CN111437235A

CN111437235A - Rice composite fermentation product with anti-aging effect and preparation method and application thereof

Info

Publication number: CN111437235A
Application number: CN202010413733.3A
Authority: CN
Inventors: 马道品; 李华发; 周广华; 陈文瀚; 陈文刚
Original assignee: Wanjing Chuangke Shandong Biotechnology Co ltd
Current assignee: Wanjing Chuangke Guangzhou Biotechnology Co ltd
Priority date: 2020-05-15
Filing date: 2020-05-15
Publication date: 2020-07-24
Anticipated expiration: 2040-05-15
Also published as: CN111437235B

Abstract

The invention provides a preparation method of a rice composite fermentation product with an anti-aging effect, which comprises the following steps: inoculating the zymophyte into a fermentation substrate consisting of rice, burdock root, silybum marianum and water for fermentation culture, and then carrying out sterilization treatment to obtain the rice composite fermentation product with the anti-aging effect. The rice composite fermentation product has good anti-aging effect and has better application prospect in the cosmetic industry.

Description

Rice composite fermentation product with anti-aging effect and preparation method and application thereof

Technical Field

The disclosure relates to the technical field of fermentation, in particular to a rice composite fermentation product with an anti-aging effect and a preparation method and application thereof.

Background

The rice is a finished product prepared by the working procedures of cleaning, hulling, milling, finishing and the like of the rice, contains nearly 64 percent of nutrient substances in the rice and more than 90 percent of nutrient elements required by a human body, and is a main food for people in most regions of China. The rice can provide abundant nutrients such as vitamins, oryzanol, protein, anthocyanin, etc. The rice has the skin care function, and the rice extract is used as a main component (which is rich in components such as y-oryzanol, rice chaff sterol, procyanidine and the like), has mild and safe properties and strong whitening effect, can supplement water missing from the skin, and has the effects of smoothing and smoothing the skin and filling elasticity. China has rich rice resources and provides favorable conditions for developing and applying rice. With the rapid development and the widening of the application range of the rice cultivation industry, the research on the nutrient components of rice and the medical care function is also gradually and deeply carried out.

Arctium lappa L, belonging to the order Campanulales, is a biennial herb of Compositae, and the root of Arctium lappa contains inulin, volatile oil, arctic acid, various polyphenol substances and aldehydes, and is rich in cellulose and amino acids, the root of Arctium lappa contains various amino acids essential to human body, and has high content, especially has high content of amino acids with special pharmacological action, such as aspartic acid with brain strengthening effect accounting for 25% -28% of total amino acids, arginine accounting for 18% -20%, and contains macroelements and microelements essential to human body such as Ca, Mg, Fe, Mn, Zn, etc., and has the effects of resisting aging, resisting inflammation, resisting virus, resisting cancer, dilating blood vessels, promoting blood circulation, etc.

Silybum marianum (L.) Gaertn is an annual or biennial herb, and has the effects of oxidation resistance, inflammation resistance, lactation promotion, liver protection and the like.

Fermentation refers to a process in which people prepare microbial cells themselves, or direct metabolites or secondary metabolites, by virtue of the life activities of microorganisms under aerobic or anaerobic conditions, and is widely used in the food industry, the biological and chemical industries. At present, most reports are made on the extraction of nutrient substances by adopting a fermentation technology, but the extraction effect is greatly different according to different fermentation substrates and fermentation process conditions. The method depends on researchers to continuously explore more fermentation substrate combinations and fermentation methods to prepare more products with better effects so as to meet the requirements of consumers.

Disclosure of Invention

The following presents a simplified summary of the disclosure in order to provide a basic understanding of some aspects of the disclosure. It should be understood that this summary is not an exhaustive overview of the disclosure. It is not intended to identify key or critical elements of the disclosure or to delineate the scope of the disclosure. Its sole purpose is to present some concepts in a simplified form as a prelude to the more detailed description that is discussed later.

In view of the above-mentioned drawbacks of the prior art, the present disclosure is directed to provide a rice complex fermentation product with anti-aging effect, and a preparation method and application thereof.

