CN112076224A - Method for preparing whole ganoderma lucidum spore oil self-emulsifying soft capsule rich in ganoderic acid - Google Patents
Method for preparing whole ganoderma lucidum spore oil self-emulsifying soft capsule rich in ganoderic acid Download PDFInfo
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- CN112076224A CN112076224A CN202010838061.0A CN202010838061A CN112076224A CN 112076224 A CN112076224 A CN 112076224A CN 202010838061 A CN202010838061 A CN 202010838061A CN 112076224 A CN112076224 A CN 112076224A
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- CN
- China
- Prior art keywords
- ganoderma lucidum
- spore oil
- lucidum spore
- whole
- whole ganoderma
- 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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- 240000008397 Ganoderma lucidum Species 0.000 title claims abstract description 234
- 235000001637 Ganoderma lucidum Nutrition 0.000 title claims abstract description 233
- 239000007901 soft capsule Substances 0.000 title claims abstract description 79
- 238000000034 method Methods 0.000 title claims abstract description 44
- 229930182735 Ganoderic acid Natural products 0.000 title claims abstract description 30
- RDMQPKIDHAFXKA-JNORPAGFSA-N Ganoderic Acid Am1 Chemical compound C([C@@]12C)C[C@H](O)C(C)(C)[C@@H]1CC(=O)C1=C2C(=O)C[C@]2(C)[C@@H]([C@@H](CC(=O)CC(C)C(O)=O)C)CC(=O)[C@]21C RDMQPKIDHAFXKA-JNORPAGFSA-N 0.000 title claims abstract description 26
- 241000222336 Ganoderma Species 0.000 claims abstract description 100
- 239000000843 powder Substances 0.000 claims abstract description 57
- 238000000605 extraction Methods 0.000 claims abstract description 38
- 238000002360 preparation method Methods 0.000 claims abstract description 24
- 238000003825 pressing Methods 0.000 claims abstract description 19
- 230000000259 anti-tumor effect Effects 0.000 claims abstract description 13
- 239000003963 antioxidant agent Substances 0.000 claims abstract description 11
- 230000003078 antioxidant effect Effects 0.000 claims abstract description 11
- 239000004064 cosurfactant Substances 0.000 claims abstract description 11
- 239000002994 raw material Substances 0.000 claims abstract description 11
- 239000004094 surface-active agent Substances 0.000 claims abstract description 11
- 239000007908 nanoemulsion Substances 0.000 claims abstract description 7
- 239000003814 drug Substances 0.000 claims abstract description 5
- 235000013305 food Nutrition 0.000 claims abstract description 4
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 27
- 239000007788 liquid Substances 0.000 claims description 21
- 238000003756 stirring Methods 0.000 claims description 21
- 108010010803 Gelatin Proteins 0.000 claims description 17
- 239000008273 gelatin Substances 0.000 claims description 17
- 229920000159 gelatin Polymers 0.000 claims description 17
- 235000019322 gelatine Nutrition 0.000 claims description 17
- 235000011852 gelatine desserts Nutrition 0.000 claims description 17
- 239000000463 material Substances 0.000 claims description 17
- 238000000926 separation method Methods 0.000 claims description 14
- 238000002156 mixing Methods 0.000 claims description 13
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 13
- 238000001035 drying Methods 0.000 claims description 11
- 235000006708 antioxidants Nutrition 0.000 claims description 10
- 238000007789 sealing Methods 0.000 claims description 10
- 238000005469 granulation Methods 0.000 claims description 9
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 8
- 230000003179 granulation Effects 0.000 claims description 8
- 239000000203 mixture Substances 0.000 claims description 8
- 239000002245 particle Substances 0.000 claims description 8
- 239000000047 product Substances 0.000 claims description 8
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- 239000008187 granular material Substances 0.000 claims description 7
- 238000010438 heat treatment Methods 0.000 claims description 7
- 239000012530 fluid Substances 0.000 claims description 6
- 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 claims description 5
- 239000002775 capsule Substances 0.000 claims description 5
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- 238000001914 filtration Methods 0.000 claims description 5
- 239000012467 final product Substances 0.000 claims description 5
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- 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 4
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 claims description 4
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- 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 claims description 2
- 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 claims description 2
- CFWRDBDJAOHXSH-SECBINFHSA-N 2-azaniumylethyl [(2r)-2,3-diacetyloxypropyl] phosphate Chemical compound CC(=O)OC[C@@H](OC(C)=O)COP(O)(=O)OCCN CFWRDBDJAOHXSH-SECBINFHSA-N 0.000 claims description 2
- ZZZCUOFIHGPKAK-UHFFFAOYSA-N D-erythro-ascorbic acid Natural products OCC1OC(=O)C(O)=C1O ZZZCUOFIHGPKAK-UHFFFAOYSA-N 0.000 claims description 2
- 229930003268 Vitamin C Natural products 0.000 claims description 2
- 229930003427 Vitamin E Natural products 0.000 claims description 2
- 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 2
- 239000000787 lecithin Substances 0.000 claims description 2
- 235000010445 lecithin Nutrition 0.000 claims description 2
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- 229940068918 polyethylene glycol 400 Drugs 0.000 claims description 2
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- LWPLEHFGBRFRKI-NBCWKOIPSA-N Ganoderic acid B Chemical compound C([C@@]12C)C[C@H](O)C(C)(C)[C@@H]1C[C@H](O)C1=C2C(=O)C[C@]2(C)[C@@H]([C@@H](CC(=O)C[C@@H](C)C(O)=O)C)CC(=O)[C@]21C LWPLEHFGBRFRKI-NBCWKOIPSA-N 0.000 abstract description 26
- LWPLEHFGBRFRKI-CQKTXKLZSA-N Ganoderic acid B Natural products C[C@H](CC(=O)C[C@H](C)C(=O)O)[C@H]1CC(=O)[C@@]2(C)C3=C(C(=O)C[C@]12C)[C@@]4(C)CC[C@H](O)C(C)(C)[C@H]4C[C@@H]3O LWPLEHFGBRFRKI-CQKTXKLZSA-N 0.000 abstract description 26
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- 239000002253 acid Substances 0.000 description 15
- 229910002092 carbon dioxide Inorganic materials 0.000 description 13
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 9
- 239000000839 emulsion Substances 0.000 description 9
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 6
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 6
- 239000013543 active substance Substances 0.000 description 6
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- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 4
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 4
- 239000000284 extract Substances 0.000 description 4
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- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 3
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- 235000010228 ethyl p-hydroxybenzoate Nutrition 0.000 description 3
- NUVBSKCKDOMJSU-UHFFFAOYSA-N ethylparaben Chemical compound CCOC(=O)C1=CC=C(O)C=C1 NUVBSKCKDOMJSU-UHFFFAOYSA-N 0.000 description 3
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- 238000010008 shearing Methods 0.000 description 3
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- 239000000126 substance Substances 0.000 description 3
- 238000003815 supercritical carbon dioxide extraction Methods 0.000 description 3
