CN115089529B - Mung bean hull fermentation product with acne removing effect, and preparation method and application thereof - Google Patents
Mung bean hull fermentation product with acne removing effect, and preparation method and application thereof Download PDFInfo
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- CN115089529B CN115089529B CN202210927333.3A CN202210927333A CN115089529B CN 115089529 B CN115089529 B CN 115089529B CN 202210927333 A CN202210927333 A CN 202210927333A CN 115089529 B CN115089529 B CN 115089529B
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- mung bean
- lactobacillus plantarum
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- acne
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- 238000000855 fermentation Methods 0.000 title claims abstract description 104
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Classifications
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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
- A61K8/9783—Angiosperms [Magnoliophyta]
- A61K8/9789—Magnoliopsida [dicotyledons]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/10—Anti-acne agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P1/00—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes
- C12P1/02—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes by using fungi
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P1/00—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes
- C12P1/04—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes by using bacteria
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P39/00—Processes involving microorganisms of different genera in the same process, simultaneously
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/225—Lactobacillus
- C12R2001/25—Lactobacillus plantarum
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/645—Fungi ; Processes using fungi
- C12R2001/85—Saccharomyces
- C12R2001/865—Saccharomyces cerevisiae
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
- Y02P60/87—Re-use of by-products of food processing for fodder production
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- Biotechnology (AREA)
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- Mycology (AREA)
- General Engineering & Computer Science (AREA)
- Dermatology (AREA)
- Genetics & Genomics (AREA)
- Biochemistry (AREA)
- Medicinal Chemistry (AREA)
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- Pharmacology & Pharmacy (AREA)
- Gerontology & Geriatric Medicine (AREA)
- Rheumatology (AREA)
- Epidemiology (AREA)
- Pain & Pain Management (AREA)
- Birds (AREA)
- Botany (AREA)
- Cosmetics (AREA)
Abstract
The invention belongs to the technical field of cosmetic raw materials, and discloses a mung bean hull fermentation product with an acne removing effect, and a preparation method and application thereof. The preparation method comprises four process steps of preparing a fermentation culture medium, fermenting, lysing and separating solid from liquid, wherein the fermentation culture medium comprises 10-100 g/kg of mung bean hull, 1-5 g/kg of soybean or soybean protein and 1-5 g/kg of glucose; the fermentation process steps are divided into lactic acid bacteria fermentation and saccharomycetes fermentation which are sequentially and stepwise completed, wherein the strain adopted by the lactic acid bacteria fermentation is lactobacillus plantarum domesticated by high-concentration gallic acid, and the strain adopted by the saccharomycetes fermentation is saccharomyces cerevisiae. The mung bean hull is obtained by separating dried mung bean after roller grinding. The fermented product has antioxidant, skin inflammation relieving and acne symptom improving effects, and has low skin irritation.
Description
Technical Field
The invention belongs to the technical field of cosmetic raw materials, and particularly relates to a mung bean hull fermentation product with an acne removing effect, and a preparation method and application thereof.
Background
The traditional Chinese medicine considers that the mung beans have the effects of clearing heat and detoxicating, treating the initial stage of carbuncles, sores and swelling, eliminating some inflammations of skin, treating eczema, miliaria and the like, and the application of adding mung bean powder into cosmetics in China and Japan. The 'materia medica outline' cloud: while the actions of mung bean for detumescence and acne are similar to that of red bean, … … with the actions of pressing heat and removing toxicity is effective in treating carbuncle, with internal support and heart protection. If the mung bean powder is directly used, the formulation of the cosmetic is greatly limited, and the cosmetic can only be applied to formulations which need cleaning after being used such as a smearing mask, for example, products described in patents CN201310269292.4, CN201010514981.3, CN201610945108.7 and preparation method of a charcoal/mung bean paste cleaning and acne removing mask containing plant ferment. More common application is to extract the active ingredient in mung beans to obtain mung bean extract, and then adding the mung bean extract into cosmetics, for example, products described in patent CN201610356277.7 mung bean ferment face-cleaning gel and CN20201142118. X. The active ingredients in mung beans can be extracted by water extraction, ethanol or polyalcohol extraction, microbial fermentation and other modes, for example, the technology described in patent CN201110300908.0, mung bean sprout extract and preparation method and application thereof, CN201711260225.0, mung bean fermentation broth for repairing damaged sebum membranes and relieving skin allergy and preparation method thereof.
