CN106479952A - Culture medium and the preparation method of Vitis secondary metabolites - Google Patents
Culture medium and the preparation method of Vitis secondary metabolites Download PDFInfo
- Publication number
- CN106479952A CN106479952A CN201610963852.XA CN201610963852A CN106479952A CN 106479952 A CN106479952 A CN 106479952A CN 201610963852 A CN201610963852 A CN 201610963852A CN 106479952 A CN106479952 A CN 106479952A
- Authority
- CN
- China
- Prior art keywords
- vitis
- culture medium
- culture
- secondary metabolites
- preparation
- 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.)
- Pending
Links
- 241000219095 Vitis Species 0.000 title claims abstract description 119
- 235000009392 Vitis Nutrition 0.000 title claims abstract description 84
- 239000001963 growth medium Substances 0.000 title claims abstract description 45
- 229930000044 secondary metabolite Natural products 0.000 title claims abstract description 42
- 238000002360 preparation method Methods 0.000 title claims abstract description 22
- 239000007640 basal medium Substances 0.000 claims abstract description 7
- 239000002537 cosmetic Substances 0.000 claims abstract description 6
- 230000003712 anti-aging effect Effects 0.000 claims abstract description 5
- 235000009754 Vitis X bourquina Nutrition 0.000 claims description 35
- 235000012333 Vitis X labruscana Nutrition 0.000 claims description 35
- 235000014787 Vitis vinifera Nutrition 0.000 claims description 35
- 239000005631 2,4-Dichlorophenoxyacetic acid Substances 0.000 claims description 20
- 108010059892 Cellulase Proteins 0.000 claims description 14
- 229940106157 cellulase Drugs 0.000 claims description 14
- 230000000694 effects Effects 0.000 claims description 14
- 108090000790 Enzymes Proteins 0.000 claims description 11
- 102000004190 Enzymes Human genes 0.000 claims description 11
- 229940088598 enzyme Drugs 0.000 claims description 11
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 8
- 239000008103 glucose Substances 0.000 claims description 8
- 238000002156 mixing Methods 0.000 claims description 5
- 235000015097 nutrients Nutrition 0.000 claims description 5
- 230000003321 amplification Effects 0.000 claims description 3
- 239000003814 drug Substances 0.000 claims description 3
- 235000013305 food Nutrition 0.000 claims description 3
- 238000003199 nucleic acid amplification method Methods 0.000 claims description 3
- 239000000047 product Substances 0.000 claims description 3
- 239000000706 filtrate Substances 0.000 claims description 2
- 239000000843 powder Substances 0.000 abstract description 27
- 230000003064 anti-oxidating effect Effects 0.000 abstract description 3
- 238000004321 preservation Methods 0.000 abstract description 3
- 230000001954 sterilising effect Effects 0.000 description 31
- 239000007788 liquid Substances 0.000 description 17
- 239000002609 medium Substances 0.000 description 12
- 238000004659 sterilization and disinfection Methods 0.000 description 12
- 239000006228 supernatant Substances 0.000 description 10
- 238000001914 filtration Methods 0.000 description 9
- 238000000034 method Methods 0.000 description 9
- 229930192334 Auxin Natural products 0.000 description 8
- 239000002363 auxin Substances 0.000 description 8
- 230000032823 cell division Effects 0.000 description 8
- SEOVTRFCIGRIMH-UHFFFAOYSA-N indole-3-acetic acid Chemical compound C1=CC=C2C(CC(=O)O)=CNC2=C1 SEOVTRFCIGRIMH-UHFFFAOYSA-N 0.000 description 8
- 239000012141 concentrate Substances 0.000 description 6
- 150000003254 radicals Chemical class 0.000 description 6
- 210000003491 skin Anatomy 0.000 description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 6
- 210000004027 cell Anatomy 0.000 description 5
- 238000005119 centrifugation Methods 0.000 description 5
- 238000000855 fermentation Methods 0.000 description 5
- 230000004151 fermentation Effects 0.000 description 5
- 150000002632 lipids Chemical class 0.000 description 5
- 238000004114 suspension culture Methods 0.000 description 5
- 239000005708 Sodium hypochlorite Substances 0.000 description 4
- 230000003035 anti-peroxidant effect Effects 0.000 description 4
- 230000008859 change Effects 0.000 description 4
- 239000003153 chemical reaction reagent Substances 0.000 description 4
- 239000008367 deionised water Substances 0.000 description 4
- 229910021641 deionized water Inorganic materials 0.000 description 4
- 230000001079 digestive effect Effects 0.000 description 4
- 238000004108 freeze drying Methods 0.000 description 4
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 4
- 238000000227 grinding Methods 0.000 description 4
- 239000012535 impurity Substances 0.000 description 4
- 239000012528 membrane Substances 0.000 description 4
- 238000010298 pulverizing process Methods 0.000 description 4
- SUKJFIGYRHOWBL-UHFFFAOYSA-N sodium hypochlorite Chemical compound [Na+].Cl[O-] SUKJFIGYRHOWBL-UHFFFAOYSA-N 0.000 description 4
- 230000032683 aging Effects 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 210000001519 tissue Anatomy 0.000 description 3
- 241000196324 Embryophyta Species 0.000 description 2
