CN105534876A - Dendrobe fermentation raw stock as well as preparation method and application thereof - Google Patents
Dendrobe fermentation raw stock as well as preparation method and application thereof Download PDFInfo
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- CN105534876A CN105534876A CN201610040588.2A CN201610040588A CN105534876A CN 105534876 A CN105534876 A CN 105534876A CN 201610040588 A CN201610040588 A CN 201610040588A CN 105534876 A CN105534876 A CN 105534876A
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- 238000000855 fermentation Methods 0.000 title claims abstract description 60
- 230000004151 fermentation Effects 0.000 title claims abstract description 60
- 238000002360 preparation method Methods 0.000 title claims abstract description 20
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 20
- 239000002537 cosmetic Substances 0.000 claims abstract description 18
- 238000000034 method Methods 0.000 claims abstract description 16
- 239000000843 powder Substances 0.000 claims abstract description 10
- 239000007788 liquid Substances 0.000 claims description 14
- 239000000047 product Substances 0.000 claims description 11
- 230000001954 sterilising effect Effects 0.000 claims description 10
- -1 DPPH free radical Chemical class 0.000 claims description 9
- OUUQCZGPVNCOIJ-UHFFFAOYSA-M Superoxide Chemical compound [O-][O] OUUQCZGPVNCOIJ-UHFFFAOYSA-M 0.000 claims description 8
- 230000002000 scavenging effect Effects 0.000 claims description 8
- 241000186660 Lactobacillus Species 0.000 claims description 7
- 229940039696 lactobacillus Drugs 0.000 claims description 7
- 150000003254 radicals Chemical class 0.000 claims description 7
- 230000003462 zymogenic effect Effects 0.000 claims description 6
- 239000012531 culture fluid Substances 0.000 claims description 4
- 239000000725 suspension Substances 0.000 claims description 4
- 230000001815 facial effect Effects 0.000 claims description 3
- 230000036541 health Effects 0.000 claims description 3
- 239000006228 supernatant Substances 0.000 claims description 3
- 238000005119 centrifugation Methods 0.000 claims description 2
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- 238000006243 chemical reaction Methods 0.000 description 14
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- 210000003491 skin Anatomy 0.000 description 6
- 241001523681 Dendrobium Species 0.000 description 5
- 150000004676 glycans Chemical class 0.000 description 5
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- 241000186672 Lactobacillus delbrueckii subsp. bulgaricus Species 0.000 description 3
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- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 2
- 108010022579 ATP dependent 26S protease Proteins 0.000 description 2
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- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- 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 description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
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- 230000003833 cell viability Effects 0.000 description 2
- 230000019522 cellular metabolic process Effects 0.000 description 2
- 229960000935 dehydrated alcohol Drugs 0.000 description 2
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- WQGWDDDVZFFDIG-UHFFFAOYSA-N pyrogallol Chemical compound OC1=CC=CC(O)=C1O WQGWDDDVZFFDIG-UHFFFAOYSA-N 0.000 description 2
- YGSDEFSMJLZEOE-UHFFFAOYSA-N salicylic acid Chemical compound OC(=O)C1=CC=CC=C1O YGSDEFSMJLZEOE-UHFFFAOYSA-N 0.000 description 2
- 230000009758 senescence Effects 0.000 description 2
- QWUWMCYKGHVNAV-UHFFFAOYSA-N 1,2-dihydrostilbene Chemical group C=1C=CC=CC=1CCC1=CC=CC=C1 QWUWMCYKGHVNAV-UHFFFAOYSA-N 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- ZZZCUOFIHGPKAK-UHFFFAOYSA-N D-erythro-ascorbic acid Natural products OCC1OC(=O)C(O)=C1O ZZZCUOFIHGPKAK-UHFFFAOYSA-N 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
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- 229940123457 Free radical scavenger Drugs 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 1
- 241000186673 Lactobacillus delbrueckii Species 0.000 description 1
- 206010067482 No adverse event Diseases 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
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- 206010040880 Skin irritation Diseases 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- 229930003268 Vitamin C Natural products 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 239000002390 adhesive tape Substances 0.000 description 1
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- KLOIYEQEVSIOOO-UHFFFAOYSA-N carbocromen Chemical compound CC1=C(CCN(CC)CC)C(=O)OC2=CC(OCC(=O)OCC)=CC=C21 KLOIYEQEVSIOOO-UHFFFAOYSA-N 0.000 description 1
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- MGJZITXUQXWAKY-UHFFFAOYSA-N diphenyl-(2,4,6-trinitrophenyl)iminoazanium Chemical compound [O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1N=[N+](C=1C=CC=CC=1)C1=CC=CC=C1 MGJZITXUQXWAKY-UHFFFAOYSA-N 0.000 description 1
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