The mung bean hull integrates most of active ingredients such as flavone, polyphenol, polysaccharide, vitamins and the like of the mung bean, but the mung bean hull byproducts are often discarded in the use of the mung bean in the food industry of China at present, so that great waste is caused. If the active ingredients in the mung bean hull are extracted and applied to cosmetics, the mung bean hull has good acne removing effect theoretically. However, mung bean polyphenols have significant irritation to human skin. Therefore, to popularize the application of mung bean hull extract in cosmetics, the irritation of mung bean polyphenol is reduced. The reduction of the pungency of mung bean polyphenols can be theoretically achieved by microbial fermentation, but no such patent technology has been found yet.
Disclosure of Invention
In order to overcome the defects and shortcomings in the prior art, the primary purpose of the invention is to provide a preparation method of mung bean hull fermentation product with acne removing effect; the method is characterized in that dry peeled mung bean hull is used as a production raw material instead of wet peeled mung bean hull, and lactobacillus plantarum domesticated by high-concentration gallic acid is used for fermentation, so that the irritation of mung bean polyphenol is greatly reduced.
The invention also aims to provide the mung bean hull fermentation product with the acne-removing effect, which is prepared by the preparation method.
The invention also aims to provide an application of the mung bean hull fermentation product with the acne-removing effect.
The aim of the invention is achieved by the following technical scheme:
the preparation method of the mung bean hull fermentation product with the acne-removing effect comprises four process steps of preparing a fermentation medium, fermenting, lysing and separating solid from liquid, wherein the fermentation medium comprises 10-100 g/kg of mung bean hull, 1-5 g/kg of soybean or soybean protein and 1-5 g/kg of glucose; the fermentation process steps are divided into lactic acid bacteria fermentation and saccharomycetes fermentation which are sequentially and stepwise completed, wherein the strain adopted by the lactic acid bacteria fermentation is lactobacillus plantarum domesticated by high-concentration gallic acid, and the strain adopted by the saccharomycetes fermentation is saccharomyces cerevisiae.
The mung bean hull is obtained by separating dried mung beans after roller grinding; the mung bean hull is obtained by dry peeling, rather than soaking mung beans in water, then rubbing and separating (wet peeling).
The lactobacillus fermentation is that the lactobacillus plantarum is inoculated and then cultured for 16 to 24 hours at the temperature of 37 ℃; the saccharomycete fermentation is carried out for 4-12 hours at 28 ℃ after inoculating the saccharomyces cerevisiae.
The lysis is to heat the fermentation liquor after the fermentation process is completed to 75 ℃ and keep the temperature for 30min, cool to 30 ℃, and then add 1g/L lysozyme for reaction for 18h.
The lactobacillus plantarum domesticated by the high-concentration gallic acid is specifically obtained by the following method: inoculating lactobacillus plantarum into MRS liquid culture medium, and culturing at 37 ℃ for 24 hours; inoculating the lactobacillus plantarum culture solution obtained in the last step into MRS liquid culture medium containing 2g/kg gallic acid, and culturing at 37 ℃ for 24 hours; inoculating the lactobacillus plantarum culture solution obtained in the last step into an MRS liquid culture medium containing 4g/kg gallic acid, and culturing for 24 hours at 37 ℃; inoculating the lactobacillus plantarum culture solution obtained in the last step into MRS liquid culture medium containing 8g/kg gallic acid, and culturing at 37 ℃ for 24 hours; culturing lactobacillus plantarum culture solution obtained in the last step on MRS solid culture medium containing 10g/kg gallic acid by streaking, and culturing at 37 ℃ for 24 hours; and (3) picking single colony on the lactobacillus plantarum solid culture obtained in the last step, inoculating the single colony into an MRS liquid culture medium containing 10g/kg gallic acid for expansion culture, and culturing at 37 ℃ for 24 hours to obtain the lactobacillus plantarum domesticated by the high-concentration gallic acid.
