CN113402626A - Nymphaea hybrid polysaccharide extract and preparation method and application thereof - Google Patents
Nymphaea hybrid polysaccharide extract and preparation method and application thereof Download PDFInfo
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- CN113402626A CN113402626A CN202110784420.3A CN202110784420A CN113402626A CN 113402626 A CN113402626 A CN 113402626A CN 202110784420 A CN202110784420 A CN 202110784420A CN 113402626 A CN113402626 A CN 113402626A
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- extract
- polysaccharide
- precipitate
- crude extract
- filtering
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Links
- 150000004676 glycans Chemical class 0.000 title claims abstract description 84
- 229920001282 polysaccharide Polymers 0.000 title claims abstract description 83
- 239000005017 polysaccharide Substances 0.000 title claims abstract description 83
- 241000209490 Nymphaea Species 0.000 title claims abstract description 53
- 239000000284 extract Substances 0.000 title claims abstract description 40
- 238000002360 preparation method Methods 0.000 title claims abstract description 14
- 238000000605 extraction Methods 0.000 title claims abstract description 11
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 28
- 239000000287 crude extract Substances 0.000 claims abstract description 27
- 239000002244 precipitate Substances 0.000 claims abstract description 25
- 238000003756 stirring Methods 0.000 claims abstract description 25
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 24
- 238000001914 filtration Methods 0.000 claims abstract description 20
- 238000000034 method Methods 0.000 claims abstract description 19
- 238000002156 mixing Methods 0.000 claims abstract description 12
- 238000001035 drying Methods 0.000 claims abstract description 10
- 230000002087 whitening effect Effects 0.000 claims abstract description 10
- 230000000844 anti-bacterial effect Effects 0.000 claims abstract description 8
- 238000000502 dialysis Methods 0.000 claims abstract description 8
- 238000004108 freeze drying Methods 0.000 claims abstract description 8
- 239000007788 liquid Substances 0.000 claims abstract description 7
- 230000003078 antioxidant effect Effects 0.000 claims abstract description 5
- 238000010298 pulverizing process Methods 0.000 claims abstract description 5
- 238000005227 gel permeation chromatography Methods 0.000 claims abstract description 4
- 239000012528 membrane Substances 0.000 claims abstract description 4
- OQUKIQWCVTZJAF-UHFFFAOYSA-N phenol;sulfuric acid Chemical compound OS(O)(=O)=O.OC1=CC=CC=C1 OQUKIQWCVTZJAF-UHFFFAOYSA-N 0.000 claims abstract description 4
- 150000002772 monosaccharides Chemical class 0.000 claims abstract description 3
- 229920001542 oligosaccharide Polymers 0.000 claims abstract description 3
- 150000002482 oligosaccharides Chemical class 0.000 claims abstract description 3
- 230000001376 precipitating effect Effects 0.000 claims abstract description 3
- 238000005406 washing Methods 0.000 claims abstract description 3
- 239000006228 supernatant Substances 0.000 claims description 8
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 6
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- 239000003963 antioxidant agent Substances 0.000 claims description 3
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- 239000004480 active ingredient Substances 0.000 claims description 2
- 230000003544 deproteinization Effects 0.000 claims description 2
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- 239000002904 solvent Substances 0.000 claims description 2
- 238000003809 water extraction Methods 0.000 claims description 2
- 235000013402 health food Nutrition 0.000 claims 2
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- 238000002835 absorbance Methods 0.000 description 6
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- 235000014469 Bacillus subtilis Nutrition 0.000 description 4
- 241000193388 Bacillus thuringiensis Species 0.000 description 4
- XUMBMVFBXHLACL-UHFFFAOYSA-N Melanin Chemical compound O=C1C(=O)C(C2=CNC3=C(C(C(=O)C4=C32)=O)C)=C2C4=CNC2=C1C XUMBMVFBXHLACL-UHFFFAOYSA-N 0.000 description 4
- 239000007864 aqueous solution Substances 0.000 description 4
- 239000002304 perfume Substances 0.000 description 4
- 239000000843 powder Substances 0.000 description 4
- 239000000047 product Substances 0.000 description 4
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 3
- RJQXTJLFIWVMTO-TYNCELHUSA-N Methicillin Chemical compound COC1=CC=CC(OC)=C1C(=O)N[C@@H]1C(=O)N2[C@@H](C(O)=O)C(C)(C)S[C@@H]21 RJQXTJLFIWVMTO-TYNCELHUSA-N 0.000 description 3
