CN110585075A - A method for preparing mushroom extract containing skin whitening factor - Google Patents
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- CN110585075A CN110585075A CN201910904277.XA CN201910904277A CN110585075A CN 110585075 A CN110585075 A CN 110585075A CN 201910904277 A CN201910904277 A CN 201910904277A CN 110585075 A CN110585075 A CN 110585075A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/49—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds
- A61K8/494—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds with more than one nitrogen as the only hetero atom
- A61K8/4946—Imidazoles or their condensed derivatives, e.g. benzimidazoles
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
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- 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
- A61K8/9728—Fungi, e.g. yeasts
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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
- 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
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Abstract
The invention provides a preparation method of a mushroom extract containing skin whitening factors, which is characterized by comprising the following steps: step 1: processing mushroom raw material to obtain mushroom powder or homogenate; step 2: mixing the mushroom powder or homogenate obtained in the step (1) with water, heating, centrifuging or filtering to obtain supernatant; and step 3: removing the solvent in the supernatant obtained in the step 2, dissolving the solvent by using an ethanol solution, standing, and centrifuging or filtering to obtain a supernatant; and 4, step 4: removing the ethanol in the supernatant obtained in the step 3 to obtain residue, and dissolving with deionized water to obtain the mushroom extract containing the skin whitening factor. The invention takes mushroom as raw material, after the processing of the process steps, the obtained extract has high content of ergothioneine and glutathione, water is used as solvent, the product has high safety, low cost, simple process steps, short period and less energy consumption, and is suitable for large-scale industrial production.
Description
Technical Field
The invention relates to a preparation method of a mushroom extract containing skin whitening factors.
Background
The world cosmetic industry is valued in the billions of dollars, but there is a continuing search for new or improved natural products as suitable raw materials in the hopes of competing with synthetic products or adding new biological activities. Natural ingredients such as plant nutrients, microbial metabolites, dairy products, minerals and animal protein ingredients are very common in skin care today. Fungi, particularly mushrooms and their preparations, their crude extracts or pure compounds are widely used in the cosmetic field to treat various common diseases related to the skin.
Mushrooms are a general name of edible mushrooms, have the functions of resisting oxidation, resisting cancer, regulating immunity, resisting inflammation, protecting cardiovascular, resisting bacteria, resisting diabetes, reducing cholesterol and the like, and the functions are not only related to proteins, polysaccharides, amino acids, dietary fibers, mineral substances and the like contained in the mushrooms, but also related to antioxidant active substances such as phenolic compounds, vitamins, terpenes, steroids and the like, so that the application range of the mushrooms is expanded from the food industry to the health-care product, medicine and cosmetic industry.
Ergothioneine (EGT) is 2-sulfydryl-histidine-trimethyl inner salt, is a colorless and tasteless natural amino acid, and has a structural formula shown in a formula (I). EGT is distributed throughout the human body and has the highest concentration in erythrocytes, liver, kidney and semen, but the human body cannot directly synthesize EGT by itself and needs to take the EGT from the outside, and the EGT is usually obtained from mushroom, red bean, oat bran and liver, wherein the content of the EGT is the highest in the mushroom. EGT has antioxidant (higher than glutathione and ascorbic acid), antiinflammatory, various radioactive injury inhibiting, cell protecting, and cell activity enhancing effects. The EGT content is related to another sulfur-containing antioxidant, namely reduced Glutathione (GSH), and the ubiquitous peptide (c-glutamylcysteinylglycine) is a sulfhydryl-containing tripeptide compound (with a structural formula shown in formula (II)) formed by polymerizing glutamic acid, cysteine and glycine, is considered to be a main intracellular antioxidant in almost all organisms, and simultaneously has the functions of detoxifying various toxins and carcinogens, regulating protein functions after translation, maintaining immune functions and the like. The synthesis of EGT depends on GSH and its precursor, and EGT can help to maintain GSH level in the presence of oxidation load through interaction with other cell defense systems, so that the two are in complementary relationship to maintain normal metabolism of cells together and ensure health of life body.