The MRS liquid culture medium comprises the following components: 10g/L of peptone, 5g/L of beef extract powder, 4g/L of yeast extract powder, 20g/L of glucose, 2g/L of monopotassium phosphate, 2g/L of triammonium citrate, 5g/L of sodium acetate, 0.2g/L of magnesium sulfate, 0.05g/L of manganese sulfate, 1g/L of tween and the balance of water.
The MRS solid culture medium comprises the following components: 10g/L of peptone, 5g/L of beef extract powder, 4g/L of yeast extract powder, 20g/L of glucose, 2g/L of monopotassium phosphate, 2g/L of triammonium citrate, 5g/L of sodium acetate, 0.2g/L of magnesium sulfate, 0.05g/L of manganese sulfate, 1g/L of tween, 15g/L of agar and the balance of water.
A fermented product of testa Phaseoli Radiati with acne removing effect is prepared by the above preparation method.
The mung bean hull fermentation product with the acne-removing effect is applied to cosmetics. The fermented product has antioxidant, skin inflammation relieving and acne symptom improving effects, and has low skin irritation.
Compared with the prior art, the invention has the following advantages and effects:
(1) The product of the invention contains mung bean active ingredients with high concentration. The mung bean hull concentrates most of active ingredients of mung beans, and the mung bean hull obtained by dry peeling contains more active ingredients than the mung bean hull obtained by wet peeling; the technical scheme is that mung bean hull obtained by dry peeling is used as a production raw material, and the obtained fermentation product has high concentration of mung bean active ingredients.
(2) The method can reduce the irritation of mung bean polyphenol. The lactobacillus plantarum is domesticated by high-concentration gallic acid, so that the lactobacillus plantarum has higher activity of expressing glycosyltransferase aiming at polyphenol. The domesticated lactobacillus plantarum is used for fermenting mung bean hull, so that the irritation of mung bean polyphenol can be greatly reduced.
(3) The fermentation product of the invention has good antioxidant activity. The method adopts a two-step fermentation process of lactobacillus and saccharomycetes, and the obtained fermentation product has higher antioxidant activity.
(4) The fermentation product of the invention has no fermented rancid taste. Lactic acid bacteria fermentation often produces sour spoilage, which disappears after absorption and transformation by yeast, leaving the characteristic odor of the plant itself.
(5) The fermentation product of the invention is rich in nutrient substances. By the treatment of the lysis process, intracellular components such as vitamins, amino acids, etc. in the microorganism are dissolved into the solution, and these nutrients can play a role in nourishing the skin.
(6) The invention has good economic benefit and environmental protection benefit. The mung bean kernel is peeled by a dry method in the food industry, and the mung bean hull is peeled by a wet method in the traditional Chinese medicine industry. The invention recycles the waste byproduct mung bean hull in the food industry to produce the cosmetic raw material with high added value, thereby having economic benefit and environmental protection benefit.
Drawings
FIG. 1 is a graph of antioxidant test results.
Figure 2 is a graph of the results of the soothing efficacy test.
Fig. 3 is a photograph of facial acne.
Detailed Description
The present invention will be described in further detail with reference to examples and drawings, but embodiments of the present invention are not limited thereto.
Example 1
This example is the preparation of mung bean hull fermentation product.
(1) Domesticating lactobacillus plantarum: inoculating lactobacillus plantarum into MRS liquid culture medium, and culturing at 37 ℃ for 24 hours; inoculating the lactobacillus plantarum culture solution obtained in the last step into MRS liquid culture medium containing 2g/kg gallic acid, and culturing at 37 ℃ for 24 hours; inoculating the lactobacillus plantarum culture solution obtained in the last step into an MRS liquid culture medium containing 4g/kg gallic acid, and culturing for 24 hours at 37 ℃; inoculating the lactobacillus plantarum culture solution obtained in the last step into MRS liquid culture medium containing 8g/kg gallic acid, and culturing at 37 ℃ for 24 hours; culturing lactobacillus plantarum culture solution obtained in the last step on MRS solid culture medium containing 10g/kg gallic acid by streaking, and culturing at 37 ℃ for 24 hours; and (3) picking single colony on the lactobacillus plantarum solid culture obtained in the last step, inoculating the single colony into an MRS liquid culture medium containing 10g/kg gallic acid for expansion culture, and culturing at 37 ℃ for 24 hours to obtain the lactobacillus plantarum domesticated by the high-concentration gallic acid.
