CN105147587A - Ganoderma lucidum fermentation protoplasm cosmetic and preparation method and application thereof - Google Patents
Ganoderma lucidum fermentation protoplasm cosmetic and preparation method and application thereof Download PDFInfo
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- CN105147587A CN105147587A CN201510639333.3A CN201510639333A CN105147587A CN 105147587 A CN105147587 A CN 105147587A CN 201510639333 A CN201510639333 A CN 201510639333A CN 105147587 A CN105147587 A CN 105147587A
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- 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
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- 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
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Abstract
The invention discloses a ganoderma lucidum fermentation protoplasm cosmetic and a preparation method and application thereof. The ganoderma lucidum fermentation protoplasm cosmetic is prepared through the following steps that dry ganoderma lucidum powder, water and ferment bacteria are mixed to obtain an initial system, and ferment cultivation is performed on the initial system to obtain ganoderma lucidum fermentation protoplasm; wine yeasts are adopted as the ferment bacteria. The invention further discloses the application of the ganoderma lucidum fermentation protoplasm cosmetic in preparing a cosmetic with the whitening and/or aging resisting functions, wherein the cosmetic specifically adopts any one of a facial mask, essence and toner. The prepared ganoderma lucidum fermentation protoplasm cosmetic does not contain any chemical ingredients, can be directly used as finished products of the facial mask or the essence or the toner, is more natural than other existing cosmetics in the market and cannot cause any negative effect on the skins; in addition, active ingredients with small molecular weight can be obtained, and therefore the ganoderma lucidum fermentation protoplasm cosmetic can be more easily absorbed by the skins.
Description
Technical field
The invention belongs to biological technical field, be specifically related to a kind of glossy ganoderma fermentation oleo stock cosmetics and preparation method thereof and application.
Background technology
Ganoderma Ganodermalucidum (Curtis:Fr.) P.Karst. is the famous and precious medicinal fungi of China.Because it can prevent or treat numerous disease, be once called as in ancient times " Herba mesonae chinensis ".Modern chemistry research proves, Ganoderma contains multiple bioactive ingredients, and as polysaccharide, triterpene, sterol, nucleic acid, protein, polypeptide, fatty acid etc., wherein ganoderan and Ganoderma triterpenoids are the crucial active ingredient of Ganoderma.Modern pharmacology research shows, the biological activity of ganoderan is very extensive, comprises the metabolism, antioxidation, anti-aging effects etc. of immunity moderation effect, antitumor action, promotion nucleic acid in vivo and protein.
What existing ganoderma active thing extraction the most often adopted is water extraction: Ganoderma dry product adds deionized water after pulverizing, and stirring and leaching a period of time after tune pH, get supernatant concentration after centrifugal, finally use alcohol settling, crushed after being dried obtains product.But the method introduces Organic substance in leaching process, not only welding, and potential potential safety hazard is there is in the process of application.
Summary of the invention
An object of the present invention is to provide a kind of preparation method of glossy ganoderma fermentation oleo stock.
The preparation method of glossy ganoderma fermentation oleo stock provided by the present invention, comprises the steps: Ganoderma dry powder, water and zymocyte bacterium liquid to be mixed to get initial system, carries out fermentation culture, obtain glossy ganoderma fermentation oleo stock to described initial system.
In above-mentioned preparation method, described Ganoderma dry powder is Ganoderma sporophore dry powder, and the size of described Ganoderma dry powder is 20-50 order, specifically can be 30 orders.
The concentration of described zymocyte bacterium liquid is 10
6-10
8cFU/mL.
The pH value of described zymocyte bacterium liquid is 6.5-7.5.
The proportioning of described zymocyte bacterium liquid, Ganoderma dry powder and water is: 18-28mL:5-10g:250g, specifically can be 22mL:8g:250g.
Described zymocyte is yeast, specifically can be wine yeast Saccharomycesellipsoideus.
The temperature of described fermentation culture is 25-35 DEG C, and specifically can be 30 DEG C, the time is 20h-28h, specifically can be 24h.
Above-mentioned preparation method also comprise to the fermentation liquid obtained after described fermentation culture carry out successively sterilizing, centrifugal, get the step that supernatant obtains glossy ganoderma fermentation oleo stock cosmetics.