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- 229930182478 glucoside Natural products 0.000 description 1
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- 239000001963 growth medium Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
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- 238000011081 inoculation Methods 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
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- 229940099596 manganese sulfate Drugs 0.000 description 1
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- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 description 1
- 231100000344 non-irritating Toxicity 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 239000004006 olive oil Substances 0.000 description 1
- 235000008390 olive oil Nutrition 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- FJKROLUGYXJWQN-UHFFFAOYSA-N papa-hydroxy-benzoic acid Natural products OC(=O)C1=CC=C(O)C=C1 FJKROLUGYXJWQN-UHFFFAOYSA-N 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- 230000003285 pharmacodynamic effect Effects 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 1
- 229920000053 polysorbate 80 Polymers 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 229940079877 pyrogallol Drugs 0.000 description 1
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- 238000011160 research Methods 0.000 description 1
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Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/99—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from microorganisms other than algae or fungi, e.g. protozoa or bacteria
-
- 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
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
-
- 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
- A61Q19/08—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
- A61K2800/85—Products or compounds obtained by fermentation, e.g. yoghurt, beer, wine
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Epidemiology (AREA)
- Birds (AREA)
- Biotechnology (AREA)
- Engineering & Computer Science (AREA)
- Dermatology (AREA)
- Mycology (AREA)
- Microbiology (AREA)
- Botany (AREA)
- Gerontology & Geriatric Medicine (AREA)
- Tropical Medicine & Parasitology (AREA)
- Cosmetics (AREA)
Abstract
The invention discloses a dendrobe fermentation raw stock as well as a preparation method and application thereof. The preparation method of the dendrobe fermentation raw stock comprises the following steps: mixing dried dendrobe powder, water and a fermentation bacterium so as to obtain a primary system, and perform fermentation culture on the primary system, thereby obtaining the dendrobe fermentation raw stock. According to the dendrobe fermentation raw stock, active substances are extracted from dendrobe by using a fermentation engineering technique, the operation is simple, and as no external substance is added, the prepared dendrobe fermentation raw stock is safe and harmless to skin, and is very high in anti-aging activity when applied to cosmetics.
Description
Technical field
The present invention relates to a kind of Herba Dendrobii fermentation oleo stock and preparation method thereof and application, belong to biological technical field.
Background technology
Herba Dendrobii (DendrobiumofficinaleKimuraetMigo) is China's rare Chinese medicine, is the orchid family Dendrobium Sw, gains the name, have the effects such as YIN nourishing of promoting the production of body fluid, improving eyesight nourishing the stomach, kidney-tonifying health-care because epidermis is ferrum green.According to the literature, the main pharmacodynamics composition in Dendrobium Sw is polysaccharide, alkaloid, glucosides, luxuriant and rich with fragrance class, bibenzyl and aminoacid etc.The content of polysaccharide in Herba Dendrobii is high, and has good antitumor, antioxidation, enhancing immunity and reduce the physiological functions such as blood glucose.
Both at home and abroad in Herba Dendrobii research, the extracting method of Herba Dendrobii active substance, based on decoction and alcohol sedimentation technique, also has the method such as Enzymatic Extraction and ultrasonic assistant extraction.
Summary of the invention
The object of this invention is to provide a kind of Herba Dendrobii fermentation oleo stock and preparation method thereof and application.The inventive method utilizes fermentation engineering to extract active substance from Herba Dendrobii, simple to operate, and owing to not adding any foreign substance, the Herba Dendrobii fermentation oleo stock prepared, to skin safe fanout free region, is applied to have very high activity of fighting against senium in cosmetics.
The preparation method of a kind of Herba Dendrobii fermentation oleo stock provided by the invention, comprises the steps: Herba Dendrobii dry powder, water and zymocyte to be mixed to get initial system, and carries out fermentation culture to described initial system, namely obtain Herba Dendrobii fermentation oleo stock.
In the present invention, described Herba Dendrobii dry powder adopts the stem of Herba Dendrobii to make.