(2) Preparing a fermentation medium: weighing 100g/kg of mung bean hull (obtained by dry peeling), 5g/kg of soy protein and 5g/kg of glucose, adding water, mixing, heating to 105 ℃, preserving heat for 5min, and sterilizing.
(3) Anaerobic fermentation of lactic acid bacteria: the lactobacillus plantarum domesticated by high-concentration gallic acid is cultivated in an MRS liquid culture medium in advance, and bacterial liquid is inoculated into a fermentation culture medium according to the inoculation amount of 3 percent under aseptic operation, and is subjected to stationary culture for 24 hours at 36 ℃.
(4) Aerobic fermentation of saccharomycetes: saccharomyces cerevisiae was cultivated in advance with a wort medium (40 g/L containing malt extract), and the broth was inoculated into the fermentation broth prepared in the previous step in an inoculum size of 2% under aseptic conditions, and cultured with shaking at 28℃for 12 hours.
(5) Lysis: and (3) heating the fermentation liquor prepared in the last step to 75 ℃, preserving heat for 30min, cooling to 30 ℃, and then adding 1g/L of lysozyme for reaction for 18h.
(6) Solid-liquid separation: filtering the fermentation lysate prepared in the last step by using a 200-mesh filter, centrifuging, and collecting supernatant by using a sterile container to obtain the mung bean hull fermentation product.
The mung bean hull fermentation product prepared in this example is yellow translucent or transparent liquid in appearance.
Example 2
This example is the preparation of mung bean hull fermentation product.
(1) Domesticating lactobacillus plantarum: the same as in example 1.
(2) Preparing a fermentation medium: weighing 10g/kg of mung bean hull (obtained by dry peeling), 1g/kg of soybean and 1g/kg of glucose, adding water, mixing, heating to 105 ℃, preserving heat for 5min, and sterilizing.
(3) Anaerobic fermentation of lactic acid bacteria: the lactobacillus plantarum domesticated by high-concentration gallic acid is cultivated in an MRS liquid culture medium in advance, and bacterial liquid is inoculated into a fermentation culture medium according to the inoculation amount of 3 percent under aseptic operation, and is subjected to stationary culture for 16 hours at 36 ℃.
(4) Aerobic fermentation of saccharomycetes: saccharomyces cerevisiae was cultivated in advance with a wort medium (40 g/L containing malt extract), and the broth was inoculated into the fermentation broth prepared in the previous step in an inoculum size of 2% under aseptic conditions, and cultured with shaking at 28℃for 4 hours.
(5) Lysis: and (3) heating the fermentation liquor prepared in the last step to 75 ℃, preserving heat for 30min, cooling to 30 ℃, and then adding 1g/L of lysozyme for reaction for 18h.
(6) Solid-liquid separation: filtering the fermentation lysate prepared in the last step by using a 200-mesh filter, centrifuging, and collecting supernatant by using a sterile container to obtain the mung bean hull fermentation product.
The mung bean hull fermentation product prepared in this example is yellow translucent or transparent liquid in appearance.
Example 3
This example is the preparation of mung bean hull fermentation product.
(1) Domesticating lactobacillus plantarum: the same as in example 1.
(2) Preparing a fermentation medium: weighing 40g/kg of mung bean hull (obtained by dry peeling), 2g/kg of soybean and 3g/kg of glucose, adding water, mixing, heating to 105 ℃, preserving heat for 5min, and sterilizing.
(3) Anaerobic fermentation of lactic acid bacteria: the lactobacillus plantarum domesticated by high-concentration gallic acid is cultivated in an MRS liquid culture medium in advance, and bacterial liquid is inoculated into a fermentation culture medium according to the inoculation amount of 3 percent under aseptic operation, and is subjected to stationary culture for 18 hours at 36 ℃.
(4) Aerobic fermentation of saccharomycetes: saccharomyces cerevisiae was cultivated in advance with a wort medium (40 g/L containing malt extract), and the broth was inoculated into the fermentation broth prepared in the previous step in an inoculum size of 2% under aseptic conditions, and cultured with shaking at 28℃for 6 hours.
