CN106890199A - Extract method and the medicine containing squalene of plant source spiny dogfish ene compositions and its preparation method and application - Google Patents
Extract method and the medicine containing squalene of plant source spiny dogfish ene compositions and its preparation method and application Download PDFInfo
- Publication number
- CN106890199A CN106890199A CN201510956807.7A CN201510956807A CN106890199A CN 106890199 A CN106890199 A CN 106890199A CN 201510956807 A CN201510956807 A CN 201510956807A CN 106890199 A CN106890199 A CN 106890199A
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- CN
- China
- Prior art keywords
- squalene
- plant source
- weight
- spiny dogfish
- separation
- 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
- BHEOSNUKNHRBNM-UHFFFAOYSA-N Tetramethylsqualene Natural products CC(=C)C(C)CCC(=C)C(C)CCC(C)=CCCC=C(C)CCC(C)C(=C)CCC(C)C(C)=C BHEOSNUKNHRBNM-UHFFFAOYSA-N 0.000 title claims abstract description 126
- PRAKJMSDJKAYCZ-UHFFFAOYSA-N dodecahydrosqualene Natural products CC(C)CCCC(C)CCCC(C)CCCCC(C)CCCC(C)CCCC(C)C PRAKJMSDJKAYCZ-UHFFFAOYSA-N 0.000 title claims abstract description 126
- 229940031439 squalene Drugs 0.000 title claims abstract description 126
- TUHBEKDERLKLEC-UHFFFAOYSA-N squalene Natural products CC(=CCCC(=CCCC(=CCCC=C(/C)CCC=C(/C)CC=C(C)C)C)C)C TUHBEKDERLKLEC-UHFFFAOYSA-N 0.000 title claims abstract description 126
- YYGNTYWPHWGJRM-UHFFFAOYSA-N (6E,10E,14E,18E)-2,6,10,15,19,23-hexamethyltetracosa-2,6,10,14,18,22-hexaene Chemical compound CC(C)=CCCC(C)=CCCC(C)=CCCC=C(C)CCC=C(C)CCC=C(C)C YYGNTYWPHWGJRM-UHFFFAOYSA-N 0.000 title claims abstract description 125
- 238000000034 method Methods 0.000 title claims abstract description 83
- 239000000203 mixture Substances 0.000 title claims abstract description 83
- 239000003814 drug Substances 0.000 title claims abstract description 23
- 238000002360 preparation method Methods 0.000 title claims abstract description 13
- 239000000284 extract Substances 0.000 title claims description 17
- 241000251778 Squalus acanthias Species 0.000 title 1
- 241000251774 Squalus Species 0.000 claims abstract description 56
- 239000002994 raw material Substances 0.000 claims abstract description 40
- 238000000926 separation method Methods 0.000 claims abstract description 32
- 229930182558 Sterol Natural products 0.000 claims abstract description 22
- 150000003432 sterols Chemical class 0.000 claims abstract description 22
- 235000003702 sterols Nutrition 0.000 claims abstract description 22
- 230000003078 antioxidant effect Effects 0.000 claims abstract description 6
- 238000000199 molecular distillation Methods 0.000 claims description 55
- 239000007788 liquid Substances 0.000 claims description 25
- 238000006243 chemical reaction Methods 0.000 claims description 24
- 239000012071 phase Substances 0.000 claims description 22
- GVJHHUAWPYXKBD-UHFFFAOYSA-N (±)-α-Tocopherol Chemical compound OC1=C(C)C(C)=C2OC(CCCC(C)CCCC(C)CCCC(C)C)(C)CCC2=C1C GVJHHUAWPYXKBD-UHFFFAOYSA-N 0.000 claims description 20
- 238000001976 enzyme digestion Methods 0.000 claims description 20
- 238000005886 esterification reaction Methods 0.000 claims description 18
- 230000032050 esterification Effects 0.000 claims description 17
- 239000003921 oil Substances 0.000 claims description 17
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 17
- 239000000463 material Substances 0.000 claims description 15
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 claims description 13
- 108090001060 Lipase Proteins 0.000 claims description 11
- 102000004882 Lipase Human genes 0.000 claims description 11
- 239000004367 Lipase Substances 0.000 claims description 10
- 229930003427 Vitamin E Natural products 0.000 claims description 10
- WIGCFUFOHFEKBI-UHFFFAOYSA-N gamma-tocopherol Natural products CC(C)CCCC(C)CCCC(C)CCCC1CCC2C(C)C(O)C(C)C(C)C2O1 WIGCFUFOHFEKBI-UHFFFAOYSA-N 0.000 claims description 10
- 235000019421 lipase Nutrition 0.000 claims description 10
- 235000019165 vitamin E Nutrition 0.000 claims description 10
- 229940046009 vitamin E Drugs 0.000 claims description 10
- 239000011709 vitamin E Substances 0.000 claims description 10
- 238000002425 crystallisation Methods 0.000 claims description 9
- 239000003960 organic solvent Substances 0.000 claims description 9
- 239000004202 carbamide Substances 0.000 claims description 8
- 230000008025 crystallization Effects 0.000 claims description 8
- 239000000546 pharmaceutical excipient Substances 0.000 claims description 7
- 230000001877 deodorizing effect Effects 0.000 claims description 6
- 239000010773 plant oil Substances 0.000 claims description 5
- 238000007445 Chromatographic isolation Methods 0.000 claims description 4
- 239000007791 liquid phase Substances 0.000 claims description 4
- 238000005809 transesterification reaction Methods 0.000 claims description 4
- 150000001336 alkenes Chemical class 0.000 claims description 3
- 239000003963 antioxidant agent Substances 0.000 claims description 3
- 235000006708 antioxidants Nutrition 0.000 claims description 3
- 239000008280 blood Substances 0.000 claims description 3
- 210000004369 blood Anatomy 0.000 claims description 3
- 238000001704 evaporation Methods 0.000 claims description 3
- 230000008020 evaporation Effects 0.000 claims description 3
- 230000036039 immunity Effects 0.000 claims description 3
- 230000009467 reduction Effects 0.000 claims description 3
- 239000007790 solid phase Substances 0.000 claims description 2
- 102000004190 Enzymes Human genes 0.000 claims 1
- 108090000790 Enzymes Proteins 0.000 claims 1
- 238000000605 extraction Methods 0.000 abstract description 14
- 235000013376 functional food Nutrition 0.000 abstract description 2
- 241000196324 Embryophyta Species 0.000 description 49
- 239000000047 product Substances 0.000 description 23
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 21
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 14
- 235000019198 oils Nutrition 0.000 description 14
- 239000006185 dispersion Substances 0.000 description 13
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 13
- 238000004519 manufacturing process Methods 0.000 description 12
- 239000003999 initiator Substances 0.000 description 11
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 10
- 239000004519 grease Substances 0.000 description 9
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 8
- 239000001301 oxygen Substances 0.000 description 8
- 229910052760 oxygen Inorganic materials 0.000 description 8
- 239000000706 filtrate Substances 0.000 description 7
- 238000001914 filtration Methods 0.000 description 7
- 230000008569 process Effects 0.000 description 7
- 239000007787 solid Substances 0.000 description 7
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 6
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 6
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 6
- 235000019441 ethanol Nutrition 0.000 description 6
- 239000000243 solution Substances 0.000 description 6
- 239000002904 solvent Substances 0.000 description 6
- WQDUMFSSJAZKTM-UHFFFAOYSA-N Sodium methoxide Chemical class [Na+].[O-]C WQDUMFSSJAZKTM-UHFFFAOYSA-N 0.000 description 5
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- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 4
