CN111588677B - Barley tremella fermentation raw stock and preparation method and application thereof - Google Patents
Barley tremella fermentation raw stock and preparation method and application thereof Download PDFInfo
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- A61K8/00—Cosmetics or similar toiletry preparations
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- 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
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Abstract
The invention provides barley tremella fermentation protoplasm and a preparation method and application thereof, wherein the preparation method comprises the following steps: mixing the mixed powder of barley malt and tremella, water and zymocyte liquid to obtain an initial system, fermenting to obtain fermentation liquor, sterilizing and centrifuging the fermentation liquor to obtain supernatant, namely the barley tremella fermentation raw stock. The invention adopts the saccharomycetes to ferment the barley malt and the tremella, retains all functional components and activity of the barley malt and the tremella, and avoids loss of active components caused by an extraction method.
Description
Technical Field
The invention relates to a fermentation product, in particular to a fermentation product with a skin care function.
Background
The barley malt is spindle-shaped, 8-12 mm long and 3-4 mm in diameter. The surface is light yellow, and the back surface is surrounded by lemma and has 5 veins; the ventral surface surrounds the palea. After removing the inner and outer seeds, the ventral surface has 1 longitudinal furrow; the radicle of the base part grows a bud and a fibrous root, and the bud is in a needle-shaped strip shape and is about 5mm long. The fiber has a plurality of strands, and is slender and bent. Hard, white cross section and powdery. Light smell, slightly sweet taste. Tremella is fruiting body of Basidiomycota fungus Tremella, and comprises more than 10 thin and multi-fold flat leaves. The white fungus fruiting body is pure white to milk white, generally in chrysanthemum or cockscomb shape, soft, white, semitransparent and elastic.
So far, in the skin care product manufacturing industry, most skin care products are not fermented skin care products, are mainly prepared by the traditional process and are most common by cooking. For example, after the medicinal materials are cooked, the medicinal materials are fished out and discarded, and ancient people drink soup, and when people use the soup on cosmetics, the soup can be filtered and added with preservative for direct use. However, the traditional process is limited by ancient thinking and is difficult to break through, and even the traditional process is used for extracting plant extracts in a plurality of skin care products, so that not only the medicinal materials are seriously consumed and the sustainable development is not realized, but also the obtained substances have only limited types and quantity and often have unknown irritability, and therefore, new technology should be considered. At present, many fermentation allegedly skin care products are actually prepared by adding only a little fermentation source components into the formula of the skin care products, so that the content of the fermentation products in the formula is only 1-2%. Many products are mainly prepared from traditional chemical substances, and various types of added plant extracts are very small in content, so that poor effects are obtained, and the appearance state is transparent and clear, but has no significance to consumers. Chinese patent (CN 109330929A) discloses a preparation method of a tremella fermentation extract and application thereof in cosmetics, and particularly discloses a tremella extract refined liquid prepared by adopting a mixture of tremella and water through the steps of high-temperature fermentation, ultrahigh-speed shearing, low-temperature fermentation and the like, and a tremella extract mask prepared further. The tremella polysaccharide molecular weight is degraded through the cooperation of two-step fermentation and high-speed shearing process, and absorption, water replenishing and moisture keeping on the surface of human skin are facilitated. The process is complex, the most important of fermentation products is the unified and pure environment, new bacteria are inevitably introduced by frequently replacing equipment after fermentation in the patent, the whole process is difficult to control, the process can only stay in a laboratory stage, and the real industrial production is difficult.
The statements in the background section are merely prior art as they are known to the inventors and do not, of course, represent prior art in this field.
Disclosure of Invention
The invention aims at one or more problems in the prior art and provides a fermentation raw stock prepared from barley malt and tremella;
the invention also aims to provide a preparation method of the fermentation raw stock prepared by the method;
the invention also aims to provide application of the fermentation protoplasm.