- 241000612118 Samolus valerandi Species 0.000 description 3
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- 150000001875 compounds Chemical class 0.000 description 3
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- 238000007254 oxidation reaction Methods 0.000 description 3
- 229910021642 ultra pure water Inorganic materials 0.000 description 3
- 239000012498 ultrapure water Substances 0.000 description 3
- IIZPXYDJLKNOIY-JXPKJXOSSA-N 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCC\C=C/C\C=C/C\C=C/C\C=C/CCCCC IIZPXYDJLKNOIY-JXPKJXOSSA-N 0.000 description 2
- 244000063299 Bacillus subtilis Species 0.000 description 2
- 241000588724 Escherichia coli Species 0.000 description 2
- 241000192125 Firmicutes Species 0.000 description 2
- WTDRDQBEARUVNC-LURJTMIESA-N L-DOPA Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C(O)=C1 WTDRDQBEARUVNC-LURJTMIESA-N 0.000 description 2
- 235000016791 Nymphaea odorata subsp odorata Nutrition 0.000 description 2
- PLXBWHJQWKZRKG-UHFFFAOYSA-N Resazurin Chemical compound C1=CC(=O)C=C2OC3=CC(O)=CC=C3[N+]([O-])=C21 PLXBWHJQWKZRKG-UHFFFAOYSA-N 0.000 description 2
- UEDUENGHJMELGK-HYDKPPNVSA-N Stevioside Chemical compound O([C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1O[C@]12C(=C)C[C@@]3(C1)CC[C@@H]1[C@@](C)(CCC[C@]1([C@@H]3CC2)C)C(=O)O[C@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O1)O)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O UEDUENGHJMELGK-HYDKPPNVSA-N 0.000 description 2
- 235000006708 antioxidants Nutrition 0.000 description 2
- 229940097012 bacillus thuringiensis Drugs 0.000 description 2
- 230000003115 biocidal effect Effects 0.000 description 2
- 238000009835 boiling Methods 0.000 description 2
- 229940046011 buccal tablet Drugs 0.000 description 2
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- 238000006243 chemical reaction Methods 0.000 description 2
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- 238000012258 culturing Methods 0.000 description 2
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- 230000018109 developmental process Effects 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- IDGUHHHQCWSQLU-UHFFFAOYSA-N ethanol;hydrate Chemical compound O.CCO IDGUHHHQCWSQLU-UHFFFAOYSA-N 0.000 description 2
- 239000008187 granular material Substances 0.000 description 2
- 238000002386 leaching Methods 0.000 description 2
- 239000000787 lecithin Substances 0.000 description 2
- 235000010445 lecithin Nutrition 0.000 description 2
- 229940067606 lecithin Drugs 0.000 description 2
- HQKMJHAJHXVSDF-UHFFFAOYSA-L magnesium stearate Chemical compound [Mg+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O HQKMJHAJHXVSDF-UHFFFAOYSA-L 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 239000011259 mixed solution Substances 0.000 description 2
- 239000013641 positive control Substances 0.000 description 2
- 229940013618 stevioside Drugs 0.000 description 2
- OHHNJQXIOPOJSC-UHFFFAOYSA-N stevioside Natural products CC1(CCCC2(C)C3(C)CCC4(CC3(CCC12C)CC4=C)OC5OC(CO)C(O)C(O)C5OC6OC(CO)C(O)C(O)C6O)C(=O)OC7OC(CO)C(O)C(O)C7O OHHNJQXIOPOJSC-UHFFFAOYSA-N 0.000 description 2
- 235000019202 steviosides Nutrition 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
- QCDWFXQBSFUVSP-UHFFFAOYSA-N 2-phenoxyethanol Chemical compound OCCOC1=CC=CC=C1 QCDWFXQBSFUVSP-UHFFFAOYSA-N 0.000 description 1
- RVBUGGBMJDPOST-UHFFFAOYSA-N 2-thiobarbituric acid Chemical compound O=C1CC(=O)NC(=S)N1 RVBUGGBMJDPOST-UHFFFAOYSA-N 0.000 description 1
- 241001391944 Commicarpus scandens Species 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 239000008868 Flower Essence Substances 0.000 description 1
- WTDRDQBEARUVNC-UHFFFAOYSA-N L-Dopa Natural products OC(=O)C(N)CC1=CC=C(O)C(O)=C1 WTDRDQBEARUVNC-UHFFFAOYSA-N 0.000 description 1
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- 241000209477 Nymphaeaceae Species 0.000 description 1
- 102000004316 Oxidoreductases Human genes 0.000 description 1
- 108090000854 Oxidoreductases Proteins 0.000 description 1
- 102000011143 Placentin Human genes 0.000 description 1
- 108050001350 Placentin Proteins 0.000 description 1
- 108010039918 Polylysine Proteins 0.000 description 1
- 238000012356 Product development Methods 0.000 description 1
- 239000004288 Sodium dehydroacetate Substances 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- 241001530121 Trollius Species 0.000 description 1
- 241000206488 Trollius laxus Species 0.000 description 1