According to one aspect of the present disclosure, there is provided a method for preparing a rice composite fermentation product having anti-aging effect, comprising: inoculating the zymophyte into a fermentation substrate consisting of rice, burdock root, silybum marianum and water for fermentation culture, and then carrying out sterilization treatment to obtain the rice composite fermentation product with the anti-aging effect.

The zymophyte adopted in the method is probiotics, so that a new skin environment can be maintained, the ecological balance and comfort of the skin can be improved, a microbial protective film is strengthened, and the skin microcirculation is promoted. Preferably, the probiotic is a lactic acid bacterium; specifically, the compound is selected from any one or more than two of the following compounds:

lactobacillus delbrueckii subsp. Bulgaricus (L actinobacillus delbruuchii) with a collection number of CGMCC1.16075, available from CGMCC;

lactobacillus buchneri (L actinobacillus buchneri) with the preservation number of CGMCC1.15607, which can be purchased from CGMCC;

bifidobacterium bifidum (CGMCC 1.5029) with preservation number can be purchased from CGMCC.

The fermentation substrate is prepared by adding a proper amount of rice, burdock root and silybum marianum into a proper amount of water, mixing and sterilizing.

The rice used in the present disclosure is finished rice grains prepared by rice cleaning, rice hulling, rice milling, finished product finishing and other processes, and may be rice flour, preferably 0.5-3%, that is, in the fermentation substrate, the rice accounts for 0.5-3% of the weight of water (e.g., 0.8%, 1%, 1.5%, 1.8%, 2.1%, 2.5%, 2.8%).

The burdock root adopted in the disclosure is the root of burdock (Arctium lappa L) of the family Compositae, the burdock root is oriented to be a vegetable with edibility and medicinal property since ancient times, the famous medical records of south-north dynasty have records about burdock medicine taking, the main medicinal part of the burdock root is burdock (burdock achene), the medicinal value of the burdock root is recorded in a plurality of medical books, such as the famous medical records, the herbal records and the herbal record, the medicinal value of the burdock root is recorded in the books, the nutritional health care and pharmacological action of the burdock root are attributed to a plurality of functional substances contained in the burdock root, and the burdock root is rich in nutrient components such as amino acid and oligosaccharide and a plurality of small molecular active components such as burdock acid, aldehydes, sulfur-containing alkynes, polyphenols and volatile oil.

The burdock extract has an inhibition effect on the level of male hormone, has a good prevention effect on diseases caused by high male hormone, and can be used for inhibiting acne and alopecia; the burdock extract has good anti-oxygen property, and the inhibition of the burdock extract on metalloprotease and the activation of cathepsin D, which shows that the burdock extract has anti-aging activity; the activation of nuclear factor kappa B receptor is inhibited, which shows that the burdock extract has anti-inflammation and can be used for preventing skin inflammation by combining the antibacterial action of the burdock extract. The contraction effect on collagen fibers shows that the collagen fibers can astringe skin and shrink pores.

The burdock root used in the present disclosure is a dry product in a sheet or ground form, preferably in an amount of 0.05-0.2%, i.e., the burdock root accounts for 0.05-0.2% (e.g., 0.08%, 0.1%, 0.12%, 0.15%, 0.18%) of the weight of water in the fermentation substrate. Too high a dosage ratio may result in a dark fermentation product, which is not suitable for cosmetic use. The burdock root is preferably crushed to pass through a 20-mesh screen.

The Silybum marianum adopted in the invention is an ecliptic fruit of Silybum marianum (L.) Gaerth of Compositae, the whole grass contains flavonoids and fumaric acid, the seeds mainly contain flavonols, silybin, isosilybin, dehydrosilybin, silydianin, silychristin, silybin polymers, cinnamic acid, myristic acid, palmitic acid, arachidic acid and the like, the Silybum marianum has strong antioxidation, can effectively remove excessive free radicals in vivo and avoid the damage of cells in vivo, on the other hand, the Silybum marianum can also neutralize the free radicals and prevent the damage of liver free radicals, the Silybum marianum can prevent skin from saccharification, keep the collagen structure in the skin unchanged, improve the smooth feeling and the compactness of the skin and reduce the wrinkle depth of the Silybum marianum, in the present disclosure, the Silybum marianum is a dry product, the preferable dosage is 0.05-0.15% of the water weight (such as 0.08%, 0.10%, 0.12%, 0.14. the dosage of the Silybum marianum is too much higher than the dosage, the product is not suitable for the synergistic effect, and the product.