The mushroom extract rich in EGT and GSH is concerned and researched by more and more scholars due to the obvious potential of antioxidation, antibiosis, anti-inflammation, cell protection and the like, and is widely applied to the cosmetic industry. It is therefore of great importance to select a simple and efficient method for the preparation of EGT and GSH in mushrooms.
Disclosure of Invention
The invention aims to provide a method with low cost, short period and simple production process for preparing the mushroom extract rich in ergothioneine and glutathione.
In order to achieve the above object, the present invention provides a method for preparing a mushroom extract containing skin whitening factors, comprising:
step 1: processing mushroom raw material to obtain mushroom powder or homogenate;
step 2: mixing the mushroom powder or homogenate obtained in the step (1) with water, heating, centrifuging or filtering to obtain supernatant;
and step 3: removing the solvent in the supernatant obtained in the step 2, dissolving the solvent by using an ethanol solution, standing, and centrifuging or filtering to obtain a supernatant;
and 4, step 4: removing the ethanol in the supernatant obtained in the step 3 to obtain residue, and dissolving with deionized water to obtain the mushroom extract containing the skin whitening factor.
The mushroom raw material is selected from shiitake, pleurotus ostreatus, agaricus bisporus, flat-bottom mushroom, ganoderma lucidum and the like which are generally selected as research objects according to the using regulations of cosmetics, fresh mushroom or dry mushroom can be selected, and the parts of pileus, stipes, mushroom folds, hypha and the like and leftovers of residual mushroom, mushroom dregs and the like can be used as extraction raw materials.
Preferably, the specific steps of processing mushroom raw material comprise: when the mushroom is dried mushroom, directly crushing the mushroom into powder, and sieving the powder by a 40-80-mesh sieve; when the mushroom is fresh mushroom, cleaning the mushroom, freeze-drying or drying the mushroom, then crushing the mushroom, and sieving the mushroom with a 40-80 mesh sieve to obtain mushroom powder, or crushing the fresh mushroom into mushroom homogenate.
Preferably, the ratio of the mushroom powder to the water in the step 2 is 1 (30-50) (m: v), and the ratio of the mushroom homogenate to the water is 1 (3-5) (m: v).
Preferably, the heating temperature in the step 2 reaches 95 +/-5 ℃ for 1-2 h.
Preferably, the centrifugal force in the steps 2 and 3 is 3000g, the centrifugal temperature is 2-4 ℃, and the time is 10-20 min.
Preferably, the solvent in the supernatant obtained in the step 2 is removed by adopting a rotary evaporation or drying method, and the rotary evaporation conditions are as follows: the temperature reaches 40 +/-5 ℃, the vacuum degree reaches-0.1 MPa, and the drying conditions are as follows: the temperature reaches 90 +/-5 ℃.
Preferably, the ethanol in the supernatant obtained in the step 3 is removed by adopting a rotary steaming or drying method, and the rotary steaming conditions are as follows: the temperature reaches 40 +/-5 ℃, the vacuum degree reaches-0.1 MPa, and the drying conditions are as follows: the temperature reaches 90 +/-5 ℃.
Preferably, the concentration of the ethanol solution in the step 3 is 70-80% (v/v).
Preferably, the standing temperature in the step 3 is 4-8 ℃, and the time is more than or equal to 2 hours.
Preferably, the content of EGT in the mushroom extract containing the skin whitening factor is 10.313-330.0 mug/g (dried mushroom), and the content of GSH is 20-400 mug/g (dried mushroom).
Compared with the prior art, the invention has the beneficial effects that:
EGT and GSH are water-soluble skin whitening factors, water is used as a solvent for extraction, the production process is safe, the extracting solution prepared by taking mushrooms as a raw material has good effects of oxidation resistance, aging resistance, whitening and the like, is safe and has no toxic or side effect, and the pursuit of people on the nature, safety and good effect of cosmetics is met. Meanwhile, most impurities such as polysaccharide, protein and the like can be removed in the standing alcohol precipitation process, higher content of EGT and GSH can be obtained in the extracting solution, the production process is simple, the preparation period is short, and the method is suitable for large-scale industrial production.