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- 150000003722 vitamin derivatives Chemical class 0.000 description 3
- XBZYWSMVVKYHQN-MYPRUECHSA-N (4as,6as,6br,8ar,9r,10s,12ar,12br,14bs)-10-hydroxy-2,2,6a,6b,9,12a-hexamethyl-9-[(sulfooxy)methyl]-1,2,3,4,4a,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b-icosahydropicene-4a-carboxylic acid Chemical compound C1C[C@H](O)[C@@](C)(COS(O)(=O)=O)[C@@H]2CC[C@@]3(C)[C@]4(C)CC[C@@]5(C(O)=O)CCC(C)(C)C[C@H]5C4=CC[C@@H]3[C@]21C XBZYWSMVVKYHQN-MYPRUECHSA-N 0.000 description 2
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 2
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
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- IEDDICKFTXIWIP-UHFFFAOYSA-N Ganoderic acid D Natural products CC(CC(=O)CCC1CC(=O)C2(C)C3=C(C(=O)CC12C)C4(C)CCC(=O)C(C)(C)C4CC3O)C(=O)O IEDDICKFTXIWIP-UHFFFAOYSA-N 0.000 description 1
- VBGDQDJVTLQGNO-JXLWEQSJSA-N Ganoderic acid E Chemical compound C([C@@]12C)CC(=O)C(C)(C)[C@@H]1CC(=O)C1=C2C(=O)C[C@]2(C)[C@@H]([C@@H](CC(=O)C[C@@H](C)C(O)=O)C)CC(=O)[C@]21C VBGDQDJVTLQGNO-JXLWEQSJSA-N 0.000 description 1
- 241001489090 Ganoderma japonicum Species 0.000 description 1
- 102000002068 Glycopeptides Human genes 0.000 description 1
- 108010015899 Glycopeptides Proteins 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 210000000683 abdominal cavity Anatomy 0.000 description 1
- 229960000583 acetic acid Drugs 0.000 description 1
- 239000000853 adhesive Substances 0.000 description 1
- 230000001070 adhesive effect Effects 0.000 description 1
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- 235000001014 amino acid Nutrition 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 239000011230 binding agent Substances 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 150000001746 carotenes Chemical class 0.000 description 1
- 235000005473 carotenes Nutrition 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000011278 co-treatment Methods 0.000 description 1
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- 238000001514 detection method Methods 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
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- 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
- 239000000945 filler Substances 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- VVXZWUASDACSNQ-UHFFFAOYSA-N ganoderic acid C Natural products CC12CCC(O)C(C)(C)C1CC(O)C1=C2C(=O)CC2(C)C(C(CC(=O)CC(C)C(O)=O)C)CC(O)C21 VVXZWUASDACSNQ-UHFFFAOYSA-N 0.000 description 1
- VBGDQDJVTLQGNO-UHFFFAOYSA-N ganoderic acid E Natural products CC12CCC(=O)C(C)(C)C1CC(=O)C1=C2C(=O)CC2(C)C(C(CC(=O)CC(C)C(O)=O)C)CC(=O)C21C VBGDQDJVTLQGNO-UHFFFAOYSA-N 0.000 description 1
- 238000003304 gavage Methods 0.000 description 1
- 210000004602 germ cell Anatomy 0.000 description 1
- 239000012362 glacial acetic acid Substances 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 150000002596 lactones Chemical class 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000035800 maturation Effects 0.000 description 1
- 238000010172 mouse model Methods 0.000 description 1
- 239000002777 nucleoside Substances 0.000 description 1
- 125000003835 nucleoside group Chemical group 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- YTJSFYQNRXLOIC-UHFFFAOYSA-N octadecylsilane Chemical compound CCCCCCCCCCCCCCCCCC[SiH3] YTJSFYQNRXLOIC-UHFFFAOYSA-N 0.000 description 1
- 230000003285 pharmacodynamic effect Effects 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 239000012088 reference solution Substances 0.000 description 1
- 239000012488 sample solution Substances 0.000 description 1
- 239000000377 silicon dioxide Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 231100000167 toxic agent Toxicity 0.000 description 1
- 239000003440 toxic substance Substances 0.000 description 1
- -1 triterpene acids Chemical class 0.000 description 1
- 210000004881 tumor cell Anatomy 0.000 description 1
- 239000013585 weight reducing agent Substances 0.000 description 1
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
- A61K36/06—Fungi, e.g. yeasts
- A61K36/07—Basidiomycota, e.g. Cryptococcus
- A61K36/074—Ganoderma
-
- 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
- A23L31/00—Edible extracts or preparations of fungi; Preparation or treatment thereof
-
- 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
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/115—Fatty acids or derivatives thereof; Fats or oils
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23P—SHAPING OR WORKING OF FOODSTUFFS, NOT FULLY COVERED BY A SINGLE OTHER SUBCLASS
- A23P10/00—Shaping or working of foodstuffs characterised by the products
- A23P10/30—Encapsulation of particles, e.g. foodstuff additives
-
- 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
- A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
- A61K31/575—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids substituted in position 17 beta by a chain of three or more carbon atoms, e.g. cholane, cholestane, ergosterol, sitosterol
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
- A61K9/4841—Filling excipients; Inactive ingredients
- A61K9/4858—Organic compounds
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- 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
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Medicinal Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Public Health (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Mycology (AREA)
- Veterinary Medicine (AREA)
- Epidemiology (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Microbiology (AREA)
- Nutrition Science (AREA)
- Biotechnology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Medical Informatics (AREA)
- Botany (AREA)
- Alternative & Traditional Medicine (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Medicines Containing Plant Substances (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
The invention discloses a preparation method of a whole ganoderma lucidum spore oil self-emulsifying soft capsule rich in ganoderic acid. The method comprises the following steps: (1) using whole ganoderma spore powder as raw material, dry-pressing and granulating, placing in supercritical CO2Extracting in an extraction kettle for supercritical CO2Extracting to obtain light yellow whole ganoderma lucidum spore oil rich in ganoderic acid;(2) the whole ganoderma lucidum spore oil is prepared into the whole ganoderma lucidum spore oil nanoemulsion together with a surfactant, a cosurfactant and an antioxidant. The whole ganoderma lucidum spore oil is rich in an effective component ganoderic acid with an anti-tumor effect, and the content of a representative component ganoderic acid B is more than 3 times of that of pure ganoderma lucidum spore oil. The invention not only expands the application resources of the ganoderma lucidum and the ganoderma lucidum spore powder, but also greatly improves the content of ganoderic acid in the ganoderma lucidum spore oil. The method provided by the invention has simple process flow and convenient operation, and is suitable for industrial production; the product has stable quality and convenient quality control, and can be used in medicine and food.
Description
Technical Field
The invention belongs to the technical field of medicines, and particularly relates to a preparation method of a whole ganoderma lucidum spore oil self-emulsifying soft capsule rich in ganoderic acid.