- TVXBFESIOXBWNM-UHFFFAOYSA-N Xylitol Natural products OCCC(O)C(O)C(O)CCO TVXBFESIOXBWNM-UHFFFAOYSA-N 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000003064 anti-oxidating effect Effects 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 239000007844 bleaching agent Substances 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- CDQSJQSWAWPGKG-UHFFFAOYSA-N butane-1,1-diol Chemical compound CCCC(O)O CDQSJQSWAWPGKG-UHFFFAOYSA-N 0.000 description 1
- 230000001914 calming 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
- 230000008859 change Effects 0.000 description 1
- 229950001002 cianidanol Drugs 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 238000012937 correction Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 230000003203 everyday effect Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000012065 filter cake Substances 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 239000003205 fragrance Substances 0.000 description 1
- 235000020510 functional beverage Nutrition 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 239000010931 gold Substances 0.000 description 1
- 229910052737 gold Inorganic materials 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 229910052742 iron Inorganic materials 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 235000019359 magnesium stearate Nutrition 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 238000010907 mechanical stirring Methods 0.000 description 1
- -1 mechanically stir Substances 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 230000008099 melanin synthesis Effects 0.000 description 1
- 201000001441 melanoma Diseases 0.000 description 1
- HEBKCHPVOIAQTA-UHFFFAOYSA-N meso ribitol Natural products OCC(O)C(O)C(O)CO HEBKCHPVOIAQTA-UHFFFAOYSA-N 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 239000012046 mixed solvent Substances 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- 238000005502 peroxidation Methods 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 229960005323 phenoxyethanol Drugs 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- 238000007747 plating Methods 0.000 description 1
- 229920000656 polylysine Polymers 0.000 description 1
- 150000004804 polysaccharides Polymers 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000002601 radiography Methods 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- 239000012488 sample solution Substances 0.000 description 1
- 229920006395 saturated elastomer Polymers 0.000 description 1
- 229940079839 sodium dehydroacetate Drugs 0.000 description 1
- 235000019259 sodium dehydroacetate Nutrition 0.000 description 1
- DSOWAKKSGYUMTF-GZOLSCHFSA-M sodium;(1e)-1-(6-methyl-2,4-dioxopyran-3-ylidene)ethanolate Chemical compound [Na+].C\C([O-])=C1/C(=O)OC(C)=CC1=O DSOWAKKSGYUMTF-GZOLSCHFSA-M 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 239000012085 test solution Substances 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 238000010257 thawing Methods 0.000 description 1
- 230000002936 tranquilizing effect Effects 0.000 description 1
- YNJBWRMUSHSURL-UHFFFAOYSA-N trichloroacetic acid Chemical compound OC(=O)C(Cl)(Cl)Cl YNJBWRMUSHSURL-UHFFFAOYSA-N 0.000 description 1
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 description 1
- 238000002137 ultrasound extraction Methods 0.000 description 1
- 230000017260 vegetative to reproductive phase transition of meristem Effects 0.000 description 1
- HEBKCHPVOIAQTA-SCDXWVJYSA-N xylitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)CO HEBKCHPVOIAQTA-SCDXWVJYSA-N 0.000 description 1
- 235000010447 xylitol Nutrition 0.000 description 1
- 239000000811 xylitol Substances 0.000 description 1
- 229960002675 xylitol Drugs 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
-
- 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
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/52—Adding ingredients
-
- 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
- A23L33/125—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives containing carbohydrate syrups; containing sugars; containing sugar alcohols; containing starch hydrolysates
-
- 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
- A61K8/72—Cosmetics or similar toiletry preparations characterised by the composition containing organic macromolecular compounds
- A61K8/73—Polysaccharides
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q17/00—Barrier preparations; Preparations brought into direct contact with the skin for affording protection against external influences, e.g. sunlight, X-rays or other harmful rays, corrosive materials, bacteria or insect stings
- A61Q17/005—Antimicrobial preparations
-