The invention takes mushroom as raw material, after the processing of the process steps, the obtained extract has high content of ergothioneine and glutathione, water is used as solvent, the product has high safety, low cost, simple process steps, short period and less energy consumption, and is suitable for large-scale industrial production.
Drawings
FIG. 1 is an HPLC plot of ergothioneine and glutathione for the samples of example 1; (EGT retention time 14.373 min; GSH retention time 15.037 min);
FIG. 2 is an HPLC plot of ergothioneine and glutathione for the samples of example 2; (EGT retention time 14.39 min; GSH retention time 15.056 min);
FIG. 3 is an HPLC plot of ergothioneine and glutathione for the samples of example 3; (EGT retention time 14.394 min; GSH retention time 15.056 min).
Detailed Description
The invention will be further illustrated with reference to the following specific examples. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. Further, it should be understood that various changes or modifications of the present invention may be made by those skilled in the art after reading the teaching of the present invention, and such equivalents may fall within the scope of the present invention as defined in the appended claims.
Each raw material used in the following examples is a commercially available product.
Example 1
A preparation method of mushroom extract containing skin whitening factor comprises the following steps:
a. pulverizing dried Lysimachia christinae Hance (removing pedicel from Lysimachia christinae Hance) with a miniature plant pulverizer, and sieving with 60 mesh sieve to obtain Lysimachia christinae Hance powder;
b. weighing 2.0g of Lysimachia christinae Hance powder, fully mixing with 100mL of deionized water, heating in a water bath at 95 ℃ for 1h, and centrifuging (3000g, 4 ℃, 15min) to obtain a supernatant;
c. concentrating the supernatant by rotary evaporation (40 deg.C, 0.1MPa) to dryness to remove solvent, dissolving with 30mL 70% (v/v) ethanol solution, placing in a reagent bottle, screwing the reagent bottle to reduce ethanol volatilization, standing at 4 deg.C for 2 hr, and centrifuging (3000g, 4 deg.C, 15min) to obtain supernatant;
d. and (3) carrying out rotary evaporation on the supernatant (at 40 ℃ and under-0.1 MPa), concentrating to remove ethanol to obtain an ethanol-free residue, and dissolving the ethanol-free residue with 5-20mL of deionized water to obtain the Lysimachia christinae Hance extract rich in ergothioneine and glutathione.
And (3) detection of a finished product by an HPLC method:
1. and (3) ergothioneine content detection: chromatographic conditions are as follows: kromasil 100-5NH2Column (250 mm. times.4.60 mm, 5 μm), mobile phase: acetonitrile-5 mmol/L ammonium acetate (80:20), flow rate 1.0mL/min, sample volume 20 μ L, column temperature 30 deg.C, detection wavelength 254nm, and ergothioneine content of 0.29 + -0.11 mg/g (dried mushroom) determined by external standard method.
2. And (3) detecting the content of glutathione: chromatographic conditions are as follows: kromasil 100-5NH2Column (250 mm. times.4.60 mm, 5 μm), mobile phase: 0.1% trifluoroacetic acid water solution-methanol (98:2, v/v), flow rate 1.0mL/min, sample amount 20 uL, column temperature 30 deg.C, detection wavelength 200nm, and glutathione content of 0.32 + -0.07 mg/g (dried mushroom) determined by external standard method.
3. HPLC profiles of the samples ergothioneine and glutathione are shown in FIG. 1.
Example 2
A preparation method of mushroom extract containing skin whitening factor comprises the following steps:
a. cleaning Log flower mushroom (removing pedicle of Log flower mushroom), freeze drying, pulverizing with miniature plant pulverizer, and sieving with 80 mesh sieve to obtain Log flower mushroom powder;
b. weighing 2.0g of raw flower mushroom powder, fully mixing with 80mL of deionized water, heating in a water bath at 98 ℃ for 1h, and centrifuging (3000g, 3 ℃, 20min) to obtain supernatant;
c. concentrating the supernatant by rotary evaporation (45 deg.C, 0.1MPa) to dryness to remove solvent, dissolving 50mL 75% (v/v) ethanol solution, placing in a reagent bottle, screwing the reagent bottle to reduce ethanol volatilization, standing at 4 deg.C for 6 hr, and centrifuging (3000g, 3 deg.C, 20min) to obtain supernatant;
d. concentrating the supernatant by rotary evaporation (45 deg.C and-0.1 MPa) to remove ethanol to obtain residue without ethanol, and dissolving with 5-20mL deionized water to obtain raw wood flower mushroom extract rich in ergothioneine and glutathione.