Background
The Ganoderma spore is germ cell continuously ejected from pileus of Ganoderma in maturation period, is seed for reproducing next generation Ganoderma, and has all genetic active substances of Ganoderma. From the beginning of the eighties of the last century, more research foundations for the anti-tumor aspect of ganoderma lucidum and ganoderma lucidum spore powder exist, in recent years, more artificial cultured ganoderma lucidum spore powder and ganoderma lucidum spore extract are researched and applied, and Chinese scientists find that lipid is the disease-resistant 'main power' of ganoderma lucidum spores. The lipid active substance extracted by supercritical carbon dioxide is Ganoderma spore oil. The main components of the ganoderma lucidum spore comprise: fatty acids, sterols, triterpenes, vitamins, carotenes, lactones, proteins and amino acids, glycopeptides, alkaloids, inorganic ions, etc., are now made using supercritical CO2The invention discloses a plurality of patents for extracting ganoderma lucidum spore oil from ganoderma lucidum spores, but the methods are all to simply obtain pure ganoderma lucidum spore oil from the ganoderma lucidum spores.
Ganoderma lucidum has medical and edible history for thousands of years in China, is a traditional famous and precious traditional Chinese medicine for strengthening body resistance, consolidating constitution, nourishing and strengthening in China, is called as the top grade by the book of the book, and is regarded as Ling Yao Xian Dan in folk; ganoderma lucidum is a general name of Ganoderma lucidum G.lucidum.karst and Ganoderma japonicum G.japonicum Lloyd of Ganoderma (Polyporaceae) in Polyporaceae of Basidiomycetes, and the main components of Ganoderma lucidum reported in literature include: triterpenes, polysaccharides, nucleosides, sterols, amino acids, vitamins, alkaloids, inorganic ions, etc. Wherein the ganoderic acid is a component with strong biological activity in ganoderma, and ganoderic acid B is one of the main representative components of ganoderic acid in ganoderma. The ganoderma lucidum triterpene component has poor water solubility, and because the ganoderma lucidum is tough and hard to break and cut, the lipid components such as the ganoderma lucidum triterpene and the like are difficult to extract by a conventional water decoction method, and the effect of the ganoderma lucidum triterpene component is difficult to be fully exerted by water decoction extraction.
Although Ganoderma contains substantially the same active substances as Ganoderma spore, actually the content of Ganoderma triterpenic acid with strong physiological activity in Ganoderma is higher than that in Ganoderma spore, especially the content of ganoderic acid BThe content is much higher than that in Ganoderma spore. Therefore, the ganoderma lucidum spore oil is extracted by combining the ganoderma lucidum and the ganoderma lucidum spores, the acquisition resources of the ganoderma lucidum spore oil are expanded, the content of the ganoderic acid B component in the obtained ganoderma lucidum spore oil is far higher than that in the pure ganoderma lucidum spore oil, and the clinical application effect is better. Chinese patent CN 200410017468.8 discloses supercritical CO of effective component in total ganoderma spore2The extraction method uses Ganoderma encarpium and Ganoderma spore pulverized into granule as raw materials, but at supercritical C02The extraction is carried out by using edible ethanol as an extracting agent, the obtained extract is in the form of ointment, contains lipid components and water-soluble polysaccharide, has a great difference with the components of the ganoderma lucidum spore oil, and the ointment is difficult to prepare into a nano-emulsion preparation. Chinese patent CN 200610035574.8 discloses a method for preparing total ganoderma spore oil, which selects most (50-100%) ganoderma spore powder, less (0-50%) ganoderma powder, some nutrient components and water as initial raw materials, and extracts supercritical CO as final raw material by inoculating ganoderma strain to culture product for up to 60 days2The extraction method not only has complex process and long time consumption, but also has difficult treatment of the extraction.
The invention fully utilizes the characteristic that the components of the ganoderma lucidum and the ganoderma lucidum spore have better complementarity, directly selects the whole ganoderma lucidum which is composed of the ganoderma lucidum with the mass percent accounting for most (51-60%, preferably 55%) and the ganoderma lucidum with the mass percent accounting for less (40-49%, preferably 45%) as the raw materials, respectively carries out superfine grinding and wall breaking treatment, then mixes the raw materials to form the whole ganoderma lucidum spore powder, then adds a small amount of adhesive to mix the whole ganoderma lucidum spore powder evenly, carries out dry pressing granulation, and adopts ultrasonic strengthening supercritical CO to carry out dry pressing granulation2Extracting the whole ganoderma lucidum spore oil by a fluid technology. Wherein the ganoderma lucidum part is subjected to superfine grinding and ultrasonic-enhanced supercritical fluid extraction, so that the triterpenic acid components of the ganoderma lucidum can be fully extracted and can be well dissolved with ganoderma lucidum spore oil extracted from ganoderma lucidum spores, and thus the lipid transparent oily liquid with high content of ganoderic acid B is obtained: whole ganoderma spore oil.
The stable whole ganoderma lucidum spore oil self-emulsifying soft capsule preparation is prepared by the obtained whole ganoderma lucidum spore oil with high content of ganoderic acid B through optimized self-emulsifying preparation conditions for the first time, not only can improve the bioavailability of the whole ganoderma lucidum spore oil, but also has quick effect, overcomes the defect of relatively low bioavailability of the common preparation of the whole ganoderma lucidum spore oil, and obviously enhances the anti-tumor effect of the whole ganoderma lucidum spore oil. Meanwhile, the product is used as a new formulation of the whole ganoderma lucidum spore oil, and can provide a new choice for consumers.
Disclosure of Invention
The invention aims to solve the problems, provides whole ganoderma lucidum spore oil with high content of ganoderic acid B, and provides a preparation method of the whole ganoderma lucidum spore oil self-emulsifying soft capsule which has high bioavailability, safe use and convenience and can obviously enhance the anti-tumor effect of the ganoderma lucidum spore oil.
In order to achieve the purpose, the invention adopts the technical scheme that:
the ganoderma lucidum (ganoderma lucidum mycelium or ganoderma lucidum fruiting body) has rich resources and contains active substances of ganoderma lucidum triterpenic acids (ganoderic acids) with strong physiological activity, but because the water solubility of the ganoderic acids is poor, the actual extraction effect of the traditional water decoction method on the ganoderic acids in the ganoderma lucidum decoction pieces is poor, most of the ganoderic acids cannot be extracted, and the resource waste is caused; even if wine extraction is used, only part of ganoderic acid components in the ganoderma lucidum decoction pieces are extracted due to the compact texture of the ganoderma lucidum, and the resource waste is also caused. Although the ganoderma lucidum spores have all the genetic active substances of the ganoderma lucidum (namely the ganoderma lucidum spores contain basically the same active substances as the ganoderma lucidum), the research finds that the content of ganoderma lucidum acid components with very strong biological activity is much higher than that of the ganoderma lucidum spores.