The purpose of the invention is realized by the following technical scheme:
a barley tremella fermented raw stock is prepared by fermenting substances including barley malt and tremella.
According to one aspect of the invention, the barley white fungus fermentation raw pulp contains 1-10mg/ml of protein, 10-40mg/ml of crude polysaccharide, 0.2-1.0mg/ml of total flavone and 0.1-0.8mg/ml of total phenol.
According to one aspect of the invention, the mass ratio of the barley malt to the tremella is (2-4): (2-4); preferably, the mass ratio of the barley malt to the tremella is 1:1.
according to one aspect of the invention, the fermentation is carried out using yellow wine yeast.
A skin care product comprises the barley tremella fermentation raw stock.
The barley white fungus fermentation protoplasm is applied as a skin care product.
A preparation method of barley white fungus fermentation raw stock comprises the steps of mixing mixed powder of barley malt and white fungus, water and fermentation bacteria liquid to obtain an initial system, fermenting to obtain fermentation liquor, sterilizing and centrifuging the obtained fermentation liquor to obtain supernatant, namely the barley white fungus fermentation raw stock.
According to one aspect of the invention, the mass ratio of the barley malt to the tremella is (2-4): (2-4); preferably, the mass ratio of the barley malt to the tremella is 1:1.
according to one aspect of the present invention, the mesh number of the barley malt and tremella mixed powder is 20-50 mesh, preferably 50 mesh.
According to one aspect of the invention, the mixed powder of barley malt and tremella: the proportion of water is (5-20) g (100-300) mL.
According to one aspect of the invention, the mixed powder of barley malt and tremella: the mass ratio of water was 10g.
According to one aspect of the invention, the concentration of the zymogen liquid is 10 5 -10 8 CFU/ml。
According to one aspect of the invention, the ratio of the zymophyte liquid to the mixed powder of the barley malt and the tremella is (10-40) ml: (5-20) g.
According to one aspect of the invention, the ratio of the zymophyte liquid to the mixed powder of the barley malt and the tremella is 20ml:10g.
The invention also provides a preparation method of the barley tremella fermentation protoplasm, wherein the fermentation temperature of the fermentation is 30-45 ℃, and preferably 37 ℃.
According to one aspect of the invention, the fermentation time of the fermentation is 10-50h, preferably 36h.
According to one aspect of the invention, the centrifugation is carried out at 4000-12000r/min for 15-30min;
according to one aspect of the invention, the centrifugation is at 4500r/min for 20min.
According to one aspect of the invention, the centrifugation radius of the centrifugation is 9-12cm.
According to one aspect of the invention, the fermentation bacteria are yellow wine yeast.
According to one aspect of the invention, the zymophyte is pretreated in advance, and the pretreatment process comprises the following steps:
activating strains: putting the bacterial colony of the zymocyte into a liquid culture medium, and then putting the culture medium into an incubator to activate the strain;
and (3) strain purification: the activated strains are subjected to gradient dilution and plating so as to obtain single colonies; and
expanding and culturing strains: inoculating the strain to be used to corresponding liquid culture medium, culturing in 30-45 deg.C incubator until OD =0.5-1.0, and the strain is in logarithmic phase, i.e. suitable inoculation concentration of 10 5 -10 8 CFU/ml; preferably, the medium is a yeast medium.
The barley tremella fermentation raw pulp provided by the invention can be used as an effective component in a skin care product, can also be independently used as a skin care product, and has an antioxidant function and a whitening function.
The preparation method of the primary fermented pulp by using barley malt and tremella provided by the invention has the following advantages:
(1) The invention adopts the saccharomycetes to ferment the barley malt and the tremella, retains all functional components and activity of the barley malt and the tremella, and avoids loss of active components caused by an extraction method.
(2) The fermentation method adopted by the invention does not add any organic reagent in the process of extracting the active ingredients of barley malt and tremella, the fermentation temperature and the fermentation pH value are mild, the structure of the active ingredients of the plants is not damaged, and the natural activity of the plants is maintained.