In the above method for preparing a rice composite fermentation product having anti-aging effect, as a preferred embodiment, the fermentation bacteria is lactobacillus delbrueckii subsp bulgaricus (L actinobacillus delbruuchisubsp. bulgaricus).

Further, the procedure for the preparation of the seed liquid of lactobacillus delbrueckii subspecies bulgaricus for inoculation into the fermentation substrate is as follows:

and (3) activation: inoculating the strain into a test tube of an MRS liquid culture medium, and activating for 20-48 h at 35-37 ℃;

and (3) purification: taking the activated liquid strain, performing gradient dilution, inoculating the liquid strain into an MRS solid culture medium plate, and performing static culture at 35-37 ℃ for 40-48 h;

liquid culture, namely inoculating 2-3 rings of single bacterial colonies in the plate into a 100m L MRS liquid culture medium, and activating for 20-48 h at 35-37 ℃;

performing amplification culture, namely measuring the bacterial liquid obtained by liquid culture by using 10% of inoculation amount, inoculating the bacterial liquid into a 100m L MRS liquid culture medium (the volume ratio of the bacterial liquid to the culture medium is 10:100), and culturing for 10-14h at the temperature of 35-37 ℃ until the bacterial amount reaches 10⁷-10¹⁰CFU/m L, obtaining seed liquid.

Wherein, the formula of the MRS liquid culture medium is as follows: 10.0g of peptone, 10.0g of beef extract, 5.0g of yeast powder, 20.0g of glucose and K₂HPO₄·7H₂O2.0 g, NaO 3H₂O5.0g, triammonium citrate 2.0g, MgSO₄·7H₂0.05g of O, 801.0 m of Tween L, pH6.2, and water is supplemented to 1000m of L. the MRS solid culture medium is prepared by adding 1.5% agar powder into the MRS liquid culture medium, and the culture medium is sterilized for 15-20min at the temperature of 115-121 ℃.

In the method for preparing the rice composite fermentation product with anti-aging effect, as a preferred embodiment, the concentration of the lactobacillus seed liquid for inoculating to the fermentation substrate is 10⁷-10¹⁰CFU/m L (e.g., 10)⁸CFU/mL、10⁹CFU/m L, etc.), and the inoculation ratio of the zymocyte, namely the volume ratio of the seed liquid to the fermentation substrate is 1-3% (such as 1.2%, 1.5%, 2%, 2.5%, 2.8%).

In the above method for preparing rice composite fermented product with antiaging effect, as a preferred embodiment, the temperature of the fermentation culture is 40-50 deg.C (such as 41 deg.C, 42 deg.C, 43 deg.C, 44 deg.C, 45 deg.C, 46 deg.C, 47 deg.C, 48 deg.C, 49 deg.C), and the time is 6-10h (such as 6.5h, 7h, 8h, 9h, 9.5 h).

In the method for preparing the rice composite fermented product with anti-aging effect, as a preferred embodiment, the temperature of the sterilization treatment is 115-121 ℃ (such as 116 ℃, 117 ℃, 118 ℃, 119 ℃, 120 ℃) for 15-20min (such as 16min, 17min, 18min, 19 min).

In the above method for preparing the rice composite fermented product with anti-aging effect, as a preferred embodiment, before the sterilization treatment, a separation treatment is further included, and the rice composite fermented product with anti-aging effect, that is, the rice composite fermented raw stock, is finally obtained.

In the preparation method of the rice composite fermented product with the anti-aging effect, as a preferred embodiment, after the sterilization treatment, a drying treatment is further included, and finally the rice composite fermented product with the anti-aging effect, namely the rice composite fermented dry powder, is obtained.

In the above method for preparing the rice composite fermentation product with anti-aging effect, as a preferred embodiment, the drying treatment may be spray drying, vacuum freeze drying, or the like.