In above-mentioned preparation method, described Herba Dendrobii dry powder, described water and described zymogenic proportioning can be 1g:(9 ~ 20) mL:(0.1 ~ 1) mL, specifically can be 1g:12mL:0.6mL or 1g:(12 ~ 20) mL:(0.6 ~ 1) mL;
Described zymogenic concentration can be (10
5~ 10
8) CFU/mL.
In above-mentioned preparation method, the temperature of described fermentation culture can be 35 ~ 50 DEG C, specifically can be 38 DEG C, 35 ~ 40 DEG C, 35 ~ 38 DEG C or 38 ~ 50 DEG C; The time of described fermentation culture can be 4.5 ~ 6.5h, specifically can be 6h, 4.5 ~ 6h or 6 ~ 6.5h.
In above-mentioned preparation method, the particle diameter of described Herba Dendrobii dry powder can be 20 ~ 50 orders, specifically can be 40 orders or 40 ~ 50 orders, and described Herba Dendrobii dry powder can be obtained by pulverizer grinding and sieving;
Described zymocyte can be lactobacillus, specifically can be Deshi Lactobacillus, more specifically can be lactobacillus delbruockii subspecies bulgaricus Lactobacillusdelbrueckiibulgaricus.
In above-mentioned preparation method, described zymocyte can the form of its culture fluid or suspension exist, and the pH value of described zymogenic culture fluid or suspension can be 4.0 ~ 7.0, specifically can be 5.0,4.0 ~ 5.0 or 5.0 ~ 7.0.
In above-mentioned preparation method, in described method after described fermentation culture, also can comprise and sterilizing, centrifugal and get the step of supernatant is carried out successively to fermentation liquid.
In above-mentioned preparation method, the condition of described sterilizing is as follows: the temperature of described sterilizing can be 115 ~ 121 DEG C, specifically can be 115 DEG C; The time of described sterilizing can be 15 ~ 25min, specifically can be 20min;
Described centrifugal condition is as follows: centrifugal rotational speed can be 3000 ~ 4000r/min, specifically can be 3800r/min or 3800 ~ 4000r/min; Centrifugation time can be 15 ~ 30min, specifically can be 20min or 20 ~ 30min; Centrifugal radius can be 9cm.
Present invention also offers the Herba Dendrobii fermentation oleo stock prepared by above-mentioned preparation method.
Invention further provides above-mentioned Herba Dendrobii fermentation oleo stock and have following 1 in preparation) and/or 2) in function product in application:
1) scavenging free radicals;
2) defying age.
In above-mentioned application, described product can be at least one in cosmetics, food and health product;
Described cosmetics can be at least one in facial film, essence and cosmetic water;
Described free radical is at least one in DPPH free radical, superoxide anion and hydroxyl radical free radical.
The present invention has following beneficial effect:
(1) preparation method of Herba Dendrobii fermentation oleo stock provided by the invention adopts fermentation engineering, by selecting suitable strain and fermentation condition, extracting and obtaining active substance and Herba Dendrobii fermentation oleo stock from Herba Dendrobii.On the one hand, the active component in plant at utmost can be retained, and relative and traditional water extraction, avoid the introducing of organic solvent, safety and environmental protection; On the other hand, do not need to add the foreign substances such as enzyme, not only save production cost, and the maximized production stage that simplifies, enable this fermentation technique realize a large amount of production, suitability for industrialized production, and fully can ensure the stability of product quality.
(2) Herba Dendrobii fermentation oleo stock provided by the invention is rich in dendrobium polysaccharide, and owing to not introducing foreign substance in preparation process, not containing any chemical composition, can directly use as the finished product of facial film, essence or cosmetic water, more natural, any negative interaction can not be caused to skin.
(3) Herba Dendrobii provided by the invention fermentation oleo stock, having stronger oxidation resistance can scavenging free radicals and superoxide anion, promotes cellular metabolism, strengthen cell viability, improve the 26S Proteasome Structure and Function of body, improve body vitality, thus delay cell senescence, there is antidotal effect.
Accompanying drawing explanation
Fig. 1 is the Herba Dendrobii fermentation oleo stock of different volumes percentage concentration and the relation curve of DPPH free radical scavenging activity.
Fig. 2 is the Herba Dendrobii fermentation oleo stock of different volumes percentage concentration and the relation curve of hydroxyl radical free radical clearance rate.
Fig. 3 is the Herba Dendrobii fermentation oleo stock of different volumes percentage concentration and the relation curve of superoxide anion clearance rate.