(3) After the fermentation is finished in the method, chemical components such as essence and the like are not added into the fermentation raw stock, so that the safety of the product to human bodies is ensured.
(4) The barley tremella fermented raw pulp provided by the invention is rich in active substances which have tyrosinase activity inhibition and melanin synthesis inhibition, so that the barley tremella fermented raw pulp has a strong skin whitening function and has a synergistic effect with components in a fermentation filtrate.
Drawings
FIG. 1 is a graph showing the scavenging effect of barley white fungus fermentation raw juice on hydroxyl radicals in an embodiment of the present invention;
FIG. 2 is a bar graph of the inhibition rate of barley white fungus fermentation raw pulp on tyrosinase activity and arbutin on tyrosinase activity in the embodiment of the invention;
FIG. 3 is a graph of long-term skin moisture data of barley white fungus fermentation broth in an embodiment of the present invention;
FIG. 4 is data of long-term transdermal water evaporation of primary fermented barley tremella pulp in an embodiment of the present invention.
Detailed Description
In the following, only certain exemplary embodiments are briefly described. As those skilled in the art will recognize, the described embodiments may be modified in various different ways, all without departing from the spirit or scope of the present invention. Accordingly, the drawings and description are to be regarded as illustrative in nature, and not as restrictive.
The following disclosure provides many different embodiments or examples for implementing different features of the invention. To simplify the disclosure of the present invention, the components and arrangements of specific examples are described below. Of course, they are merely examples and are not intended to limit the present invention. Moreover, the present invention may repeat reference numerals and/or reference letters in the various examples, which have been repeated for purposes of simplicity and clarity and do not in themselves dictate a relationship between the various embodiments and/or configurations discussed. In addition, the present invention provides examples of various specific processes and materials, but one of ordinary skill in the art will recognize the application of other processes and/or the use of other materials.
The following description of the preferred embodiments of the present invention is provided for the purpose of illustration and description, and is in no way intended to limit the invention.
In the following examples, the fermentation tubes used were pretreated in advance, and the pretreatment process included:
activating strains: putting the bacterial colony of the zymophyte into a liquid culture medium, and then putting the culture medium into an incubator to activate the strain;
and (3) strain purification: the activated strains are subjected to gradient dilution and plating so as to obtain single colonies; and
expanding culture of strains: inoculating the strain to be used into corresponding liquid culture medium, culturing in 30-45 deg.C incubator until OD value =0.5-1.0, and the strain is in logarithmic phase, i.e. suitable inoculation concentration of 10 5 -10 8 CFU/ml; the culture medium is a yeast culture medium.
The specific process of strain activation and purification is the conventional technology.
Example 1:
the concentration of zymophyte in the culture medium is 10 5 -10 8 20mL of CFU/mL zymophyte liquid is inoculated into 50 meshes of 10g of barley malt and tremella mixed powder (the mass ratio of barley malt to tremella is 1).
The barley and tremella fermented puree cosmetic prepared in the embodiment is viscous liquid in appearance and is colorless, transparent and brown in color. pH value 4.5-6.8, viscosity 100-800cP, soluble solid content 1.0-5.0%, colony count less than 50CFU/ml, no pathogenic bacteria detection. Meets the quality requirement of cosmetics.
Performing component analysis on the barley and tremella fermented raw pulp cosmetic: the barley tremella fermentation protoplasm cosmetic prepared by the invention contains 1-10mg/ml of protein, 10-40mg/ml of crude polysaccharide, 0.2-1.0mg/ml of total flavone and 0.1-0.8mg/ml of total phenol.
Analyzing the antioxidant effect of the barley and tremella fermentation raw pulp:
referring to table 1, using 10mL, the test tube was set up with 3 parallel tubes for each concentration of sample tube (T) for each sample of sample tube (T), sample background (T0), blank tube (C), control tube (C0).