- 102000006587 Glutathione peroxidase Human genes 0.000 description 2
- 108700016172 Glutathione peroxidases Proteins 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 description 2
- 102000019197 Superoxide Dismutase Human genes 0.000 description 2
- 108010012715 Superoxide dismutase Proteins 0.000 description 2
- 230000003078 antioxidant effect Effects 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- PSFDQSOCUJVVGF-UHFFFAOYSA-N harman Chemical compound C12=CC=CC=C2NC2=C1C=CN=C2C PSFDQSOCUJVVGF-UHFFFAOYSA-N 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 239000002207 metabolite Substances 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 239000003960 organic solvent Substances 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- PFTAWBLQPZVEMU-DZGCQCFKSA-N (+)-catechin Chemical compound C1([C@H]2OC3=CC(O)=CC(O)=C3C[C@@H]2O)=CC=C(O)C(O)=C1 PFTAWBLQPZVEMU-DZGCQCFKSA-N 0.000 description 1
- PFTAWBLQPZVEMU-ZFWWWQNUSA-N (+)-epicatechin Natural products C1([C@@H]2OC3=CC(O)=CC(O)=C3C[C@@H]2O)=CC=C(O)C(O)=C1 PFTAWBLQPZVEMU-ZFWWWQNUSA-N 0.000 description 1
- PFTAWBLQPZVEMU-UKRRQHHQSA-N (-)-epicatechin Chemical compound C1([C@H]2OC3=CC(O)=CC(O)=C3C[C@H]2O)=CC=C(O)C(O)=C1 PFTAWBLQPZVEMU-UKRRQHHQSA-N 0.000 description 1
- XFZJEEAOWLFHDH-UHFFFAOYSA-N (2R,2'R,3R,3'R,4R)-3,3',4',5,7-Pentahydroxyflavan(48)-3,3',4',5,7-pentahydroxyflavan Natural products C=12OC(C=3C=C(O)C(O)=CC=3)C(O)CC2=C(O)C=C(O)C=1C(C1=C(O)C=C(O)C=C1O1)C(O)C1C1=CC=C(O)C(O)=C1 XFZJEEAOWLFHDH-UHFFFAOYSA-N 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 240000006053 Garcinia mangostana Species 0.000 description 1
- 235000017048 Garcinia mangostana Nutrition 0.000 description 1
- 102000004317 Lyases Human genes 0.000 description 1
- 108090000856 Lyases Proteins 0.000 description 1
- WSMYVTOQOOLQHP-UHFFFAOYSA-N Malondialdehyde Chemical compound O=CCC=O WSMYVTOQOOLQHP-UHFFFAOYSA-N 0.000 description 1
- CWEZAWNPTYBADX-UHFFFAOYSA-N Procyanidin Natural products OC1C(OC2C(O)C(Oc3c2c(O)cc(O)c3C4C(O)C(Oc5cc(O)cc(O)c45)c6ccc(O)c(O)c6)c7ccc(O)c(O)c7)c8c(O)cc(O)cc8OC1c9ccc(O)c(O)c9 CWEZAWNPTYBADX-UHFFFAOYSA-N 0.000 description 1
- MOJZMWJRUKIQGL-FWCKPOPSSA-N Procyanidin C2 Natural products O[C@@H]1[C@@H](c2cc(O)c(O)cc2)Oc2c([C@H]3[C@H](O)[C@@H](c4cc(O)c(O)cc4)Oc4c3c(O)cc(O)c4)c(O)cc(O)c2[C@@H]1c1c(O)cc(O)c2c1O[C@@H]([C@H](O)C2)c1cc(O)c(O)cc1 MOJZMWJRUKIQGL-FWCKPOPSSA-N 0.000 description 1
- 241000973497 Siphonognathus argyrophanes Species 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- SRPWHXRCOLBHSL-UHFFFAOYSA-N astragaloside III Natural products CC(C)(O)C1COC(C)(C1)C2C(O)CC3(C)C4CC(OC5OCC(O)C(O)C5OC6OC(CO)C(O)C(O)C6O)C7C(C)(C)C(O)CCC78CC48CCC23C SRPWHXRCOLBHSL-UHFFFAOYSA-N 0.000 description 1
- FVFSMBDVZVUETN-PTCJJXKDSA-N astragalosideiii Chemical compound O1[C@H](C(C)(O)C)CC[C@@]1(C)[C@@H]1[C@@]2(C)CC[C@]34C[C@]4(CC[C@H](O[C@H]4[C@@H]([C@@H](O)[C@H](O)CO4)O[C@H]4[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O4)O)C4(C)C)[C@H]4[C@@H](O)C[C@H]3[C@]2(C)C[C@@H]1O FVFSMBDVZVUETN-PTCJJXKDSA-N 0.000 description 1
- 230000003197 catalytic effect Effects 0.000 description 1
- ADRVNXBAWSRFAJ-UHFFFAOYSA-N catechin Natural products OC1Cc2cc(O)cc(O)c2OC1c3ccc(O)c(O)c3 ADRVNXBAWSRFAJ-UHFFFAOYSA-N 0.000 description 1
- 235000005487 catechin Nutrition 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- 229950001002 cianidanol Drugs 0.000 description 1
- 238000004737 colorimetric analysis Methods 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 238000004132 cross linking Methods 0.000 description 1
- 239000003431 cross linking reagent Substances 0.000 description 1
- 210000000172 cytosol Anatomy 0.000 description 1
- 230000007123 defense Effects 0.000 description 1
- 210000004207 dermis Anatomy 0.000 description 1
- 238000007599 discharging Methods 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- LPTRNLNOHUVQMS-UHFFFAOYSA-N epicatechin Natural products Cc1cc(O)cc2OC(C(O)Cc12)c1ccc(O)c(O)c1 LPTRNLNOHUVQMS-UHFFFAOYSA-N 0.000 description 1
- 235000012734 epicatechin Nutrition 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000004744 fabric Substances 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- LNTHITQWFMADLM-UHFFFAOYSA-N gallic acid Chemical compound OC(=O)C1=CC(O)=C(O)C(O)=C1 LNTHITQWFMADLM-UHFFFAOYSA-N 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- RBTARNINKXHZNM-UHFFFAOYSA-K iron trichloride Chemical compound Cl[Fe](Cl)Cl RBTARNINKXHZNM-UHFFFAOYSA-K 0.000 description 1
- 238000002386 leaching Methods 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 229940118019 malondialdehyde Drugs 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 238000005502 peroxidation Methods 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 230000003389 potentiating effect Effects 0.000 description 1
- HGVVOUNEGQIPMS-UHFFFAOYSA-N procyanidin Chemical compound O1C2=CC(O)=CC(O)=C2C(O)C(O)C1(C=1C=C(O)C(O)=CC=1)OC1CC2=C(O)C=C(O)C=C2OC1C1=CC=C(O)C(O)=C1 HGVVOUNEGQIPMS-UHFFFAOYSA-N 0.000 description 1
- 229920002414 procyanidin Polymers 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000009758 senescence Effects 0.000 description 1