Detailed description of the invention
The experimental technique used in following embodiment if no special instructions, is conventional method.
Material used in following embodiment, reagent etc., if no special instructions, all can obtain from commercial channels.
Fluid medium in following embodiment is by solute and solvent composition, solvent is water, concentration in solute and liquid medium within thereof is: peptone 10.0g, Carnis Bovis seu Bubali cream 10.0g, yeast extract 5.0g, diammonium hydrogen citrate 2.0g, glucose 20.0g, Tween 80 1.0mL, sodium acetate (CH3COONa3H2O) 5.0g, dipotassium hydrogen phosphate (K2HPO43H2O) 2.0g, magnesium sulfate (MgSO47H2O) 0.58g, manganese sulfate (MnSO4H2O) 0.25g, agar 18.0g, distilled water 1000mL, pH6.2 ~ 6.6.
Herba Dendrobii (stem) in following embodiment, from Yiwu, Zhejiang Xian He company limited, pulverized 40 mesh sieves.
The ATCC of the lactobacillus delbruockii subspecies bulgaricus (Lactobacillusdelbrueckiibulgaricus) in following embodiment is numbered 11842, can buy obtain from ATCC.
Zymocyte bacterium liquid in following embodiment obtains in accordance with the following steps:
1, the activation of strain
Picking lactobacillus delbruockii subspecies bulgaricus (Lactobacillusdelbrueckiibulgaricus) bacterium colony one ring, in fluid medium, puts into shaking table by actication of culture, obtains activating rear lactobacillus delbrueckii subspecies bulgaricus liquid.
2, the purification of strain
Bacterium liquid gradient dilution bed board after activation step 1 obtained, to obtain single bacterium colony, obtains Deshi Lactobacillus after purification.
3, the amplification culture of strain
Strain inoculation after purification step 2 obtained, in MRS culture medium (pH value is 7.0), is cultivated in the shaking table of 25 DEG C, and as OD value=0.5-1.0, (strain is in logarithmic (log) phase, and concentration is 10 to obtain zymocyte bacterium liquid
5~ 10
8cFU/mL).
Embodiment 1, prepare Herba Dendrobii fermentation oleo stock
Prepare Herba Dendrobii fermentation oleo stock in accordance with the following steps:
(1) be 10 by the 9mL concentration of 20mL
5~ 10
8the zymocyte bacterium liquid of CFU/mL is inoculated in the Herba Dendrobii dry powder of 15g and the water of 180mL, obtains fermentation system;
(2) above-mentioned fermentation system is fermented 6 hours in 38 DEG C of shaking tables, obtain tunning;
(3) by sterilizing 20min at above-mentioned tunning 115 DEG C, make bacterium inactivation, obtain the tunning after sterilizing; By the tunning after sterilizing at 3800r/min, centrifugal radius is centrifugal 20min under the condition of 9cm, abandons precipitation, collects supernatant, namely obtains Herba Dendrobii fermentation oleo stock.
Embodiment 2, Herba Dendrobii fermentation oleo stock is as the application in cosmetics
One, the character of Herba Dendrobii proferment pulp cosmetic
By the fermented product extract of gained prepared by the embodiment of the present invention 1 through activated carbon decolorizing, de-taste after, obtained Herba Dendrobii proferment pulp cosmetic.Its outward appearance is thick liquid, color is faint yellow to brown color.PH value 5.0-6.1, viscosity 200-500cP, solubility is admittedly containing thing content 2.0-6.0%, and total plate count is less than 50CFU/mL, detects without pathogenic bacterium.According to hygienic standards for cosmetics GB7916-87, cosmetics total number of bacteria is not higher than 1000CFU/ml, so this fermentation oleo stock meets cosmetics quality requirement.
Measure the main composition dendrobium polysaccharide content of this Herba Dendrobii proferment pulp cosmetic, detection method is with reference to GB/T5009.8-2008, and recording dendrobium polysaccharide content is 428.6g/kg.
Two, the safety of Herba Dendrobii fermentation oleo stock detects
Human body patch test is mainly used for the zest detecting cosmetics finished product or raw material.Human closed formula patch test is carried out to the Herba Dendrobii fermentation oleo stock obtained in the embodiment of the present invention 1, is intended to assess its potential skin irritation.
1, subjects
Select suitable volunteer 30 people, the range of age is at 18-60 year Stochastic choice.