Precisely transferring 3mL of ferrous sulfate solution into a test tube, sequentially adding 3mL of hydrogen peroxide solution and 3mL of salicylic acid solution, uniformly mixing, and heating in a water bath at 37 ℃ for 15min; respectively adding 1mL of sample solutions with different concentrations as T, and continuously heating for 15min; in the same way, the blank tube (C) and the comparison tube (C0) replace the sample with distilled water with the same volume, and the sample background (T0) and the comparison tube (C0) replace the hydrogen peroxide reaction system with distilled water with the same volume. The absorbance was measured at 510 nm.
TABLE 1 sample addition requirement
A curve of action of barley tremella fermentation raw stock on scavenging hydroxyl free radical is shown in figure 1.
As can be seen from figure 1, the barley tremella protoplasm has obvious effect of removing hydroxyl radicals, and the capacity of removing the hydroxyl radicals is gradually enhanced along with the increase of the concentration of the barley tremella protoplasm. The 30% barley tremella protoplasm can eliminate about 38.67% hydroxyl free radical, and the IC50 is 38.074%.
And (3) analyzing the whitening effect of the barley and tremella fermentation primary pulp:
tyrosinase is a key enzyme in melanogenesis, which controls the process of melanogenesis, and its degree of activity plays a major role in pigment deposition. Many whitening and freckle-removing products sold in the market at present achieve the whitening effect by inhibiting tyrosinase, so the strength of the tyrosinase inhibition effect is a main index for evaluating whitening cosmetics.
The whitening function of the sample is evaluated by measuring the influence of the sample on tyrosinase, and the specific method comprises the following steps:
prepare the solution as in table 2:
TABLE 2 solution preparation List
| Unit (mL) | C 1 | C 2 | T 1 | T 2 |
| L-tyrosine | 2 | 2 | 2 | 2 |
| Sample(s) | 0 | 0 | 2 | 2 |
| PBS | 4 | 5 | 2 | 3 |
| |
1 | 0 | 1 | 0 |
| |
7 | 7 | 7 | 7 |
Note: c 1 And T 1 Adding 1mL of tyrosinase, wherein the enzyme activity is 100U/mL.
(1) After the C2 tube is prepared and shaken up, the mixture is heated in a water bath kettle at 37 ℃ for 10min, and the zero setting is carried out under the wavelength of 475 nm.
(2) Mixing the solution in the C1 tube, shaking, performing water bath at 37 deg.C for 10min, adding 1ml tyrosinase, continuing water bath for 10min, and measuring the absorbance value of C1.
(3) The absorbance value of T1 was measured by zeroing T2 in the same manner as in (1) and (2).
(4) The inhibition rate T (%) of the sample on tyrosinase activity was calculated. T (%) = (C1-T1)/C1X 100%
The 1% arbutin is a positive control of a tyrosinase activity inhibition experiment, and the inhibition rate of the arbutin on the tyrosinase activity is 75%.30% of barley tremella primary pulp can inhibit 25.56% of tyrosinase activity, so the barley tremella primary pulp has the effect of inhibiting the tyrosinase activity and has a certain whitening effect.
Moisture retention efficacy analysis of barley white fungus fermentation raw pulp:
the moisture retention capacity is an important index for judging the performance of the product. Skin moisture content can be increased after use of the product and can remain higher than unapplied for extended periods of time, which is recognized as a good moisturizing capability. Can moisten and soften skin.
The experimental method comprises the following steps: firstly, measuring areas are made on the left arm and the right arm of a subject (3 people), the area of each area is 3cmX3cm, and a measuring sample is coated for a single time according to the dosage of (2.0 +/-1.0) mg/cm < 2 >. The initial values for each test area were determined (before the sample was used, then after a set time the moisture content and water loss of the test and control areas were determined. Volunteers chose and tested according to methods QL-SOP-YF-3-01 and QL-SOP-YF-3-02.