- 239000002002 slurry Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/0018—Culture media for cell or tissue culture
- C12N5/0025—Culture media for plant cell or plant tissue culture
-
- 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
- A61K31/33—Heterocyclic compounds
- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/35—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
- A61K31/352—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom condensed with carbocyclic rings, e.g. methantheline
- A61K31/353—3,4-Dihydrobenzopyrans, e.g. chroman, catechin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/49—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds
- A61K8/4973—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds with oxygen as the only hetero atom
- A61K8/498—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds with oxygen as the only hetero atom having 6-membered rings or their condensed derivatives, e.g. coumarin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/08—Anti-ageing preparations
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P17/00—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms
- C12P17/16—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms containing two or more hetero rings
- C12P17/162—Heterorings having oxygen atoms as the only ring heteroatoms, e.g. Lasalocid
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2500/00—Specific components of cell culture medium
- C12N2500/30—Organic components
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2500/00—Specific components of cell culture medium
- C12N2500/30—Organic components
- C12N2500/34—Sugars
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2500/00—Specific components of cell culture medium
- C12N2500/30—Organic components
- C12N2500/40—Nucleotides, nucleosides, bases
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Biotechnology (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Animal Behavior & Ethology (AREA)
- Biomedical Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Epidemiology (AREA)
- Pharmacology & Pharmacy (AREA)
- General Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Gerontology & Geriatric Medicine (AREA)
- Dermatology (AREA)
- Botany (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Cell Biology (AREA)
- Birds (AREA)
- Cosmetics (AREA)
Abstract
The present invention relates to the preparation method of cell field, more particularly to culture medium and OPC.The culture medium includes 2,4 D, 6 BA and basal medium.Secondary metabolites in the Vitis culture obtained by the culture medium contains OPC, and its secondary metabolites is collected to carry out being lyophilized preservation.Have anti-oxidation efficacy using OPC, the purpose of anti-aging is reached, the freeze-dried powder can be applied in cosmetics.
Description
Technical field
The present invention relates to cell field, the more particularly to preparation method of culture medium and Vitis secondary metabolites.
Background technology
Grape pip procyanidin is covalent by C4-C6 or C4-C8 key by catechin, epicatechin and its gallate
The polymer being connected to form.Due to electron rich hydroxylic moiety in the molecule of this kind of material, making it have stronger antioxygen
Change, remove the ability of free radical and anti-lipid peroxidation, be the most potent Astragaloside Ⅲ and lipid having now been found that
One of peroxidating inhibitor.
Aging is the extremely complex physiological phenomenon necessarily occurred in life process, at present most representational senescence
One of say it is free radical theory that Harman in 1956 is proposed.The research of Das etc. shows exist in human body and remove the anti-of free radical
Imperial system, including superoxide dismutase (superoxide dismutase, SOD), glutathione peroxidase
Antioxidant Enzymes such as (glutathione peroxidase, GSH-Px).With advancing age, the conjunction of anti-oxidative defense material
Become and activity decrease, a large amount of metabolite MDA (malondialdehyde, MDA) is produced, body produces and remove free radical
Balance be destroyed, free radical pile up, accelerate body aging.
The OPC of grape all uses chemical leaching test at present.There are the following problems：1. chemical synthesis process complex process.
2. security is low, and used in production process, a large amount of organic solvents, easily remain in finished product.3. the Growing season of grape is limited to
Section, affects the yield of finished product.4. cell mass propgation has concentration working substance.
Content of the invention
In view of this, the present invention provides the preparation method of a kind of culture medium and Vitis secondary metabolites OPC.
The present invention is using the secondary metabolites in Vitis culture (in synthesizing in Vitis incubation and discharging to nutrient solution
The general name of tool bioactivator) contain OPC, its secondary metabolites is collected to carry out being lyophilized preservation.Using OPC
Tool anti-oxidation efficacy, reaches the purpose of anti-aging, can apply to the freeze-dried powder in cosmetics.