- 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
-
- 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
- A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening the skin
-
- 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
-
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/805—Corresponding aspects not provided for by any of codes A61K2800/81 - A61K2800/95
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/84—Products or compounds obtained by lyophilisation, freeze-drying
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Veterinary Medicine (AREA)
- Chemical & Material Sciences (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Dermatology (AREA)
- Polymers & Plastics (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Molecular Biology (AREA)
- Nutrition Science (AREA)
- Materials Engineering (AREA)
- Medicinal Chemistry (AREA)
- Biochemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Organic Chemistry (AREA)
- Sustainable Development (AREA)
- Gerontology & Geriatric Medicine (AREA)
- Birds (AREA)
- Epidemiology (AREA)
- Mycology (AREA)
- Medicines Containing Plant Substances (AREA)
- Cosmetics (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses a nymphaea hybrid polysaccharide extract and a preparation method and application thereof. Pulverizing nymphaea hybrid, adding water for extraction, and concentrating the extract to obtain extract; adding ethanol solution into the extract, stirring, filtering, and freeze drying the precipitate to obtain polysaccharide crude extract I; dissolving polysaccharide crude extract I in water, deproteinizing by Sevag method, washing out precipitate, filtering, and drying precipitate to obtain polysaccharide crude extract II; removing monosaccharide and oligosaccharide: dissolving polysaccharide crude extract II in water, dialyzing with 8000-14000D dialysis membrane with cut-off molecular weight, adding ethanol solution into dialyzed polysaccharide crude extract liquid, stirring until precipitate is sucked out, filtering, and drying precipitate to obtain polysaccharide crude extract III; purifying by using a gel column: purifying the polysaccharide crude extract III with gel chromatography column, tracking the fraction with sulfuric acid-phenol method, collecting main peak, mixing, adding ethanol solution, stirring, filtering, precipitating, and freeze drying to obtain the polysaccharide extract of herba Eichhorniae. It has antibacterial, skin whitening, moisture keeping, and antioxidant effects.
Description
The technical field is as follows:
the invention belongs to the field of plant extraction, and particularly relates to a nymphaea hybrid polysaccharide extract and a preparation method and application thereof.
Background art:
the nymphaea hybrid is large-scale nymphaea hybrid, and belongs to perennial aquatic perennial root herbaceous plants of Nymphaeaceae. The perfume lotus has light fragrance and long flowering phase, and is the highest grade variety of urban garden waterscape, courtyard radiography scenery and potted ornamental precious aquatic flowers at home and abroad. In the last 70 th century, the American globeflower was introduced into Taiwan of China, and on the basis of the Taiwan, nine major color systems such as gold, yellow, purple, blue, red, tea, green, red, white and the like are developed through cultivation, so that the Taiwan is also named as 'Jiupin globeflower'. The water lily is suitable for tropical to temperate climates, so that the water lily can be widely planted in places such as Guangdong, Hunan, Zhejiang, Jiangsu, Hainan and the like. The flowers of the nymphaea hybrid have very remarkable ornamental property and are high-grade aquatic ornamental aquatic plant species in China.
The flower of the lotus tea can be directly used for making fresh perfume lotus tea, can be used for raw eating, tea making, wine soaking, stewing and boiling food and the like, and can be used as a new resource plant with homology of medicine and food. In addition, the nymphaea hybrid can be prepared into a nine-grade nymphaea hybrid tea by sterilizing and baking at the high temperature of 120 ℃, and has the summer heat relieving and health care effects. The nymphaea hybrid flowers contain plant placentin and natural pollen, have good tranquilizing and calming effects, and also have the effects of softening skin and removing dead skin. In addition, the nymphaea hybrid also has the effects of beautifying, enhancing immunity and promoting metabolism. Currently, the perfume lotus has formally become the etiquette flower of the olympic games.
