.000 claims description 2
- 241000283707 Capra Species 0.000 claims description 2
- 241000283074 Equus asinus Species 0.000 claims description 2
- 241000283073 Equus caballus Species 0.000 claims description 2
- 241000699666 Mus <mouse, genus> Species 0.000 claims description 2
- 241001494479 Pecora Species 0.000 claims description 2
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 2
- 241000282898 Sus scrofa Species 0.000 claims description 2
- 210000001557 animal structure Anatomy 0.000 claims description 2
- 229910052799 carbon Inorganic materials 0.000 claims description 2
- 238000001704 evaporation Methods 0.000 claims description 2
- 230000008020 evaporation Effects 0.000 claims description 2
- 244000005700 microbiome Species 0.000 claims description 2
- MTCFGRXMJLQNBG-UHFFFAOYSA-N Serine Natural products OCC(N)C(O)=O MTCFGRXMJLQNBG-UHFFFAOYSA-N 0.000 claims 1
- 125000001095 phosphatidyl group Chemical group 0.000 claims 1
- 239000000284 extract Substances 0.000 description 123
- 150000003904 phospholipids Chemical class 0.000 description 76
- 238000000605 extraction Methods 0.000 description 53
- JZNWSCPGTDBMEW-UHFFFAOYSA-N Glycerophosphorylethanolamin Natural products NCCOP(O)(=O)OCC(O)CO JZNWSCPGTDBMEW-UHFFFAOYSA-N 0.000 description 35
- 239000000047 product Substances 0.000 description 34
- 238000005194 fractionation Methods 0.000 description 30
- 230000007935 neutral effect Effects 0.000 description 24
- 150000001875 compounds Chemical class 0.000 description 23
- 150000008104 phosphatidylethanolamines Chemical class 0.000 description 19
- -1 PI PS PE Chemical class 0.000 description 17
- 150000003905 phosphatidylinositols Chemical class 0.000 description 15
- 239000000843 powder Substances 0.000 description 12
- 235000010469 Glycine max Nutrition 0.000 description 7
- 238000012545 processing Methods 0.000 description 7
- 239000012141 concentrate Substances 0.000 description 6
- 235000013345 egg yolk Nutrition 0.000 description 6
- 210000002969 egg yolk Anatomy 0.000 description 6
- 239000007788 liquid Substances 0.000 description 6
- 102100036962 5'-3' exoribonuclease 1 Human genes 0.000 description 5
- 101000804879 Homo sapiens 5'-3' exoribonuclease 1 Proteins 0.000 description 5
- 229940106189 ceramide Drugs 0.000 description 5
- 239000012530 fluid Substances 0.000 description 5
- WTJKGGKOPKCXLL-RRHRGVEJSA-N phosphatidylcholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCCC=CCCCCCCCC WTJKGGKOPKCXLL-RRHRGVEJSA-N 0.000 description 5
- 239000000126 substance Substances 0.000 description 5
- PORPENFLTBBHSG-MGBGTMOVSA-N 1,2-dihexadecanoyl-sn-glycerol-3-phosphate Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP(O)(O)=O)OC(=O)CCCCCCCCCCCCCCC PORPENFLTBBHSG-MGBGTMOVSA-N 0.000 description 4
- YDNKGFDKKRUKPY-JHOUSYSJSA-N C16 ceramide Natural products CCCCCCCCCCCCCCCC(=O)N[C@@H](CO)[C@H](O)C=CCCCCCCCCCCCCC YDNKGFDKKRUKPY-JHOUSYSJSA-N 0.000 description 4
- 102000002322 Egg Proteins Human genes 0.000 description 4
- 108010000912 Egg Proteins Proteins 0.000 description 4
- 241000239366 Euphausiacea Species 0.000 description 4
- 244000068988 Glycine max Species 0.000 description 4
- CRJGESKKUOMBCT-VQTJNVASSA-N N-acetylsphinganine Chemical compound CCCCCCCCCCCCCCC[C@@H](O)[C@H](CO)NC(C)=O CRJGESKKUOMBCT-VQTJNVASSA-N 0.000 description 4
- ZVEQCJWYRWKARO-UHFFFAOYSA-N ceramide Natural products CCCCCCCCCCCCCCC(O)C(=O)NC(CO)C(O)C=CCCC=C(C)CCCCCCCCC ZVEQCJWYRWKARO-UHFFFAOYSA-N 0.000 description 4
- VVGIYYKRAMHVLU-UHFFFAOYSA-N newbouldiamide Natural products CCCCCCCCCCCCCCCCCCCC(O)C(O)C(O)C(CO)NC(=O)CCCCCCCCCCCCCCCCC VVGIYYKRAMHVLU-UHFFFAOYSA-N 0.000 description 4
- 238000000926 separation method Methods 0.000 description 4
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 3
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 3
- 241000196324 Embryophyta Species 0.000 description 3
- 229930186217 Glycolipid Natural products 0.000 description 3
- 229930183167 cerebroside Natural products 0.000 description 3
- 150000001784 cerebrosides Chemical class 0.000 description 3
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 3
- 210000001519 tissue Anatomy 0.000 description 3
- JQWAHKMIYCERGA-UHFFFAOYSA-N (2-nonanoyloxy-3-octadeca-9,12-dienoyloxypropoxy)-[2-(trimethylazaniumyl)ethyl]phosphinate Chemical compound CCCCCCCCC(=O)OC(COP([O-])(=O)CC[N+](C)(C)C)COC(=O)CCCCCCCC=CCC=CCCCCC JQWAHKMIYCERGA-UHFFFAOYSA-N 0.000 description 2
- PZNPLUBHRSSFHT-RRHRGVEJSA-N 1-hexadecanoyl-2-octadecanoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCCCC(=O)O[C@@H](COP([O-])(=O)OCC[N+](C)(C)C)COC(=O)CCCCCCCCCCCCCCC PZNPLUBHRSSFHT-RRHRGVEJSA-N 0.000 description 2
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- 241000237536 Mytilus edulis Species 0.000 description 2
- 230000036995 brain health Effects 0.000 description 2
- 210000004027 cell Anatomy 0.000 description 2
- 239000008344 egg yolk phospholipid Substances 0.000 description 2
- 229940068998 egg yolk phospholipid Drugs 0.000 description 2
- 239000000469 ethanolic extract Substances 0.000 description 2
- 230000002349 favourable effect Effects 0.000 description 2
- 239000012467 final product Substances 0.000 description 2
- 239000007789 gas Substances 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- 235000020638 mussel Nutrition 0.000 description 2
- 235000016709 nutrition Nutrition 0.000 description 2
- 239000003960 organic solvent Substances 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 238000011084 recovery Methods 0.000 description 2
- 239000008347 soybean phospholipid Substances 0.000 description 2
- 235000014268 sports nutrition Nutrition 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- JLPULHDHAOZNQI-ZTIMHPMXSA-N 1-hexadecanoyl-2-(9Z,12Z-octadecadienoyl)-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCC\C=C/C\C=C/CCCCC JLPULHDHAOZNQI-ZTIMHPMXSA-N 0.000 description 1
- IIZPXYDJLKNOIY-JXPKJXOSSA-N 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCC\C=C/C\C=C/C\C=C/C\C=C/CCCCC IIZPXYDJLKNOIY-JXPKJXOSSA-N 0.000 description 1
- HVCOBJNICQPDBP-UHFFFAOYSA-N 3-[3-[3,5-dihydroxy-6-methyl-4-(3,4,5-trihydroxy-6-methyloxan-2-yl)oxyoxan-2-yl]oxydecanoyloxy]decanoic acid;hydrate Chemical compound O.OC1C(OC(CC(=O)OC(CCCCCCC)CC(O)=O)CCCCCCC)OC(C)C(O)C1OC1C(O)C(O)C(O)C(C)O1 HVCOBJNICQPDBP-UHFFFAOYSA-N 0.000 description 1
- 241001062872 Cleyera japonica Species 0.000 description 1
- 201000004624 Dermatitis Diseases 0.000 description 1
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 1
- 208000001145 Metabolic Syndrome Diseases 0.000 description 1
- 102000007474 Multiprotein Complexes Human genes 0.000 description 1
- 108010085220 Multiprotein Complexes Proteins 0.000 description 1
- 241001537211 Perna canaliculus Species 0.000 description 1
- 229910000831 Steel Inorganic materials 0.000 description 1
- 238000003811 acetone extraction Methods 0.000 description 1
- PNEYBMLMFCGWSK-UHFFFAOYSA-N aluminium oxide Inorganic materials [O-2].[O-2].[O-2].[Al+3].[Al+3] PNEYBMLMFCGWSK-UHFFFAOYSA-N 0.000 description 1
- 230000001093 anti-cancer Effects 0.000 description 1
- 230000002924 anti-infective effect Effects 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- 206010003246 arthritis Diseases 0.000 description 1
- 208000010668 atopic eczema Diseases 0.000 description 1
- 238000010923 batch production Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 229910002092 carbon dioxide Inorganic materials 0.000 description 1
- 230000036996 cardiovascular health Effects 0.000 description 1
- 230000030833 cell death Effects 0.000 description 1
- 230000024245 cell differentiation Effects 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 230000005754 cellular signaling Effects 0.000 description 1
- 150000001783 ceramides Chemical class 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 238000005112 continuous flow technique Methods 0.000 description 1
- 235000005911 diet Nutrition 0.000 description 1
- 230000000378 dietary effect Effects 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000027721 electron transport chain Effects 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 238000011067 equilibration Methods 0.000 description 1
- 238000002481 ethanol extraction Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 235000019197 fats Nutrition 0.000 description 1
- 229930195729 fatty acid Natural products 0.000 description 1
- 239000000194 fatty acid Substances 0.000 description 1
- 150000004665 fatty acids Chemical class 0.000 description 1
- 235000013350 formula milk Nutrition 0.000 description 1
- 210000004907 gland Anatomy 0.000 description 1
- 150000002327 glycerophospholipids Chemical class 0.000 description 1
- 244000005709 gut microbiome Species 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 229930195733 hydrocarbon Natural products 0.000 description 1
- 150000002430 hydrocarbons Chemical class 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 239000012212 insulator Substances 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 125000001449 isopropyl group Chemical group [H]C([H])([H])C([H])(*)C([H])([H])[H] 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 229940067606 lecithin Drugs 0.000 description 1
- 239000000787 lecithin Substances 0.000 description 1
- 235000010445 lecithin Nutrition 0.000 description 1
- 238000004811 liquid chromatography Methods 0.000 description 1
- 230000008018 melting Effects 0.000 description 1
- 238000002844 melting Methods 0.000 description 1
- 235000021243 milk fat Nutrition 0.000 description 1
- 210000001700 mitochondrial membrane Anatomy 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 210000000663 muscle cell Anatomy 0.000 description 1
- 210000004165 myocardium Anatomy 0.000 description 1
- 208000015122 neurodegenerative disease Diseases 0.000 description 1
- 210000002569 neuron Anatomy 0.000 description 1
- 230000000704 physical effect Effects 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000002390 rotary evaporation Methods 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 230000019491 signal transduction Effects 0.000 description 1
- 210000002027 skeletal muscle Anatomy 0.000 description 1