And (3) detection of a finished product by an HPLC method: 1. and (3) ergothioneine content detection: chromatographic conditions are as follows: kromasil 100-5NH2Column (250 mm. times.4.60 mm, 5 μm), mobile phase: acetonitrile-5 mmol/L ammonium acetate (80:20), flow rate 1.0mL/min, sample volume 20 μ L, column temperature 30 deg.C, detection wavelength 254nm, and ergothioneine content of 0.98 + -0.15 mg/g (dried mushroom) determined by external standard method.
2. And (3) detecting the content of glutathione: chromatographic conditions are as follows: kromasil 100-5NH2Column (250 mm. times.4.60 mm, 5 μm), mobile phase: 0.1% trifluoroacetic acid water solution-methanol (98:2, v/v), flow rate 1.0mL/min, sample amount 20 uL, column temperature 30 deg.C, detection wavelength 200nm, and glutathione content of 0.27 + -0.02 mg/g (dried mushroom) determined by external standard method.
3. HPLC profiles of the samples ergothioneine and glutathione are shown in FIG. 2.
Example 3
A preparation method of mushroom extract containing skin whitening factor comprises the following steps:
a. cleaning Agaricus bisporus (pedicel-removed Agaricus bisporus), and sufficiently crushing to obtain an Agaricus bisporus homogenate;
b. weighing 20.0g of agaricus bisporus homogenate, fully mixing with 80mL of deionized water, heating in a water bath at 95 +/-2 ℃ for 1h, and centrifuging (3000g, 4 ℃, 15min) to obtain a supernatant;
c. concentrating the supernatant by rotary evaporation (40 deg.C, 0.1MPa) to dryness to remove solvent, dissolving 30mL 80% (v/v) ethanol solution, placing in a reagent bottle, screwing the reagent bottle to reduce ethanol volatilization, standing at 5 deg.C for 8 hr, and centrifuging (3000g, 4 deg.C, 15min) to obtain supernatant;
d. and (3) carrying out rotary evaporation on the supernatant (at 40 ℃ and under-0.1 MPa), concentrating to remove ethanol to obtain an ethanol-free residue, and dissolving the ethanol-free residue with 5-20mL of deionized water to obtain the agaricus bisporus extracting solution rich in ergothioneine and glutathione.
And (3) detection of a finished product by an HPLC method: 1. and (3) ergothioneine content detection: chromatographic conditions are as follows: kromasil 100-5NH2Column (250 mm. times.4.60 mm, 5 μm), mobile phase: acetonitrile-5 mmol/L ammonium acetate (80:20, v/v), flow rate 1.0mL/min, sample volume 20 μ L, column temperature 30 deg.C, detection wavelength 254nm, and ergothioneine content of 1.02 + -0.21 mg/g (dried mushroom) determined by external standard method.
2. And (3) detecting the content of glutathione: chromatographic conditions are as follows: kromasil 100-5NH2Column (250 mm. times.4.60 mm, 5 μm), mobile phase: 0.1% trifluoroacetic acid water solution-methanol (98:2, v/v), flow rate 1.0mL/min, sample amount 20 uL, column temperature 30 deg.C, detection wavelength 200nm, and glutathione content of 0.49 + -0.13 mg/g (dried mushroom) determined by external standard method.
3. HPLC profiles of the samples ergothioneine and glutathione are shown in FIG. 3.
Claims (10)
1. A method for preparing mushroom extract containing skin whitening factors is characterized by comprising the following steps:
step 1: processing mushroom raw material to obtain mushroom powder or homogenate;
step 2: mixing the mushroom powder or homogenate obtained in the step (1) with water, heating, centrifuging or filtering to obtain supernatant;
and step 3: removing the solvent in the supernatant obtained in the step 2, dissolving the solvent by using an ethanol solution, standing, and centrifuging or filtering to obtain a supernatant;
and 4, step 4: removing the ethanol in the supernatant obtained in the step 3 to obtain residue, and dissolving with deionized water to obtain the mushroom extract containing the skin whitening factor.