In order to realize foregoing invention purpose, the present invention provides technical scheme below：
The invention provides a kind of culture medium, including 2,4-D, 6-BA and basal medium.
In some specific embodiments of the present invention, the concentration of 2,4-D in the culture medium described in the culture medium
For 0.2～2.0mg/L；Concentration of the 6-BA in the culture medium is 0.5～2.0mg/L.Some in the present invention are concrete
In embodiment, the culture medium also includes glucose.
In some specific embodiments of the present invention, concentration of the glucose in the culture medium described in the culture medium
For 1.0～5.0g/L.
In some specific embodiments of the present invention, basal medium described in the culture medium is MS culture medium.
Present invention also offers the culture medium prepares the application in secondary metabolites in culture Vitis.
In some specific embodiments of the present invention, the secondary metabolites is former blue and white element.
Present invention also offers a kind of preparation method of Vitis secondary metabolites, comprises the steps：
Step 1：Obtain Vitis；
Step 2：Take step 1 acquisition the culture medium that provides with the present invention of Vitis mix after tentatively cultivate, then with
Amplification Culture after the culture medium mixing that invention is provided, collects nutrient solution.
In some specific embodiments of the present invention, the preparation side of Vitis described in the preparation method step 1
Method is digested for taking after grape pomace is mixed with basal medium, cellulase and pectase, is filtered, is collected filtrate.
In some specific embodiments of the present invention, the addition of cellulase described in the preparation method is described
The 0.2～1.0% of grape pomace quality, preferably 0.5～1.0%；The addition of the pectase is the uva cortex
The 0.1～0.5% of amount.
In some specific embodiments of the present invention, the enzyme activity of the cellulase is 30000U/mg, the pectase
Enzyme activity be 50000U/mg.
In some specific embodiments of the present invention, the volume ratio 2 of cellulase and pectase:1.
In some specific embodiments of the present invention, the time of the enzymolysis is 1.5 hours.
In some specific embodiments of the present invention, the condition of the preliminary culture is：Take Vitis addition I to go out
Vitis strain is configured to 5～10 × 10 by bacterium culture medium5The Vitis liquid of/ml density, takes the addition of 100ml Vitis liquid
In the aseptic conical flask of 250ml, on rotary shaker, with 90～150rpm/min, 25～26 DEG C, cultivate 14 days.
The formula of I sterilising medium is as follows：
The formula (in 1L MS culture medium) of I culture medium
| Title | Content | Title | Content |
| 2,4-D | 0.2～1.0mg/L | 6-BA | 1.0～2.0mg/L |
Note：2,4-D is auxin, and 6-BA is the basic element of cell division.
In some specific embodiments of the present invention, the condition of the Amplification Culture is：After preliminary culture, 250g
Centrifugation 10min removes supernatant, and the grape for adding No. II sterilising medium that Vitis are configured to 5-8x106/ml density is thin
Cytosol is placed in fermentation tank carries out suspension culture, 25-50rpm/min, 25-26 DEG C, is passed through air filtering, and speed is 0.001-
0.005m3/ s, cultivates 7 days.
The formula of No. II sterilising medium is as follows:
The formula (in 1L MS culture medium) of No. II sterilising medium
| Title | Content | Title | Content |
| 2,4-D | 1.0～2.0mg/L | 6-BA | 0.5～1.5mg/L |
| Glucose | 1.0～5.0g/L |
Note：2,4-D is auxin, and 6-BA is the basic element of cell division.
In some specific embodiments of the present invention, after step 2 collects nutrient solution, also include to filter, concentrate, collect
Degerming after concentrate, lyophilized step.
In some specific embodiments of the present invention, described being lyophilized is that vacuum is 50～200Pa in -20～-35 DEG C
Under conditions of, it is lyophilized 24～36 hours.
Present invention also offers Vitis secondary metabolites obtained in described preparation method.
In some specific embodiments of the present invention, the Vitis secondary metabolites is OPC.
Present invention also offers the Vitis secondary metabolites prepare the medicine of anti peroxidation of lipid, food and/
Or the application in cosmetics.
Additionally, present invention also offers the Vitis secondary metabolites prepare the medicine of anti-aging, food and/or
Application in cosmetics.
The invention provides a kind of culture medium, including 2,4-D, 6-BA and basal medium.By the culture medium and originally
Secondary metabolites in Vitis culture obtained in the method that invention is provided (synthesizes in Vitis incubation and discharges
Have the general name of bioactivator to nutrient solution) contain OPC, its secondary metabolites is collected to carry out being lyophilized preservation.Profit
Have anti-oxidation efficacy with OPC, the purpose of anti-aging is reached, the freeze-dried powder can be applied in cosmetics.
Specific embodiment
The invention discloses the preparation method of a kind of culture medium and Vitis secondary metabolites OPC, this area skill
Art personnel can use for reference present disclosure, be suitably modified technological parameter realization.Specifically, all similar replacements and
Change is apparent to those skilled in the art, and they are considered as including in the present invention.The method of the present invention and
Application is described by preferred embodiment, and related personnel substantially can be without departing from present invention, spirit and scope
Interior method described herein and application are modified or suitably change and combine, realize and apply the technology of the present invention.