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- 235000008390 olive oil Nutrition 0.000 description 4
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- 238000010992 reflux Methods 0.000 description 4
- OILXMJHPFNGGTO-UHFFFAOYSA-N (22E)-(24xi)-24-methylcholesta-5,22-dien-3beta-ol Natural products C1C=C2CC(O)CCC2(C)C2C1C1CCC(C(C)C=CC(C)C(C)C)C1(C)CC2 OILXMJHPFNGGTO-UHFFFAOYSA-N 0.000 description 3
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 3
- 241001465754 Metazoa Species 0.000 description 3
- 108010084311 Novozyme 435 Proteins 0.000 description 3
- 230000008859 change Effects 0.000 description 3
- 239000012141 concentrate Substances 0.000 description 3
- 239000013078 crystal Substances 0.000 description 3
- 238000004332 deodorization Methods 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
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- 229910052757 nitrogen Inorganic materials 0.000 description 3
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- 239000011780 sodium chloride Substances 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- OQMZNAMGEHIHNN-UHFFFAOYSA-N 7-Dehydrostigmasterol Natural products C1C(O)CCC2(C)C(CCC3(C(C(C)C=CC(CC)C(C)C)CCC33)C)C3=CC=C21 OQMZNAMGEHIHNN-UHFFFAOYSA-N 0.000 description 2
- XKRFYHLGVUSROY-UHFFFAOYSA-N Argon Chemical compound [Ar] XKRFYHLGVUSROY-UHFFFAOYSA-N 0.000 description 2
- 241000251730 Chondrichthyes Species 0.000 description 2
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 2
- OYHQOLUKZRVURQ-HZJYTTRNSA-N Linoleic acid Chemical compound CCCCC\C=C/C\C=C/CCCCCCCC(O)=O OYHQOLUKZRVURQ-HZJYTTRNSA-N 0.000 description 2
- 108010048733 Lipozyme Proteins 0.000 description 2
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 2
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 2
- 239000005864 Sulphur Substances 0.000 description 2
- HZYXFRGVBOPPNZ-UHFFFAOYSA-N UNPD88870 Natural products C1C=C2CC(O)CCC2(C)C2C1C1CCC(C(C)=CCC(CC)C(C)C)C1(C)CC2 HZYXFRGVBOPPNZ-UHFFFAOYSA-N 0.000 description 2
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- 239000007864 aqueous solution Substances 0.000 description 2
- LGJMUZUPVCAVPU-UHFFFAOYSA-N beta-Sitostanol Natural products C1CC2CC(O)CCC2(C)C2C1C1CCC(C(C)CCC(CC)C(C)C)C1(C)CC2 LGJMUZUPVCAVPU-UHFFFAOYSA-N 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 239000010495 camellia oil Substances 0.000 description 2
- 239000001569 carbon dioxide Substances 0.000 description 2
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- 150000002148 esters Chemical class 0.000 description 2
- XJFYWGIWEYQMPK-UHFFFAOYSA-N ethanol;urea Chemical compound CCO.NC(N)=O XJFYWGIWEYQMPK-UHFFFAOYSA-N 0.000 description 2
- 235000019197 fats Nutrition 0.000 description 2
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- OYHQOLUKZRVURQ-IXWMQOLASA-N linoleic acid Natural products CCCCC\C=C/C\C=C\CCCCCCCC(O)=O OYHQOLUKZRVURQ-IXWMQOLASA-N 0.000 description 2
- FCCDDURTIIUXBY-UHFFFAOYSA-N lipoamide Chemical compound NC(=O)CCCCC1CCSS1 FCCDDURTIIUXBY-UHFFFAOYSA-N 0.000 description 2
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- 239000011347 resin Substances 0.000 description 2
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- SQGYOTSLMSWVJD-UHFFFAOYSA-N silver(1+) nitrate Chemical compound [Ag+].[O-]N(=O)=O SQGYOTSLMSWVJD-UHFFFAOYSA-N 0.000 description 2
- HCXVJBMSMIARIN-PHZDYDNGSA-N stigmasterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)/C=C/[C@@H](CC)C(C)C)[C@@]1(C)CC2 HCXVJBMSMIARIN-PHZDYDNGSA-N 0.000 description 2
- 235000016831 stigmasterol Nutrition 0.000 description 2
- 229940032091 stigmasterol Drugs 0.000 description 2
- BFDNMXAIBMJLBB-UHFFFAOYSA-N stigmasterol Natural products CCC(C=CC(C)C1CCCC2C3CC=C4CC(O)CCC4(C)C3CCC12C)C(C)C BFDNMXAIBMJLBB-UHFFFAOYSA-N 0.000 description 2
- -1 triterpene compound Chemical class 0.000 description 2
- 235000015112 vegetable and seed oil Nutrition 0.000 description 2
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- LXEKPEMOWBOYRF-QDBORUFSSA-N AAPH Chemical compound Cl.Cl.NC(=N)C(C)(C)\N=N\C(C)(C)C(N)=N LXEKPEMOWBOYRF-QDBORUFSSA-N 0.000 description 1
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- OILXMJHPFNGGTO-NRHJOKMGSA-N Brassicasterol Natural products O[C@@H]1CC=2[C@@](C)([C@@H]3[C@H]([C@H]4[C@](C)([C@H]([C@@H](/C=C/[C@H](C(C)C)C)C)CC4)CC3)CC=2)CC1 OILXMJHPFNGGTO-NRHJOKMGSA-N 0.000 description 1
- SGNBVLSWZMBQTH-FGAXOLDCSA-N Campesterol Natural products O[C@@H]1CC=2[C@@](C)([C@@H]3[C@H]([C@H]4[C@@](C)([C@H]([C@H](CC[C@H](C(C)C)C)C)CC4)CC3)CC=2)CC1 SGNBVLSWZMBQTH-FGAXOLDCSA-N 0.000 description 1
- 241000222122 Candida albicans Species 0.000 description 1
- 206010007134 Candida infections Diseases 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
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- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
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- 108010022535 Farnesyl-Diphosphate Farnesyltransferase Proteins 0.000 description 1
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- BTEISVKTSQLKST-UHFFFAOYSA-N Haliclonasterol Natural products CC(C=CC(C)C(C)(C)C)C1CCC2C3=CC=C4CC(O)CCC4(C)C3CCC12C BTEISVKTSQLKST-UHFFFAOYSA-N 0.000 description 1
- 235000019484 Rapeseed oil Nutrition 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 241001122767 Theaceae Species 0.000 description 1
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- 150000001298 alcohols Chemical class 0.000 description 1
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- OILXMJHPFNGGTO-ZAUYPBDWSA-N brassicasterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)/C=C/[C@H](C)C(C)C)[C@@]1(C)CC2 OILXMJHPFNGGTO-ZAUYPBDWSA-N 0.000 description 1
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- SGNBVLSWZMBQTH-PODYLUTMSA-N campesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CC[C@@H](C)C(C)C)[C@@]1(C)CC2 SGNBVLSWZMBQTH-PODYLUTMSA-N 0.000 description 1
- 235000000431 campesterol Nutrition 0.000 description 1
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- 229940079593 drug Drugs 0.000 description 1
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- IDGUHHHQCWSQLU-UHFFFAOYSA-N ethanol;hydrate Chemical compound O.CCO IDGUHHHQCWSQLU-UHFFFAOYSA-N 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 229930195729 fatty acid Natural products 0.000 description 1
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- 150000004671 saturated fatty acids Chemical class 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 239000010686 shark liver oil Substances 0.000 description 1
- 229940069764 shark liver oil Drugs 0.000 description 1
- 238000010898 silica gel chromatography Methods 0.000 description 1
- 229910001961 silver nitrate Inorganic materials 0.000 description 1
- GFNHODBBCUPTMB-UHFFFAOYSA-N silver;methanol;nitrate Chemical compound [Ag+].OC.[O-][N+]([O-])=O GFNHODBBCUPTMB-UHFFFAOYSA-N 0.000 description 1
- NLQLSVXGSXCXFE-UHFFFAOYSA-N sitosterol Natural products CC=C(/CCC(C)C1CC2C3=CCC4C(C)C(O)CCC4(C)C3CCC2(C)C1)C(C)C NLQLSVXGSXCXFE-UHFFFAOYSA-N 0.000 description 1
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- 239000004328 sodium tetraborate Substances 0.000 description 1
- 150000003431 steroids Chemical class 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-N sulfuric acid group Chemical class S(O)(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 1