- 230000036559 skin health Effects 0.000 description 1
- 239000011343 solid material Substances 0.000 description 1
- 229940083466 soybean lecithin Drugs 0.000 description 1
- 150000003408 sphingolipids Chemical class 0.000 description 1
- 239000010959 steel Substances 0.000 description 1
- 238000000194 supercritical-fluid extraction Methods 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- UFTFJSFQGQCHQW-UHFFFAOYSA-N triformin Chemical compound O=COCC(OC=O)COC=O UFTFJSFQGQCHQW-UHFFFAOYSA-N 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
- 235000021119 whey protein Nutrition 0.000 description 1
- 230000029663 wound healing Effects 0.000 description 1
Classifications
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- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
- C11B7/00—Separation of mixtures of fats or fatty oils into their constituents, e.g. saturated oils from unsaturated oils
- C11B7/0008—Separation of mixtures of fats or fatty oils into their constituents, e.g. saturated oils from unsaturated oils by differences of solubilities, e.g. by extraction, by separation from a solution by means of anti-solvents
- C11B7/005—Separation of mixtures of fats or fatty oils into their constituents, e.g. saturated oils from unsaturated oils by differences of solubilities, e.g. by extraction, by separation from a solution by means of anti-solvents in solvents used at superatmospheric pressures
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J7/00—Phosphatide compositions for foodstuffs, e.g. lecithin
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
- A23L5/20—Removal of unwanted matter, e.g. deodorisation or detoxification
- A23L5/23—Removal of unwanted matter, e.g. deodorisation or detoxification by extraction with solvents
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C2240/00—Use or particular additives or ingredients
- A23C2240/05—Milk products enriched with milk fat globule membrane
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
Description
WO 2007/123424 PCT/NZ2007/000087 PRODUCT AND PROCESS FIELD OF INVENTION 5 This invention relates to a separation process. More particularly it relates to a process for separating lipid materials containing phospholipids and/or glycolipids, including for example phosphatidyl serine, gangliosides, cardiolipin, sphingomyelin, plasmalogens, alkylacylphospholipids, phosphonolipids, cerebrosides or a combination thereof. BACKGROUND 10 Phospholipids are a major component of all biological membranes, and include phosphoglycerides (phosphatidyl choline (PC), phosphatidyl ethanolamine (PE), phosphatidyl inositol (PI), cardiolipin (CL), phosphatidyl serine (PS)), plasmalogens (PL), phosphonolipids (PP), alkylacylphospholipids (ALP); and sphingolipids such as 15 sphingomyelin (SM) and ceramide aminoethylphosphonate (CAEP). Gangliosides are glycolipid components in the cell plasma membrane, which modulate cell signal transductions events. They are implicated as being important in immunology and neurodegenerative disorders. Cerebrosides are important components in animal muscle and nerve cell membranes. 20 Both phospholipids and gangliosides are involved in cell signalling events leading to, for example, cell death (apoptosis), cell growth, cell proliferation, and cell differentiation. Reasonable levels of some of these components can be found in milk, soy products, eggs, animal glands and organs, marine animals, plants and other sources. A source of these components is the bovine milk fat globule membrane (MFGM) which is known to contain 25 useful quantities of sphingomyelin, ceramides, gangliosides, and phosphatidyl serine. Another source of these components is the green-shell mussel, which is known to contain useful quantities of plasmalogens, alkylacylphospholipids and ceramide aminoethylphosphonate Both phospholipids and gangliosides have been implicated in conferring a number of health 30 benefits including brain health, skin health, eczema treatment, anti-infection, wound healing, gut microbiota modifications, anti-cancer activity, alleviation of arthritis, improvement of
I
WO 2007/123424 PCT/NZ2007/000087 cardiovascular health, and treatment of metabolic syndromes. They can also be used in sports nutrition. Cardiolipin is an important component of the inner mitochondrial membrane. It is typically present in metabolically active cells of the heart and skeletal muscle. It serves as an 5 insulator and stabilises the activity of protein complexes important to the electron transport chain. Existing methods for isolation of these compounds rely on the use of chromatographic techniques, which are slow and costly processes to operate. These techniques can also require the use of solvents that are unsuitable and/or undesirable in products for nutritional 10 or human use. For example, Palacios and Wang [1] describe a process for extraction of phospholipids from egg yolks using acetone and ethanol extractions, followed by a methanol/chloroform separation. Kang and Row [2] describe a liquid chromatography process for separation of soybean derived PC from PE and PI. This process may be expensive to carry out on an industrial scale, and also uses hexane, methanol, and isopropyl 15 alcohol as solvents. Kearns et al [3] describe a process for purification of egg yolk derived PC from PE using mixtures of acetonitrile, hydrocarbons, and fluorocarbons. Again, these solvents are undesirable for nutritional or pharmaceutical use. Supercritical fluid extraction processes using CO 2 are becoming increasingly popular because of a number of processing and consumer benefits. CO 2 can be easily removed from 20 the final product by reducing the pressure, whereupon the CO 2 reverts to a gaseous state, giving a completely solvent free product. The extract is considered to be more 'natural' than extracts produced using other solvents, and the use of CO 2 in place of conventional organic solvents also confers environmental benefits through reduced organic solvent use. The disadvantage of supercritical CO 2 processing is that the solubility of many compounds in 25 CO 2 is low, and only neutral lipids can be extracted. It is known that the use of CO 2 with organic co-solvents such as ethanol allows extraction of some phosphatidyl choline and to a much lesser extent phosphatidyl ethanolamine. For example, Teberikler et al [4] describe a process for extraction of PC from a soybean lecithin. Using 10% ethanol in CO 2 at 60'C they found that PC was easily extracted, while PE and PI 30 were extracted to a very low extent. Extraction at 12.5 % ethanol at 80 0 C gave a four-fold increase in solubility of PC. Montanari et al [5] describe a process for extracting phospholipids from soybean flakes. After first extracting neutral lipids using only CO 2 at 320 bar, they found that using 10 % ethanol co-solvent at pressures of 194 to 689 bar resulted in 2 WO 2007/123424 PCT/NZ2007/000087 some extraction of PC, PE, PI, and phosphatidic acid (PA). PC is selectively extracted under some conditions, but at higher temperatures and pressures some extraction of PE and PI was achieved. The pressures required to achieve good extraction were impractically high for industrial application, and the high temperatures used (80 0 C) could cause polyunsaturated 5 fatty acids to be degraded. Taylor et al [6] describe a process in which soybean flakes are first extracted using only CO 2 , followed by CO 2 with 15% ethanol at 80 0 C and 665 bar. A mixture of phospholipids is obtained which were fractionated by alumina column. Again, the temperatures and pressures are too high for practical application. In these works, the soybean-derived feed materials do not contain detectable levels of SM, CL, GS or PS. 10 Tanaka and Sakaki [7] describe a method for extracting phospholipids from waste tuna shavings using CO 2 and ethanol as a co-solvent. They describe extraction of DHA containing phospholipids using 5 % ethanol in CO 2 , and by presoaking the tuna flakes in straight ethanol and then extracting using CO 2 . The phospholipids obtained in this process are not specified and no fractionation of the different phospholipids is described. In addition, 15 the phospholipids fraction makes up a relatively small proportion of the total processed material, requiring use of large pressure vessels to produce a small yield of phospholipids. Bulley et al [8] describe extraction of frozen egg yolks using CO 2 and 3 % ethanol, and CO 2 with up to 5 % methanol. Higher rates of triglyceride extraction were obtained with the use of the co-solvent. Extraction of small amounts of phospholipids, up to 17% concentration in 20 the extract, was also achieved. Fractionation of the phospholipids is not described. In this specification where reference has been made to patent specifications, other external documents, or other sources of information, this is generally for the purpose of providing a context for discussing the features of the invention. Unless specifically stated otherwise, reference to such external documents or such sources of information is not to be construed as 25 an admission that such documents or such sources of information, in any jurisdiction, are prior art or form part of the common general knowledge in the art. It is an object of this invention to provide a process for producing a product that contains desirable levels of particular phospholipids and/or gangliosides and/or cerebrosides, or at least to offer the public a useful choice. 