The invention provides a kind of preparation method of mangosteen cell secondary metabolites, comprises the steps：
Obtain Vitis strain：
Take the fresh grape containing grape pip of 1kg and sterilization is carried out, be immersed in 20 points in 15%～20% sodium hypochlorite of 10L
Clock.Liquor natrii hypochloritis is gone, aseptically, is cleaned using 2L aseptic deionized water every time, totally three times, cleaned every time
It is gently agitated in journey.
Under sterile conditions, the grape pomace after sterilization is removed, and grape is pulverized with the grinding after sterilizing.After pulverizing
Grape is transferred in the beaker of sterile clean, adds 10ml MS (Murashige and Skoog) culture per 1.0g grape pomace
Base, adds 0.2～1.0% (w/w) cellulase and 0.1～0.5% (w/w) pectase, digests 1.5 hours.
Digestive juice is filtered with 200 mesh aseptic mesh screen, impurity is removed, obtain Vitis strain.
Vitis mass propgation：
Add I sterilising medium that Vitis strain is configured to 5～10 × 105The Vitis liquid of/ml density, takes
100ml Vitis liquid is added in the aseptic conical flask of 250ml, on rotary shaker, with 90-150rpm/min, 25-26
DEG C, cultivate 14 days.
After preliminary culture, 250g centrifugation 10min removes supernatant, adds No. II sterilising medium to join Vitis
Make 5-8 × 106The Vitis liquid of/ml density is placed in fermentation tank carries out suspension culture, 25-50rpm/min, 25-26 DEG C,
Air filtering was passed through, speed is 0.001-0.005m3/ s, cultivates 7 days.
The preparation of Vitis secondary metabolites freeze-dried powder：
Vitis culture is collected, which is filtered with 0.45 μM of filter membrane, concentrate, the concentration supernatant for obtaining is used
0.22 μM of filtration sterilization.
Frozen dried is carried out after degerming, freeze temperature is -20～-35 DEG C, vacuum is 50～200Pa, freeze-drying time is
24～36 hours, that is, obtain the secondary metabolites freeze-dried powder of Vitis.Freeze-dried powder is distributed into 1ml/ prop up.
The present invention adopts culture plant cell method to extract the blue and white element of the original in grape.And the OPC to extracting is carried out
Frozen dried, it is easy to preserve and apply, increase stability.
1. extraction process is stable, and active ingredient concentration is big, and can preserve its activity.
2. safe, not residual organic solvent.
3. the season of growth impact of grape is not only restricted to.
The present invention provide culture medium and Vitis secondary metabolites OPC preparation method in raw materials used and
Reagent all can be buied by market.
With reference to embodiment, the present invention is expanded on further：
Embodiment 1
Obtain Vitis strain：
Take the fresh grape containing grape pip of 1kg and sterilization is carried out, be immersed in 20 points in 15%～20% sodium hypochlorite of 10L
Clock.Liquor natrii hypochloritis is gone, aseptically, is cleaned using 2L aseptic deionized water every time, totally three times, cleaned every time
It is gently agitated in journey.
Under sterile conditions, the grape pomace after sterilization is removed, and grape is pulverized with the grinding after sterilizing.After pulverizing
Grape is transferred in the beaker of sterile clean, adds 10ml MS (Murashige and Skoog) culture per 1.0g grape pomace
Base, adds 0.2% (w/w) cellulase and 0.3% (w/w) pectase, digests 1.5 hours.The enzyme activity of cellulase is
30000U/mg, the enzyme activity of pectase is 50000U/mg.
Digestive juice is filtered with 200 mesh aseptic mesh screen, impurity is removed, obtain Vitis strain.
Vitis mass propgation：
Add I sterilising medium that Vitis strain is configured to 5-10 × 105The Vitis liquid of/ml density, takes
100ml Vitis liquid is added in the aseptic conical flask of 250ml, on rotary shaker, with 90-150rpm/min, 25-26
DEG C, cultivate 14 days.
After preliminary culture, 250g centrifugation 10min removes supernatant, adds No. II sterilising medium to join Vitis
Make 5-8 × 106The Vitis liquid of/ml density is placed in fermentation tank carries out suspension culture, 25-50rpm/min, 25-26 DEG C,
Air filtering was passed through, speed is 0.001-0.005m3/s, cultivates 7 days.
The formula (in 1L MS culture medium) of 1 I culture medium of table
| Title | Content | Title | Content |
| 2,4-D | 0.2mg/L | 6-BA | 2.0mg/L |
Note：2,4-D is auxin, and 6-BA is the basic element of cell division
The formula (in 1L MS culture medium) of 2 No. II sterilising mediums of table
| Title | Content | Title | Content |
| 2,4-D | 1.0mg/L | 6-BA | 1.5mg/L |
| Glucose | 3.0g/L |
Note：2,4-D is auxin, and 6-BA is the basic element of cell division
The preparation of Vitis secondary metabolites freeze-dried powder：
Vitis culture is collected, which is filtered with 0.45 μM of filter membrane, concentrate, the concentration supernatant for obtaining is used
0.22 μM of filtration sterilization.
Frozen dried is carried out after degerming, freeze temperature is -20 DEG C, vacuum is 120Pa, freeze-drying time is 24 hours, i.e.,
Obtain the secondary metabolites freeze-dried powder of Vitis.Freeze-dried powder is distributed into 1ml/ prop up.
Embodiment 2
Obtain Vitis strain：
Take the fresh grape containing grape pip of 1kg and sterilization is carried out, be immersed in 20 points in 15%～20% sodium hypochlorite of 10L
Clock.Liquor natrii hypochloritis is gone, aseptically, is cleaned using 2L aseptic deionized water every time, totally three times, cleaned every time
It is gently agitated in journey.