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Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/01—Hydrocarbons
-
- 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
Abstract
The present invention relates to functional food field, method and the medicine containing squalene of extraction plant source spiny dogfish ene compositions and its preparation method and application are disclosed.The method for extracting plant source spiny dogfish ene compositions includes：The raw material that will be enriched in squalene is digested and concentration and separation successively, wherein, the condition of the concentration and separation is controlled so that the squalene containing 35-80 weight %, the sterol of squalene and 10-40 weight % containing 5-30 weight % in the raw material rich in squalene in the plant source spiny dogfish ene compositions for obtaining.The new plant source spiny dogfish ene compositions that obtain being extracted using the method for the present invention, the squalene (purity is usually above 90%) of high-purity that is provided with prior art quite even preferably antioxidant effect is provided on the premise of it need not further purify.
Description
Technical field
The present invention relates to functional food field, in particular it relates to a kind of extract plant source spiny dogfish ene compositions
Method, a kind of preparation method of medicine containing squalene and by the method prepare containing squalene
Medicine and its application.
Background technology
Squalene is a kind of highly undersaturated hydrocarbon compound, is a kind of water white oil with special odor
Shape liquid, its systematic naming method be 2,6,10,15,19,23- hexamethyl -2, the alkene of 6,10,14,18,22- tetracosa carbon six,
Molecular formula is C30H50.Squalene belongs to a kind of highly undersaturated straight chain triterpene compound, structural formula
For：
Squalene has anti-oxidant, promotion skin health, promotes cardiovascular health, improves human body resist oxygen lack
Many effects such as ability, are widely used in medicine, cosmetic field, in the last few years in field of health care products
Using also progressively extending.
Presently commercially available squalene product is and extracts what is obtained from eepwater shark liver oil, with to deep-sea
The animal protection cry of shark is surging, and European Union member countries limited deep-sea in 2006 in Atlantic Ocean northeastward
Fishing for for shark is increasingly deficient which results in the squalene resource with hydrogenated hydro carbons grease as raw material.
Scientific research personnel also attempts to expand by chemistry or the method for biosynthesis the market supply of squalene,
But the technological process of chemical reaction is long, and use poisonous chemical agent, production technology distance industry more
Change large-scale production and also there is a certain distance, and chemical synthesis squalene double bond structure and natural angle
There is difference in the alltrans structure of MF59.
In addition, CN102257149B discloses a kind of method for preparing purifying yeast, produced using excess
The yeast of squalene prepares the squalene of high-purity, for preparing oil-in-water emulsion as immunologic adjuvant.
CN103266137B is related to a kind of production method of squalene, and squalene synthetase gene is cloned
And be co-expressed in Escherichia coli, the method so as to construct the recombinant bacterial strain that can synthesize squalene.But,
Biosynthesis is all also undesirable at the aspect such as yield of strain and purpose thing, also in the presence of some limitation, away from
With a certain distance from industrialization large-scale production also.
Because squalene is also widely present in plant, there is the research work to be with olive oil or deodorization distillate
Raw material carries out extraction research to the squalene of plant source, because squalene content is relatively low in raw material, it is contemplated that
Production cost, squalene carries out purification and does not have industrialized production and commercialized condition, therefore plants
The mechanisms such as research institute and colleges and universities are confined to the work of material resource squalene more.
The purification production of plant source squalene because material system component is complicated and involved with compared with
The squalene product of high-purity (more than 90%) is target, and prior art is required to using multi-step process
It is likely to be breached the purpose for isolating and purifying, and the process step for using is more, experiment flow is long, solvent and energy
Amount consumption is big, low production efficiency.
The plant oil deodorizing distillate for producing during vegetable oil deodorized at present, especially olive oil deodorization
Squalene is extracted in distillate turns into an effective method, and according to China's national situation, most of olive oil is all
It is to rely on import, lacks the stable resources of olive oil deodorization distillate.Therefore from the existing grease money of China
The technique that design separation squalene is developed in source is a kind of economically viable processing mode.
CN102146014A discloses the method for extracting squalene as raw material with tea seed, its extracting method
Technological process be：Extraction, concentration, esterification, complexant-borax reaction, macroreticular resin chromatogram point
Continue to concentrate after purifying, concentration, extraction and washing, obtain carrying out again after squalene crude oil overcritical
Carbon dioxide abstraction, obtains the squalene product that content is more than 80%.However, the method process route
Complexity, solvent energy consumption consumption is big, and chromatographic isolation has that dead absorption, filler need regeneration,
Increased production cost.
CN1284752C discloses a kind of extracting method of plant squalene, by the benevolence or kind of Momordica grosvenori
Seed is crushed, and liposoluble substance is gone out with organic solvent Soakage extraction, removes organic solvent, and Momordica grosvenori angle is obtained
MF59 crude product；Momordica grosvenori squalene crude product is crossed into silica gel column chromatography, is eluted with organic solvent, collect wash-out
Liquid colourless part, organic solvent is removed with distillation under vacuum, and Momordica grosvenori squalene fine work is obtained.However,
It is low to there is raw material squalene content in the method, extraction cost shortcoming high.
It is auxiliary agent that CN103483305A is disclosed and used urea and solvent, makes itself and grease deodorized distillate
Dissolving mixing, carries out vacuum outgas and seals, then made using super-pressure saturated fatty acid in distillate,
The rapid crystallization such as aliphatic hydrocarbon and sterol condenses, and press filtration obtains filter residue and filtrate after separating, can after desolvation
Obtain the mixture of the VE, squalene and polyunsaturated fatty acid of higher degree.This technique also uses super
High pressure, and vacuum outgas is needed, increased the difficulty of energy consumption and subsequent treatment.