3 WO 2007/123424 PCT/NZ2007/000087 SUMMARY OF INVENTION Accordingly the present invention provides a process for separating a feed material into soluble and insoluble components, comprising: (a) providing a feed material comprising one or more of: 5 (i) at least 1% by mass phosphatidyl serine (ii) at least 1% by mass sphingomyelin (iii) at least 0.3 % by mass acylalkylphospholipids and/or plasmalogens (iv)at least 0.5 % by mass aminoethylphosphonate and/or other phosphonolipids (v) at least 1% by mass cardiolipin 10 (vi) at least 0.3% by mass gangliosides (b) providing a solvent comprising: (i) supercritical or near-critical CO 2 , and (ii) a co-solvent comprising one or more Ci-C 3 monohydric alcohols, and water wherein the co-solvent makes up at least 10% by mass of the CO 2 , and the water content 15 of the co-solvent is 0 to 40 % by mass (c) contacting the feed material and the solvent and subsequently separating the solvent containing the soluble components from the insoluble components (d) optionally separating the soluble components and the solvent. Preferably the feed material comprises greater than 1% phosphatidyl serine. More 20 preferably the feed material comprises greater than 2% phosphatidyl serine. Most preferably the feed material comprises greater than 5% phosphatidyl serine. Alternatively the feed material comprises greater than 1% sphingomyelin. More preferably the feed material comprises greater than 5% sphingomyelin. Most preferably the feed material comprises greater than 15% sphingomyelin. 4 WO 2007/123424 PCT/NZ2007/000087 Alternatively the feed material comprises greater than 1% cardiolipin. More preferably the feed material comprises greater than 2% cardiolipin. Most preferably the feed material comprises greater than 5% cardiolipin. Alternatively the feed material comprises greater than 0.3% gangliosides. More preferably 5 the feed material comprises greater than 1% gangliosides. Most preferably the feed material comprises greater than 2% gangliosides. Alternatively the feed material comprises greater than 0.5% acylalkyphospholipids and/or plasmalogens. More preferably the feed material comprises greater than 2% acylalkyphospholipids and/or plasmalogens. Most preferably the feed material comprises 10 greater than 10% acylalkyphospholipids and/or plasmalogens. Alternatively the feed material comprises greater than 0.5% aminoethylphosphonate and/or other phosphonolipids. More preferably the feed material comprises greater than 5% aminoethylphosphonate and/or other phosphonolipids. Most preferably the feed material comprises greater than 20% aminoethylphosphonate and/or other phosphonolipids. 15 The present invention also provides a process for separating a feed material into soluble and insoluble components, comprising (a) providing a feed material comprising one or more of: (i) at least 1% by mass phosphatidyl serine, (ii) at least 1% by mass sphingomyelin, 20 (iii) at least 0.3 % by mass acylalkylphospholipids and/or plasmalogens (iv) at least 0.5 % by mass aminoethylphosphonate and/or other phosphonolipids (v) at least 1% by mass cardiolipin, or (vi) at least 0.3% by mass gangliosides (b) providing a first solvent comprising supercritical or near-critical CO 2 25 (c) contacting the feed material and the first solvent and subsequently separating the first solvent containing the first soluble components from the first insoluble components (d) optionally separating the first soluble components and the first solvent 5 WO 2007/123424 PCT/NZ2007/000087 (e) providing a second solvent comprising: (i) supercritical or near-critical C0 2 , and (ii) a co-solvent comprising one or more C 1
-C
3 monohydric alcohols, and water wherein the co-solvent makes up at least 10% by mass of the C0 2 , and the water content 5 of the co-solvent is 0 to 40% by mass (f) contacting the first insoluble components and the second solvent and subsequently separating the second solvent containing the second soluble components from the second insoluble components (g) optionally separating the second soluble components and the second solvent. 10 Preferably the first solvent comprises a mixture of supercritical or near-critical CO 2 and less than 10% C 1
-C
3 monohydric alcohol. The feed material preferably comprises greater than 1% phosphatidyl serine. More preferably the feed material comprises greater than 2% phosphatidyl serine. Most preferably the feed material comprises greater than 5% phosphatidyl serine. 15 Alternatively the feed material comprises greater than 1% sphingomyelin. Preferably the feed material comprises greater than 5% sphingomyelin. More preferably the feed material comprises greater than 15% sphingomyelin. Alternatively the feed material comprises greater than 1% cardiolipin. Preferably the feed material comprises greater than 2% cardiolipin. More preferably the feed material comprises 20 greater than 5% cardiolipin. Alternatively the feed material comprises greater than 0.3% gangliosides. Preferably the feed material comprises greater than 1% gangliosides. More preferably the feed material comprises greater than 2% gangliosides. Alternatively the feed material comprises greater than 0.5% acylalkyphospholipids and/or 25 plasmalogens. Preferably the feed material comprises greater than 2% acylalkyphospholipids and/or plasmalogens. More preferably the feed material comprises greater than 10% acylalkyphospholipids and/or plasmalogens. 6 WO 2007/123424 PCT/NZ2007/000087 Alternatively the feed material comprises greater than 0.5% aminoethylphosphonate and/or other phosphonolipids. Preferably the feed material comprises greater than 5% aminoethylphosphonate and/or other phosphonolipids. More preferably the feed material comprises greater than 20% aminoethylphosphonate and/or other phosphonolipids. 5 The feed material of the present invention may be derived from terrestrial animals, marine animals, terrestrial plants, marine plants, or micro-organisms such as microalgae, yeast and bacteria. Preferably the feed material is derived from sheep, goat, pig, mouse, water buffalo, camel, yak, horse, donkey, llama, bovine or human. Optionally the feed material is selected from: tissue, a tissue fraction, organ, an organ 10 fraction, milk, a milk fraction, colostrum, a colostrum fraction, blood and a blood fraction. Preferably the feed material is derived from dairy material, soy material, eggs, animal tissue, animal organ or animal blood. More preferably the feed material is selected from: a composition comprising dairy lipids, a composition comprising egg lipids, and a composition comprising marine lipids. 15 Most preferably the feed material used in the process of the present invention is a bovine milk fraction. Preferably the feed material is selected from: buttermilk, a buttermilk fraction, beta serum, a beta serum fraction, butter serum, a butter serum fraction, whey, a whey fraction, colostrum, and a colostrum fraction. The feed material may comprise milk fat globule membrane. 20 Preferably, the feed material is in solid form. When solid, the feed material may be cryomilled before contact with the solvent. The solvent of the present invention preferably comprises: (a) an alcohol selected from: methanol, ethanol, n-propanol, isopropanol and mixtures thereof; and 25 (b) 0 - 40% v/v water More preferably the solvent comprises between 0 and 20% v/v water. Most preferably the solvent comprises between 1 and 10% v/v water. Preferably the alcohol is ethanol. 7 WO 2007/123424 PCT/NZ2007/000087 Preferably the solvent used in the process of the present invention comprises 95% aqueous ethanol. Preferably the mass fraction of the co-solvent in CO 2 is between 5% and 60%. More preferably the mass fraction is between 20% and 50%. Most preferably the mass fraction is 5 between 25% and 30%. Preferably the contacting temperature between the feed material and solvent is between 10 0 C and 80 0 C. More preferably the contacting temperature is between 55 0 C and 65 0 C. Most preferably the contacting pressure is between 100 bar and 500 bar. Preferably the contacting pressure is between 200 bar and 300 bar. More preferably the ratio 10 of the co-solvent to feed material is in the range 10:1 to 200:1. Most preferably the ratio of the co-solvent to feed material is in the range 15:1 to 50:1. Preferably the separating pressure is between atmospheric pressure and 90 bar. More preferably the separating pressure is between 40 bar and 60 bar. Preferably the co-solvent is recycled for further use. 15 Preferably the CO 2 is recycled for further use. The co-solvent may be removed by evaporation under vacuum. Preferably the feed material is contacted with a continuous flow of solvent. Preferably the feed material is contacted with one or more batches of solvent. The lipid and solvent streams may be fed continuously. 20 Optionally, the feed material and co-solvent streams may be mixed prior to contacting with
CO
2 . The invention also provides products produced by the process of the invention, both the insoluble components remaining after contact with the solvent (also referred to herein as the "residue"); and the soluble components that are dissolved in the solvent after contact with 25 the feed material (also referred to herein as the "extract"). Where the feed material is contacted with more than one batch of solvent, or the solvent is cooled in a number of steps, there will be multiple "extract" products. 8 WO 2007/123424 PCT/NZ2007/000087 Preferably the product contains more sphingomyelin than the feed material. More preferably the product comprises greater than 3% sphingomyelin. Even more preferably the product comprises greater than 10% sphingomyelin. Most preferably the product comprises greater than 15% sphingomyelin. 5 Preferably the product contains more phosphatidyl serine than the feed material. More preferably the product comprises greater than 5% phosphatidyl serine. Even more preferably the product comprises greater than 30% phosphatidyl serine. Most preferably the product comprises greater than 70% phosphatidyl serine. Preferably the product contains more gangliosides than the feed material. More preferably 10 the product comprises greater than 2% gangliosides. Even more preferably the product comprises greater than 4% gangliosides. Most preferably the product comprises greater than 6% gangliosides. Preferably the product contains more cardiolipin than the feed material. More preferably the product comprises greater than 5% cardiolipin. Even more preferably the product comprises 15 greater than 10% cardiolipin. Most preferably the product comprises greater than 25% cardiolipin. Preferably the product contains more acylalkyphospholipids and/or plasmalogens than the feed material. More preferably the product comprises greater than 5% acylalkyphospholipids and/or plasmalogens. Even more preferably the product comprises 20 greater than 10% acylalkyphospholipids and/or plasmalogens. Most preferably the product comprises greater than 25% acylalkyphospholipids and/or plasmalogens. Preferably the product contains more aminoethylphosphonate and/or other phosphonolipids than the feed material. More preferably the product comprises greater than 5% aminoethylphosphonate and/or other phosphonolipids. Even more preferably the product 25 comprises greater than 10% aminoethylphosphonate and/or other phosphonolipids. Most preferably the product comprises greater than 25% aminoethylphosphonate and/or other phosphonolipids. 