In the above method for preparing a rice composite fermentation product with anti-aging effect, as a preferred embodiment, the separation treatment is a centrifugation treatment; more preferably, the centrifugation treatment is carried out at 3500-8000r/min (e.g., 4000r/min, 4500r/min, 5000r/min, 5500r/min, 6500r/min, 7000r/min, 7500r/min) for 20-60min (e.g., 25min, 30min, 40min, 50min, 55 min).

According to still another aspect of the disclosure, a rice composite fermentation dry powder prepared by the preparation method is also provided.

According to another aspect of the disclosure, the rice composite fermentation raw pulp prepared by the preparation method is also provided.

According to another aspect of the disclosure, the application of the rice composite fermentation raw pulp or the rice composite fermentation dry powder in preparing cosmetics is also provided.

According to another aspect of the disclosure, the application of the rice composite fermentation protoplasm in preparing cosmetics is further provided.

The cosmetic can be facial mask, essence or toner.

According to the technical scheme of the embodiment of the disclosure, the novel rice composite fermentation raw pulp is prepared by carrying out composite fermentation on the rice, the burdock roots and the silybum marianum, and has a good anti-aging effect.

These and other advantages of the present disclosure will become more apparent from the following detailed description of the preferred embodiments of the present disclosure when taken in conjunction with the accompanying drawings.

Drawings

The disclosure may be better understood by reference to the following description taken in conjunction with the accompanying drawings. The accompanying drawings, which are incorporated in and form a part of this specification, illustrate preferred embodiments of the present disclosure and, together with the detailed description, serve to explain the principles and advantages of the disclosure. Wherein:

FIG. 1 shows the results of skin moisture content (abbreviated as "water content") and skin moisture loss (i.e., percutaneous water loss, abbreviated as "water dispersion") tests using the rice complex fermentation broth prepared in example 1 and 0.1% hyaluronic acid (0.1% HA);

FIG. 2 shows the results of skin moisture content (abbreviated as "moisture content") test using the rice complex fermentation broth prepared in example 1 and 0.1% hyaluronic acid (0.1% HA), wherein A represents the variation trend of percentage of moisture content before use at different time points and each area after use, and B represents the median variation trend of moisture content at different time points and different areas;

FIG. 3 shows the results of skin water loss (i.e., percutaneous water loss, abbreviated as "water dispersion") tests using the rice complex fermentation broth prepared in example 1 and 0.1% hyaluronic acid (0.1% HA), wherein A represents the change percentage of the TEW L value at different time points after use and before use in each area, and B represents the median change of the TEW L value in different areas at different time points;

FIG. 4 shows the results of the transdermal absorption experiment using the rice complex fermentation broth prepared in example 1;

FIG. 5 shows the results of ABTS free radical test for antioxidant scavenging using rice composite fermentation broth prepared in example 1;

fig. 6 shows photographs taken by volunteers for skin wrinkle and roughness tests after using the rice composite fermentation broth prepared in example 1, wherein the photographs are taken from top to bottom for analyzing pores, fine lines and wrinkles, and from left to right for 5min, 20min and 60min before and after using the sample;

FIG. 7 is a bar graph showing the skin pore number of the volunteer of FIG. 6 before and after using the rice composite fermentation broth prepared in example 1;

FIG. 8 is a bar graph showing skin fine line depth at the pre-and post-test sites of the volunteers of FIG. 6 using rice composite fermentation broth prepared in example 1;

fig. 9 is a bar graph showing skin wrinkle depth at the pre-and post-test sites of the volunteers of fig. 6 using the rice composite fermentation broth prepared in example 1.

Detailed Description

Exemplary embodiments of the present disclosure will be described hereinafter with reference to the accompanying drawings.

The experimental procedures used in the following examples are all conventional procedures unless otherwise specified.

Materials, reagents and the like used in the following examples are commercially available unless otherwise specified.

The rice in the following examples is commercially available.

The burdock root in the following examples is a dry product in a sheet form produced by agriculture subsidiary product purchasing limited of Hoodia city, Bozhou, Anhui province.

Silybum marianum in the following examples is from dried lean fruit produced by commercial agricultural byproducts of Hoodian, Bozhou, Anhui province.