Application No. 201410375211.3 prior art discloses a kind of technique of complex extractions squalene,
The benevolence of tea oil tree is crushed, plus petroleum ether carries out ultrasonic extraction, filtering, filtrate decompression concentration, then
Gained camellia oil liposoluble substance concentrate is mixed with strong alkali alcosol, petroleum ether is used after being heated to reflux
Extract, being subsequently adding silver nitrate methanol aqueous solution carries out complex reaction, then proceedes to be stripped with petroleum ether
Squalene petroleum ether organic phase after being dissociated；By the squalene petroleum ether organic phase NaCl aqueous solution
Vacuum distillation is carried out after cleaning, is cleaned with distilled water again, be vacuum dried, that is, obtain end-product squalene
Fine work.However, silver nitrate is introduced in the technological process of the prior art carries out complex reaction, significantly
Increased process costs, and generate the possibility of chemical contamination, be unfavorable for industrial popularization.
The content of the invention
The purpose of the present invention is the defect for overcoming prior art, is not reducing the effect of the product containing squalene
A kind of new plant source spiny dogfish ene compositions are provided on the premise of fruit, and a kind of technological process letter is provided
The method of the acquisition new plant source spiny dogfish ene compositions of single, low cost and suitable industrialized production,
The new plant source spiny dogfish ene compositions for obtaining are extracted using the method for the present invention need not further carried
The squalene of high-purity that is there is provided with prior art is obtained with the premise of pure, and (purity is usually above
90%) suitable even preferably antioxidant effect.
The present inventor is based on completion technical scheme considered below：The angle of plant source
There is very big difference in MF59 raw material, the content of some components is in animal sources with the squalene raw material of animal sources
In relatively fewer, and then rich content in the raw material of plant source, by taking stigmasterol as an example, its molecular weight is
412.69, it is difficult to be separated with squalene with the molecular weight (410.72) of squalene closely.
If inventor's consideration can be moderate by simple technological operation acquisition squalene content and can be retained
The composition of other the various active materials in plant source squalene raw material, then can greatly save and be produced into
The green production of the product of sheet and realization containing squalene.Based on this, inventor has carried out a series of creation
Journal of Sex Research, and it was found that the squalene content obtained by simple technological operation is moderate and retains plant source
The composition of other the various active materials in squalene raw material in effect with high-purity (more than 90%)
Squalene quite, it is or even better.
In a first aspect, the present invention provides a kind of method for extracting plant source spiny dogfish ene compositions, the method bag
Include：The raw material that will be enriched in squalene is digested and concentration and separation successively, wherein, control the enrichment point
From condition cause the squalene containing 35-80 weight %, institute in the plant source spiny dogfish ene compositions for obtaining
State the sterol of the squalene containing 5-30 weight % and 10-40 weight % in the raw material rich in squalene.
Second aspect, the present invention provides a kind of preparation method of the medicine containing squalene, and the method includes：
(1) plant source spiny dogfish ene compositions are extracted：Using the extraction plant source squalene that the present invention is foregoing
The method of composition extracts plant source spiny dogfish ene compositions；
(2) medicine is prepared：The medicine of the plant source spiny dogfish ene compositions that will be obtained containing step (1)
Active component is learned to be mixed with pharmaceutic adjuvant.
The third aspect, the present invention provides the medicine containing squalene prepared by preceding method of the invention
Thing.
Fourth aspect, the present invention provides the foregoing medicine containing squalene for strengthen immunity, anti-oxidant
With the application in the product of reduction blood fat density.
The plant source spiny dogfish obtained by the method for extraction plant source spiny dogfish ene compositions involved in the present invention
Also contain sterol isoreactivity material in ene compositions in addition to the squalene containing 35-80 weight %, by
The method of the present invention extract the plant source spiny dogfish ene compositions that obtain can keep it is suitable with prior art very
To more preferable pharmaceutical active.And, the method technique letter for extracting plant source spiny dogfish ene compositions of the invention
Single, low production cost is suitable as industrial method and is promoted.
Other features and advantages of the present invention will be described in detail in subsequent specific embodiment part.
Brief description of the drawings
Accompanying drawing is, for providing a further understanding of the present invention, and to constitute the part of specification, with
Following specific embodiment is used to explain the present invention together, but is not construed as limiting the invention.
In accompanying drawing：
Fig. 1 is the result schematic diagram of the test case that the present invention is provided.
Specific embodiment
Specific embodiment of the invention is described in detail below.It should be appreciated that this place is retouched
The specific embodiment stated is merely to illustrate and explain the present invention, and is not intended to limit the invention.
In a first aspect, the invention provides a kind of method for extracting plant source spiny dogfish ene compositions, the method
Including：The raw material that will be enriched in squalene is digested and concentration and separation successively, wherein, control the enrichment
The condition of separation causes the squalene containing 35-80 weight % in the plant source spiny dogfish ene compositions for obtaining,
The sterol of squalene and 10-40 weight % containing 5-30 weight % in the raw material rich in squalene.
Preferably, the condition of the concentration and separation is controlled to cause to contain in the plant source spiny dogfish ene compositions for obtaining
There is the squalene of 40-70 weight %.
Under preferable case, squalene containing 10-30 weight % in the raw material rich in squalene and
The sterol of 10-40 weight %.
Further contain the sterol of 0.5-10 weight % in the plant source spiny dogfish ene compositions；More preferably
Ground, the sterol of 0.5-5 weight % is further contained in the plant source spiny dogfish ene compositions.
Sterol of the present invention is in phytosterol, including stigmasterol, campesterol and brassicasterol
It is at least one.
A small amount of other hydro carbons, alcohols can also be contained in the raw material rich in squalene of the present invention
With Ester etc..
In the present invention, the raw material rich in squalene can be various conventional to meet above-mentioned composition
Raw material.In a preferred embodiment, the raw material rich in squalene can be by including following step
Rapid technique is obtained：
(1) plant oil deodorizing distillate is carried out into esterification and transesterification successively, and will be obtained
Product carry out the first Crystallization Separation, obtain the first solid phase and first liquid phase；
(2) gained first liquid is mutually carried out into the first molecular distillation, is obtained rich in vitamin E and spiny dogfish
The cut of alkene；And
(3) the resulting cut rich in vitamin E and squalene is carried out into the first chromatographic isolation successively
And evaporation, obtain the vitamin E and the raw material rich in squalene of high-purity.
In the present invention, it is preferred to the plant oil deodorizing distillate is produced for vegetable oil during refining and deodorizing
Raw accessory substance.
In the present invention, it is preferred to aliphatic acid containing 10-80 weight % in the plant oil deodorizing distillate,
The glyceride of 10-30 weight %, the vitamin E of 0.5-20 weight %, the phytosterol of 0.5-20 weight %,
The phytosterin ester of 0.5-20 weight % and the squalene of 0.5-30 weight %.
In the present invention, in order to obtain the raw material rich in squalene, Publication No. can be used
Method described in the patent application of CN105016956A is obtained.The present invention will not be repeated here.