In the extraction and purification process of the nymphaea hybrid polysaccharide, the existing technology relies on macroporous resin adsorption, and the prepared polysaccharide has obvious color, so that the color is easy to change after long-term storage, and the subsequent development of related products, particularly whitening products, is disturbed. At present, the extraction of the nymphaea hybrid polysaccharide generally adopts a high-temperature and ultrasonic-assisted extraction method, however, the polysaccharide structure is easy to break and denature under the action of high temperature or ultrasonic waves.
Meanwhile, the extraction of the nymphaea hybrid polysaccharide is mainly concentrated on a crude extract or a high molecular weight polysaccharide, and the content of the polysaccharide is not high or the property of the polysaccharide cannot meet the requirement of subsequent product development.
The invention content is as follows:
based on the defects of the current extraction method of the nymphaea hybrid polysaccharide, the invention aims to provide the nymphaea hybrid polysaccharide extract and the preparation method thereof, so that the nymphaea hybrid polysaccharide with high purity, stable property, white color and medium molecular weight (the molecular weight is mainly 80-360KD) is efficiently extracted, and the nymphaea hybrid polysaccharide is more conveniently absorbed through the body surface of the skin on the basis of keeping the effects of moisture preservation, whitening, antibiosis, antioxidation and the like.
The preparation method of the nymphaea hybrid polysaccharide extract comprises the following steps:
A. pulverizing nymphaea hybrid, adding water for extraction, and concentrating the extract to obtain extract;
B. adding ethanol solution into the extract, stirring, filtering, and freeze drying the precipitate to obtain polysaccharide crude extract I;
C. dissolving polysaccharide crude extract I in water, deproteinizing by Sevag method, washing out precipitate, filtering, and drying precipitate to obtain polysaccharide crude extract II;
D. removing monosaccharide and oligosaccharide: dissolving polysaccharide crude extract II in water, dialyzing with 8000-14000D dialysis membrane with cut-off molecular weight, adding ethanol solution into dialyzed polysaccharide crude extract liquid, stirring until precipitate is sucked out, filtering, and drying precipitate to obtain polysaccharide crude extract III;
E. purifying by using a gel column: purifying the polysaccharide crude extract III with gel chromatography column, tracking the fraction with sulfuric acid-phenol method, collecting main peak, mixing, adding ethanol solution, stirring, filtering, precipitating, and freeze drying to obtain the polysaccharide extract of herba Eichhorniae.
The nymphaea hybrid is dry or fresh flowers.
The pulverization is to 20 meshes so as to ensure that the polysaccharide can be quickly extracted.
The water extraction is to soak the smashed nymphaea hybrid with 5-10 times of water, mechanically stir, extract for 3-5 times in water bath for 1-12 hours each time, then filter and combine the filtrate. Extracting at room temperature to 50 deg.C to ensure stable property of polysaccharide; in addition, the amount of water is 5-10 times of the amount of perfume.
The Sevag method deproteinization is to take polysaccharide crude extract I to be dissolved in water, and chloroform with the volume ratio of 5: 1 is added: and mixing the n-butanol with a solvent, stirring, layering, recovering the supernatant, adding an ethanol solution into the supernatant, stirring until a precipitate is separated out, filtering, and drying the precipitate to obtain the polysaccharide crude extract II.
The dialysis is dialysis for 2 days.
The method has the advantages that the extraction at room temperature or low temperature ensures that the original structure of the nymphaea hybrid polysaccharide is not degraded, and the prepared polysaccharide is pure white in color, high in purity (94% -99%), stable in property and easy to dissolve in water. The prepared nymphaea hybrid polysaccharide has controllable quality, high active ingredient content (94% -99%), and pharmacological functions of resisting bacteria, whitening skin, keeping moisture, resisting oxidation, etc. In addition, the preparation process is convenient to operate, and the product can be widely applied to cosmetics, beverages and health-care foods.
The specific implementation mode is as follows:
the following will further illustrate the essence and effect of the present invention with reference to the embodiment, which is only used to illustrate the process and result of the present invention, but not to limit the present invention.