Under sterile conditions, the grape pomace after sterilization is removed, and grape is pulverized with the grinding after sterilizing.After pulverizing
Grape is transferred in the beaker of sterile clean, adds 10ml MS (Murashige and Skoog) culture per 1.0g grape pomace
Base, adds 1.0% (w/w) cellulase and 0.1% (w/w) pectase, digests 1.5 hours.The enzyme activity of cellulase is
30000U/mg, the enzyme activity of pectase is 50000U/mg.
Digestive juice is filtered with 200 mesh aseptic mesh screen, impurity is removed, obtain Vitis strain.
Vitis mass propgation：
Add I sterilising medium that Vitis strain is configured to 5-10 × 105The Vitis liquid of/ml density, takes
100ml Vitis liquid is added in the aseptic conical flask of 250ml, on rotary shaker, with 90-150rpm/min, 25-26
DEG C, cultivate 14 days.
After preliminary culture, 250g centrifugation 10min removes supernatant, adds No. II sterilising medium to join Vitis
Make 5-8 × 106The Vitis liquid of/ml density is placed in fermentation tank carries out suspension culture, 25-50rpm/min, 25-26 DEG C,
Air filtering was passed through, speed is 0.001-0.005m3/ s, cultivates 7 days.
The formula (in 1L MS culture medium) of 3 I culture medium of table
| Title | Content | Title | Content |
| 2,4-D | 1.0mg/L | 6-BA | 1.0mg/L |
Note：2,4-D is auxin, and 6-BA is the basic element of cell division
The formula (in 1L MS culture medium) of 4 No. II sterilising mediums of table
| Title | Content | Title | Content |
| 2,4-D | 2.0mg/L | 6-BA | 1.0mg/L |
| Glucose | 1.0g/L |
Note：2,4-D is auxin, and 6-BA is the basic element of cell division
The preparation of Vitis secondary metabolites freeze-dried powder：
Vitis culture is collected, which is filtered with 0.45 μM of filter membrane, concentrate, the concentration supernatant for obtaining is used
0.22 μM of filtration sterilization.
Frozen dried is carried out after degerming, freeze temperature is -35 DEG C, vacuum is 200Pa, freeze-drying time is 36 hours, i.e.,
Obtain the secondary metabolites freeze-dried powder of Vitis.Freeze-dried powder is distributed into 1ml/ prop up.
Embodiment 3
Obtain Vitis strain：
Take the fresh grape containing grape pip of 1kg and sterilization is carried out, be immersed in 20 points in 15%～20% sodium hypochlorite of 10L
Clock.Liquor natrii hypochloritis is gone, aseptically, is cleaned using 2L aseptic deionized water every time, totally three times, cleaned every time
It is gently agitated in journey.
Under sterile conditions, the grape pomace after sterilization is removed, and grape is pulverized with the grinding after sterilizing.After pulverizing
Grape is transferred in the beaker of sterile clean, adds 10ml MS (Murashige and Skoog) culture per 1.0g grape pomace
Base, adds 0.5% (w/w) cellulase and 0.5% (w/w) pectase, digests 1.5 hours.The enzyme activity of cellulase is
30000U/mg, the enzyme activity of pectase is 50000U/mg.
Digestive juice is filtered with 200 mesh aseptic mesh screen, impurity is removed, obtain Vitis strain.
Vitis mass propgation：
Add I sterilising medium that Vitis strain is configured to 5-10 × 105The Vitis liquid of/ml density, takes
100ml Vitis liquid is added in the aseptic conical flask of 250ml, on rotary shaker, with 90-150rpm/min, 25-26
DEG C, cultivate 14 days.
After preliminary culture, 250g centrifugation 10min removes supernatant, adds No. II sterilising medium to join Vitis
Make 5-8 × 106The Vitis liquid of/ml density is placed in fermentation tank carries out suspension culture, 25-50rpm/min, 25-26 DEG C,
Air filtering was passed through, speed is 0.001-0.005m3/s, cultivates 7 days.
The formula (in 1L MS culture medium) of 5 I culture medium of table
| Title | Content | Title | Content |
| 2,4-D | 0.6mg/L | 6-BA | 1.5mg/L |
Note：2,4-D is auxin, and 6-BA is the basic element of cell division
The formula (in 1L MS culture medium) of 6 No. II sterilising mediums of table
| Title | Content | Title | Content |
| 2,4-D | 1.5mg/L | 6-BA | 0.5mg/L |
| Glucose | 5.0g/L |
Note：2,4-D is auxin, and 6-BA is the basic element of cell division
The preparation of Vitis secondary metabolites freeze-dried powder：
Vitis culture is collected, which is filtered with 0.45 μM of filter membrane, concentrate, the concentration supernatant for obtaining is used
0.22 μM of filtration sterilization.
Frozen dried is carried out after degerming, freeze temperature is -28 DEG C, vacuum is 50Pa, freeze-drying time is 30 hours, obtains final product
Secondary metabolites freeze-dried powder to Vitis.Freeze-dried powder is distributed into 1ml/ prop up.
The measure of the blue and white element in 4 secondary metabolites freeze-dried powder Central Plains of embodiment
Using molysite catalytic colorimetry.