As shown in fig. 3: compared with the uncoated blank control area, the barley white fungus fermentation raw juice has obvious advantages of long moisture retention; the moisture content of skin of the subject is always higher than that of the uncoated blank control area after the barley tremella is used for fermenting the primary pulp for 1 day, 7 days and 14 days.
As shown in fig. 4: compared with the uncoated blank control area, the barley tremella protoplasm can reduce the water loss of the skin through the epidermis and keep moisture for a long time. After the test subject uses the barley tremella primary pulp for 14 days, the water locking capacity is increased by 12.5%, the water evaporation prevention capacity of the skin is always stronger than that of an uncoated blank control area, namely the water evaporation amount is remarkably lower, and the barrier capacity is continuously enhanced.
And (3) evaluating the safety of the barley tremella fermentation primary pulp:
the patch test is mainly used for detecting the irritation of the final product or raw material of the cosmetic. The invention carries out a human body closed patch test on the barley and tremella fermentation protoplasm cosmetic and evaluates the potential skin irritation of the barley and tremella fermentation protoplasm cosmetic.
Suitable volunteers were selected for 30 persons, and were randomly selected in the age range of 18-60 years. 0.020g to 0.025g of solid or semi-solid sample is weighed into a plaque test device for use. The liquid sample, 0.2mL to 0.025mL, was dropped onto the filter paper sheet, which was then placed in the plaque tester. A blank control is set for each sample and an equal amount of sample solvent, such as distilled water or olive oil, is added to the control chamber.
The test part is selected as the back of a human body, and the spot tester is fixedly attached to the back of the testee by using a non-irritating adhesive tape. The test period lasted 36h. In order to ensure the accuracy, credibility and science of test results, the volunteer cannot remove the spot tester or make the tested part contact water according to the requirements during the test.
And after 36h, removing the plaque tester, standing for 30min, waiting for the indentation to disappear, and observing the reaction of the skin. If the test result is negative, the test needs to be observed once more at 36h and 48h after the patch test.
The patch test results are shown in table 3:
TABLE 3 Patch test results
* "-" = negative reaction;
"±" = suspicious reaction: only faint erythema;
"+" = weak positive reaction (erythema reaction): erythema, infiltration, edema, and possibly pimples;
"++" = strong positive reaction (herpetic response); erythema, infiltration, edema, pimples, herpes; the reaction may be beyond the test area;
"+ + + +" = very strong positive reaction (fusogenic herpes response); obvious erythema, severe infiltration, edema, and fusional herpes; the reaction goes beyond the test area.
As can be seen from Table 3, the barley white fungus fermentation broth provided in example 1 produced no more than 1 of grade 2 reactions in the experiment. The barley tremella fermentation raw stock is judged to be safe and does not bring adverse reaction to human bodies.
Example 2:
the concentration of zymophyte in the culture medium is 10 5 -10 8 40mL of CFU/mL zymocyte liquid is inoculated into 50 meshes of 5g of barley malt and tremella mixed powder (the mass ratio of barley malt to tremella is 1Centrifuging for 15min, and collecting supernatant to obtain the barley tremella fermentation raw pulp cosmetic provided by the invention.
Example 3:
the concentration of zymophyte in the culture medium is 10 5 -10 8 Inoculating 10mL of CFU/mL zymocyte liquid into 50 meshes of 20g of barley malt and tremella mixed powder (the mass ratio of barley malt to tremella is 2.
The methods and results of analyzing the components of the products of examples 2 and 3 are the same as those of example 1, and therefore will not be described again.
The products of examples 2 and 3 were analyzed for antioxidant effect, whitening effect and moisturizing effect in the same manner as in example 1, and the results were substantially the same as in example 1. And therefore will not be reiterated.
The safety evaluation methods and results of the products of examples 2 and 3 are the same as those of example 1, and therefore will not be described.