Example 1 preparation of polysaccharide extract of nymphaea hybrid
Taking 100g of freeze-dried nymphaea hybrid, crushing the 100g of nymphaea hybrid by using a crusher, and filtering the crushed powder by using a 20-mesh iron sieve to obtain nymphaea hybrid micropowder. Adding 10 times volume (1L) of water into the nymphaea hybrid micropowder, and leaching in a water bath with mechanical stirring (400 r/min) at room temperature for 4-5 h. Primarily filtering the leaching solution through gauze, and collecting filtrate. Extracting the residue with the same method for 2 times, and mixing the filtrates for 3 times. The filtrate was concentrated under reduced pressure by means of a rotary evaporator to 1/10(100mL) in the original volume, and a 95 vol% ethanol solution (130mL) was added to the concentrated solution and stirred to effect precipitation, and when the concentration of the added ethanol reached 60% (by volume), a large amount of flocculent precipitates appeared. The precipitate was filtered through gauze and dried under vacuum to give crude polysaccharide extract I (27 g).
Crude polysaccharide extract I was dissolved in water (100mL) and purified by adding chloroform in a 5: 1 volume ratio: and (3) mixing a n-butanol mixed solvent (100mL), fully stirring, demixing, recovering a supernatant, adding an ethanol aqueous solution (130mL) with the volume fraction of 95% into the supernatant, stirring until a precipitate is separated out, filtering, and drying the precipitate to obtain the polysaccharide crude extract II (16 g).
Dissolving the crude polysaccharide extract II in water (100mL), dialyzing with dialysis membrane with molecular weight cut-off of 8-14KD for 2 days, adding 95% ethanol water solution (130mL) with volume fraction into the dialyzed crude polysaccharide extract liquid, stirring, sucking out the precipitate, filtering, and drying the precipitate to obtain crude polysaccharide extract III (11 g);
purifying the polysaccharide crude extract III with gel chromatography column, tracking the fraction with sulfuric acid-phenol method, collecting main peak, and mixing. Mixing the eluates, concentrating under pressure at room temperature with a rotary evaporator, concentrating to 50mL, adding 95% ethanol water (80mL) at one time, stirring, and filtering. Freeze-drying the filter cake for 12h to obtain the nymphaea hybrid polysaccharide extract (7 g). The lotus polysaccharide is white solid powder, the purity is 94% -99%, and the molecular weight is mainly 80-360 KD. The lotus polysaccharide can be rapidly dissolved in water at room temperature (10min), and can be placed in a sealed manner at room temperature for 30 days without color and physicochemical property changes.
Example 2: evaluation of antibacterial Activity of Compound
In this example, the Minimum Inhibitory Concentration (MIC) of the antibacterial activity of a sample is determined by the Resazurin color development method. The experimental strains are gram-positive bacteria or gram-negative bacteria such as Methicillin-resistant Staphylococcus aureus (MRSA), Staphylococcus Aureus (SA), Bacillus cereus (BC, Bacillus cereus), B.subtilis (BS, Bacillus subtilis), B.thuringiensis (BT, Bacillus thuringiensis) or Escherichia coli (EC, Escherichia coli).
The assay will use a 96-well plate dilution titer technique, with the Minimum Inhibitory Concentration (MIC) of the various substances being determined simultaneously. Firstly, 7.5mL of indicator solution (100 mu g/mL of resazurin aqueous solution) and 5mL of bacteria solution to be tested (10)8CFU/mL) and 100 μ L of the mixed bacteria solution was added to each of all the test wells of columns 1 to 8. Then 100. mu.L of the aqueous solution (32mg/mL) of the sample to be tested was sequentially added to each well of the first row, after uniform mixing, 100. mu.L of the solution was taken out and transferred to the corresponding well of the second row, and the solution was diluted to the 8 th row in the same manner. Finally, the well plate with the added sample is placed into a constant temperature incubator and cultured for 10-12h at 37 ℃. The bacteria liquid turns red into no bacteriostatic activity, blue is bacteriostatic activity, and the lowest dilution concentration of the bacteria liquid maintaining blue is considered as the lowest bacteriostatic concentration of the compound to be detected. Each sample was made in 3 replicates.