9 WO 2007/123424 PCT/NZ2007/000087 BRIEF DESCRIPTION OF THE DRAWINGS The invention may be more fully understood by having reference to the accompanying drawings wherein: Figure 1 is scheme drawing illustrating a preferred process of the current invention. 5 Figure 2 is a scheme drawing illustrating a second preferred process of the current invention Figure 3 is a scheme drawing illustrating a third preferred process of the current invention Figure 4 is a scheme drawing illustrating a fourth preferred process of the current invention 10 ABBREVIATIONS AND ACRONYMS In this specification the following are the meanings of the abbreviations or acronyms used. "CL" means cardiolipin "PC" means phosphatidyl choline "PI" means phosphatidyl inositol 15 "PS" means phosphatidyl serine "PE" means phosphatidyl ethanolamine "PA" means phosphatidic acid "PL" means plasmalogen "PP" means phosphonolipid 20 "ALP" means alkylacylphospholipid "SM" means sphingomyelin "CAEP" means ceramide aminoethylphosphonate "GS" means ganglioside "N/D" means not detected 25 "CO 2 " means carbon dioxide 10 WO 2007/123424 PCT/NZ2007/000087 GENERAL DESCRIPTION OF THE INVENTION As discussed in the Background, it is known that supercritical CO 2 with up to 12.5% ethanol as a co-solvent can extract the phospholipids PC, and to a much lesser extent, PE and PI 5 from soy or egg. Surprisingly, we have found that the phospholipids PS, CAEP and CL; and gangliosides are virtually insoluble in CO 2 and a Ci-C 3 monohydric alcohol co-solvent, and that SM, ALP, PL and PP are soluble. Therefore it is possible to separate the soluble phospholipids from the insoluble phospholipids and gangliosides to achieve fractions enriched in one or other of the desired components. 10 There are a number of factors affecting the operation of the process: * Feed material and feed preparation * Extraction temperature and pressure 15 * Co-solvent concentration * Total solvent throughput * Solvent flow rate and contacting conditions It is advantageous to start with a feed material containing at least 5 % by mass of lipids, and 20 ideally at least 2 % by mass of phospholipids, particularly PS, SM, CL, ALP, PL, PP, CAEP and/or gangliosides. The feed material can be processed using pure CO 2 before the co-solvent is introduced to remove much or all of neutral lipids. This reduces the neutral lipid content in the C0 2 +co 25 solvent extract leading to an extract enriched in soluble phospholipids and/or gangliosides. The form of the feed material depends on the source of the lipids and its lipid composition. For example dairy lipid extracts high in phospholipids may be substantially solid even at elevated temperatures. Egg yolk and marine lipids in comparison have a lower melting point. 30 The presence of neutral lipids also tends to produce a more fluid feed material. To promote good contacting it may be beneficial to prepare the feed material. Solid materials containing lipids may be able to be cryomilled. Lipid feed materials can also be made more fluid by the inclusion of some ethanol or water. 11 WO 2007/123424 PCT/NZ2007/000087 Changing the processing conditions of temperature, pressure, co-solvent concentration, and total solvent usage, influences the amount of material extracted, the purity of the final product, and the recovery (or efficiency) of the process. For example, the virtually insoluble lipids such as PS, GS, CAEP and CL, have very slight solubilities so that excessive use of 5 solvent, or very favourable extraction conditions, can result in small losses of PS, GS and CL from the residual fraction. A high purity product may be achieved, but with a reduced yield. Conversely the enrichment of soluble lipids will be greater if smaller amounts of the other lipids are co-extracted, but the total yield will be lower. Processing economics, and the relative values of the products, will determine where this balance lies. A further option to 10 obtain multiple enriched fractions is to carry out extractions under progressively more favourable extraction conditions, such as increasing the temperature. We have found that co-solvent concentrations below about 10% produce very little extract of phospholipids and/or gangliosides. At higher concentrations the rate of material extracted 15 increases rapidly. We have found the co-solvent concentrations of at least 20%, and more preferably 30% achieve high levels of extraction of PC, PE, SM, ALP, PL, PP and PI, while the lipids PS, CL and GS remain virtually insoluble. Every substance has its own "critical" point at which the liquid and vapour state of the 20 substance become identical. Above but close to the critical point of a substance, the substance is in a fluid state that has properties of both liquids and gases. The fluid has a density similar to a liquid, and viscosity and diffusivity similar to a gas. The term "supercritical" as used herein refers to the pressure-temperature region above the critical point of a substance. The term "subcritical" as used herein refers to the pressure-temperature 25 region equal to or above the vapour pressure for the liquid, but below the critical temperature. The term "near-critical" as used herein encompasses both "supercritical" and "subcritical" regions, and refers to pressures and temperatures near the critical point. Percentages unless otherwise indicated are on a w/w solids basis. 30 The term "comprising" as used in this specification means "consisting at least in part of". When interpreting each statement in this specification that includes the term "comprising", features other than that or those prefaced by the term may also be present. Related terms such as "comprise" and "comprises" are to be interpreted in the same manner. 35 12 WO 2007/123424 PCT/NZ2007/000087 The invention consists in the foregoing and also envisages constructions of which the following gives examples only. EXAMPLES 5 The experimental process is described, with reference to figure 1, as follows. A measured mass of feed material containing lipids to be fractionated was placed in basket BK1 with a porous sintered steel plate on the bottom. Basket BK1 was placed in a 300 mL extraction vessel EX1. The apparatus was suspended in heated water bath WB1 and 10 maintained at a constant temperature through use of a thermostat and electric heater. In the continuous extraction mode of operation, liquid CO 2 from supply bottle B 1 was pumped using pump P1 into extraction vessel EX1 until the pressure reached the desired operating pressure, after which valve V 1 was operated to maintain a constant pressure in the 15 extraction vessel. After passing through valve V1, the pressure was reduced to the supply cylinder pressure of 40 to 60 bar, which caused the CO 2 to be converted to a lower density fluid and lose its solvent strength. Precipitated material was captured in separation vessel SEP1, and the CO 2 exited fr-om the top of separator SEP1 and was recycled back to the feed pump through coriolis mass flow meter FM1 and cold trap CT1 operated at -5 0 C. Extracted 20 material was collected periodically from separator SEP1 by opening valve V2. The extraction was optionally carried out using CO 2 only until all of the compounds soluble in
CO
2 only, such as neutral lipids, were extracted. When no further extract was produced by
CO
2 extraction, ethanol co-solvent with or without added water was added to the CO 2 at the desired flow ratio from supply bottle B2 using pump P2. Ethanol and further extracted 25 material were separated from the CO 2 in separator SEP1 and periodically removed through valve V2. After the desired amount of ethanol had been added the ethanol flow was stopped and the CO 2 flow continued alone until all the ethanol had been recovered from the system. The remaining CO 2 was vented and the residual material in basket BK1 was removed and dried under vacuum. The extract fraction was evaporated to dryness by rotary evaporation. 30 In the batch extraction mode of operation CO 2 alone was optionally passed continuously through the apparatus, as for the continuous flow mode of operation, until all CO 2 alone extractable material was removed. The CO 2 flow was then stopped and valve V1 closed to maintain the pressure. Approximately 140g of ethanol was pumped from supply bottle B2 13 WO 2007/123424 PCT/NZ2007/000087 through pump P2 into extraction vessel EX1. The system was left for 15 minutes to allow the system to equilibrate, after which time the CO 2 flow was started and valve V1 opened to maintain a constant pressure and allow ethanol and dissolved compounds to flow through to separator SEP1. This process was repeated twice more, after which the CO 2 was vented and 5 the residual material in basket BK1 was removed and dried under vacuum. Extract and residue fractions were analysed for phospholipid content and profile by 3 1
P