(5) Lysis: and (3) heating the fermentation liquor prepared in the last step to 75 ℃, preserving heat for 30min, cooling to 30 ℃, and then adding 1g/L of lysozyme for reaction for 18h.
(6) Solid-liquid separation: filtering the fermentation lysate prepared in the last step by using a 200-mesh filter, centrifuging, and collecting supernatant by using a sterile container to obtain the mung bean hull fermentation product.
The mung bean hull fermentation product prepared in this example is yellow translucent or transparent liquid in appearance.
Example 4
This example shows the preparation of a fermentation product of mung bean hull which deviates from the technical scheme of the invention.
The preparation method comprises the following steps: the procedure of example 3 was repeated except that mung bean hull (dry dehulled) in the fermentation medium was changed to mung bean hull (wet dehulled) in the same manner as in example 3.
The mung bean hull fermentation product prepared in this example is yellow translucent or transparent liquid in appearance.
Example 5
This example shows the preparation of a fermentation product of mung bean hull which deviates from the technical scheme of the invention.
The preparation method comprises the following steps: the procedure of example 3 was followed except that the acclimatized Lactobacillus plantarum of step (1) was omitted and the lactic acid bacteria fermentation was performed by using the non-acclimatized Lactobacillus plantarum of step (3).
The mung bean hull fermentation product prepared in this example is yellow translucent or transparent liquid in appearance.
Example 6
This example shows the measurement of the active ingredients of the fermented mung bean hull products prepared in examples 3, 4 and 5.
The mung bean active ingredient takes total flavone and total polyphenol as markers. The method for measuring the total flavone is according to the spectrophotometry of the method for measuring the total flavone content in propolis of national standard GB/T20574-2006, and the method for measuring the total polyphenol is according to the spectrophotometry of the method for measuring the total polyphenol content in plant extracts and products thereof of industrial standard T/AHFIA 005-2018.
The results of the active ingredient testing are shown in table 1. The results show that the mung bean hull fermentation product produced by using mung bean hull (dry peeling) contains more mung bean active ingredients.
TABLE 1 active ingredient test results
| Example 3 sample | Example 4 sample | Example 5 sample | |
| Total flavone (mg/L) | 205.09 | 9.65 | 178.66 |
| Total polyphenol (mg/L) | 660.21 | 100.50 | 406.87 |
Example 7
This example shows the antioxidant efficacy test of the mung bean hull fermentation products prepared in examples 1, 2 and 3.
Antioxidant efficacy is characterized by DPPH free radical scavenging rate, and higher DPPH free radical scavenging rate indicates better antioxidant efficacy, and the measurement method is in accordance with the industry standard "T-SHRH 006-2018 cosmetic-free radical (DPPH) scavenging test method".
Test group: mung bean hull fermentation products prepared in examples 1, 2 and 3.
Control group: a commercial cosmetic raw material mung bean fermentation broth.
The results of the antioxidant test are shown in FIG. 1. The result shows that the mung bean hull fermentation product prepared by the method has good antioxidant activity.
Example 8
This example is a test of the soothing efficacy of the mung bean hull fermentation products prepared in examples 1, 2, and 3.
The test method of the soothing efficacy is according to the industry standard TSHRH 033-2020 cosmetic soothing efficacy test-in vitro TNF-alpha inflammatory factor content determination lipopolysaccharide induced macrophage RAW264.7 test method.
Test group: mung bean hull fermentation products prepared in examples 1, 2 and 3.
Bid control group: a commercial cosmetic raw material mung bean fermentation broth.
Positive control group: dipotassium glycyrrhizinate 100ug/mL.
Negative control group: and (3) the macrophage induced by lipopolysaccharide without adding medicine.
Blank control group: macrophages not induced by lipopolysaccharide.
The results of the soothing efficacy test for the test group and the control group are shown in FIG. 2, and the lower the TNF-. Alpha.inflammatory factor content, the better the soothing efficacy. The result shows that the mung bean hull fermentation product prepared by the method has a relieving effect and is superior to the mung bean fermentation liquid and the positive control on the market.