Because the resources of the ganoderma lucidum spores are limited, in order to expand the resource sources of the ganoderma lucidum spore oil and improve the content and the physiological activity of the ganoderma lucidum acid components in the ganoderma lucidum spore oil, according to the existence proportion of the ganoderma lucidum and the ganoderma lucidum spores in the nature, the invention fully utilizes the characteristic that the components of the ganoderma lucidum and the ganoderma lucidum spores have better complementarity, selects the ganoderma lucidum and the ganoderma lucidum spores with certain mass percentagePulverizing Ganoderma lucidum spore powder and breaking cell wall respectively, mixing to obtain Ganoderma lucidum spore powder, adding small amount of binder, mixing, granulating under dry pressure, and performing supercritical CO treatment under optimized conditions2Extracting to obtain a lipid transparent oily liquid with high content of ganoderic acid B: whole ganoderma spore oil. The whole ganoderma lucidum spore oil is different from the general ganoderma lucidum spore oil, and the content of ganoderic acid B is far higher than that in ganoderma lucidum spores. The specific preparation scheme and steps of the whole ganoderma lucidum spore oil and the nanoemulsion thereof are as follows:
a method for preparing whole ganoderma lucidum spore oil self-emulsifying soft capsules rich in ganoderic acid comprises the following steps:
(1) using whole ganoderma spore powder as raw material, dry-pressing and granulating, placing in supercritical CO2Extracting in an extraction kettle for supercritical CO2Extracting to obtain light yellow whole ganoderma lucidum spore oil rich in ganoderic acid; the obtained light yellow whole ganoderma lucidum spore oil is rich in ganoderma lucidum triterpenic acid with anti-tumor effect, wherein the content of the representative component ganoderma lucidum acid B is more than or equal to 75mg/100g and is more than 3 times of the content of pure ganoderma lucidum spore oil ganoderma lucidum acid B25 mg/100 g.
(2) The whole ganoderma lucidum spore oil is prepared into the whole ganoderma lucidum spore oil nanoemulsion together with a surfactant, a cosurfactant and an antioxidant.
Preferably, in the above method for preparing whole ganoderma lucidum spore oil self-emulsifying soft capsules, the whole ganoderma lucidum consists of 51-60% of ganoderma lucidum fruiting body and 40-49% of ganoderma lucidum spore powder; the whole ganoderma lucidum spore powder is prepared by the following method: pulverizing Ganoderma fruiting body into superfine powder of 150 meshes or more, and breaking cell wall of Ganoderma spore powder to obtain cell wall-broken Ganoderma spore powder with cell wall breaking rate of 95% or more, and mixing the two powders to obtain the final product.
Preferably, in the above method for preparing the whole ganoderma lucidum spore oil self-emulsifying soft capsule, the step (1) is: firstly, adding 0.3-1.0% of starch or dextrin into the whole ganoderma lucidum spore powder raw material, uniformly mixing, preparing granules with the particle size of 0.1-0.3 cm by adopting a dry pressing method of a granulator in one step, and selecting the main pressure of the granulator to be 5-9 during granulationMPa, the lateral pressure is 0.5-0.9 Mpa, and then the tube is placed in supercritical CO2And (5) extracting in an extraction kettle.
Preferably, in the above method for preparing whole ganoderma lucidum spore oil self-emulsifying soft capsules, the supercritical CO is2Adopts supercritical CO with an ultrasonic extraction device2Fluid extraction apparatus combines ultrasonic extraction with supercritical fluid extraction, the supercritical fluid being CO2The extraction temperature is 23-45 ℃, the extraction pressure is 10-34 MPa, the extraction time is 1-3 hours, the extraction flow is 32-58 kg/h, the ultrasonic frequency is 30-60 kHz, and the power is 100-800W; two-stage separation is adopted, and the separation process conditions are as follows: the pressure of the primary analyzer is 9-12 MPa, and the separation temperature is 37-43 ℃; the pressure of the secondary analyzer is 4-6 Mpa, and the separation temperature is 27-33 ℃; light yellow whole ganoderma spore oil rich in ganoderic acid.
Preferably, in the above preparation method of the whole ganoderma lucidum spore oil self-emulsifying soft capsule, the whole ganoderma lucidum spore oil, the surfactant, the cosurfactant and the antioxidant are as follows by mass percent:
45-70% of whole ganoderma lucidum spore oil, 20-55% of surfactant, 0-20% of cosurfactant and 0.1-1.0% of antioxidant.
Preferably, in the above preparation method of the whole ganoderma lucidum spore oil self-emulsifying soft capsule, the surfactant is one or more of soybean lecithin, egg yolk lecithin, liquid lecithin and/or cephalin.
Preferably, in the above method for preparing the whole ganoderma lucidum spore oil self-emulsifying soft capsule, the cosurfactant is one or two of glycerol, ethanol and/or polyethylene glycol 400.
Preferably, in the above preparation method of the whole ganoderma lucidum spore oil self-emulsifying soft capsule, the antioxidant is one or two of vitamin E and vitamin C.
The preparation method of the whole ganoderma lucidum spore oil self-emulsifying soft capsule comprises the following steps:
the method comprises the steps of taking whole ganoderma lucidum spore oil, adding a surfactant, a cosurfactant and an antioxidant at the temperature of 20-60 ℃, fully and uniformly stirring, and filtering to obtain a transparent whole ganoderma lucidum spore oil self-emulsifying mixture;
adding gelatin into a gelatin dissolving tank, adding water with the temperature of 60-80 ℃, sealing and heating simultaneously, stirring until the gelatin is completely melted and swelled, adding glycerol, sealing the gelatin preparing tank, closing the stirring, starting a vacuum pump, vacuumizing, and completely pumping bubbles to obtain the soft capsule material liquid, and preserving heat at the temperature of 50-70 ℃ for later use;
thirdly, taking the self-emulsified mixture of the capsule wall material liquid of the soft capsule and the whole ganoderma lucidum spore oil, loading the mixture into a soft capsule machine, preparing the soft capsule by adopting a pressing method, and drying, washing, drying again, sorting and packaging to obtain the finished product.
The application of the whole ganoderma lucidum spore oil self-emulsifying soft capsule prepared by the preparation method in preparing anti-tumor food, health-care food or medicine. Compared with the pure ganoderma spore oil self-emulsifying system preparation, the whole ganoderma spore oil self-emulsifying soft capsule has obviously enhanced anti-tumor effect.
The ganoderic acid components are ingredients with strong activity in ganoderma lucidum and ganoderma lucidum spores, triterpenoid ganoderic acid components such as ganoderic acid B, ganoderic acid C, ganoderic acid E and the like are detected in the ganoderma lucidum and the ganoderma lucidum spores, but the content of the triterpenoid ganoderic acid components is greatly different, the content of the ganoderic acid components in the ganoderma lucidum is obviously higher than that of the ganoderma lucidum spores, and the ganoderic acid components in the ganoderma lucidum are difficult to extract by using a traditional water decoction method, so that the resource waste is caused; because the content difference of ganoderic acid components in ganoderma lucidum and ganoderma lucidum spore has strong complementarity, according to the existence proportion of ganoderma lucidum and ganoderma lucidum spore in nature, the whole ganoderma lucidum composed of ganoderma lucidum and ganoderma lucidum spore powder with a certain mass percentage is selected as raw material, and is respectively undergone the processes of ultramicropulverization and wall-breaking treatment, and mixed to form whole ganoderma lucidum spore powder, and then the whole ganoderma lucidum spore powder is made up by using ultrasonic strong supercritical CO2The extraction method extracts the whole ganoderma lucidum spore oil, so that the ganoderma lucidum acid components in the ganoderma lucidum are easy to extract, and the effect of the weight reduction is achieved.