Preferably, the step of enzymolysis includes：In the presence of lipase, described squalene will be rich in
Raw material enzyme digestion reaction is carried out in water；Then the mixture that will be obtained after enzyme digestion reaction using organic solvent
Extracted, and collected oil phase.
Preferably, the temperature of the enzyme digestion reaction is 20-80 DEG C, and the time is 1-48h.It is further preferred that
The temperature of the enzyme digestion reaction is 30-50 DEG C, and the time is 15-25h.
Preferably, on the basis of the gross weight of the raw material rich in squalene, the enzyme digestion reaction is carried out
Lipase consumption be 0.1-5 weight %, more preferably 0.5-1 weight %.
Preferably, it is 0.1-3 to carry out the water-oil factor volume ratio in the enzyme digestion reaction：1, more preferably
0.5-2：1.
Preferably, the organic solvent for being extracted after enzyme digestion reaction for example can be n-hexane.This hair
The bright oil phase for obtaining that will can also collect is washed in such as ethanol-water solution of 10 weight %
To neutral, the sample of pending concentration and separation is obtained.The lipase of the invention can derive from aspergillus
One or more of bacterium, cylindrical candidiasis, pseudomonad and head mold.For example, the lipase
Lipase LVK, the wild public affairs in Japanese day that including but not limited to Shenzhen Lv Weikang bioengineering Co., Ltd is produced
Produced G50, Lipase F-AP15 the and Lipase AY 30G of department and Novozymes Company
Novozym435, Lipozyme RM IM and Lipozyme TL IM etc..
Preferably, the step of concentration and separation is carried out using three-level molecular distillation or rectification under vacuum.
Preferably, the condition of the first order molecular distillation in the three-level molecular distillation includes：Pressure is
5-25Pa, temperature is 120-140 DEG C, and knifing rotating speed is 200-300rpm；The bar of second level molecular distillation
Part includes：Pressure is 5-25Pa, and temperature is 110-125 DEG C, and knifing rotating speed is 200-300rpm；3rd
The condition of level molecular distillation includes：Pressure is 5-25Pa, and temperature is 100-120 DEG C, and knifing rotating speed is
200-300rpm.In the case of more preferably, the temperature of the third level molecular distillation is than second fraction
The temperature of son distillation is low 5-10 DEG C.It was found by the inventors of the present invention that controlling the third level molecular distillation
Temperature is lower 5-10 DEG C than the temperature of the second level molecular distillation, and causes the third level molecular distillation
Temperature and the second level molecular distillation temperature in above range of the invention when, enable to
The antioxidation of the plant source spiny dogfish ene compositions for arriving is stronger.
Preferably, the condition of the rectification under vacuum includes：Pressure is 100-2000Pa, and temperature is
180-250 DEG C, theoretical cam curve is 15-30.It is further preferred that the condition of the rectification under vacuum includes：
Pressure is 300-600Pa, and temperature is 200-230 DEG C, and theoretical cam curve is 15-25.It is of the invention described
Rectification under vacuum can be carried out in the presence of such as nitrogen is as protective gas, and light component cut therein is steamed
Go out, wax and alkanes substance of the cut obtained by bottom also containing part heavy can be by subsequently carrying out
Urea clathrate treatment or Freeze crystallization are removed.
Another preferred embodiment of the invention, the method for the present invention includes：The richness
The step of collection is separated is carried out using three-level molecular distillation, and before the concentration and separation is carried out, first will
Esterification is carried out by the product obtained after the enzymolysis.Preferably, the condition bag of the esterification
Include：Temperature is 70-90 DEG C, and the time is 1-10h.
Another preferred embodiment of the invention, the method for the present invention includes：The richness
The step of collection is separated is carried out using rectification under vacuum, and before the concentration and separation is carried out, will first be passed through
The product obtained after the enzymolysis carries out esterification；And enter the product obtained after the concentration and separation
The treatment of row urea clathrate and/or Freeze crystallization.Preferably, the condition of the esterification includes：Temperature
It is 70-90 DEG C to spend, and the time is 1-10h.Preferably, the step of urea clathrate is processed can be：1)
Compound concentration is the urea-ethanol solution of 0.01-7g/mL, then to adding institute in the urea-ethanol solution
The product obtained after concentration and separation is stated, and is heated to reflux 0.5-8h and included；2) by step 1)
To product be cooled to room temperature, carry out crystallisation by cooling under conditions of being placed in subzero 20 DEG C to 10 DEG C above freezing
3-48h, is then filtered；3) to step 2) gained filtrate in add organic solvent (for example just oneself
Alkane), and the filter residue obtained after filtering is washed using organic solvent, after merging organic phase successively
Washed with such as 5 weight % sodium chloride solutions and water, collect organic phase and concentrated.Preferably,
The step of freeze-drying can be：The product obtained after the concentration and separation is dissolved in ethanol in proper amount
In, it is heated to being completely dissolved；0.5-5h is placed in being subsequently placed in 0-35 DEG C, lower temperature is then transferred to
Placed 1-10 hours under (second temperature), there are a large amount of crystal to separate out；After filtering, then filtrate is placed in
Second temperature left overnight, carries out secondary cold analysis；In order to promote crystalline growth, can be added in system
Diatomite carries out rush crystals growth, filtering and collecting filter liquid and solvent evaporated.
Second aspect, the invention provides a kind of preparation method of the medicine containing squalene, the method includes：
(1) plant source spiny dogfish ene compositions are extracted：Plant source angle is extracted using the foregoing method of the present invention
MF59 composition；
(2) medicine is prepared：The medicine of the plant source spiny dogfish ene compositions that will be obtained containing step (1)
Active component is learned to be mixed with pharmaceutic adjuvant.
In the second aspect of the present invention, about the method for extracting plant source spiny dogfish ene compositions as originally
Described in the aforementioned first aspect of invention, in order to avoid repeating, the present invention will not be repeated here.
Preferably, in step (1), 0.5-10 is further contained in the plant source spiny dogfish ene compositions
The sterol of weight %；It is further preferred that further containing 0.5-5 in the plant source spiny dogfish ene compositions
The sterol of weight %.
The plant source spiny dogfish ene compositions that step (1) of the present invention obtains are before it need not further purify
Put just directly can be mixed to prepare medicine with pharmaceutic adjuvant.This greatlys save production cost,
The reservation of other active components being also beneficial in the raw material rich in squalene of the invention.
Pharmaceutic adjuvant of the present invention can be conventional use of various pharmaceutic adjuvants in the art, for example
Gelatin, glycerine etc..Preferably, in the preparation method of the medicine containing squalene of the present invention, this
Art personnel can determine that the pharmacy activity component is auxiliary with medicinal according to the conventional amount used of this area
The usage ratio of material, such as described pharmacy activity component can be 1 with the consumption weight ratio of pharmaceutic adjuvant：
0.1-1000。
The third aspect, the invention provides the medicine containing squalene prepared by preceding method.
Fourth aspect, the invention provides the foregoing medicine containing squalene for strengthen immunity, antioxygen
Change the application in the product with reduction blood fat density.
Below will the present invention will be described in detail by embodiment.
In case of no particular description, various raw materials used below are all from commercially available.
Water used below is the homemade deionized water in laboratory.