2, test method
Taking 0.020g-0.025g solid sample or semisolid sample, to put into speckle examination device for subsequent use.Fluid sample 0.2mL to 0.025mL is dripped on filter paper, then filter paper is placed in speckle examination device.Each sample all arranges blank, in contrast speckle examination device hole, add the sample solvent with sample equivalent, as distilled water or olive oil (the present embodiment employing distilled water).
Human body back is elected at test position as, utilizes non-irritating adhesive tape speckle to be tried device and fixedly sticks in experimenter back.Test period continues 24h.In order to the accurate, credible of result of the test and science, test period volunteer as requested, speckle examination device can not be removed, recipient site also can not be made to contact water.Remove speckle examination device after 24h, after leaving standstill 30min, wait for that impression disappears, observe the reaction of skin.If result of the test is negative, then 24h and 48h after patch test is needed to observe once respectively again.
3, result of the test
Patch test results is as shown in table 1, and the implication that in table 1, each symbol represents is as follows: "-"=negative reaction; " ± "=suspicious reaction: only have faint erythema; "+"=weak positive reaction (erythematous response): erythema, infiltration, edema, can pimple be had; " ++ "=strong positive reaction (herpes reaction); Erythema, infiltration, edema, pimple, herpes; Reaction can exceed tested district; " +++ "=extremely strong positive reaction (reaction of amalgamation herpes); Obvious erythema, severe infiltration, edema, amalgamation herpes; Reaction exceeds tested district.
Criterion: according to cosmetics health specification 2007, occur in 30 routine experimenters that the number of 1 grade of skin adverse reaction is more than 5 examples, or occur that the number of secondary skin adverse reaction is more than 2 examples, or when there is any 1 example more than three grades or three grades skin adverse reactions, then judge that tested material has untoward reaction to human body, otherwise, be then considered as having no adverse reaction to human body.
The patch test results of the Herba Dendrobii fermentation oleo stock that table 1, the embodiment of the present invention 1 obtain
As can be seen from Table 1: the Herba Dendrobii fermentation oleo stock that the embodiment of the present invention 1 obtains does not have untoward reaction to occur, and illustrates that Herba Dendrobii fermented product of the present invention all has safety, can not bring untoward reaction to human body.
Three, the antioxygenic property of Herba Dendrobii fermentation oleo stock detects
1, DPPH radical scavenging activity detects
DPPH is a kind of organic free radical of early stage synthesis, be commonly used to the hydrogen supply capacity assessing polyphenoils, it is highly stable in organic solvent, in purple, and have a characteristic absorption peak at 517nm place, when running into free radical scavenger, the lone pair electrons of DPPH are paired and make it fade, and namely diminish at the light absorption value of maximum absorption wave strong point.Therefore, the change by measuring light absorption value carrys out the elimination effect of assess sample to DPPH free radical.
The specific experiment step of DPPH free radical scavenging experiment is:
(1) liquid to be measured and 2 × 10 of equal-volume (being generally 3mL) is got
-4the DPPH solution mixing (A of mol/L
1pipe);
(2) isopyknic dehydrated alcohol (determinand solvent) and 2 × 10 is got
-4the DPPH solution mixing (A of mol/L
2pipe);
(3) isopyknic dehydrated alcohol is got and liquid to be measured mixes (A
3pipe);
(4), after reacting 30min, under 517nm, A is surveyed
1, A
2, A
3pipe absorbance.
Clearance rate computing formula is: clearance rate (%)=[(A
2+ A
3)-A
1]/A
2(1)
With the concentration expressed in percentage by volume of Herba Dendrobii fermentation oleo stock liquid to be measured for abscissa, clearance rate is vertical coordinate, makes Herba Dendrobii fermentation oleo stock to DPPH free radical scavenging effect curves, as shown in Figure 1.
As shown in Figure 1, obtained in the embodiment of the present invention 1 Herba Dendrobii fermentation oleo stock is to the IC of DPPH scavenging action
50=6.01mg/mL, illustrating that Herba Dendrobii of the present invention fermentation oleo stock has stronger oxidation resistance can scavenging free radicals, promotes cellular metabolism, strengthen cell viability, improve the 26S Proteasome Structure and Function of body, improve body vitality, thus delay cell senescence, play its antidotal effect.