The fermentation bacteria in the following examples are lactic acid bacteria, specifically Lactobacillus delbrueckii subspecies bulgaricus (L actinobacillus delbrueckii subsp. Bulgaricus) with the collection number of CGMCC1.16075, which is purchased from CGMCC, those skilled in the art can also use Lactobacillus buchneri (L actinobacillus buchneri with the collection number of CGMCC1.15607, which can be purchased from CGMCC), Bifidobacterium bifidum (Bifidobacterium bifidum with the collection number of CGMCC1.5029, which can be purchased from CGMCC), or any two or three of the three.

In the following examples, the formulation of MRS liquid medium was: 10.0g of peptone, 10.0g of beef extract, 5.0g of yeast powder, 20.0g of glucose and K₂HPO₄·7H₂O2.0 g, NaO 3H₂O5.0g, triammonium citrate 2.0g, MgSO₄·7H₂O0.05g, Tween 801.0 m L, pH6.2, water content 1000m L, adding 1.5% agar powder into MRS liquid culture medium as MRS solid culture medium, and sterilizing at 115 deg.C for 20min after preparing the culture medium components.

Example 1 preparation of Rice Compound fermentation puree

1. Activation of the bacterial species Lactobacillus delbrueckii subspecies bulgaricus (L actinobacillus delbruuchisuubp. Bulgaricus) colony 2 was selected and loop-inoculated into a test tube of 100m L MRS liquid medium and activated for 24h at 35 ℃.

2. And (3) strain purification, namely taking the liquid strain, performing gradient dilution, inoculating 200 mu L, coating the liquid strain into an MRS solid culture medium plate, and performing standing culture at 35 ℃ for 48 hours.

3. Liquid culture, taking the single colony 3 rings in the plate, inoculating in 100m L MRS culture medium, activating for 48h at 35 ℃.

4. Expanding culture, namely measuring the liquid test tube by 10 percent of inoculation amount, inoculating the liquid test tube into 100m L MRS culture medium, culturing for 12h at 37 ℃, and when the bacterial mass reaches 10⁸CFU/m L, obtaining seed liquid.

5. Preparing a fermentation substrate: adding 0.5g of burdock root (crushed and sieved by a 20-mesh sieve), 5g of rice and 0.5g of silybum marianum into 500g of water to obtain a fermentation substrate; wherein, the burdock root accounts for 0.1 percent, the rice accounts for 1 percent, and the silybum marianum accounts for 0.1 percent. Sterilizing the fermentation substrate at 115 deg.C for 20min, and cooling.

6. The rice composite fermentation raw stock is obtained by inoculating 15m L of zymocyte liquid obtained in the step 4 into 506g of fermentation substrate obtained in the step 5 to obtain a fermentation system, fermenting the fermentation system in an incubator at 45 ℃ for 8 hours to obtain a fermentation product, centrifuging the fermentation product at 5000r/min for 30min, removing precipitate, collecting supernatant, sterilizing at 115 ℃ for 20min to inactivate bacteria, and obtaining the sterilized fermentation product, namely the rice composite fermentation raw stock.

The rice composite fermentation raw stock prepared in the example 1 is viscous liquid in appearance and is light white to light yellow in color. The pH value is 5.1-6.3, the viscosity is 200-700cP, the content of soluble solid is 1.5-5.0%, the total number of colonies is less than 50CFU/ml, and no pathogenic bacteria are detected. According to the cosmetic hygiene standard GB7916-87, the total number of bacteria in the cosmetic is not higher than 1000CFU/ml, so that the fermented extract meets the quality requirement of the cosmetic.

Performing component analysis on the rice composite fermentation primary pulp, wherein a protein detection method refers to GB5009.5-2010, a crude polysaccharide detection method refers to GB/T5009.8-2008, a flavone detection method refers to GB/T5009.124-2003, and a total phenol detection method refers to GB/T8313-2008; the results obtained were as follows: the rice composite fermentation raw pulp prepared in the embodiment 1 contains 1.560g/kg of protein, 6.543g/kg of crude polysaccharide, 0.051g/kg of total flavone (counted by rutin) and 0.063g/kg of total phenol.