The results show that: the nymphaea hybrid polysaccharide extract has obvious antibacterial activity on gram-positive bacteria, and almost has obvious inhibition effects on Methicillin-resistant Staphylococcus aureus (MRSA), Methicillin-resistant Staphylococcus Aureus (SA), Bacillus cereus (BC, Bacillus cereus), B.subtilis (BS, Bacillus subtilis) and B.thuringiensis (BT, Bacillus thuringiensis) when the concentration is more than 2 mg/mL. The results show that the nymphaea hybrid polysaccharide extract has good application prospect in the aspects of antibiosis and antiphlogosis. The specific experimental results are shown in table 1 below.
TABLE 1 in vitro antibacterial Activity (MIC) of the nymphaea hybrid polysaccharides
Example 3: determination of lipid peroxidation inhibition Activity
0.50mL of lecithin solution was weighed and added to 1.0mL of PBS buffer with pH 7.0. 1.0mL of sample solutions containing different concentrations was mixed uniformly with 1.0mL of an aqueous solution containing 2.5mmol/L of EDTA-Fe (II), which was then added to the aforementioned PBS mixed solution of lecithin. After mixing, the mixture is placed in a water bath at 37 ℃ for reaction for 45 min. Then, 2.0mL of 28% (w/v) trichloroacetic acid and 1.0mL of 1% (w/v) thiobarbituric acid were added successively. After mixing uniformly, the mixed solution is placed in a boiling water bath at 100 ℃ for heating for 10min, and after cooling, the absorbance is measured at 532 nm. The absorbance A0 was measured by zeroing with PBS buffer and replacing the sample with PBS buffer for blank tubes. 3 replicates of each sample were taken and averaged for A1. The lipid peroxidation inhibition was calculated as follows: inhibition ratio (%) [ (a0-a1)/a0] × 100 in the formula: a0 is the absorbance without sample; a1 is the absorbance of the sample.
The results show that: the nymphaea hybrid polysaccharide extract has obvious inhibitory activity on lipid peroxidation. The results show that the nymphaea hybrid polysaccharide extract has certain potential in the aspect of being used as an antioxidant. The specific experimental results are shown in table 2 below.
TABLE 2 in vitro anti-lipid peroxidation Activity of Nelumbo Nucifera polysaccharide
Example 4: activity measurement of trollius chinensis polysaccharide for inhibiting tyrosinase in cells
Adjusting the number of melanoma cells in mice to 5 × 104One cell/mL, 100. mu.L of cell fluid was added to each well of a 96-well plate, 100. mu.L of the culture solution was supplemented, and after plating for 12 hours, the supernatant was discarded. Thereafter, 200. mu.L of culture medium containing samples of different mass concentrations (mass concentration gradient of 0.1, 0.2, 0.4, 0.8, 1.6, 3.2mg/mL) was added to the system at 37 ℃ with a volume fraction of 5% CO2Culturing under saturated wet condition, setting 6 multiple wells per mass concentration, culturing for 48h with cells cultured without adding sample as blank control, and changing liquid every day. Tyrosinase activity was measured by the dopa oxidation method. Discarding supernatant in the well plate, rinsing with PBS buffer solution, adding 1% Triton-X10040 μ L into each well, rapidly placing into-80 deg.C refrigerator, thawing at 37 deg.C after 30min, disrupting cells, and adding 1% L-DOAnd (3) performing correction on 10 mu L/hole of PA solution by using L-DOPA as a background, performing reaction at 37 ℃ for 30min by using cells cultured without adding a whitening agent as a control, and measuring the A450 value at 450nm by using an enzyme-labeling instrument. The inhibition rate of the test solution (including the positive control) on the tyrosinase is calculated by using the following formula with the corresponding negative control as a reference during the determination. Inhibition (%) - (a-B)/a × 100% where a is the absorbance of the standard control and B is the absorbance of the positive control. Each experiment was done in 3 replicates. The high inhibition rate indicates that the inhibition intensity of the compound on tyrosinase activity is high.
The results show that: the nymphaea hybrid polysaccharide extract has obvious inhibitory activity on tyrosinase, particularly shows good anti-tyrosinase activity when the concentration is more than 1.6 mg/mL, and the specific experimental results are shown in the following table 3. Tyrosinase is an oxidase and is the rate-limiting enzyme in the regulation of melanin production. It is present in plant and animal tissues and catalyzes the production of melanin and other pigments from the oxidation of tyrosine. Tyrosinase inhibitors are generally excellent whitening agents, and can inhibit the production of skin melanin. The results show that the nymphaea hybrid polysaccharide extract can be widely applied as a whitening agent and used for developing whitening products.