Compared with the prior art, the invention has the following beneficial effects:
1. the whole ganoderma lucidum spore is prepared from ganoderma lucidum and ganoderma lucidum spore in a reasonable proportion, so that the waste of ganoderma lucidum resources is reduced, and the application resources of ganoderma lucidum spore are expanded, thereby saving the production cost;
2. the main triterpene ganoderic acid chemical components in the whole ganoderma lucidum spore oil obtained by the invention are clear in composition, and the quality of the product is easy to control by HPLC content determination of ganoderic acid B;
3. the invention adopts a dry-pressing one-step granulation method, solves the problem of supercritical CO2The safety of the material mixing during extraction and incomplete extraction are solved, the granulating method is simple, the heating and drying processes are not needed, and the possibility of damaging the components caused by heating is avoided.
4. The invention adopts ultrasonic strong supercritical CO2When the fluid extraction method is used for preparing the whole ganoderma lucidum spore oil, ultrasonic extraction and supercritical fluid extraction technologies are combined, so that the extraction time can be shortened and the extraction efficiency can be increased under the same extraction temperature and pressure.
5. The obtained whole ganoderma lucidum spore oil and the self-emulsifying system thereof not only have the similar effect as the pure ganoderma lucidum spore oil, but also have the content of the ganoderic acid B in the whole ganoderma lucidum spore oil which is more than 3 times of the content in the pure ganoderma lucidum spore oil through the determination of the chemical components of the ganoderic acid and the comparison of the anti-tumor activity, and the anti-tumor action strength of the whole ganoderma lucidum spore oil and the self-emulsifying system thereof is obviously higher than that of the pure ganoderma lucidum spore oil, thereby producing satisfactory beneficial effect.
The technical solution of the present invention is further illustrated by the following examples.
Detailed Description
Example 1 method for determining content of Ganoderic acid B in Whole or pure Ganoderma spore oil
The Ganoderma spore oil contains multiple chemical components, wherein Ganoderma triterpene acids (Ganoderma acids) compound is one of main effective components, and has various pharmacological effects of resisting tumor, protecting liver, removing toxic substance, and resisting aging; the whole ganoderma lucidum spore oil contains all fat-soluble components contained in ganoderma lucidum and ganoderma lucidum spores, and at least more than twenty ganoderma lucidum acid components are found in the oil at present. At present, a High Performance Liquid Chromatography (HPLC) method is mostly adopted for more accurate determination of ganoderic acid components in ganoderma lucidum spore oil.
Determining the content of ganoderic acid B in the whole ganoderma lucidum spore oil or the pure ganoderma lucidum spore oil: measuring chromatographic conditions and system applicability by high performance liquid chromatography (0512 in the chapter of the first division of the Chinese pharmacopoeia 2015) with octadecylsilane chemically bonded silica as filler; gradient elution was carried out using acetonitrile (A) -as mobile phase A and 0.1% glacial acetic acid solution as mobile phase B as specified in the following table, and the detection wavelength was 254 nm. The number of theoretical plates is not less than 2000 calculated by ganoderic acid B.
| Time (minutes) | Mobile phase A (%) | Mobile phase B (%) |
| 0 | 30 | 70 |
| 0～20 min | 30→40 | 70→60 |
Preparation of control solution A proper amount of ganoderic acid B control was precisely weighed, added with methanol to obtain a solution containing 0.08mg of ganoderic acid B per lml, and shaken well to obtain a control solution.
Preparation of test solution Whole Ganoderma spore oil or pure Ganoderma spore oil 1g is precisely weighed, dissolved with petroleum ether 10, added to treated silica gel column (8 g,18mm × 400 mm), and added with 100ml ethyl acetate: eluting with 25:75 mixed solution of petroleum ether (60-90 ℃), discarding eluent, and adding 120ml ethyl acetate to silica gel column: methanol 80: and (3) eluting the mixed solution of 20, collecting eluent, recovering the solvent until the solvent is dry, dissolving the residue with methanol, transferring the residue into a measuring flask, and fixing the volume to obtain the test solution.
The determination method comprises precisely sucking 10 μ l of reference solution and sample solution, respectively, injecting into liquid chromatograph, determining, and calculating ganoderic acid B content in whole or pure Ganoderma spore oil.
Example 2 Whole ganoderma spore oil and one of the methods for preparing self-emulsifying soft capsules thereof
1. Extraction of whole ganoderma spore oil
Taking 4.3kg of lucid ganoderma, and crushing the lucid ganoderma into 210-mesh superfine powder by using a superfine crusher to obtain about 3.8kg of lucid ganoderma superfine powder; and 3.5kg of ganoderma lucidum spore powder is taken, and is subjected to freezing ultramicro wall breaking treatment to obtain about 3.1kg of wall-broken ganoderma lucidum spore powder with the wall breaking rate of about 99.2%. Taking the obtained ganoderma lucidum superfine powder and the wall-broken ganoderma lucidum spore powder, adding 55g of starch, uniformly mixing, directly performing dry pressing granulation by using a dry pressing granulator, wherein the main pressure is 6MPa, and the lateral pressure is 0.6MPa, preparing into granules of 0.2-0.3 cm, sieving to remove fine powder of more than 60 meshes, and performing granulation by the same method on the fine powder. Placing the obtained Ganoderma spore powder in supercritical carbon dioxide extraction kettle equipped with ultrasonic extraction device, C02Pressurizing by a high-pressure pump, and circularly extracting. The pressure of the extraction kettle is 28MPa, the temperature is 38 ℃, when the indicated temperature and pressure are reached, the ultrasonic equipment is started, the frequency of ultrasonic waves is 35kHz, and the power is 200W; the first stage separation is 10MPa and 42 ℃, and the second stage separation is 6MPa and 31 ℃. After 2 hours of continuous extraction, the spore oil was collected in a separator to obtain 784.7g of light yellow whole ganoderma spore oil with specific smell of ganoderma spores. The obtained Ganoderma spore oil is determined by the above method (high performance liquid chromatography) for determining content of ganoderic acid B in Ganoderma spore oil or pure Ganoderma spore oil, and the determined content of ganoderic acid B is 97.5mg/100g, which is 3.85 times of content of ganoderic acid B in pure Ganoderma spore oil of 25.3mg/100 g.