Standard group：By former blue and white element be configured to concentration be 0,20,40,60,80, the standard liquid of 100mg/ml
Sample sets 1～3：Freeze-dried powder prepared by 1～embodiment of embodiment 3 is dissolved into liquid.
Mixed liquor：By volume 85:5:0.4 n-butanol, concentrated hydrochloric acid, Fe3+Ar ion mixing liquid.
Sample and titer respectively take 1ml, all plus 9ml mixed liquor, in 60 DEG C of bath temperature, react 60min, cold after taking-up
But, its absorption photometric value A is surveyed under 550nm wavelength, draw its former blue and white element concentration.The results are shown in Table 7.
Table 7
| Group | Former blue and white cellulose content (mg/ml) |
| Sample sets 1 (prepared by embodiment 1) | 86.4 |
| Sample sets 2 (prepared by embodiment 2) | 86.7 |
| Sample sets 3 (prepared by embodiment 3) | 85.7 |
Conclusion：The culture plant cell method of this patent can successfully extract the blue and white element of abundant original, and between different batches
Content nothing significant difference (P<0.05), it was demonstrated that method is stable.
Impact of the 5 grape secondary metabolites freeze-dried powder of embodiment to MDA growing amount in the homogenate of big white mouse skin histology
After free radical is formed, its metabolite MDA (MDA) content substantially increases, and MDA is as lively as a cricket crosslinking agent, it
Can make dermis that macromolecules cross-linking occurs, increase corium fabric distortion thick disorderly, the skin day by day aging in appearance of people is made,
The height of MDA content can characterize anti peroxidation of lipid (the i.e. anti-MDA is generated) activity of material indirectly, and MDA content is less, shows thing
The anti peroxidation of lipid ability of matter is stronger；Conversely, then anti peroxidation of lipid ability is weaker.
40 big white mouse are divided into 4 groups, per 10 are organized, its back are gone to by hair, exposed area about 6cm.,
Take with Vitis secondary metabolites freeze-dried powder obtained in a batch of embodiment 1, will match with which before smearing
2ml lyase is poured in freeze-dried powder completely, fully mixes, 2ml mixture is all equably applied to the exposed back of mouse, per
Smear once within two days.
First group, put to death after continuously smearing Vitis secondary metabolites freeze-dried powder obtained in embodiment 15 times, take back
Skin histology is homogenized, and is made as 10% homogenate.Second group, continuously smear Vitis secondary metabolites obtained in embodiment 1
Put to death after freeze-dried powder 10 times, back skin tissues homogenate is taken, is made as 10% homogenate.3rd group, continuously smear embodiment 1 and make
Vitis secondary metabolites freeze-dried powder 15 times after put to death, take back skin tissues homogenate, be made as 10% homogenate.The
Four groups are not smeared Vitis secondary metabolites freeze-dried powder obtained in embodiment 1, put to death, take back skin tissues even after 30 days
Slurry, is made as 10% homogenate.
Standard pipe take 0.2ml 10mol/ml standard items, standard blank tube take 0.2ml absolute ethyl alcohol, determine pipe take 0.2ml
Test sample, then three pipes all addition 0.2ml reagents one, after mixing, all add 3ml reagent two and 1ml reagent three.
Swirl mixing device is mixed, and test tube mouth tightened with antistaling film, pierces an aperture, 95 DEG C of water-baths 40 minutes, is flowed after taking-up
Water cooling, is centrifuged 10 minutes by then 3500 revs/min.Supernatant is taken, at 532mm, 1cm optical path, distilled water return to zero, and colorimetric surveys each pipe
Absorbance, detects the MAD content of each group.The results are shown in Table 8.
Table 8
Note：* data are shown between control group there were significant differences (P<0.05)
Conclusion：Prove that the blue and white element of the original in the Vitis secondary metabolites freeze-dried powder that the present invention is provided has obvious antioxygen
Change acts on (P<0.05), with the increase using number of days, effect is obviously improved.
Vitis secondary metabolites freeze-dried powder prepared by Example 2, embodiment 3 carries out above-mentioned experiment, experimental result
The experimental result of the Vitis secondary metabolites freeze-dried powder prepared with embodiment 1 is close, with which nothing significant difference (P ＞
0.05).
The above is only the preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, under the premise without departing from the principles of the invention, some improvements and modifications can also be made, these improvements and modifications also should
It is considered as protection scope of the present invention.
Claims (10)
1. a kind of culture medium, it is characterised in that including 2,4-D, 6-BA and basal medium.
2. culture medium according to claim 1, it is characterised in that the concentration of 2, the 4-D in the culture medium is 0.2
～2.0mg/L；Concentration of the 6-BA in the culture medium is 0.5～2.0mg/L.
3. culture medium according to claim 1 and 2, it is characterised in that also include glucose.
4. culture medium according to claim 3, it is characterised in that including following component：The glucose is in the culture
Concentration in base is 1.0～5.0g/L.
5. the culture medium according to any one of Claims 1-4 prepares the application in secondary metabolites in culture Vitis.
6. a kind of preparation method of Vitis secondary metabolites, it is characterised in that comprise the steps：
Step 1：Obtain Vitis；
Step 2：Take step 1 acquisition Vitis mix with the culture medium as described in claim 1 or claim 2 after tentatively
Culture, then Amplification Culture after mixing with the culture medium as described in claim 3 or claim 4, collect nutrient solution.
7. preparation method according to claim 6, it is characterised in that the preparation method of Vitis described in step 1 is
Take after grape pomace is mixed with basal medium, cellulase and pectase and digest, filter, collect filtrate.