TABLE 3 in vitro antityrosinase Activity of nymphaea hybrid polysaccharides
Example 5: preparation method of nymphaea hybrid polysaccharide essence with whitening, antibacterial, moisturizing and antioxidant functions
Dissolving 200mg of the trollius chinensis bunge polysaccharide extract in 100mL of ultrapure water, stirring for 30min, and sequentially adding phenoxyethanol (0.5mL), butanediol (0.2mL), catechin (1.0g) and orange flower essence (0.2 mL). And fully and uniformly stirring to obtain the nymphaea hybrid polysaccharide essence similar to gel.
Example 6: preparation method of nymphaea hybrid polysaccharide beverage
Dissolving 200mg of the trollius chinensis bunge polysaccharide extract in 100mL of ultrapure water, stirring for 30min, and sequentially adding polylysine (0.5g), sodium dehydroacetate (0.5g), sucrose (2.0g), stevioside (0.2g) and orange juice (10 mL). After fully and uniformly stirring, obtaining the nymphaea hybrid polysaccharide functional beverage with the taste similar to orange juice.
Example 7: preparation of nymphaea hybrid polysaccharide buccal tablet
Taking 1.0g of the trollius chinensis bunge polysaccharide extract, 4.0g of xylitol, 50mg of stevioside, 50mg of orange juice powder and 50mg of magnesium stearate, stirring uniformly, and crushing into micro powder with the granularity of more than 300 meshes by a crusher. After adding 3mL of ultrapure water and stirring the mixture uniformly, the mixture was granulated (about 25 granules). Freeze-drying the prepared granules, and tabletting by a tabletting machine to obtain the functional trollius chinensis polysaccharide buccal tablet.
Claims (9)
1. The preparation method of the nymphaea hybrid polysaccharide extract is characterized by comprising the following steps:
A. pulverizing nymphaea hybrid, adding water for extraction, and concentrating the extract to obtain extract;
B. adding ethanol solution into the extract, stirring, filtering, and freeze drying the precipitate to obtain polysaccharide crude extract I;
C. dissolving polysaccharide crude extract I in water, deproteinizing by Sevag method, washing out precipitate, filtering, and drying precipitate to obtain polysaccharide crude extract II;
D. removing monosaccharide and oligosaccharide: dissolving polysaccharide crude extract II in water, dialyzing with 8000-14000D dialysis membrane with cut-off molecular weight, adding ethanol solution into dialyzed polysaccharide crude extract liquid, stirring until precipitate is sucked out, filtering, and drying precipitate to obtain polysaccharide crude extract III;
E. purifying by using a gel column: purifying the polysaccharide crude extract III with gel chromatography column, tracking the fraction with sulfuric acid-phenol method, collecting main peak, mixing, adding ethanol solution, stirring, filtering, precipitating, and freeze drying to obtain the polysaccharide extract of herba Eichhorniae.
2. The method of claim 1, wherein the nymphaea hybrid is a dried or fresh flower.
3. The method of claim 1, wherein the pulverizing is to 20 mesh.
4. The preparation method of claim 1, wherein the water extraction is carried out by soaking the pulverized nymphaea hybrid with 5-10 times of water, mechanically stirring, extracting for 1-12 hours each time for 3-5 times in water bath, filtering, and combining the filtrates.
5. The method of claim 1, wherein the Sevag deproteinization is carried out by dissolving crude polysaccharide extract I in water, adding chloroform at a volume ratio of 5: 1: and mixing the n-butanol with a solvent, stirring, layering, recovering the supernatant, adding an ethanol solution into the supernatant, stirring until a precipitate is separated out, filtering, and drying the precipitate to obtain the polysaccharide crude extract II.
6. The method of claim 1, wherein the dialysis is dialysis for 2 days.
7. A nymphaea hybrid polysaccharide extract prepared according to the preparation method of claim 1, 2, 3, 4, 5 or 6.
8. The use of the nymphaea hybrid polysaccharide extract according to claim 7 for preparing cosmetics, beverages or health foods with antibacterial, whitening, moisturizing and antioxidant effects.
9. An antibacterial, whitening, moisturizing and antioxidant cosmetic, beverage or health food, characterized by containing the nymphaea hybrid polysaccharide extract according to claim 7 as an active ingredient.
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