2. The preparation of the self-emulsifying soft capsule of the whole ganoderma lucidum spore oil comprises the following steps:
preparing a self-emulsifying system of the whole ganoderma lucidum spore oil: mixing soybean lecithin 30.0g and glycerol 9.4g, stirring twice with shearing machine (8000 rpm/min), the first 5 min and the second 3 min to disperse uniformly, adding warm whole Ganoderma spore oil 60g and vitamin E0.6 g, stirring for 3 times, each for 5 min, and filtering to obtain uniform liquid, i.e. whole Ganoderma spore oil self-emulsifying system.
Preparing the soft capsule material liquid: adding 200g of gelatin into a gelatin dissolving tank, adding 240g of water with the temperature of 60-80 ℃, adding 0.2g of preservative ethylparaben, adding an opacifier and a colorant if necessary, sealing and heating at the same time, stirring until the materials are completely melted and swelled, adding glycerol, sealing the gelatin preparing tank, closing the stirring, starting a vacuum pump, vacuumizing until bubbles are completely pumped out, and obtaining the soft capsule wall material liquid. Keeping the temperature at 50-70 ℃ for later use.
Preparing a finished self-emulsifying soft capsule product: loading the capsule wall material liquid and the whole Ganoderma spore oil into a soft capsule machine, preparing soft capsule by pressing, drying, washing, drying, sorting, and packaging to obtain the final product.
And fourthly, detecting the self-emulsifying effect, namely adding 0.5ml of the self-emulsifying system of the whole ganoderma spore oil into lOml artificial gastric juice, and gently stirring by using a glass rod to quickly form an emulsion with blue opalescence. The particle size of the emulsion was measured by a TSM ultrafine particle size analyzer, and the average particle size of the emulsion droplets was about 0.25. mu.m.
Example 3 Whole Ganoderma lucidum spore oil and preparation method of self-emulsifying Soft Capsule
1. Extraction of whole ganoderma spore oil
Taking 5.2kg of lucid ganoderma, and crushing the lucid ganoderma into superfine powder of 200 meshes by a superfine crusher to obtain about 4.6kg of lucid ganoderma superfine powder; and 3.5kg of ganoderma lucidum spore powder is taken, and is subjected to freezing ultramicro wall breaking treatment to obtain about 3.1kg of wall-broken ganoderma lucidum spore powder with the wall breaking rate of about 98.8%. And (3) adding 62g of dextrin into the obtained ganoderma lucidum superfine powder and the wall-broken ganoderma lucidum spore powder, uniformly mixing, directly performing dry pressing granulation by using a dry pressing granulator, wherein the main pressure is 7MPa, and the lateral pressure is 0.6MPa, preparing into granules of 0.2-0.3 cm, sieving to remove fine powder of more than 60 meshes, and granulating the fine powder by the same method. Placing the obtained Ganoderma spore powder in supercritical carbon dioxide extraction kettle equipped with ultrasonic extraction device, C02Pressurizing by a high-pressure pump, and circularly extracting. The pressure of the extraction kettle is30MPa, 40 ℃, when the indicated temperature and pressure are reached, starting the ultrasonic equipment, wherein the frequency of ultrasonic waves is 45kHz, and the power is 250W; the first stage separation is 10MPa and 40 ℃, and the second stage separation is 5MPa and 29 ℃. After continuous extraction for 1.8 hours, the spore oil was collected in a separator to obtain 792.5g of light yellow whole ganoderma spore oil with specific smell of ganoderma spores. The obtained Ganoderma spore oil is determined by the above method (high performance liquid chromatography) for determining content of ganoderic acid B in Ganoderma spore oil or pure Ganoderma spore oil, and the determined content of ganoderic acid B is 112.2mg/100g, which is 4.43 times of content of ganoderic acid B in pure Ganoderma spore oil of 25.3mg/100 g.
2. The preparation of the self-emulsifying soft capsule of the whole ganoderma lucidum spore oil comprises the following steps:
preparing a self-emulsifying system of the whole ganoderma lucidum spore oil: mixing soybean lecithin 25.0g and polyethylene glycol 4009.5 g, stirring twice with shearing machine (8000 rpm) for 5 minutes for the first time and 4 minutes for the second time to disperse uniformly, adding warm whole Ganoderma spore oil 65g and vitamin E0.5 g, stirring for 3 times, each for 5 minutes, filtering to obtain uniform liquid, i.e. whole Ganoderma spore oil self-emulsifying system.
Preparing the soft capsule material liquid: adding 200g of gelatin into a gelatin dissolving tank, adding 240g of water with the temperature of 60-80 ℃, adding 0.2g of preservative ethylparaben, adding an opacifier and a colorant if necessary, sealing and heating at the same time, stirring until the materials are completely melted and swelled, adding glycerol, sealing the gelatin preparing tank, closing the stirring, starting a vacuum pump, vacuumizing until bubbles are completely pumped out, and obtaining the soft capsule wall material liquid. Keeping the temperature at 50-70 ℃ for later use.
Preparing a finished self-emulsifying soft capsule product: loading the capsule wall material liquid and the whole Ganoderma spore oil into a soft capsule machine, preparing soft capsule by pressing, drying, washing, drying, sorting, and packaging to obtain the final product.
And fourthly, detecting the self-emulsifying effect, namely adding 0.5ml of the self-emulsifying system of the whole ganoderma spore oil into lOml artificial gastric juice, and gently stirring by using a glass rod to quickly form an emulsion with blue opalescence. The particle size of the emulsion was measured by a TSM ultrafine particle size analyzer, and the average particle size of the emulsion droplets was about 0.28. mu.m.
Example 4 preparation of Whole Ganoderma spore oil and self-emulsifying Soft Capsule
1. Extraction of whole ganoderma spore oil
Taking 3.8kg of lucid ganoderma, and crushing the lucid ganoderma into superfine powder of 220 meshes by a superfine crusher to obtain about 3.3kg of lucid ganoderma superfine powder; and 3.5kg of ganoderma lucidum spore powder is taken, and is subjected to freezing ultramicro wall breaking treatment to obtain about 3.1kg of wall-broken ganoderma lucidum spore powder with the wall breaking rate of about 98.3%. Taking the obtained ganoderma lucidum superfine powder and the wall-broken ganoderma lucidum spore powder, adding 51g of dextrin, uniformly mixing, directly performing dry pressing granulation by using a dry pressing granulator, wherein the main pressure is 8MPa, and the lateral pressure is 0.7MPa, preparing into granules of 0.1-0.2 cm, sieving to remove fine powder of more than 60 meshes, and performing granulation by the same method on the fine powder. Placing the obtained Ganoderma spore powder in supercritical carbon dioxide extraction kettle equipped with ultrasonic extraction device, C02Pressurizing by a high-pressure pump, and circularly extracting. The pressure of the extraction kettle is 33MPa, the temperature is 30 ℃, when the indicated temperature and pressure are reached, the ultrasonic equipment is started, the frequency of ultrasonic waves is 35kHz, and the power is 300W; the primary separation is 11MPa and 41 ℃, and the secondary separation is 4MPa and 29 ℃. After 2.1 hours of continuous extraction, the spore oil was collected in a separator to obtain 778.9g of light yellow whole ganoderma spore oil with specific smell of ganoderma spores. The obtained Ganoderma spore oil is determined by the above method (high performance liquid chromatography) for determining content of ganoderic acid B in Ganoderma spore oil or pure Ganoderma spore oil, and the determined content of ganoderic acid B is 88.3mg/100g, which is 3.49 times of content of ganoderic acid B in pure Ganoderma spore oil of 25.3mg/100 g.