The rate of recovery (%) in the present invention is accounted in raw material according to the weight of finally obtained pure material
The percentage composition of pure material gross weight is calculated.
Following squalene is analyzed by HPLC, analysis condition be using XunionC18 posts (5um,
4.6mm × 150mm), mobile phase volume ratio is acetonitrile：Methyl alcohol=60：40, flow velocity is 2mL/min,
Detection UV absorption at 210nm, it is 25 DEG C to control column temperature.
Sterol is measured using GC, and analysis condition is that split ratio is 39：1, nitrogen flow is
40mL/min, injection port and detector (fid detector) temperature is 300 DEG C.Using temperature programming,
Initial temperature is 210 DEG C, and 275 DEG C are risen to 10 DEG C/min, retains 25min.
Preparation example 1 is used to illustrate the source of the raw material rich in squalene of the invention.
Embodiment 1-7 is used for the method for illustrating extraction plant source spiny dogfish ene compositions of the invention.
Test case 1 is used to illustrate the application of the composition containing squalene of the invention.
Preparation example 1
(as shown in table 1, remaining is miscellaneous for composition and relevant parameter to take 1000g rapeseed oil deodorized distillates
Matter), add the 35g concentrated sulfuric acids and 300g methyl alcohol that esterification is carried out at 70 DEG C, by esterification reaction product
Wash and dehydration by evaporation is to below the weight % of moisture 0.4, be subsequently adding 40g sodium methoxides and 300g
Methyl alcohol carries out transesterification at a temperature of 70 DEG C, and transesterification product then is carried out into the first crystallization point
From.15 DEG C are cooled to the rate of temperature fall of 8 DEG C/h in the first Crystallization Separation step, this is maintained
Temperature growing the grain 10 hours, then carries out first time separation of solid and liquid；The liquid that first time separation of solid and liquid is obtained
Body carries out the second cooling, and rate of temperature fall is 4 DEG C/h, is cooled to 5 DEG C, maintains this temperature growing the grain 10 small
When, carry out second separation of solid and liquid.Separation of solid and liquid gained solid (i.e. thick phytosterol solid) is passed through twice
The phytosterol 118.14g that purity is 94.8 weight % is obtained after ethanol cleaning, recrystallization, drying, is returned
Yield 71.79%.
Second separation of solid and liquid gained filtrate carries out the first molecular distillation, the wherein pressure of first order molecular distillation
Power is 15Pa, and temperature is 120 DEG C, and knifing rotating speed is 280rpm；The pressure of second level molecular distillation is
20Pa, vapo(u)rizing temperature is 160 DEG C, and knifing rotating speed is 280rpm；The pressure of third level molecular distillation is
15Pa, vapo(u)rizing temperature is 200 DEG C, and knifing rotating speed is 280rpm；
First order molecular distillation and second level molecular distillation light phase cut are collected, fatty acid ester is obtained about
750g；
Collecting third level molecular distillation gained light phase cut simultaneously carries out the first chromatographic isolation, filler using it is cloudy from
Sub-exchange resin, mobile phase uses absolute ethyl alcohol, is rinsed with mobile phase after loading, collects unadsorbed
Material, that is, obtain the raw material rich in squalene.
The composition and content of the resulting raw material rich in squalene are respectively the weight % of squalene 15, steroid
The weight % of alcohol 28.
Desorbed toward pouring carbon dioxide in chromatographic column again, rinsed with mobile phase, collecting to be desorbed
The liquid phase rich in the raw table E of dimension is mutually, the liquid phase that will obtain being enriched with vitamin E is concentrated, then
Refined, obtained the vitamin E product 43.19g that purity is 90.3 weight %, the rate of recovery 90.70%.
Table 1
| Component | Content (weight %) |
| Aliphatic acid and glyceride | 77.01 |
| Content of phytosterol | 15.60 |
| Content of vitamin E | 4.30 |
| Squalene content | 2.01 |
Embodiment 1
Extract plant source spiny dogfish ene compositions：The present embodiment is extracted using the technique of enzymolysis-molecular distillation plants
Material resource spiny dogfish ene compositions.Specifically：
Enzymolysis：Take the raw material 20g rich in squalene, water 20g and 100mg that preparation example 1 is prepared
Day open country lipase ay 30 be placed in flask, at 50 DEG C, according to water oil volume ratio be 1.5：1 enters
Row enzyme digestion reaction, the reaction time is 30h；Then obtained after the n-hexane extraction enzyme digestion reaction for using 20mL × 3
The mixture for arriving, and oil phase is collected, and the concentration oil phase obtains grease.
Molecular distillation：The grease that will be obtained after enzymolysis carries out molecular distillation, is carried by three-level molecular distillation
It is pure to obtain spiny dogfish ene compositions, wherein,
The pressure of first order molecular distillation is 10Pa, and temperature is 125 DEG C, and knifing rotating speed is 250rpm；
The pressure of second level molecular distillation is 10Pa, and temperature is 115 DEG C, and knifing rotating speed is 250rpm；
The pressure of third level molecular distillation is 10Pa, and temperature is 108 DEG C, and knifing rotating speed is 250rpm.
As a result the content of squalene is 49.98%, the content of sterol in third level molecular distillation gained light phase
It is 2.4 weight %.
Embodiment 2
Extract plant source spiny dogfish ene compositions：The present embodiment is carried using the technique of enzymolysis -ester-molecular distillation
Take plant source spiny dogfish ene compositions.Specifically：
Enzymolysis：Take the raw material 20g rich in squalene, water 20g and 200mg that preparation example 1 is prepared
Lipase (wherein, the use of AY30 and Novozyme435 containing AY30 and Novozyme435
Amount weight ratio is 1：1) it is placed in flask, is 0.5 according to water oil volume ratio at 65 DEG C:1 ratio
Enzyme digestion reaction is carried out, the reaction time is 25h；Then with after the n-hexane extraction enzyme digestion reaction of 20mL × 3
The mixture for obtaining, and oil phase is collected, and the concentration oil phase obtains grease.
Esterification：To the 22g that the sodium methoxide dissolved with 1.1g is added in the product obtained after above-mentioned enzyme digestion reaction
Methyl alcohol in, in 70 DEG C flow back 1.5h；Then added containing the dense sulphur of 1.5g in reaction mixture refluxed
The methyl alcohol of the 10mL of acid, 70 DEG C of keeping temperature flows back 2 hours；Then products therefrom is washed with water
Wash until pH value is 7.0；Collect oil phase；
Molecular distillation：The grease that will be obtained after esterification carries out molecular distillation, is carried by three-level molecular distillation
It is pure to obtain spiny dogfish ene compositions, wherein,
The pressure of first order molecular distillation is 10Pa, and temperature is 125 DEG C, and knifing rotating speed is 260rpm；
The pressure of second level molecular distillation is 12Pa, and temperature is 118 DEG C, and knifing rotating speed is 260rpm；
The pressure of third level molecular distillation is 10Pa, and temperature is 108 DEG C, and knifing rotating speed is 250rpm.
Analyzed by HPLC, as a result the content of squalene is in third level molecular distillation gained light phase
51.01%, the content of sterol is 2.3 weight %.