The condition of described sterilizing is: sterilising temp is 110-121 DEG C, is specially 115 DEG C, and sterilization time is 15-30min, specifically can be 20min.
Described centrifugal condition is: centrifugal rotational speed is 4500-6000r/min, and be specially 5000r/min, centrifugation time is 20-30min, be specially 25min, centrifugal radius is 8-12cm, is specially 9cm.
The preparation-obtained glossy ganoderma fermentation oleo stock of the present invention also belongs to protection scope of the present invention.
The present invention's preparation-obtained glossy ganoderma fermentation oleo stock is being prepared the application that has in the cosmetics of following at least one function or is directly also belonging to protection scope of the present invention as the application had in the cosmetics of following at least one function:
1) whitening;
2) defying age;
3) scavenging free radicals;
4) restraint of tyrosinase.
In above-mentioned application, described cosmetics are facial film, essence or cosmetic water.
The invention provides a kind of preparation method of glossy ganoderma fermentation oleo stock cosmetics, the method selects more suitable strain, make product more natural, and do not need to add the foreign substances such as enzyme, not only save production cost, and maximizedly simplify production stage, thus enable this fermentation technique realize a large amount of production, suitability for industrialized production, and fully can ensure the stability of product quality.Prove by experiment: glossy ganoderma fermentation oleo stock prepared by the present invention, not containing any chemical composition, can directly use as the finished product of facial film or essence or cosmetic water, more natural than existing other products on the market, can not cause any negative interaction to skin; And the little active component of molecular weight can be obtained, more easily fully absorbed by skin.
The present invention advantageously (1) the present invention adopts wine yeast to carry out whole plant fermentation to Ganoderma, remains whole functional component and the activity thereof of plant, avoids the active component loss that extracting method causes.(2) fermentation method of the present invention, in effective component of glossy ganoderma leaching process, does not add any organic reagent, and fermentation temperature and fermentation pH gentleness, active components of plants structure is not destroyed, and keeps the natural activity of plant.(3), after cosmetics provided by the invention have fermented, do not add essence etc. to chemical composition in fermentation oleo stock, guarantee the safety of product to human body.(4) material such as polysaccharide, albumen be rich in glossy ganoderma fermentation oleo stock provided by the invention, has synergism with the probiotics fermention product component in ferment filtrate, thus has antioxidation and anti-aging effects significantly.
Accompanying drawing explanation
Fig. 1 is the active polypeptide molecular weight distribution HPLC collection of illustrative plates of glossy ganoderma fermentation oleo stock cosmetics.
Fig. 2 is the removing DPPH free radical result schematic diagram of glossy ganoderma fermentation oleo stock cosmetics.
Fig. 3 is the active schematic diagram of restraint of tyrosinase of glossy ganoderma fermentation oleo stock cosmetics.
Detailed description of the invention
Below by specific embodiment, the present invention will be described, but the present invention is not limited thereto.
The experimental technique used in following embodiment if no special instructions, is conventional method; Reagent used in following embodiment, material etc., if no special instructions, all can obtain from commercial channels.
The fluid medium adopted in following embodiment is by solute and solvent composition, and solvent is water, and the concentration in solute and liquid medium within thereof is: glucose 20g/L, peptone 20g/L, yeast extract 10g/L, pH value is 7.0.
Adopt in following embodiment YPD culture medium by solute and solvent composition, solvent is water, concentration in solute and liquid medium within thereof is: yeast extract 1% (mass fraction), tryptone 2% (mass fraction), glucose 2% (mass fraction), pH value is 7.0.
The Ganoderma adopted in following embodiment is purchased from Kang Pu bio tech ltd, Qinghai.
The ATCC of the wine yeast Saccharomycesellipsoideus adopted in following embodiment is numbered 4108, can obtain from ATCC.
The preparation of embodiment 1, glossy ganoderma fermentation oleo stock
1, the activation of strain
Picking wine yeast (Saccharomycesellipsoideus) bacterium colony one ring, in fluid medium, puts into shaking table by actication of culture, obtains activating rear wine yeast liquid.