2. The preparation of the self-emulsifying soft capsule of the whole ganoderma lucidum spore oil comprises the following steps:
preparing a self-emulsifying system of the whole ganoderma lucidum spore oil: mixing soybean lecithin 32.0g, glycerol 5g, and polyethylene glycol 4007.5 g, stirring twice with shearing machine (8000 rpm/min), 5 min for the first time and 6 min for the second time to disperse uniformly, adding warm whole Ganoderma spore oil 55g and vitamin E0.8 g, stirring for 3 times, 5 min each time, and filtering to obtain uniform liquid, i.e. whole Ganoderma spore oil self-emulsifying system.
Preparing the soft capsule material liquid: adding 200g of gelatin into a gelatin dissolving tank, adding 240g of water with the temperature of 60-80 ℃, adding 0.2g of preservative ethylparaben, adding an opacifier and a colorant if necessary, sealing and heating at the same time, stirring until the materials are completely melted and swelled, adding glycerol, sealing the gelatin preparing tank, closing the stirring, starting a vacuum pump, vacuumizing until bubbles are completely pumped out, and obtaining the soft capsule wall material liquid. Keeping the temperature at 50-70 ℃ for later use.
Preparing a finished self-emulsifying soft capsule product: loading the capsule wall material liquid and the whole Ganoderma spore oil into a soft capsule machine, preparing soft capsule by pressing, drying, washing, drying, sorting, and packaging to obtain the final product.
And fourthly, detecting the self-emulsifying effect, namely adding 0.5ml of the self-emulsifying system of the whole ganoderma spore oil into lOml artificial gastric juice, and gently stirring by using a glass rod to quickly form an emulsion with blue opalescence. The particle size of the emulsion was measured by a TSM ultrafine particle size analyzer, and the average particle size of the emulsion droplets was found to be about 0.27. mu.m.
Example 5 tumor inhibition rate experiment of Whole ganoderma lucidum spore oil and its self-emulsified soft capsule, pure ganoderma lucidum spore oil and its self-emulsified soft capsule
The effect of the whole ganoderma lucidum spore oil and the self-emulsifying system thereof provided by the invention and the effect of the pure ganoderma lucidum spore oil and the self-emulsifying system thereof on the anti-tumor effect are compared through a pharmacodynamic test.
1. Test sample
Preparing whole ganoderma spore oil and a self-emulsifying system thereof: taking the whole ganoderma lucidum spore oil and the whole ganoderma lucidum spore oil self-emulsifying system prepared in the example 2 as the whole ganoderma lucidum spore oil and the test sample of the self-emulsifying system; preparing pure ganoderma spore oil and a self-emulsifying system thereof: pure ganoderma lucidum spore oil and a pure ganoderma lucidum spore oil self-emulsifying system serving as pure ganoderma lucidum spore oil and a sample of the self-emulsifying system are respectively prepared from wall-broken pure ganoderma lucidum spore powder with the wall-breaking rate of 98% according to the conditions and the method of the example 2.
2. Grouping of experimental animals and experimental conditions
90 mice of Kunming species with male closed groups, weighing 18-22 g, were prepared into EAC model mice, which were randomly divided into A, B, C groups of 30 mice each. Respectively as group A: a saline control group; group B: a pure ganoderma spore oil self-emulsifying system (prepared under the conditions and methods of example 2) treatment group; c: whole Ganoderma lucidum spore oil self-emulsifying System (prepared in example 2) treatment group. The experimental temperature is 23-25 ℃, and the humidity is 58-60%.
Preparation of S180 ascites tumor model mouse
Well-grown S180 ascites tumor was taken under aseptic conditions, diluted with physiological saline to prepare a tumor cell suspension of 1.O X106 cells/mL, and inoculated into the abdominal cavity of 90 mice, each of which was inoculated with 0.2 mL.
4. Experimental methods
4.1 group a saline control group: the administration of O.9% physiological saline 0.15m1 by intragastric administration to each mouse is carried out once a day for 12 days continuously.
4.2 group B pure Ganoderma spore oil self-emulsifying soft capsule treatment group: the gavage administration of pure Ganoderma spore oil from emulsifying system 0.15m1 is carried out once a day for 12 days.
4.3 group C Whole ganoderma lucidum spore oil self-emulsifying soft capsule treatment group: perfect intragastric administration of Ganoderma lucidum spore oil from emulsifying system 0.15m1 for each mouse, and intragastric administration once a day for 12 days.
4.4 animal feeding and handling
The mice were fed regular chow and water was normally drunk during the dosing period. After 12 days, the mice were sacrificed by cutting the cone, ascites tumor bodies were separated, epidermal tissues were removed, and accurate weighing was performed, and the tumor inhibition rate was calculated.
4.5 tumor inhibition Rate calculation
Tumor inhibition rate = (A-B)/A × 100%
(A is the mean tumor weight of the normal saline control group, B is the mean tumor weight of the treatment group)
Comparing whether the tumor inhibition rates of the normal saline control group, the pure ganoderma lucidum spore oil self-emulsifying soft capsule treatment group and the whole ganoderma lucidum spore oil self-emulsifying soft capsule treatment group and the tumor inhibition rates of the pure ganoderma lucidum spore oil self-emulsifying soft capsule treatment group and the whole ganoderma lucidum spore oil self-emulsifying soft capsule treatment group have significant difference or not.
5 results of the experiment
After continuous administration for 12 days, the average tumor weight of the normal saline control group is 3.92 g, the average tumor weight of the pure ganoderma lucidum spore oil self-emulsifying soft capsule treatment group is 1.99 g, and the inhibition rate is 49.23%; the average tumor weight of the whole ganoderma lucidum spore oil self-emulsifying soft capsule treatment group is 0.91 g, and the inhibition rate is 76.79%. Compared with a normal saline control group, the tumor inhibition rates of the pure ganoderma lucidum spore oil self-emulsifying soft capsule treatment group and the whole ganoderma lucidum spore oil self-emulsifying soft capsule treatment group are obviously different (P is less than 0.01); the tumor inhibition rates of the pure ganoderma lucidum spore oil self-emulsifying soft capsule treatment group and the whole ganoderma lucidum spore oil self-emulsifying soft capsule treatment group are also obviously different (P is less than 0.01), and the tumor inhibition rate of the whole ganoderma lucidum spore oil self-emulsifying soft capsule is about 1.56 times of the tumor inhibition rate of the pure ganoderma lucidum spore oil self-emulsifying soft capsule.