NMR. The phospholipid mass fractions reported here are for phosphatidylcholine (PC), phosphatidylinositol (PI), phosphatidylethanolamine (PE), plasmalogens (PL), 10 phosphonolipids (PP), alkylacylphospholipids (ALP), sphingomyelin (SM), ceramide aminoethylphosphonate (CAEP), phosphatidylserine (PS), and cardiolipin (CL). The process option illustrated in Figure 1 is for a batch process while the processing options illustrated in Figures 2-4 are for a continuous flow process. 15 Example 1: Fractionation of dairy lipid extract A, ethanol mass fraction 25% Lipid extract A is a total lipid extract obtained by a processes disclosed in PCT international applications PCT/NZ2005/000262 (published as WO 2006/041316). 20 40g of dairy lipid extract A, with composition shown in Table 1 (feed), was extracted using the continuous extraction mode of operation at 60 0 C and 300 bar. The 'other compounds' consist mainly of neutral lipids. 44% of the feed material was extracted (extract 1) using CO 2 only. This extract contained no phospholipids, and was entirely neutral lipids. A further 31% of the feed material (extract 2) was extracted using 95% aqueous ethanol at a concentration 25 in CO 2 of 25%. The total ethanol and water added was 880g. The composition of the fraction extracted with CO 2 and ethanol (extract 2), and the composition of the residual fraction are shown in Table 1. The extract is enriched in phosphatidylcholine (PC) and sphingomyelin (SM) which are more soluble in CO 2 and ethanol, while the residual fraction is substantially enriched in phosphatidylserine (PS). Phosphatidylserine levels are virtually undetectable in 30 the extract phase indicating very low solubility in CO 2 and ethanol, and almost complete recovery of phosphatidylserine in the residue phase. 14 WO 2007/123424 PCT/NZ2007/000087 Table 1 Composition, % Yield Other Other compounds % of feed PC PI PS PE SM Phospholipids Feed 11.2 2.8 4.3 13.2 7.8 2.2 58.3 Extract 2 31 28.2 0.0 0.2 14.4 15.4 4.9 37.0 Residue 25 6.5 10.5 15.6 30.8 10.2 3.6 22.8 5 Example 2: Fractionation of dairy lipid extract A, ethanol mass fraction 31% 41g of dairy lipid extract A, with composition as for example 1 was extracted using the 10 continuous extraction mode of operation at 60 0 C and 300 bar as for example 1, using firstly
CO
2 alone to extract 50 % of the feed material (extract 1), which is neutral lipids only, and then using 95% aqueous ethanol at a concentration in CO 2 of 31%. 33% of the feed material was extracted (extract 2). The total ethanol and water added was 1150g. The composition of the residual fraction is shown in Table 2. The higher ethanol concentration gives a more 15 complete extraction of lipids and the concentration of phosphatidylserine in the residue fraction is higher than found in example 1 at 19.3 %. Table 2 Composition, % Yield Other Other compounds % of feed PC PI PS PE SM Phospholipids Feed 11.2 2.8 4.3 13.2 7.8 2.2 58.3 Extract 2 33 - - - - Residue 17 4.4 12.6 19.3 27.1 8.5 2.5 25.5 20 Example 3: Fractionation of dairy lipid extract A, ethanol mass fraction 43% 40g of dairy lipid extract A, with composition as for example 1 was extracted using the 25 continuous extraction mode of operation at 60 0 C and 300 bar as for example 1, using firstly
CO
2 alone to extract 41 % of the feed material (extract 1), which is neutral lipids only, and then using 95% aqueous ethanol at a concentration in CO 2 of 43% to extract 32 % of the feed (extract 2). The total ethanol and water added was 960g. The composition of extract 2 and residual fractions are shown in Table 3. The concentration of phosphatidylserine in the 30 residue fraction is higher than found in example 1 and example 2 at 20.7 %. The 15 WO 2007/123424 PCT/NZ2007/000087 concentration of SM in the extract, at 12.5 % by mass, is enriched relative to the feed, at 7.8 % by mass, even though it also contains a high level of neutral lipids. Table 3 Composition, % Yield Other Other compounds % of feed PC PI PS PE SM Phospholipids Feed 11.2 2.8 4.3 13.2 7.8 2.2 58.3 Extract 2 32 21.1 0.0 0.5 13.3 12.5 3.5 49.1 Residue 27 4.2 13.6 20.7 26.7 7.8 1.9 25.0 5 Example 4: Fractionation of dairy lipid extract A, 40 0 C 39g of dairy lipid extract A, with composition as for example 1 was extracted using the continuous extraction mode of operation at 300 bar using firstly CO 2 alone to extract 54 % of 10 the feed material (extract 1), which is neutral lipids only, and then using 95% aqueous ethanol at a concentration in CO 2 of 30 % to extract 12 % of the feed (extract 2). The temperature in this example was 40 0 C. The total ethanol and water added was 975g. The composition of the extracted and residual fractions are shown in Table 5. The degree of extraction of SM is lower than for examples 1 to 3 at 60 0 C, but the concentration in the 15 extract is higher. The concentration of PS in the residue, at 12.4 %, is lower than examples 1 to 3. Table 4 Composition, % Yield Other Other compounds % of feed PC PI PS PE SM Phospholipids Feed 11.2 2.8 4.3 13.2 7.8 2.2 58.3 Extract 2 12 27.9 0.0 0.3 16.7 15.3 4.9 34.9 Residue 34 9.9 8.1 12.4 25.3 12.2 3.6 28.5 20 Example 5: Fractionation of dairy phospholipid concentrate 40g of a dairy phospholipid concentrate with composition as shown in Table 5 (feed) was extracted using the continuous extraction mode of operation at 300 bar and 60 0 C without the prior CO 2 only extraction step. The ethanol (95% aqueous ethanol) mass fraction in CO 2 was 25 30%. The total ethanol and water added was 1026g. The composition of the extracted and residual fractions are shown in Table 5. Only 11% of the feed lipid was extracted, so the enrichment ofphosphatidylserine in the residue is not significant, but the concentration did increase from 8% to 8.8%. The poor degree of extraction in this example is due to the 16 WO 2007/123424 PCT/NZ2007/000087 physical properties of the solid feed material limiting mass transfer. In comparison, the dairy lipid extract in examples 1 through 4, is liquid at the processing temperature and better extraction rates are observed. 5 Different feed preparation methods and/or longer equilibration times and/or greater solvent quantities are expected to increase the amount of extractable material. Table 5 Composition, % Yield Other Other compounds % of feed PC PI PS PE SM Phospholipids Feed 15.4 5.3 8.0 21.6 15.1 0.3 34.3 Extract 11 Residue 89 13.0 5.9 8.8 21.4 10.9 2.8 37.2 10 Example 6: Fractionation of dairy phospholipid concentrate using the batch extraction process 19g of a dairy phospholipid concentrate with composition as described in example 5 was extracted using the batch extraction mode of operation at 300 bar and 60 0 C. A total of 22% 15 of the feed mass was extracted in three sequential extractions each consisting of 140g of ethanol (95% aqueous ethanol) in 300mL of CO 2 . The composition of the extracted and final residual fractions are shown in Table 6. In this example 22% of the feed lipid was extracted, significantly higher than that obtained in the continuous extraction example (example 5) and using a lower total quantity of ethanol co-solvent. The phosphatidylserine concentration in 20 the residue has increased from 8% to 11.2%; and the sphingomyelin concentration in the extract has increased from 15.1 to 16.7 %. This example shows the increase in total extracted material by allowing a greater contacting time to more completely dissolve the soluble fraction. 25 Table 6 YieldComposition, % Yield Other Other compounds % of feed PC PI PS PE SM Phospholipids Feed 15.4 5.3 8.0 21.6 15.1 0.3 34.3 Extract 22 32.4 0.5 0.4 17.7 16.7 4.7 27.5 Residue 78 13.6 7.4 11.2 26.6 13.3 2.9 25.0 17 WO 2007/123424 PCT/NZ2007/000087 Example 7: Fractionation of dairy lipid extract B, ethanol mass fraction 10% This example relates to extraction of dairy lipid extract B, a total lipid extract obtained from 5 high fat whey protein concentrate processes disclosed in PCT international applications PCT/NZ2004/000014 (published as WO WO2004/066744). with composition shown in Table 7 (feed). The 'other compounds' listed include 2-3% gangliosides and about 3% lactose, both absent in dairy lipid extract A. In this example 42g of dairy lipid extract B was extracted using the continuous extraction mode of operation at 10 300 bar and 60 0 C. 52% of the feed mass was extracted using CO 2 alone (extract 1). Only 3% of the feed lipid was further extracted using 460g of 95% aqueous ethanol (extract 2), and the extract contained less than 10% phospholipids. The extraction of phospholipids does not occur to any significant extent for ethanol mass fractions of 10% or lower. The ethanol does however extract some additional neutral lipid that is not extracted using CO 2 alone. In this 15 case, both the PS and SM are enriched in the residue. Table 7 Composition, % Yield Other Other compounds % of feed PC PI PS PE SM Phospholipids Feed 7.4 2.5 3.9 10.3 5.7 1.3 69.0 Extract 2 3 4.5 0.0 0.0 1.6 1.0 0.3 92.6 Residue 45 15.0 6.1 8.7 21.8 12.0 5.9 30.7 20 Example 8: Fractionation of dairy lipid extract B, ethanol mass fraction 30% In this example 40g of dairy lipid extract B was extracted using the continuous extraction 25 mode of operation at 300 bar and 60 0 C. 51% of the feed mass was extracted using CO 2 alone (extract 1). A further 7% of the feed material was extracted using 760g of 95% aqueous ethanol at a mass concentration of 30% in CO 2 (extract 2). Phospholipid profiles for the extract and residual fractions are shown in Table 8. Both PS and SM are enriched in the residue 18 WO 2007/123424 PCT/NZ2007/000087 Table 8 Composition, % Yield Other Other compounds % of feed PC PI PS PE SM Phospholipids Feed 7.4 2.5 3.9 10.3 5.7 1.3 69.0 Extract (2) 7 22.5 0.5 0.4 14.0 11.2 3.3 48.2 -Residue 41 12.0 5.5 8.5 20.2 10.0 2.4 41.5 Example 9: Fractionation of dairy lipid extract A, ethanol mass 5 fraction 3% This example shows that when the co-solvent concentration is below 10% by mass, no phospholipids are extracted. 10 In this example 27g of dairy lipid extract A, as described in example 1, was extracted using the continuous extraction mode of operation at 300 bar and 60 0 C, using 98% ethanol at 3 % by mass ratio with C0 2 , without the CO 2 only extraction step. 62% of the feed mass was extracted. No detectable phospholipids were extracted. This extract represents 90% of the neutral lipid present in the feed material. The rate of extraction of neutral lipid from the feed 15 material was substantially faster using the ethanol co-solvent than using CO 2 only. The extract material was substantially extracted using less than the total of 150g of ethanol in 4850g of CO 2 used, while typically 10 kg of CO 2 alone is required for extraction of neutral lipids, as in example 1. 20 Example 10: Fractionation of egg yolk lecithin This example relates to fractionation of a commercially available egg yolk lecithin, with phospholipid profile shown in Table 9. No phosphatidylserine was detected in the feed lipid, indicating concentration levels <0.5%. In this example 34g of the feed material was 25 extracted using the continuous extraction mode of operation at 300 bar and 60 0 C, and 95% aqueous ethanol at a concentration of 25%. 45% of the feed mass was extracted as neutral lipids using CO 2 alone. A further 49% of the feed material was extracted using ethanol and