8. preparation method according to claim 7, it is characterised in that the addition of the cellulase is the uva
The 0.2～1.0% of cortex amount, the addition of the pectase are the 0.1～0.5% of the grape pomace quality；
The enzyme activity of the cellulase is 30000U/mg, and the enzyme activity of the pectase is 50000U/mg.
9. Vitis secondary metabolites obtained in the preparation method according to any one of claim 6 to 8.
10. Vitis secondary metabolites according to claim 9 is preparing the medicine of anti-aging, food and/or cosmetic
Application in product.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610963852.XA CN106479952A (en) | 2016-11-04 | 2016-11-04 | Culture medium and the preparation method of Vitis secondary metabolites |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610963852.XA CN106479952A (en) | 2016-11-04 | 2016-11-04 | Culture medium and the preparation method of Vitis secondary metabolites |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN106479952A true CN106479952A (en) | 2017-03-08 |
Family
ID=58271883
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610963852.XA Pending CN106479952A (en) | 2016-11-04 | 2016-11-04 | Culture medium and the preparation method of Vitis secondary metabolites |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106479952A (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104099287A (en) * | 2014-07-07 | 2014-10-15 | 福建省农业科学院农业工程技术研究所 | Acquiring and subculture maintaining method for high yield oligomeric proanthocyanidins vitis davidii callus |
| CN104628698A (en) * | 2015-02-11 | 2015-05-20 | 广州赛莱拉干细胞科技股份有限公司 | Preparation method and application of proanthocyanidin extract in grapes |
-
2016
- 2016-11-04 CN CN201610963852.XA patent/CN106479952A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104099287A (en) * | 2014-07-07 | 2014-10-15 | 福建省农业科学院农业工程技术研究所 | Acquiring and subculture maintaining method for high yield oligomeric proanthocyanidins vitis davidii callus |
| CN104628698A (en) * | 2015-02-11 | 2015-05-20 | 广州赛莱拉干细胞科技股份有限公司 | Preparation method and application of proanthocyanidin extract in grapes |
Non-Patent Citations (2)
| Title |
|---|
| CORMIER ET AL: "Anthocyanin Production in Selected Cell Lines of Grape (Vitis vinifera L.)", 《PLANT》 * |
| 傅术琳等: "木立芦荟愈伤组织的诱导及快速繁殖", 《植物生理学通讯》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103145780A (en) | Preparation method of blueberry anthocyanin and application of blueberry anthocyanin prepared | |
| CN102584411A (en) | Agar protococcus biologic fertilizer and preparation method thereof | |
| KR101148513B1 (en) | Method for preparing producing an enrichment composition of ergothioneine from gastrodiae elata | |
| EP3033073A1 (en) | Preparation for protection against extrinsic and intrinsic skin aging | |
| CN108353906A (en) | The application of indole -3-formaldehyde and its derivative in preventing the plant disease caused by plant pathogenic fungi | |
| KR20180020571A (en) | Cosmetic compositon for removing heavy metal or fine dust containing extracts of ficus religosa | |
| KR20180020573A (en) | Cosmetic compositon for removing heavy metal or fine dust containing extracts of ficus panda and epipremnum aureum | |
| WO2021093299A1 (en) | Astragalus-paecilomyces cicadae fermented fungal substance, preparation method and application thereof | |
| CN106479952A (en) | Culture medium and the preparation method of Vitis secondary metabolites | |
| CN104706554B (en) | A kind of preparation method and applications of lotus leaf cell secondary metabolites freeze-dried powder | |
| CN105362140A (en) | Non-cosmetics preservative composition with anticorrosion effect and application of non-cosmetics preservative composition | |
| KR101045011B1 (en) | Method for preparing composition comprising fermented by using of hippophae rhamnoides l | |
| CN108553487A (en) | A kind of preparation method of fermentation cordyceps Cs-4 medicine materical crude slice | |
| CN112353734A (en) | Fermentation method of centella fermented product and application of centella fermented product in repair-efficacy cosmetics | |
| CN112006947B (en) | Seaweed fermentation liquor with multiple skin care effects and preparation method thereof | |
| CN103877586B (en) | Medicinal hard capsule shell material containing eucheuma gelatinae polysaccharide and preparation method | |
| CN112852572A (en) | Method for preparing selenium-rich, strontium-rich and zinc-rich procyanidine SOD yellow wine | |
| CN106497860A (en) | Culture medium and the preparation method of Garcinia mangostana cell secondary metabolitess | |
| CN102676397B (en) | Artificial cultivation method of Paecilomyces tenuipes (Peck) Samson and pharmaceutical use of Paecilomyces tenuipes (Peck) Ssamson | |
| CN109793699A (en) | A kind of preparation method of Rhizoma Gastrodiae facial mask cream | |
| KR20110067813A (en) | Mixed tea using leaf, root bark and fruit of mulberry tree | |
| CN115058347B (en) | Preparation and application of lepista sordida and non-saccharide extract thereof | |
| CN114891852B (en) | Method for obtaining high-content cyclamen saponin fermentation liquor, fermentation liquor and application thereof | |
| CN115286459B (en) | Plant fertilizer containing polysaccharide complex elements and preparation method thereof | |
| CN110818811B (en) | Method for extracting and separating high-viscosity narcissus polysaccharide from narcissus bulb |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20170308 |