2, the purification of strain
Bacterium liquid gradient dilution bed board after activation step 1 obtained, to obtain single bacterium colony, obtains wine yeast after purification.
3, the amplification culture of strain
Strain inoculation after purification step 2 obtained, in YPD culture medium (pH value is 7.0), is cultivated in the shaking table of 25 DEG C, and as OD value=0.6-1.0, (strain is in logarithmic (log) phase, and concentration is 10 to obtain zymocyte bacterium liquid
6-10
8cFU/ml).
4, the acquisition of glossy ganoderma fermentation oleo stock
Zymocyte bacterium liquid 22mL step 3 obtained is inoculated in the Ganoderma dry powder of 8g and the water of 250g, obtains fermentation system (in fermentation system, the proportioning of bacterium liquid and Ganoderma and water is 22mL:8g:250g); Fermentation system is fermented 24 hours in 30 DEG C of shaking tables, obtains tunning; By tunning 115 DEG C of autoclaving 20min, make bacterium inactivation, obtain the tunning after sterilizing; By the centrifugal 25min under 5000r/min, centrifugal radius are the condition of 9cm of the tunning after sterilizing, abandon precipitation, collect supernatant, be glossy ganoderma fermentation oleo stock.
Embodiment 2, glossy ganoderma fermentation oleo stock are in the application as cosmetics:
One, the character of glossy ganoderma fermentation oleo stock:
The outward appearance that this embodiment prepares the glossy ganoderma fermentation oleo stock (supernatant) of gained is thick liquid, color is faint yellow to brown color.PH value 5.2-6.3, viscosity 200-500cP, solubility is admittedly containing thing content 1.5-5.0%, 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.
Carry out component analysis to this glossy ganoderma fermentation oleo stock, protein detection method is with reference to GB5009.5-2010; Crude polysaccharides detection method is with reference to GB/T5009.8-2008; Flavone detection method is with reference to GB/T5009.124-2003; Total phenols detection method is with reference to GB/T8313-2008, and acquired results is as follows:
The glossy ganoderma fermentation oleo stock that the present invention obtains is containing protein 2.93g/kg, crude polysaccharides 9.91g/kg, total flavones 0.045g/kg (in rutin), total phenols 0.054g/kg.
Two, the safety of glossy ganoderma fermentation oleo stock detects
Human body patch test is mainly used for the zest detecting cosmetics finished product or raw material.The present invention carries out human closed formula patch test to the glossy ganoderma fermentation oleo stock that embodiment 1 obtains, and 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
If solid sample or semisolid sample, then take 0.020g-0.025g be placed in speckle examination device for subsequent use; If fluid sample, then measure 0.025mL-0.2mL and drip on filter paper, then filter paper is placed in speckle examination device.The present invention adopts the latter, and each sample all arranges blank, in contrast speckle examination device hole, add the sample solvent with sample equivalent, as distilled water or olive oil.
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:
"-"=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.
According to cosmetics health specification 2007, human body patch test criterion is: occur in 30 routine experimenters that the number of 1 grade of skin adverse reaction is more than 5 examples, or 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.
As can be seen from Table 1: the glossy ganoderma fermentation oleo stock that embodiment 1 obtains does not produce suspicious reaction, illustrate that Ganoderma fermented product provided by the invention all has safety, untoward reaction can not be brought to human body.
The patch test results of the glossy ganoderma fermentation oleo stock that table 1, embodiment 1 obtain
Three, the molecular size range of glossy ganoderma fermentation oleo stock detects
Molecular size range checks the key factor whether can carrying out Cutaneous permeation.The present invention measures the molecular size range of glossy ganoderma fermentation oleo stock prepared by embodiment 1 by high performance liquid chromatography (HPLC).Concrete steps are as follows:
1, the reagent of test and the preparation of solution
(1) mobile phase: acetonitrile: water: trifluoroacetic acid=30:70:0.1 (v/v/v).
(2) sample preparation: the sample take mobile phase as solvent compound concentration being 5mg/ml, then for sample introduction after filtering with microporous membrane (0.45 μm).