CO
2 with a total ethanol flow of 640g. Phospholipid profiles for the extract and residual fractions are shown in Table 9. In this example, the phosphatidylserine levels in the residual 30 material are substantially enriched compared with non-detectable levels in the feed material. 19 WO 2007/123424 PCT/NZ2007/000087 Table 9 Composition, % Yield Other Other compounds % of feed PC PI PS PE SM Phospholipids Feed 56.4 N/D N/D 6.4 2.0 5.7 29.4 Extract 49 43.5 N/D N/D 9.2 2.6 2.1 42.5 Residue 6 17.4 8.0 5.9 19.1 3.8 3.8 42.0 Example 11: Fractionation of egg yolk phospholipid extract 5 This example relates to fractionation of an egg yolk phospholipid fraction with phospholipid profile shown in Table 9. In this example 40g of the feed material was extracted using the continuous extraction mode of operation at 300 bar and 60'C, and 95% aqueous ethanol at a concentration of 28%. 50% of the feed mass was extracted as neutral lipids using CO 2 alone. A further 46% of the feed material was extracted using ethanol and CO 2 with a total ethanol 10 flow of 800g. Phospholipid profiles for the extract and residual fractions are shown in Table 10. In this example, the phosphatidylserine levels in the residual material are substantially enriched compared with levels in the feed material, while sphingomyelin is enriched in the extract relative to the feed. 15 Table 10 Composition, % Yield Other Other compounds % of feed PC PI PS PE SM Phospholipids Feed 21.2 0.6 0.4 5.2 1.6 0.9 70.1 Extract 46 65.6 0.3 N/D 6.3 2.8 2.3 22.8 Residue 4 12.9 11.2 8.2 27.6 2.8 8.2 29.2 Example 12: Fractionation of Hoki head lipid extract This example relates to fractionation of a Hoki head lipid extract with phospholipid profile 20 shown in Table 11. In this example 25g of the feed material was extracted using the continuous extraction mode of operation at 300 bar and 60 0 C, and 95% aqueous ethanol at a concentration of 31%. 1% of the feed mass was extracted as neutral lipids using CO 2 alone. A further 72% of the feed material was extracted using ethanol and CO 2 with a total ethanol flow of 940g. Phospholipid profiles for the extract and residual fractions are shown in Table 25 11. In this example, the phosphatidylserine levels in the residual material are substantially enriched compared with levels in the feed material. Some PS is also observed in the extract phase. The alkylacylphosphatidylcholine (AAPC), a type of alkylacylphospholipid, is completely extracted. 20 WO 2007/123424 PCT/NZ2007/000087 Table 11 Composition, % Yield Other Other compounds % of feed PC PI PS PE SM AAPC phosph Feed 1 9.2 1.1 1.4 4.8 0.5 1.1 1.8 80.8 Extract 72 14.2 0.0 0.7 5.3 0.5 1.6 0.6 71.2 Residue 27 14.3 7.1 7.6 13.9 0.0 0.0 6.2 47.7 Example 13: Fractionation of bovine heart lipid extract 5 This example relates to fractionation of a bovine heart phospholipid lipid extract with phospholipid profile shown in Table 9. In this example 40g of the feed material was extracted using the continuous extraction mode of operation at 300 bar and 60 0 C, and 95% aqueous ethanol at a concentration of 33% in CO 2 . No lipid was extracted using CO 2 alone. 10 79% of the feed material was extracted using ethanol and CO 2 with a total ethanol flow of 960g. Phospholipid profiles for the extract and residual fractions are shown in Table 12. The phosphatidylserine levels in the residual material are substantially enriched compared with levels in the feed material. Cardiolipin is also significantly enriched in the residue. 15 Table 12 Composition, wt% Yield Other wt% of Phospholipid Other compounds feed CL PC PI PS PE SM s Feed 16.8 13.4 3.2 1.5 12.3 3.6 15.3 33.9 Extract 79 8.2 - 18.6 0.8 0.4 8.6 3.5 13.1 46.7 Residue 21 42.2 2.8 14.1 4.7 23.4 12.8 0.0 Example 14: Fractionation of dairy lipid extract A with propan-2-ol co-solvent 20 In this example 39g of the dairy lipid extract A, as described in example 1, was extracted using the continuous extraction mode of operation at 300 bar and 60oC, and 95% aqueous propan-2-ol at a mass concentration of 35% in CO 2 . 48% of the feed material was extracted as neutral lipids using CO 2 alone. 23% of the feed material was further extracted using the 25 propan-2-ol co-solvent and CO 2 with a total propanol mass of 810g. Phospholipid profiles for the extract and residual fractions are shown in Table 13. The phosphatidylserine levels in the residual material are substantially enriched, and the result is comparable to results for examples 1 and 2. A slightly lower total PS level is achieved than for example 2 using a 21 WO 2007/123424 PCT/NZ2007/000087 comparable concentration of ethanol. The levels of PS observed in the extracted fraction is also higher suggesting the propan-2-ol is not as selective as ethanol. On this basis alone ethanol would be the preferred co-solvent. 5 Table 13 _Composition, % Yield Other Other compounds % of feed PC PI PS PE SM Phospholipids Feed 11.2 2.8 4.3 13.2 7.8 2.2 58.3 Extract 23 27.9 0.8 1.3 19.5 14.0 4.2 32.4 Residue 29 10.7 8.6 13.0 23.8 15.5 3.4 25.0 Example 15: Fractionation of soy lecithin This example relates to fractionation of a soy lecithin (Healtheries Lecithin natural dietary 10 supplement, Healtheries of New Zealand Limited) with composition shown in Table 9. In this example 42g of feed material was extracted using the continuous extraction mode of operation at 300 bar and 60 0 C, and 95% aqueous ethanol at a concentration of 33% in CO 2 . No lipid was extracted using CO 2 alone. 91% of the feed material was extracted using ethanol and CO 2 with a total ethanol flow of 520g. Phospholipid profiles for the extract and 15 residual fractions are shown in Table 14. PC and PE are preferentially extracted and are significantly enriched in the extract. There are no detectable levels of PS or SM in this example. Table 14 Composition, % Yield Other Other compounds % of feed PC PI PS PE SM Phospholipids Feed 22.2 12.3 0.0 17.4 0.0 11.7 36.4 Extract 9 31.9 0.7 0.0 9.9 0.0 6.1 51.4 Residue 91 20.7 13.2 0.0 18.4 0.0 12.4 35.2 20 Example 16: Continuous fractionation of egg yolk lipids This example relates to fractionation of an egg yolk lipid extract containing 15% 25 phospholipids and the balance mostly neutral lipids by HPLC analysis. The phospholipid fraction contains 55% PC, 29% PE, and 14% PI. The feed lipid was pumped into the top of a 10L pressure vessel, and contacted with CO 2 containing 8.7 % of 98% aqueous ethanol flowing upwards through the vessel at 300 bar pressure and temperature of 60 0 C. An extract phase was continuously taken from the top of the contacting vessel, and a raffinate phase 22 WO 2007/123424 PCT/NZ2007/000087 was periodically withdrawn from the bottom of the vessel. The lipid feed rate was 1.5 kg/hr. The C0 2 + co-solvent flow rate was 27 kg/hr. The extract phase was predominantly neutral lipids but contained 20% of the phospholipids present in the feed stream. The phospholipids in the extract fraction consisted of between 5 70% and 100% PC, with the balance mostly PE. This represents a preferential extraction of PC over other phospholipids. In a second experiment, feed lipid was premixed with 98% ethanol (with 2 % water) at a concentration of 10.2% lipid. This mixture was pumped into the top of the pressure vessel and contacted with CO 2 in upflow. The overall concentration of ethanol in CO 2 under steady 10 state processing conditions was 5.9%. In this case 50% of the mass of phospholipids in the feed were extracted. The composition of the extract phase consisted of between 60% and 70% PC, with the balance mostly PE. The presence of PI and other phospholipids in the extract was not detectable by the HPLC method used. Example 17: Fractionation of green-lipped mussel lipid extract 15 This example relates to fractionation of a green-lipped mussel lipid extract with phospholipid profile shown in Table 11. In this example 12.2g of the feed material was extracted using a batch stirred tank method at 250 bar and 60oC using CO 2 and ethanol (containing 5 % water) at a concentration of 30.5 %. The lipid was placed in the stirred tank, CO 2 was added to give 20 the desired pressure and then the 95 % ethanol was added in during constant stirring. 65 % of the feed material was then extracted using CO 2 and ethanol after stirring for 1 hour by sampling the extract phase at constant pressure. Phospholipid profiles for the extract and residual fractions are shown in Table 15. In this example, the CAEP levels in the residual material are substantially enriched compared with levels in the feed material. The 25 alkylacylphosphatidylcholine (AAPC), a type of alkylacylphospholipid, is partially extracted. Table 15 Yield Composition, % Yield Other compounds %of feed PC PI PS PE SM AAPC CAEP Feed 1.89 0.0 0.0 1.60 0.0 0.8 2.0 92.2 Extract 65.3 1.97 0.0 0.0 0.0 0.0 0.9 0.71 96.0 Residue 34.7 3.77 0.0 0.0 3.67 0.0 1.3 3.22 84.0 23 WO 2007/123424 PCT/NZ2007/000087 Example 18: Fractionation of krill lipids This example shows the fractionation of krill lipids from krill powder and demonstrates concentration of AAPC in the extract, and AAPE in the residue. 5619.9 g of freeze-dried krill powder containing 21.4 % lipid and corresponding phospholipids concentrations shown 5 in table 16 was extracted continuously with supercritical CO 2 at 300 bar and 313 K until no further extract was obtained. This extract (extract 1) contained no phospholipids, and was substantially all neutral lipids. A total of 650 g of this extract was obtained, and 66.41 kg of