Embodiment 3
Extract plant source spiny dogfish ene compositions：The present embodiment is using enzymolysis -ester-rectification under vacuum-urea clathrate
Technique extract plant source spiny dogfish ene compositions.Specifically：
Enzymolysis：Take the raw material 20g rich in squalene, water 20g and 400mg that preparation example 1 is prepared
Lipase LVK and Lipase F-AP15 (weight ratio be 1：1) it is placed in flask, at 60 DEG C,
It is 2 according to water oil volume ratio：1 ratio carries out enzyme digestion reaction, and the reaction time is 30h；Then 20mL is used
The mixture obtained after × 3 n-hexane extraction enzyme digestion reaction, and oil phase is collected, and concentrate the oil
Mutually obtain grease.
Esterification：To the 25g that the sodium methoxide dissolved with 1.2g is added in the product obtained after above-mentioned enzyme digestion reaction
Methyl alcohol in, in 80 DEG C flow back 2.5h；Then added containing the dense sulphur of 1.2g in reaction mixture refluxed
The methyl alcohol of the 12mL of acid, 80 DEG C of backflow 3h of keeping temperature；Then products therefrom is washed with water
Until pH value is 7.0；Collect oil phase.
Rectification under vacuum：Product after esterification is carried out into rectification under vacuum separation, rectifying is equivalent to 20 column plates
On the tower of height, carried out under the vacuum of 300Pa, by the tower that sample implantation temperature after esterification is 210 DEG C
Top, while be filled with nitrogen being protected, the cut of light component is steamed, and obtains bottom cut.
Urea clathrate：60mL ethanol, 15g urea are heated to reflux 30min at 80 DEG C, are added while hot
Enter resulting material after above-mentioned rectification under vacuum, continue to be heated to reflux 60min, then put in 5 DEG C of cold baths
3h is put, is filtered, gained filtrate is mixed with water, then with 50mL n-hexane extractions 3 times, merging is just
Hexane phase, is washed 3 times with the NaCl solution of 5 weight %, then is washed with deionized 3 times.Gained
Organic phase rotates at 55 DEG C and is evaporated to solvent volatilization and finishes, and obtains grease, and resulting material proceeds the
Secondary inclusion, obtains the weight % of squalene 58.63, and sterol content is 2.3 weight %.
Embodiment 4
Extract plant source spiny dogfish ene compositions：The present embodiment is using enzymolysis -ester-rectification under vacuum-freezing and crystallizing
Technique extract plant source spiny dogfish ene compositions.Specifically：
Enzymolysis：Carried out using method same as Example 3；
Esterification：Carried out using method same as Example 3；
Rectification under vacuum：Carried out using method same as Example 3；
Freezing and crystallizing：Rectification under vacuum bottom resulting material is dissolved in 60mL ethanol and is heated to 45 DEG C of holdings
15min；Then placed 5 hours at 0 DEG C after products therefrom being cooled into room temperature, there are a large amount of crystal to separate out；
After filtering, then filtrate is placed in -5 DEG C of left overnights, carries out secondary cold analysis；Filtering and collecting filter liquid,
Solvent evaporated obtains oily mater, wherein obtaining the weight % of squalene 59.45, sterol content is 2.3 weights
Amount %.
Embodiment 5
Extract plant source spiny dogfish ene compositions：The present embodiment is extracted using the technique of enzymolysis-molecular distillation plants
Material resource spiny dogfish ene compositions.Specifically：
Enzymolysis：Carried out using method same as Example 1；
Molecular distillation：Carried out using method similar to Example 1, except that：In the present embodiment
The temperature of second level molecular distillation is 110 DEG C.Remaining is in the same manner as in Example 1.
As a result the content of squalene is 48.46 weight % in third level molecular distillation gained light phase, sterol
Content is 2.4 weight %.
Embodiment 6
Extract plant source spiny dogfish ene compositions：The present embodiment is extracted using the technique of enzymolysis-molecular distillation plants
Material resource spiny dogfish ene compositions.Specifically：
Enzymolysis：Carried out using method same as Example 1；
Molecular distillation：Carried out using method similar to Example 1, except that：In the present embodiment
The temperature of second level molecular distillation is 120 DEG C.Remaining is in the same manner as in Example 1.
As a result the content of squalene is 48.67 weight % in third level molecular distillation gained light phase, sterol
Content is 2.4 weight %.
Embodiment 7
Extract plant source spiny dogfish ene compositions：The present embodiment is extracted using the technique of enzymolysis-molecular distillation plants
Material resource spiny dogfish ene compositions.Specifically：
Enzymolysis：Carried out using method same as Example 1；
Molecular distillation：Carried out using method similar to Example 1, except that：In the present embodiment
The temperature of second level molecular distillation is 108 DEG C, and the temperature of third level molecular distillation is 101 DEG C.Remaining is equal
It is in the same manner as in Example 1.
As a result the content of squalene is 48.23 weight % in third level molecular distillation gained light phase, sterol
Content is 2.4 weight %.
Test case 1
The anti-of the composition containing squalene that embodiments of the invention are prepared is tested using following methods
Oxidation.
Prepare micella dispersion liquid：Quantitative linoleic acid is dissolved in a small amount of chloroform, then is blown chloroform with argon gas
It is dry, make linoleic acid that thin film is formed in bottle wall, 10min is vacuumized to remove the chloroform of possible remaining,
It is subsequently adding the 50mmol phosphate buffers (pH value is 7.0) containing surfactant, buffer solution
To be complexed trace metal impurities that may be present, the mixture solution exists the EDTA of middle addition 0.1mmol
5min is vibrated in CQ250 type ultrasonic oscillators and forms transparent micella dispersion liquid.
Oxygen absorption is determined with SP-2 type oxygen absorptions instrument, the current potential YEW3066 type electronics of oxygen electrode
Potential difference meter is recorded, and sensitivity is 5 microvolts, and the device can measure 10-8The oxygen of mol/L, during measure,
The above-mentioned micella dispersion liquid for preparing of the invention is placed in the reactor with electromagnetic agitation, it is close
Close, lucifuge, 37 DEG C of constant temperature, oxygen content is determined with oxygen electrode, then add AAPH to draw with injector
Hair agent (18mmol/L) triggers linoleic peroxidating, or adds initiator (18mmol/L) simultaneously
The composition (10 μm of ol/L) containing squalene prepared with above-described embodiment, continuously records oxygen content
Change with time, as a result as shown in fig. 1, wherein, in Fig. 1,
A is represented not to addition initiator in the micella dispersion liquid and/or the feelings of the composition containing squalene
Condition；
B is represented to the situation that initiator is added in the micella dispersion liquid；
C is represented to the composition containing squalene that initiator and embodiment 1 are added in the micella dispersion liquid
Situation；
D is represented to the composition containing squalene that initiator and embodiment 2 are added in the micella dispersion liquid
Situation；
E is represented to the composition containing squalene that initiator and embodiment 3 are added in the micella dispersion liquid
Situation；
F is represented to the composition containing squalene that initiator and embodiment 4 are added in the micella dispersion liquid
Situation；
G is represented to the composition containing squalene that initiator and embodiment 5 are added in the micella dispersion liquid
Situation；
H is represented to the composition containing squalene that initiator and embodiment 6 are added in the micella dispersion liquid
Situation；
I is represented to the composition containing squalene that initiator and embodiment 7 are added in the micella dispersion liquid
Situation；
J is represented to addition initiator and the implementation in CN105016956A in the micella dispersion liquid
The situation of the squalene that example 1 is prepared.