(3) preparation of standard sample: bovine serum albumin (Mr67000), B12 (Mr1335), oxidized form of glutathione (Mr614) are made into mixed mark, and the concentration of often kind of material is 5mg/ml.
2, test method
Standard substance by three kinds of known molecular amounts: bovine serum albumin (Mw67000), vitamin B
12(Mw1335) and oxidized form of glutathione (Mw614) by high performance liquid chromatograph (Waters company) reference literature " Wang Chengzhong etc. the research of polypeptide molecular weight distribution in protease hydrolyzed wheat germ protein and enzymolysis solution. grain processing; 2012:37 (2) " in method carry out HPLC analysis, draw the standard curve of molecular weight logarithm and elution time.According to gel column permeation chromatography principle, the material that polypeptide molecular weight is large is first by eluting out, and the logarithm of elution time and molecular weight is linear.After the same method the glossy ganoderma fermentation oleo stock of preparation in embodiment 1 is carried out HPLC analysis again, obtain the elution time of respective peaks, elution time is substituted into standard curve, the molecular size range of glossy ganoderma fermentation oleo stock prepared by embodiment 1 can be obtained.
Chromatographic condition: chromatographic column: TsKgel2000SWXL300mm × 7.8mm; Determined wavelength: UV280nm; Flow velocity: 1ml/min; Column temperature: 30 DEG C.
3, result of the test
The molecular weight of standard substance and the as shown in table 2 of elution time, be figure according to the logarithm of molecular weight and elution time and obtain standard curve: y=-2.4738x+17.656, R
2=0.9998.Wherein y is elution time (min); X is molecular weight logarithm.
The molecular weight of table 2, standard substance and elution time
Embodiment 1 prepare glossy ganoderma fermentation oleo stock HPLC collection of illustrative plates as shown in Figure 1: as can be seen from Figure 1,1 component peaks is mainly comprised in the collection of illustrative plates of glossy ganoderma fermentation oleo stock, elution time is 11.74min, elution time is substituted into standard curve: y=-2.4738x+17.656, the molecular weight calculating peak is 247.22Da, and accounts for 97.26% according to calculated by peak area glossy ganoderma fermentation oleo stock.It is generally acknowledged that molecular weight about 500Da bioactive peptide composition is more easily absorbed by the skin, so there is a large amount of absorbable active polypeptide in Ganoderma fermented product.
Four, the antioxygenic property of glossy ganoderma fermentation oleo stock 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
Make glossy ganoderma fermentation oleo stock (supernatant) to DPPH free radical scavenging effect curves, see accompanying drawing 2.
As shown in Figure 2, embodiment 1 gained glossy ganoderma fermentation oleo stock is to the IC50=22.23% of DPPH scavenging action, illustrating that glossy ganoderma fermentation oleo stock has stronger oxidation resistance can scavenging free radicals, promote 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.
Five, the white-skinned face function analysis of glossy ganoderma fermentation oleo stock
Tryrosinase is melanogenic key enzyme, which control the forming process of melanocyte, and the deposition of its level of activity to pigment plays a major role.Many whitenings of selling in the market, speckle dispelling product are all reach whitening function with restraint of tyrosinase, therefore are evaluate the leading indicator of whitening cosmetic to the power of tyrosinase inhibitory action.
Carried out the whitening function of assess sample by the impact of working sample on tryrosinase, concrete grammar is as follows:
Solution is configured by table 3:
Table 3, solution preparation list
| Unit (mL) | C 1 | C 2 | T 1 | T 2 |
| TYR | 2 | 2 | 2 | 2 |
| Sample | 0 | 0 | 2 | 2 |
| PBS | 4 | 5 | 2 | 3 |
| Tryrosinase | 1 | 0 | 1 | 0 |
| Cumulative volume | 7 | 7 | 7 | 7 |
Note: C
1and T
1add 1mL tryrosinase, enzyme is lived as 100U/mL.
(1) C
2after pipe prepares and shakes up, heating in water bath 10min in 37 DEG C of water-baths, returns to zero under wavelength 475nm.
(2) C
1pipe solution prepares and shakes up, and after 37 DEG C of water-bath 10min, adds tryrosinase 1ml, continues water-bath 10 minutes, measures C
1absorbance.