CO
2 was used. The residual powder was then extracted with CO 2 and absolute ethanol, using a mass ratio of ethanol to CO 2 of 11 %. The CO 2 and ethanol extract phase was passed 10 through two sequential separators in which the pressure was 95 and 60 bar respectively. The bulk of the phospholipids-rich extract (extract 2) was obtained in the first separator, and the bulk of the co-solvent in the second separator (extract 3). The composition of extract 2 and residual powder are shown in table 16. The alkylacylphosphatidylcholine (AAPC), a type of alkylacylphospholipid, is highly enriched in the concentrated phospholipids-rich extract, 15 whilst alkylacylphosphatidylethanolamine (AAPE), another type of alkylacylphospholipid, is not extracted to any great degree. Table 16 Composition, % Yield Other compounds %of feed PC PI PS PE CL AAPC AAPE Feed 6.6 0.0 0.0 0.4 0.1 0.6 0.1 78.6 Extract 2 4.3 39.8 0.0 0.0 0.3 0.2 4.6 0.2 53.7 Residue 79.2 3.6 0.0 0.0 0.3 0.2 0.5 0.1 93.4 Example 18: Fractionation of dairy lipids from beta-serum powder 20 This example shows the fractionation of dairy lipids from beta-serum powder (a milk fat globular membrane concentrate powder) and demonstrates concentration of PS in the residual powder, and concentration of SM in the extract obtained using supercritical CO 2 + ethanol. 5835.3 grams of beta-serum powder containing phospholipids in the concentrations shown in table 17, was extracted continuously with supercritical CO 2 at 300 bar and 313 K 25 until no further extract was obtained. This extract contained no phospholipids, and was substantially all neutral lipids. 1085.6 g of this extract (extract 1) was obtained using 94.42 kg of CO 2 . 2906.3 grams of the residual powder was then re-extracted with CO 2 and anhydrous ethanol at 300 bar and 323 K, using a mass ratio of ethanol to CO 2 of 25 %. The powder was extracted with this mixture for 90 minutes (7.82 kg ethanol). The CO 2 and 24 WO 2007/123424 PCT/NZ2007/000087 ethanol extract phase was passed through two sequential separators in which the pressure was 100 (extract 2) and 54 bar (extract 3) respectively. The extract was split between both separators. A total of 262.2 g of extract was obtained. The composition of the combined extract (extract 2 and 3) and residual powder are shown in table 17. The extract is highly 5 enriched in sphingomyelin, whilst the residue is enriched in phosphatidylserine. Table 17 Composition, % Yield Other Other compounds % of feed PC PI PS PE SM Phospholipids Feed 4.9 1.5 2.3 5.6 4.3 0.1 81.3 Extract 9.02 2+3 49.6 0.0 0.0 12.4 30.1 0.7 7.1 Residue 71.14 0.3 2.0 3.0 3.0 0.5 0.1 91.1 INDUSTRIAL APPLICATION The present invention has utility in providing products with high levels of particular 10 phospholipids and/or glycolipids including cardiolipin and phosphatidyl serine, and sphingomyelin. The described compositions and methods of the invention may be employed in a number of applications, including infant formulas, brain health, sports nutrition and dermatological compositions. REFERENCES 15 1. Palacios, L.E., Wang, T., Extraction of egg-yolk lecithin, JAOCS 82,8,2005 2. Kang, D.H., Row, K.H., Fractionation of soybean phospholipids by preparative high performance liquid chromotography with sorbents of various particle size, Journal of Chromatography A, 949, 2002 20 3. Kearns, J.J., Tremblay, P.A., Robey, R.J., Sunder, S., Process for purification of phospholipids, US patent 4814111, 1989 4. Teberikler, L., Koseoglu, S., Akgerman, A., Selective extraction of phosphatidylcholine from lecithin by supercritical carbon dioxide/ethanol mixture, JAOCS 78,2,2001 25 WO 2007/123424 PCT/NZ2007/000087 5. Montanari, L., Fantozzi, P., Snyder, J.M., King, J.W., Selective extraction of phospholipids from soybeans with supercritical carbon dioxide and ethanol, J Supercritical Fluids, 14, 1999 6. Taylor, S.L., King, J.W., Montanari, L., Fantozzi, P., Blanco, M.A., Enrichment and 5 fractionation of phospholipid concentrates by supercritical fluid extraction and chromatography, Ital J Food Sci, 1,12,2000 7. Tanaka, Y., Sakaki, I., Extraction of phospholipids from unused natural resources with supercritical carbon dioxide and an entrainer, Journal of oleo science, 54, 11, 2005 10 8. Bulley, N.R., Labay, L., Arntfield, S.D., Extraction/Fractionation of egg yolk using supercritical CO2 and alcohol entrainers, J supercritical fluids, 5, 1992. 26
Claims (70)
1. A process for separating a feed material into soluble and insoluble components, comprising 5 (e) providing a feed material comprising one or more of: (i) at least 1% by mass phosphatidyl serine (ii) at least 1% by mass sphingomyelin (iii) at least 0.3 % by mass acylalkylphospholipids and/or plasmalogens (iv)at least 0.5 % by mass aminoethylphosphonate and/or other phosphonolipids 10 (v) at least 1% by mass cardiolipin (vi) at least 0.3% by mass gangliosides (f) providing a solvent comprising: (i) supercritical or near-critical CO 2 , and (ii) a co-solvent comprising one or more C 1 -C 3 monohydric alcohols, and water 15 wherein the co-solvent makes up at least 10% by mass of the C0 2 , and the water content of the co-solvent is 0 to 40 % by mass (g) contacting the feed material and the solvent and subsequently separating the solvent containing the soluble components from the insoluble components (h) optionally separating the soluble components and the solvent.
20. 2. The process of claim 1 wherein the feed material comprises greater than 1% phosphatidyl serine. 3. The process of claim 1 wherein the feed material comprises greater than 2% phosphatidyl serine. 4. The process of claim 1 wherein the feed material comprises greater than 5% 25 phosphatidyl serine. 27 WO 2007/123424 PCT/NZ2007/000087 5. The process of claim 1 wherein the feed material comprises greater than 1% sphingomyelin. 6. The process of claim 1 wherein the feed material comprises greater than 5% sphingomyelin. 5 7. The process of claim 1 wherein the feed material comprises greater than 15% sphingomyelin. 8. The process of claim 1 wherein the feed material comprises greater than 1% cardiolipin. 9. The process of claim 1 wherein the feed material comprises greater than 2% cardiolipin. 10. The process of claim 1 wherein the feed material comprises greater than 5% cardiolipin. 10 11. The process of claim 1 wherein the feed material comprises greater than 0.3% gangliosides. 12. The process of claim 1 wherein the feed material comprises greater than 1% gangliosides. 13. The process of claim 1 wherein the feed material comprises greater than 2% 15 gangliosides. 14. The process of claim 1 wherein the feed material comprises greater than 0.5% acylalkyphospholipids and/or plasmalogens. 15. The process of claim 1 wherein the feed material comprises greater than 2% acylalkyphospholipids and/or plasmalogens. 20 16. The process of claim 1 wherein the feed material comprises greater than 10% acylalkyphospholipids and/or plasmalogens. 17. The process of claim 1 wherein the feed material comprises greater than 0.5% aminoethylphosphonate and/or other phosphonolipids. 18. The process of claim 1 wherein the feed material comprises greater than 5% 25 aminoethylphosphonate and/or other phosphonolipids. 19. The process of claim 1 Wherein the feed material comprises greater than 20% aminoethylphosphonate and/or other phosphonolipids. 28 WO 2007/123424 PCT/NZ2007/000087 20. A process for separating a feed material into soluble and insoluble components, comprising (h) providing a feed material comprising one or more of: (i) at least 1% by mass phosphatidyl serine, 5 (ii) at least 1% by mass sphingomyelin, (iii) at least 0.3 % by mass acylalkylphospholipids and/or plasmalogens (iv) at least 0.5 % by mass aminoethylphosphonate and/or other phosphonolipids (v) at least 1% by mass cardiolipin, or (vi) at least 0.3% by mass gangliosides 10 (i) providing a first solvent comprising supercritical or near-critical CO 2 (j) contacting the feed material and the first solvent and subsequently separating the first solvent containing the first soluble components from the first insoluble components (k) optionally separating the first soluble components and the first solvent (1) providing a second solvent comprising: 15 (iii) supercritical or near-critical CO 2 , and (iv)a co-solvent comprising one or more C 1 I-C 3 monohydric alcohols, and water wherein the co-solvent makes up at least 10% by mass of the CO 2 , and the water content of the co-solvent is 0 to 40% by mass (min) contacting the first insoluble components and the second solvent and subsequently 20 separating the second solvent containing the second soluble components from the second insoluble components (n) optionally separating the second soluble components and the second solvent.
21. The process of claim 20 wherein the first solvent comprises a mixture of supercritical or near-critical CO 2 and less than 10% C 1 -C 3 monohydric alcohol. 29 WO 2007/123424 PCT/NZ2007/000087
22. The process of claim 20 or claim 21 wherein the feed material comprises greater than 1% phosphatidyl serine.
23. The process of claim 20 or claim 21 wherein the feed material comprises greater than 2% phosphatidyl serine. 5 24. The process of claim 20 or claim 21 wherein the feed material comprises greater than 5% phosphatidyl serine.
25. The process of claim 20 or claim 21 wherein the feed material comprises greater than 1% sphingomyelin.
26. The process of claim 20 or claim 21 wherein the feed material comprises greater than 5% 10 sphingomyelin.
27. The process of claim 20 or claim 21 wherein the feed material comprises greater than 15% sphingomyelin.
28. The process of claim 20 or claim 21 wherein the feed material comprises greater than 1% cardiolipin. 15 29. The process of claim 20 or claim 21 wherein the feed material comprises greater than 2% cardiolipin.