Example 9
This example shows the irritation test of the mung bean hull fermentation products prepared in examples 1, 2 and 3.
The irritation is characterized by the rate of hemolysis, and the Test method is designed according to the erythrocyte hemolysis Test document EURL ECVAM DB-ALM Method Summary n DEG 37:Red Blood Cell (RBC) Test-Summary, EURL ECVAM DB-ALM Protocol n DEG 37:Red Blood Cell (RBC) Test System, published by European alternative validation center (EURL ECVAM). Reagents were added and the procedure was performed in the order from top to bottom according to table 2. Then the haemolysis rate was calculated:
TABLE 2 procedure for measurement of hemolysis ratio
Test group: mung bean hull fermentation products prepared in examples 1, 2 and 3.
Control group: fermentation substrates of examples 1, 2, 3.
The results of the hemolysis test are shown in Table 3, and the higher the hemolysis rate, the more irritating the sample. The results show that the hemolysis rate of the fermentation product prepared by the invention is very low, and the hemolysis rate of the fermentation substrate is very high, which indicates that the irritation of mung bean polyphenol after fermentation is reduced.
TABLE 3 hemolysis test results
Example 10
This example is a human body efficacy test of mung bean acne-removing gel prepared by using the mung bean hull fermentation product prepared in example 3.
The mung bean acne-removing gel takes mung bean hull fermentation products as unique acne-removing functional components, and comprises the following formula: 86.6% of water, 5% of mung bean hull fermentation product, 3% of glycerol, 0.6% of xanthan gum, 4% of propylene glycol, 0.4% of p-hydroxyacetophenone and 0.4% of hexanediol.
The testing method comprises the following steps:
(1) Detection instrument: canfield VISIA facial skin analyzer, delfin SkinColorCatch skin color measuring instrument, CBS-805 intelligent skin detector.
(2) Test conditions: test period: day 0, day 7, day 21; number of test persons: 7 people; test part: facial acne parts; the using method comprises the following steps: the mung bean acne-removing gel is applied to the acne part every day.
Test results:
(1) After the mung bean acne-removing gel was used, the MI value of melanin at the acne part was increased by 1.97% on day 7, but decreased by-0.69% from the initial value on day 21 (Table 4). The mung bean acne-removing gel has the effect of reducing the content of melanin in acne marks after long-term use.
(2) After the mung bean acne-removing gel was used, the color a value of the acne part was reduced by 5.37%, the heme EI value was reduced by 0.80% on day 7, the a value was reduced by 7.94% and the EI value was reduced by 1.50% on day 21 (tables 5 and 6). The mung bean acne-removing gel has the effect of reducing acne inflammation.
(3) After the mung bean acne-removing gel was used, the color L of the acne part was reduced by 1.53% and the ITA ° value was reduced by-17.46% on day 7, but the value of ITA ° was increased by 2.32% and 34.31% on day 21 as compared with the initial value L (tables 7 and 8). The mung bean acne-removing gel has the effect of improving the skin color of acne parts after long-term use.
(4) Comparing the facial acne photos of day 0, day 7 and day 21, the volunteers can intuitively see that the acne symptoms of the two subjects (with codes of LDM and YZQ respectively) are improved after the mung bean acne-removing gel is used.
(FIG. 3)
(5) In conclusion, the mung bean acne-removing gel has the effect of improving acne symptoms.
TABLE 4 melanin MI values for acne sites
Table 5 color a values for acne sites
| Color a value of acne part | Rate of change compared to day 0 | |
| Day 0 | 16.76±2.35 | - |
| For 7 days | 15.86±2.44 | -5.37％ |
| 21 days | 15.43±3.11 | -7.94％ |
TABLE 6 heme EI value for acne sites
| Heme EI value at acne site | Rate of change compared to day 0 | |
| Day 0 | 482.76±8.28 | - |
| For 7 days | 478.90±9.74 | -0.80％ |
| 21 days | 475.52±13.26 | -1.50％ |
Table 7 color L values of acne sites
| Color L value of acne part | Rate of change compared to day 0 | |
| Day 0 | 53.10±2.34 | - |
| For 7 days | 52.29±2.59 | -1.53％ |
| 21 days | 54.33±3.51 | 2.32％ |
Table 8 acne sites ITA values
| ITA degree value of acne part | Rate of change compared to day 0 | |
| Day 0 | 8.19±6.40 | - |
| For 7 days | 6.76±6.62 | -17.46％ |
| 21 days | 11.00±8.34 | 34.31％ |
The above examples are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above examples, and any other changes, modifications, substitutions, combinations, and simplifications that do not depart from the spirit and principle of the present invention should be made in the equivalent manner, and the embodiments are included in the protection scope of the present invention.