It can be seen that the present invention provide the composition containing squalene antioxidation have it is excellent
The antioxidant effect of the squalene of the high-purity provided in prior art.
The preferred embodiment of the present invention described in detail above, but, the present invention is not limited to above-mentioned reality
The detail in mode is applied, in range of the technology design of the invention, can be to technical side of the invention
Case carries out various simple variants, and these simple variants belong to protection scope of the present invention.
It is further to note that each particular technique described in above-mentioned specific embodiment is special
Levy, in the case of reconcilable, can be combined by any suitable means, in order to avoid need not
The repetition wanted, the present invention is no longer separately illustrated to various possible combinations.
Additionally, can also be combined between a variety of implementation methods of the invention, as long as its
Without prejudice to thought of the invention, it should equally be considered as content disclosed in this invention.
Claims (11)
1. a kind of method for extracting plant source spiny dogfish ene compositions, the method includes：Will be enriched in squalene
Raw material digested successively and concentration and separation, wherein, control the condition of the concentration and separation to cause to obtain
Plant source spiny dogfish ene compositions in the squalene containing 35-80 weight %, the original rich in squalene
The sterol of squalene and 10-40 weight % containing 5-30 weight % in material.
2. method according to claim 1, wherein, control the condition of the concentration and separation to cause
Squalene containing 40-70 weight % in the plant source spiny dogfish ene compositions for obtaining.
3. method according to claim 1, wherein, the raw material rich in squalene is by bag
The technique for including following steps is obtained：
(1) plant oil deodorizing distillate is carried out into esterification and transesterification successively, and will be obtained
Product carry out the first Crystallization Separation, obtain the first solid phase and first liquid phase；
(2) gained first liquid is mutually carried out into the first molecular distillation, is obtained rich in vitamin E and spiny dogfish
The cut of alkene；And
(3) the resulting cut rich in vitamin E and squalene is carried out into the first chromatographic isolation successively
And evaporation, respectively obtain the vitamin E and the raw material rich in squalene of high-purity.
4. method according to claim 1, wherein, include the step of the enzymolysis：In fat
In the presence of enzyme, the raw material rich in squalene is carried out into enzyme digestion reaction in water；Then using organic
Solvent is extracted the mixture obtained after enzyme digestion reaction, and collects oil phase；Preferably
The temperature of the enzyme digestion reaction is 20-80 DEG C, and the time is 1-48h；More preferably
On the basis of the gross weight of the raw material rich in squalene, the lipase of the enzyme digestion reaction is carried out
Consumption be 0.1-5 weight %；And the water oil volume ratio in enzyme digestion reaction is 0.1-3：1.
5. the method according to any one in claim 1-4, wherein, the concentration and separation
Step is carried out using three-level molecular distillation or rectification under vacuum；Preferably
The condition of the first order molecular distillation in the three-level molecular distillation includes：Pressure is 5-25Pa, temperature
It is 120-140 DEG C to spend, and knifing rotating speed is 200-300rpm；The condition of second level molecular distillation includes：Pressure
Power is 5-25Pa, and temperature is 110-125 DEG C, and knifing rotating speed is 200-300rpm；Third level molecular distillation
Condition include：Pressure is 5-25Pa, and temperature is 100-120 DEG C, and knifing rotating speed is 200-300rpm；
Preferably
The condition of the rectification under vacuum includes：Pressure is 200-1000Pa, and temperature is 200-230 DEG C, reason
It is 15-30 by the number of plates.
6. method according to claim 5, wherein, three-level is used the step of the concentration and separation
Molecular distillation is carried out, and the method is further included：Before the concentration and separation is carried out, first will be through
Crossing the product obtained after the enzymolysis carries out esterification；Preferably
The condition of the esterification includes：Temperature is 70-90 DEG C, and the time is 1-10h.
7. method according to claim 5, wherein, vacuum is used the step of the concentration and separation
Rectifying is carried out, and the method is further included：Before the concentration and separation is carried out, first will be by institute
Stating the product obtained after enzymolysis carries out esterification；And carry out the product obtained after the concentration and separation
Urea clathrate treatment and/or Freeze crystallization.
8. a kind of preparation method of the medicine containing squalene, the method includes：
(1) plant source spiny dogfish ene compositions are extracted：Using described in any one in claim 1-7
Method extracts plant source spiny dogfish ene compositions；
(2) medicine is prepared：The medicine of the plant source spiny dogfish ene compositions that will be obtained containing step (1)
Active component is learned to be mixed with pharmaceutic adjuvant.
9. method according to claim 8, wherein, in step (1), the plant source angle
Further contain the sterol of 0.5-10 weight % in MF59 composition；Preferably
Further contain the sterol of 0.5-5 weight % in the plant source spiny dogfish ene compositions.
10. the medicine containing squalene that the method as described in claim 8 or 9 is prepared.
The medicine containing squalene described in 11. claims 10 for strengthen immunity, it is anti-oxidant and
Application in the product of reduction blood fat density.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107474093A (en) * | 2017-08-23 | 2017-12-15 | 福建省格兰尼生物工程股份有限公司 | A kind of deodorization distillate continuous production VE, sterol, methyl esters, glycerine, the method for squalene and high-boiling components |
| WO2019214410A1 (en) * | 2018-05-07 | 2019-11-14 | 宜春大海龟生命科学有限公司 | Plant squalene composition and preparation method therefor and application thereof, and product applying same |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102089263A (en) * | 2008-07-07 | 2011-06-08 | 索菲姆公司 | Process for the extraction of squalene, sterols and vitamin e contained in condensates of physical refining and/or in distillates of deodorization of plant oils |
| CN105016956A (en) * | 2014-04-23 | 2015-11-04 | 中粮营养健康研究院有限公司 | Squalene extracting method |
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2015
- 2015-12-18 CN CN201510956807.7A patent/CN106890199A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102089263A (en) * | 2008-07-07 | 2011-06-08 | 索菲姆公司 | Process for the extraction of squalene, sterols and vitamin e contained in condensates of physical refining and/or in distillates of deodorization of plant oils |
| CN105016956A (en) * | 2014-04-23 | 2015-11-04 | 中粮营养健康研究院有限公司 | Squalene extracting method |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107474093A (en) * | 2017-08-23 | 2017-12-15 | 福建省格兰尼生物工程股份有限公司 | A kind of deodorization distillate continuous production VE, sterol, methyl esters, glycerine, the method for squalene and high-boiling components |
| CN107474093B (en) * | 2017-08-23 | 2019-07-23 | 福建省格兰尼生物工程股份有限公司 | A kind of method of deodorization distillate continuous production VE, sterol, methyl esters, glycerol, squalene and high-boiling components |
| WO2019214410A1 (en) * | 2018-05-07 | 2019-11-14 | 宜春大海龟生命科学有限公司 | Plant squalene composition and preparation method therefor and application thereof, and product applying same |
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Application publication date: 20170627 |