30. The process of claim 20 or claim 21 wherein the feed material comprises greater than 5% cardiolipin.
31. The process of claim 20 or claim 21 wherein the feed material comprises greater than 20 0.3% gangliosides.
32. The process of claim 20 or claim 21 wherein the feed material comprises greater than 1% gangliosides.
33. The process of claim 20 or claim 21 wherein the feed material comprises greater than 2% gangliosides. 25 34. The process of claim 20 or claim 21 wherein the feed material comprises greater than 0.5% acylalkyphospholipids and/or plasmalogens. 30 WO 2007/123424 PCT/NZ2007/000087
35. The process of claim 20 or claim 21 wherein the feed material comprises greater than 2% acylalkyphospholipids and/or plasmalogens.
36. The process of claim 20 or claim 21 wherein the feed material comprises greater than 10% acylalkyphospholipids and/or plasmalogens. 5 37. The process of claim 20 or claim 21 wherein the feed material comprises greater than 0.5% aminoethylphosphonate and/or other phosphonolipids.
38. The process of claim 20 or claim 21 wherein the feed material comprises greater than 5% aminoethylphosphonate and/or other phosphonolipids.
39. The process of claim 20 or claim 21 wherein the feed material comprises greater than 10 20% aminoethylphosphonate and/or other phosphonolipids.
40. The process of any one of claim 1 to 39 wherein the feed material is derived from terrestrial animals, marine animals, terrestrial plants, marine plants, or micro-organisms such as microalgae, yeast and bacteria.
41. The process of claim 40 wherein the feed material is derived from sheep, goat, pig, 15 mouse, water buffalo, camel, yak, horse, donkey, llama, bovine or human.
42. The process of claim 40 or claim 41 wherein the feed material is selected from: tissue, a tissue fraction, organ, an organ fraction, milk, a milk fraction, colostrum, a colostrum fraction, blood and a blood fraction.
43. The process of claim 40 wherein the feed material is derived from dairy material, soy 20 material, eggs, animal tissue, animal organ or animal blood.
44. The process of claim 40 wherein the feed material is selected from: a composition comprising dairy lipids, a composition comprising egg lipids, and a composition comprising marine lipids.
45. The process of any one of claims 1 to 44 wherein the feed material is a bovine milk 25 fraction.
46. The process of claim 45 wherein the feed material is selected from: buttermilk, a buttermilk fraction, beta serum, a beta serum fraction, butter serum, a butter serum fraction, whey, a whey fraction, colostrum, and a colostrum fraction. 31 WO 2007/123424 PCT/NZ2007/000087
47. The process of any one of claims 1 to 46 wherein the feed material comprises milk fat globule membrane.
48. The process of any one of claims 1 to 47 wherein the feed material comprises at least: (a) 1% phosphatidyl serine, and 5 (b) 0.3% gangliosides.
49. The process of claim 48 wherein the feed material comprises at least: (a) 1% phosphatidyl serine, (b) 1% sphingomyelin, and (c) 0.3% gangliosides. 10 50. The process of claim 48 wherein the feed material comprises at least: (a) 1% phosphatidyl serine, (b) 1% sphingomyelin, (c) 1% cardiolipin, and (d) 0.3% gangliosides. 15 51. The process of any one of claims 1 to 50 wherein the feed material has been genetically modified.
52. The process of any one of claims 1 to 51 wherein the feed material is in solid form.
53. The process of claim 52 wherein the feed material is cryomilled before contact with a solvent. 20 54. The process of any one of claims 1 to 53 wherein the co-solvent comprises: (a) an alcohol selected from: methanol, ethanol, n-propanol, isopropanol and mixtures thereof; and (b) 0-40% by mass water. 32 WO 2007/123424 PCT/NZ2007/000087
55. The process of claim 54 wherein the co-solvent comprises between 0 and 20% by mass water.
56. The process of claim 54 wherein the co-solvent comprises between 1 and 10% by mass water. 5 57. The process of any one of claims 54 to 56 wherein the alcohol is ethanol.
58. The process of any one of claims 1 to 57 wherein the co-solvent is 95% aqueous ethanol.
59. The process of any one of claims 1 to 58 wherein the mass fraction of the co-solvent in CO 2 is between 5% and 60%.
60. The process of claim 59 wherein the mass fraction is between 20% and 50%. 10 61. The process of claim 59 wherein the mass fraction is between 25% and 30%.
62. The process of any one of claims 1 to 61 wherein the contacting temperature between the feed material and solvent is between 10 0 C and 80 0 C.
63. The process of claim 62 wherein the contacting temperature is between 55 0 C and 65 0 C.
64. The process of any one of claims 1 to 63 wherein the contacting pressure is between 100 15 bar and 500 bar.
65. The process of claim 64 wherein the contacting pressure is between 200 bar and 300 bar.
66. The process of any one of claims 1 to 65 wherein the ratio of the co-solvent to feed material is in the range 10:1 to 200:1.
67. The process of claim 66 wherein the ratio of the co-solvent to feed material is in the 20 range 15:1 to 50:1.
68. The process of any one of claims 1 to 67 wherein the separating pressure is between atmospheric pressure and 90 bar.
69. The process of claim 68 wherein the separating pressure is between 40 bar and 60 bar.
70. The process of any one of claims 1 to 69 wherein the co-solvent is recycled for further 25 use.
71. The process of any one of claims 1 to 70 wherein the CO 2 is recycled for further use. 33 WO 2007/123424 PCT/NZ2007/000087
72. The process of any one of claims 1 to 71 wherein the co-solvent is removed by evaporation under vacuum.
73. The process of any one of claims 1 to 72 wherein the feed material is contacted with a continuous flow of solvent. 5 74. The process of any one of claims 1 to 72 wherein the feed material is contacted with one or more batches of solvent.
75. The process of any one of claims 1 to 73 wherein the lipid and solvent streams are fed continuously.
76. The process of any one of claims 1 to 75 wherein the feed material and co-solvent 10 streams are mixed prior to contacting with CO 2 .
77. A product produced by the process of any one of claims 1 to 76.
78. The product of claim 77 wherein the product contains more sphingomyelin than the feed material.
79. The product of claim 77 wherein the product comprises greater than 3% sphingomyelin. 15 80. The product of claim 77 wherein the product comprises greater than 10% sphingomyelin.
81. The product of claim 77 wherein the product comprises greater than 15% sphingomyelin.
82. The product of claim 77 wherein the product contains more phosphatidyl serine than the feed material.
83. The product of claim 77 wherein the product comprises greater than 5% phosphatidyl 20 serine.
84. The product of claim 77 wherein the product comprises greater than 30% phosphatidyl serine.
85. The product of claim 77 wherein the product comprises greater than 70% phosphatidyl serine. 25 86. The product of claim 77 wherein the product contains more gangliosides than the feed material. 34 WO 2007/123424 PCT/NZ2007/000087
87. The product of claim 77 wherein the product comprises greater than 2% gangliosides.
88. The product of claim 77 wherein the product comprises greater than 4% gangliosides.
89. The product of claim 77 wherein the product comprises greater than 6% gangliosides.
90. The product of claim 77 wherein the product contains more cardiolipin than the feed 5 material.
91. The product of claim 77 wherein the product comprises greater than 5% cardiolipin.
92. The product of claim 77 wherein the product comprises greater than 10% cardiolipin.
93. The product of claim 77 wherein the product comprises greater than 25% cardiolipin.
94. The product of claim 77 wherein the product contains more acylalkyphospholipids 10 and/or plasmalogens than the feed material.
95. The product of claim 77 wherein the product comprises greater than 5% acylalkyphospholipids and/or plasmalogens.
96. The product of claim 77 wherein the product comprises greater than 10% acylalkyphospholipids and/or plasmalogens. 15 97. The product of claim 77 wherein the product comprises greater than 25% acylalkyphospholipids and/or plasmalogens.
98. The product of claim 77 wherein the product contains more aminoethylphosphonate and/or other phosphonolipids than the feed material.
99. The product of claim 77 wherein the product comprises greater than 5% 20 aminoethylphosphonate and/or other phosphonolipids.
100. The product of claim 77 wherein the product comprises greater than 10% aminoethylphosphornate and/or other phosphonolipids.
101. The product of claim 77 wherein the product comprises greater than 25% aminoethylphosphonate and/or other phosphonolipids. 25 35
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US20170119015A1 (en) * | 2015-10-30 | 2017-05-04 | Stephen J. Adolphson | Method for Defatting Whey Protein Concentrate and Producing Whey Protein Isolate |
TWI815840B (en) | 2017-12-22 | 2023-09-21 | 香港商法瑪林克國際有限公司 | Lipid combinations |
US11425915B2 (en) | 2018-05-02 | 2022-08-30 | Land O'lakes, Inc. | Methods of concentrating phospholipids |
CN110938091B (en) * | 2019-12-30 | 2022-10-28 | 青岛佰福得科技有限公司 | High-efficiency extraction method of egg yolk lecithin |
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JPH03133991A (en) * | 1989-10-19 | 1991-06-07 | Green Cross Corp:The | Method for purifying lecithin |
DE19854807A1 (en) * | 1998-11-27 | 2000-05-31 | Krupp Uhde Gmbh | Process for obtaining lecithin, in particular, from dry egg |
KR100451647B1 (en) * | 2001-02-01 | 2004-10-08 | 주식회사 고센바이오텍 | Method for extracting functional substance from yolk by supercritical fluid extraction process |
US20050170063A1 (en) * | 2004-01-29 | 2005-08-04 | Lalit Chordia | Production of powder and viscous material |
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2007
- 2007-04-20 TW TW096114188A patent/TW200811280A/en unknown
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WO2007123424A1 (en) | 2007-11-01 |
TW200811280A (en) | 2008-03-01 |
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