2. The method of preparing mushroom extract containing skin whitening factors according to claim 1, wherein the specific steps of treating mushroom material comprise: when the mushroom is dried mushroom, directly crushing the mushroom into powder, and sieving the powder by a 40-80-mesh sieve; when the mushroom is fresh mushroom, cleaning the mushroom, freeze-drying or drying the mushroom, then crushing the mushroom, and sieving the mushroom with a 40-80 mesh sieve to obtain mushroom powder, or crushing the fresh mushroom into mushroom homogenate.
3. The method of preparing mushroom extract containing skin whitening factors according to claim 1, wherein the ratio of the mushroom powder to water in step 2 is 1: 30-50 (m/v), and the ratio of the mushroom homogenate to water is 1: 3-5 (m/v).
4. The method of preparing mushroom extract containing skin whitening factors according to claim 1, wherein the heating temperature in the step 2 reaches 95 ± 5 ℃ for 1-2 hours.
5. The method of preparing mushroom extract containing skin whitening factors according to claim 1, wherein the centrifugal force in the steps 2 and 3 is 3000g, the centrifugal temperature is 2-4 ℃, and the time is 10-20 min.
6. The method of preparing mushroom extract containing skin whitening factors according to claim 1, wherein the removal of the solvent from the supernatant obtained in the step 2 is performed by a rotary steaming or oven-drying method, wherein the rotary steaming conditions are as follows: the temperature reaches 40 +/-5 ℃, the vacuum degree reaches-0.1 MPa, and the drying conditions are as follows: the temperature reaches 90 +/-5 ℃.
7. The method of preparing mushroom extract containing skin whitening factors according to claim 1, wherein the removal of ethanol from the supernatant obtained in the step 3 is performed by a rotary steaming or oven-drying method, wherein the rotary steaming conditions are as follows: the temperature reaches 40 +/-5 ℃, the vacuum degree reaches-0.1 MPa, and the drying conditions are as follows: the temperature reaches 90 +/-5 ℃.
8. The method of preparing mushroom extract containing skin whitening factors according to claim 1, wherein the concentration of the ethanol solution in the step 3 is 70-80% (v/v).
9. The method of preparing mushroom extract containing skin whitening factors according to claim 1, wherein the temperature of the standing in the step 3 is 4 to 8 ℃ for 2 hours or more.
10. The method of preparing the skin whitening factor-containing mushroom extract according to claim 1, wherein the content of EGT in the skin whitening factor-containing mushroom extract is 10.313-330.0 μ g/g (dried mushroom), and the content of GSH is 20-400 μ g/g (dried mushroom).
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113181085A (en) * | 2021-04-29 | 2021-07-30 | 广州睿森生物科技有限公司 | Composition matrix for improving skin problems after makeup and application of composition matrix in cosmetics |
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| CN106831597A (en) * | 2017-03-06 | 2017-06-13 | 浙江尖峰健康科技有限公司 | A kind of method that utilization mushroom leftover bits and pieces prepares natural erythrothioneine |
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Patent Citations (3)
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| JP2007300916A (en) * | 2006-04-10 | 2007-11-22 | Manabu Nukina | Method for producing ergothioneine |
| CN106831597A (en) * | 2017-03-06 | 2017-06-13 | 浙江尖峰健康科技有限公司 | A kind of method that utilization mushroom leftover bits and pieces prepares natural erythrothioneine |
| CN109293572A (en) * | 2018-09-26 | 2019-02-01 | 上海市农业科学院 | A kind of method of erythrothioneine and polysaccharide in extraction Pleurotus citrinopileatus |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN113181085A (en) * | 2021-04-29 | 2021-07-30 | 广州睿森生物科技有限公司 | Composition matrix for improving skin problems after makeup and application of composition matrix in cosmetics |
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