2, hydroxyl radical free radical Scavenging activity detects
The FeSO of 3ml2mmol/L is added successively in 25ml color comparison tube
4, then add the salicylic acid of 3ml6mmol/L, after shaking up, then add the H of 3ml1mmol/L
2o
2solution, shakes up, and starts reaction.37
0take out after C heating in water bath 15min, survey its absorbance A
0.Then add 1ml at certain density night to be measured respectively, shake up, continue heating in water bath 15min, take out and survey its absorbance A
x.Blank group replaces night to be measured with 1ml deionized water.The system absorbance caused for common 1ml night to be measured of adding after eliminating and deionized water is worth reducing, and method is the same, surveys its absorbance A after constant temperature 15min
00, add 1ml deionized water, at survey once its absorbance A after shaking up
xX, A
reduce=A
00-A
xX, 3 repetitions are done in each process respectively, hydroxyl radical free radical clearance rate (%)=100% (A
0-A
x-A
reduce)/A
0, experimental result as shown in Figure 2.
As shown in Figure 2, increase with Herba Dendrobii fermentation oleo stock concentration, it constantly strengthens the Scavenging activity of hydroxyl radical free radical, illustrates that Herba Dendrobii of the present invention fermentation oleo stock has the ability removing hydroxyl radical free radical.
3, to the mensuration of superoxide anion scavenging action
Get the Tris-HCL buffer 4.5ml of 0.05mol/LpH8.2 in test tube, be placed in 250C water-bath preheating 20min, 1ml liquid to be measured and 0.4ml is added respectively after taking-up, the pyrogallol solution of 25mmol/L, shake up and survey its absorbance under 299nm, an absorbance is surveyed every 30s, correct to 5min, with Tris-HCL buffer reference, blank replaces night to be measured with 1ml deionized water, vitamin C does positive control, and 3 repetitions are done in each process, ultra-oxygen anion free radical clearance rate (%)=(F
0-F
x)/F
0× 100, F
0for the absorbance rate over time of blank solution, F
xfor liquid to be measured rate over time, result as shown in Figure 3.
As shown in Figure 3, Herba Dendrobii oleo stock has very strong Scavenging activity to superoxide anion, IC50=27.35mg/ml.Excessive superoxide anion can produce toxicity and damage dna to body in vivo, proves that the ability of the removing superoxide anion that Herba Dendrobii oleo stock of the present invention is outstanding shows to have certain anti-senescence function.
Claims (10)
1. a preparation method for Herba Dendrobii fermentation oleo stock, comprises the steps: Herba Dendrobii dry powder, water and zymocyte to be mixed to get initial system, and carries out fermentation culture to described initial system, namely obtains Herba Dendrobii fermentation oleo stock.
2. method according to claim 1, is characterized in that: described Herba Dendrobii dry powder, described water and described zymogenic proportioning are 1g:(9 ~ 20) mL:(0.1 ~ 1) mL;
Described zymogenic concentration is (10
5~ 10
8) CFU/mL.
3. method according to claim 1 and 2, is characterized in that: the temperature of described fermentation culture is 40 ~ 50 DEG C;
The time of described fermentation culture is 4.5 ~ 6.5.
4. the method according to any one of claim 1-3, is characterized in that: the particle diameter of described Herba Dendrobii dry powder is 20 ~ 50 orders;
Described zymocyte is lactobacillus.
5. the method according to any one of claim 1-4, is characterized in that: described zymocyte exists with the form of its culture fluid or suspension;
The pH value of described zymogenic culture fluid or suspension is 4.0 ~ 7.0.
6. the method according to any one of claim 1-5, is characterized in that: in described method after described fermentation culture, also comprises and carries out sterilizing, centrifugal and get the step of supernatant successively to fermentation liquid.
7. the method according to any one of claim 1-6, is characterized in that: the temperature of described sterilizing is 115 ~ 121 DEG C; The time of described sterilizing is 15 ~ 25min;
Described centrifugal condition is as follows: centrifugal rotational speed is 3000 ~ 4000r/min; Centrifugation time is 15 ~ 30min; Centrifugal radius is 9cm.
8. the Herba Dendrobii fermentation oleo stock that the preparation method according to any one of claim 1-7 is obtained.
9. Herba Dendrobii according to claim 8 fermentation oleo stock has following 1 in preparation) and/or 2) in function product in application:
1) scavenging free radicals;
2) defying age.
10. application according to claim 9, is characterized in that: described product is at least one in cosmetics, food and health product;
Described cosmetics are at least one in facial film, essence and cosmetic water;
Described free radical is at least one in DPPH free radical, superoxide anion and hydroxyl radical free radical.
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