Finally, it should be noted that: although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that changes may be made in the embodiments and/or equivalents thereof without departing from the spirit and scope of the invention. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (20)
1. The application of the barley tremella fermentation raw pulp in preparing whitening and moisturizing skin care products; the preparation method of the barley tremella fermentation protoplasm comprises the following steps: mixing the mixed powder of barley malt and tremella, water and a zymophyte liquid to obtain an initial system, fermenting to obtain a fermentation liquid, sterilizing and centrifuging the obtained fermentation liquid to obtain a supernatant, namely the barley tremella fermentation raw stock; the mass ratio of the barley malt to the tremella in the mixed powder is (2-4): (2-4); the zymocyte is yellow wine yeast.
2. Use according to claim 1, characterized in that: the mass ratio of the barley malt to the tremella is 1:1.
3. use according to claim 1, characterized in that: the mesh number of the mixed powder of the barley malt and the tremella is 20-50 meshes.
4. Use according to claim 3, characterized in that: the mesh number of the mixed powder of the barley malt and the tremella is 50 meshes.
5. Use according to claim 1, characterized in that: the mixed powder of barley malt and tremella: the proportion of water is (5-20) g (100-300) mL.
6. Use according to claim 5, characterized in that: the mixed powder of barley malt and tremella: the proportion of water was 10g.
7. Use according to claim 1, characterized in that: the concentration of the zymophyte liquid is 10 5 -10 8 CFU/ml。
8. Use according to claim 1, characterized in that: the proportion relation of the zymophyte liquid to the mixed powder of the barley malt and the tremella is (10-40) ml: (5-20) g.
9. Use according to claim 8, characterized in that: the proportion relation between the zymophyte liquid and the mixed powder of the barley malt and the tremella is 20ml:10g.
10. Use according to claim 1, characterized in that: the fermentation temperature of the fermentation is 30-45 ℃.
11. Use according to claim 10, characterized in that: the fermentation temperature of the fermentation was 37 ℃.
12. Use according to claim 1, characterized in that: the fermentation time of the fermentation is 10-50h.
13. Use according to claim 12, characterized in that: the fermentation time of the fermentation is 36h.
14. Use according to claim 1, characterized in that: the centrifugation is carried out for 15-30min at 4000-12000 r/min.
15. Use according to claim 14, characterized in that: the centrifugation was carried out at 4500r/min for 20min.
16. Use according to claim 14 or 15, characterized in that: the centrifugation radius of the centrifugation is 9-12cm.
17. Use according to claim 1, characterized in that: the zymophyte is pretreated in advance, and the pretreatment process comprises the following steps:
activating strains: putting the bacterial colony of the zymocyte into a liquid culture medium, and then putting the culture medium into an incubator to activate the strain;
and (3) strain purification: the activated strains are subjected to gradient dilution and plating so as to obtain single colonies; and
expanding culture of strains: inoculating the strain to be used to corresponding liquid culture medium, culturing in 30-45 deg.C incubator until OD =0.5-1.0, and the strain is in logarithmic phase, i.e. suitable inoculation concentration of 10 5 -10 8 CFU/ml。
18. Use according to claim 17, characterized in that: the culture medium is a yeast culture medium.
19. Use according to claim 1, characterized in that: the barley white fungus fermentation raw pulp contains 1-10mg/ml of protein, 10-40mg/ml of crude polysaccharide, 0.2-1.0mg/ml of total flavone and 0.1-0.8mg/ml of total phenol.
20. Use according to claim 1, characterized in that: the pH value of the barley tremella fermentation protoplasm is 4.5-6.8.
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| CN107137318A (en) * | 2017-05-31 | 2017-09-08 | 华子昂 | A kind of ferment facial mask for improving skin moisture function and preparation method thereof |
| CN110772468A (en) * | 2019-12-12 | 2020-02-11 | 山东福瑞达生物科技有限公司 | Oat and tremella fermentation product filtrate and preparation method thereof |
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