6 conclusion and analysis
6.1 compared with the control group, the tumor inhibition rate of the pure ganoderma lucidum spore oil self-emulsifying soft capsule treatment group is very obviously different, which shows that the pure ganoderma lucidum spore oil self-emulsifying soft capsule has better tumor inhibition effect. Because the pure ganoderma spore oil is self-emulsified, the nanoemulsion is formed in the stomach of a mouse after the intragastric administration, the absorption is easy, and the activity and the drug effect are improved compared with the pure ganoderma spore oil which is not self-emulsified.
6.2 similarly, compared with the control group, the whole ganoderma lucidum spore oil self-emulsifying soft capsule treatment group has very obvious difference in tumor inhibition rate, which shows that the whole ganoderma lucidum spore oil self-emulsifying soft capsule has good tumor inhibition effect. Because the ganoderma lucidum spore oil is self-emulsified, after the administration by intragastric administration, the nanoemulsion is formed in the stomach of a mouse, the absorption is easy, and the activity and the drug effect are greatly improved compared with the non-self-emulsified ganoderma lucidum spore oil.
6.3 compared with the pure ganoderma spore oil self-emulsifying soft capsule treatment group, the whole ganoderma spore oil self-emulsifying soft capsule treatment group has very obvious difference in tumor inhibition rate, which shows that the whole ganoderma spore oil self-emulsifying soft capsule has very good tumor inhibition effect. Because the pure ganoderma lucidum spore oil contains the ganoderma lucidum acid component with the traditional anti-tumor effect and the ganoderma lucidum acid component ganoderma lucidum acid B with the anti-tumor effect in the whole ganoderma lucidum spore oil is increased by double, the activity and the drug effect of the whole ganoderma lucidum spore oil are greatly improved.
Claims (10)
1. A method for preparing a whole ganoderma lucidum spore oil self-emulsifying soft capsule rich in ganoderic acid is characterized by comprising the following steps:
(1) using whole ganoderma spore powder as raw material, dry-pressing and granulating, placing in supercritical CO2Extracting in an extraction kettle for supercritical CO2Extracting to obtain light yellow whole ganoderma lucidum spore oil rich in ganoderic acid;
(2) the whole ganoderma lucidum spore oil is prepared into the whole ganoderma lucidum spore oil nanoemulsion together with a surfactant, a cosurfactant and an antioxidant.
2. The method for preparing whole ganoderma lucidum spore oil self-emulsifying soft capsules according to claim 1, wherein the whole ganoderma lucidum consists of 51% -60% of ganoderma lucidum fruiting bodies and 40% -49% of ganoderma lucidum spore powder; the whole ganoderma lucidum spore powder is prepared by the following method: pulverizing Ganoderma fruiting body into superfine powder of 150 meshes or more, and breaking cell wall of Ganoderma spore powder to obtain cell wall-broken Ganoderma spore powder with cell wall breaking rate of 95% or more, and mixing the two powders to obtain the final product.
3. The method for preparing the whole ganoderma lucidum spore oil self-emulsifying soft capsule as claimed in claim 1, wherein the step (1) is as follows: firstly, adding 0.3-1.0% of starch or dextrin into the whole ganoderma lucidum spore powder raw material, uniformly mixing, preparing granules with the particle size of 0.1-0.3 cm by adopting a dry pressing method of a granulator in one step, selecting the main pressure of the granulator to be 5-9 MPa and the lateral pressure to be 0.5-0.9 MPa during granulation, and then placing the granules in a supercritical CO2And (5) extracting in an extraction kettle.
4. The method for preparing whole ganoderma lucidum spore oil self-emulsifying soft capsules according to claim 1 or 3, wherein the supercritical CO is2Adopts supercritical CO with an ultrasonic extraction device2Fluid extraction apparatus combines ultrasonic extraction with supercritical fluid extraction, the supercritical fluid being CO2The extraction temperature is 23-45 ℃, the extraction pressure is 10-34 MPa, the extraction time is 1-3 hours, the extraction flow is 32-58 kg/h, the ultrasonic frequency is 30-60 kHz, and the power is 100-800W;two-stage separation is adopted, and the separation process conditions are as follows: the pressure of the primary analyzer is 9-12 MPa, and the separation temperature is 37-43 ℃; the pressure of the secondary analyzer is 4-6 Mpa, and the separation temperature is 27-33 ℃; light yellow whole ganoderma spore oil rich in ganoderic acid.
5. The method for preparing the whole ganoderma lucidum spore oil self-emulsifying soft capsule as claimed in claim 1, wherein the whole ganoderma lucidum spore oil, the surfactant, the cosurfactant and the antioxidant are as follows by mass percent:
45-70% of whole ganoderma lucidum spore oil, 20-55% of surfactant, 0-20% of cosurfactant and 0.1-1.0% of antioxidant.
6. The method for preparing whole ganoderma lucidum spore oil self-emulsifying soft capsules according to claim 1, wherein the surfactant is one or more of soybean lecithin, egg yolk lecithin, liquid lecithin and/or cephalin.
7. The method for preparing whole ganoderma lucidum spore oil self-emulsifying soft capsules according to claim 1, wherein the cosurfactant is one or two of glycerol, ethanol and/or polyethylene glycol 400.
8. The method for preparing the whole ganoderma lucidum spore oil self-emulsifying soft capsule as claimed in claim 1, wherein the antioxidant is one or two of vitamin E and vitamin C.
9. The method for preparing the whole ganoderma lucidum spore oil self-emulsifying soft capsule as claimed in claim 1, which is characterized by comprising the following steps:
the method comprises the steps of taking whole ganoderma lucidum spore oil, adding a surfactant, a cosurfactant and an antioxidant at the temperature of 20-60 ℃, fully and uniformly stirring, and filtering to obtain a transparent whole ganoderma lucidum spore oil self-emulsifying mixture;
adding gelatin into a gelatin dissolving tank, adding water with the temperature of 60-80 ℃, sealing and heating simultaneously, stirring until the gelatin is completely melted and swelled, adding glycerol, sealing the gelatin preparing tank, closing the stirring, starting a vacuum pump, vacuumizing, and completely pumping bubbles to obtain the soft capsule material liquid, and preserving heat at the temperature of 50-70 ℃ for later use;
thirdly, taking the self-emulsified mixture of the capsule wall material liquid of the soft capsule and the whole ganoderma lucidum spore oil, loading the mixture into a soft capsule machine, preparing the soft capsule by adopting a pressing method, and drying, washing, drying again, sorting and packaging to obtain the finished product.
10. The use of whole ganoderma lucidum spore oil self-emulsifying soft capsules prepared by the preparation method of claim 1 in the preparation of anti-tumor food, health food or medicaments.
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| CN116726070A (en) * | 2022-08-03 | 2023-09-12 | 江西鑫康健科技发展有限公司 | Selfheal seed oil self-emulsifying soft capsule with anti-tumor effect and preparation method and application thereof |
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