(3) by (1) (2) same method, with T
2zeroing measures T
1absorbance.
(4) calculation sample is to the maximum inhibition T (%) of tryrosinase.T (%)=(C
1-T
1)/C
1× 100% experiment prove the glossy ganoderma fermentation oleo stock of dilution 10 times to the suppression ratio of tyrosinase activity be the arbutin of 35.47%, 1% to the suppression ratio 99.52% of tyrosinase activity, see accompanying drawing 3.Namely the whitening effect diluting the glossy ganoderma fermentation oleo stock of ten times is equivalent to the effect of the arbutin of 0.356%, has certain white-skinned face function.
Claims (10)
1. a glossy ganoderma fermentation oleo stock preparation method, comprises the steps: Ganoderma dry powder, water and zymocyte bacterium liquid to be mixed to get initial system, carries out fermentation culture, obtain glossy ganoderma fermentation oleo stock to described initial system.
2. preparation method according to claim 1, is characterized in that: the proportioning of described zymocyte bacterium liquid and Ganoderma dry powder and water is: 18-28mL:5-10g:250g, and described zymocyte bacterial concentration is 10
6-10
8cFU/mL.
3. preparation method according to claim 1 and 2, is characterized in that: the temperature of described fermentation culture is 25-35 DEG C; Time is 20h-28h.
4. the preparation method according to any one of claim 1-3, is characterized in that: described zymocyte is yeast, is specially wine yeast (Saccharomycesellipsoideus);
Described Ganoderma dry powder is Ganoderma sporophore dry powder.
5. the preparation method according to any one of claim 1-4, is characterized in that: the size of described Ganoderma dry powder is 20-50 order.
6. the preparation method according to any one of claim 1-5, is characterized in that: described preparation method also comprise to gained fermentation liquid after described fermentation culture carry out successively sterilizing, centrifugal, get the step that supernatant obtains glossy ganoderma fermentation oleo stock.
7. preparation method according to claim 6, is characterized in that: the condition of described sterilizing is as follows: sterilising temp is 110-121 DEG C, and sterilization time is 15-30min;
Described centrifugal condition is as follows: centrifugal rotational speed is 4500-6000r/min, centrifugation time is 20-30min, and centrifugal radius is 8-12cm.
8. the glossy ganoderma fermentation oleo stock that the preparation method according to any one of claim 1-7 obtains.
9. glossy ganoderma fermentation oleo stock according to claim 8 prepare the application that has in the cosmetics of following at least one function or directly as the application had in the cosmetics of following at least one function:
1) whitening;
2) defying age;
3) scavenging free radicals;
4) restraint of tyrosinase.
10. application according to claim 9, is characterized in that: described cosmetics are facial film, essence or cosmetic water.
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| CN109260067A (en) * | 2018-09-26 | 2019-01-25 | 广东盛美化妆品有限公司 | A kind of myogenic honey of the ferment filtrate containing Ganoderma Sinense and preparation method thereof |
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| CN111135136A (en) * | 2020-03-26 | 2020-05-12 | 广州七遇生物科技有限公司 | Anti-aging skin care composition and preparation method thereof |
| CN111973498A (en) * | 2020-09-25 | 2020-11-24 | 广州环亚化妆品科技有限公司 | Ganoderma lucidum fermentation product and preparation method and application thereof |
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| CN105769667A (en) * | 2016-03-18 | 2016-07-20 | 无限极（中国）有限公司 | Lucid ganoderma fermentation liquor preparation method and application of lucid ganoderma fermentation liquor |
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| CN110638697B (en) * | 2019-11-01 | 2022-08-16 | 上海浅娇生物科技有限公司 | Preparation and application of ganoderma lucidum lactobacillus fermentation extract |
| CN110638697A (en) * | 2019-11-01 | 2020-01-03 | 上海浅娇生物科技有限公司 | Preparation and application of ganoderma lucidum lactobacillus fermentation extract |
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| CN112245343B (en) * | 2020-09-27 | 2021-08-10 | 北京工商大学 | Method for fermenting burdock root by lucid ganoderma, method for compositely fermenting burdock root, fermented product and application |
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