Claims (6)
1. The preparation method of the mung bean hull fermentation product with the acne-removing effect comprises four process steps of preparing a fermentation medium, fermenting, lysing and separating solid from liquid, and is characterized in that: the fermentation medium comprises 10-100 g/kg of mung bean hull, 1-5 g/kg of soybean or soybean protein and 1-5 g/kg of glucose; the fermentation process steps are divided into lactic acid bacteria fermentation and saccharomycetes fermentation which are sequentially and stepwise completed, wherein the strain adopted by the lactic acid bacteria fermentation is lactobacillus plantarum domesticated by high-concentration gallic acid, and the strain adopted by the saccharomycetes fermentation is saccharomyces cerevisiae;
the mung bean hull is obtained by separating dried mung beans after roller grinding;
the lysis is to heat the fermentation liquor after the fermentation process is completed to 75 ℃ and keep the temperature for 30min, cool to 30 ℃, and then add 1g/L lysozyme for reaction for 18h;
the lactobacillus plantarum domesticated by the high-concentration gallic acid is specifically obtained by the following method: inoculating lactobacillus plantarum into MRS liquid culture medium, and culturing at 37 ℃ for 24 hours; inoculating the lactobacillus plantarum culture solution obtained in the last step into MRS liquid culture medium containing 2g/kg gallic acid, and culturing at 37 ℃ for 24 hours; inoculating the lactobacillus plantarum culture solution obtained in the last step into an MRS liquid culture medium containing 4g/kg gallic acid, and culturing for 24 hours at 37 ℃; inoculating the lactobacillus plantarum culture solution obtained in the last step into MRS liquid culture medium containing 8g/kg gallic acid, and culturing at 37 ℃ for 24 hours; culturing lactobacillus plantarum culture solution obtained in the last step on MRS solid culture medium containing 10g/kg gallic acid by streaking, and culturing at 37 ℃ for 24 hours; and (3) picking single colony on the lactobacillus plantarum solid culture obtained in the last step, inoculating the single colony into an MRS liquid culture medium containing 10g/kg gallic acid for expansion culture, and culturing at 37 ℃ for 24 hours to obtain the lactobacillus plantarum domesticated by the high-concentration gallic acid.
2. The method of manufacturing according to claim 1, characterized in that: the lactobacillus fermentation is carried out for 16-24 hours at 37 ℃ after lactobacillus plantarum inoculation; the saccharomycete fermentation is carried out for 4-12 hours at 28 ℃ after the inoculation of the saccharomyces cerevisiae.
3. The method of manufacturing according to claim 1, characterized in that: the MRS liquid culture medium comprises the following components: 10g/L of peptone, 5g/L of beef extract powder, 4g/L of yeast extract powder, 20g/L of glucose, 2g/L of monopotassium phosphate, 2g/L of triammonium citrate, 5g/L of sodium acetate, 0.2g/L of magnesium sulfate, 0.05g/L of manganese sulfate, 1g/L of tween and the balance of water.
4. The method of manufacturing according to claim 1, characterized in that: the MRS solid culture medium comprises the following components: 10g/L of peptone, 5g/L of beef extract powder, 4g/L of yeast extract powder, 20g/L of glucose, 2g/L of monopotassium phosphate, 2g/L of triammonium citrate, 5g/L of sodium acetate, 0.2g/L of magnesium sulfate, 0.05g/L of manganese sulfate, 1g/L of tween, 15g/L of agar and the balance of water.
5. A mung bean hull fermentation product with acne-removing effect prepared by the preparation method of any one of claims 1-4.
6. The use of the mung bean hull fermentation product with acne-removing effect according to claim 5 in the preparation of cosmetics.
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