Example 2 application of Rice Compound fermentation protoplasm as cosmetic

Safety detection of rice composite fermentation raw stock

The human body patch test is mainly used for detecting the irritation of the final cosmetic product or raw materials. The closed patch test of human body was performed on the rice composite fermentation raw pulp obtained in example 1 according to the cosmetic hygiene norm (2015) in order to evaluate the potential skin irritation thereof.

1. Test subjects:

in the experiment, 30 suitable volunteers are selected, and the age range is 18-60 years.

2. The test method comprises the following steps:

the method comprises the steps of dropwise adding 0.02m L-0.025 m L liquid samples (100% rice composite fermentation raw stock without dilution) onto a filter paper sheet, placing the filter paper sheet into a spot tester, setting a blank control (water) for each sample, adding sample solvent distilled water which is equal to the sample in the holes of the spot tester, continuing the test period for 24 hours, and removing the spot tester after 24 hours for 30 minutes of standing (waiting for the indentation to disappear) for the purpose of accuracy, credibility and scientificity of test results, wherein the spot tester cannot be removed by a volunteer during the test period according to requirements, and the tested part cannot be contacted with the water, and observing the reaction of the skin for 24 hours and 48 hours, wherein the adverse reaction grading standard of the skin in the spot test of a human body is shown in table 1.

TABLE 1 grading Standard of adverse skin reactions

3. And (3) test results:

see table 2. As can be seen from the table: the rice composite fermentation raw pulp obtained in the embodiment 1 is used for negative reaction, which shows that the rice composite fermentation raw pulp provided by the invention has safety and does not bring adverse reaction to human body.

Table 2, test results of spot-sticking of rice composite fermentation broth obtained in example 1

Second, testing water content and water dispersion of rice composite fermentation raw stock

The moisturizing effect of the rice composite fermentation raw stock obtained in example 1, specifically, a skin moisture content test and a skin moisture loss test, Hyaluronic Acid (HA) is generally regarded as a positive control sample having a good moisturizing effect, and a comparison experiment was performed using 10% of rice composite fermentation raw stock (denoted as 10% sample, namely, an aqueous solution having a volume concentration of 10% prepared from the rice composite fermentation raw stock prepared in example 1) and 0.1% of hyaluronic acid (denoted as 0.1% HA, namely, an aqueous solution having a volume concentration of 0.1% prepared from hyaluronic acid).

1. And (3) testing environment: the temperature is 22 +/-1 ℃; humidity is 50-60%.

2. Test area: skin moisture content test, skin moisture loss test: the left and right forearms.

3. Testing time points: skin moisture content test, skin moisture loss test: before use; it is administered for 5min, 20min, and 60 min.

4. An experimental instrument: cornemeter, CM825, Tewameter, TM 300.

5. The test method comprises the following steps:

1)30 qualified volunteers were tested in a test place without direct light and wind, at room temperature of 22-24 deg.C and humidity of 40-60%, before testing, the forearms on both sides were cleaned with facial cleanser, rested for 30min, the inner sides of the forearms on both sides of the testee were drawn with a marker pen to form normal skin with an area of 3.5 × 3.5.5 cm, and the moisture content and the moisture loss of the skin before use were measured in sequence.

2) Cutting the facial masks soaked with the samples into 3 × 3cm size, respectively sticking on corresponding marks of forearm, taking down after 15min, lightly wetting the un-dried essence on the tested part with cosmetic cotton, and timing.

3) The water content of the stratum corneum and the TEW L value of 3 sites were tested at 5min, 20min and 60min respectively, and 3 measurements were taken for each site.

6. And (3) testing results:

fig. 1 is a dispersion graph of skin water content and water dispersion of 30 volunteers in arm tests, and it can be seen from fig. 1 that the water content and water dispersion trend of 10% sample (i.e. 10% rice composite fermentation raw stock) and 0.1% HA are similar, and the water content 5min after use is significantly higher than background (sample p is 0.004, vs background; HA p is 0.01, vs background) by analyzing the significance of T-Test, and the water content and the background have no significant difference at other time points.

The data were further analyzed and in FIG. 2, A is a plot of the percentage change trend for the water content of the 10% sample and 0.1% HA and B is a plot of the median change trend for the water content of the 10% sample and 0.1% HA. It can be seen from fig. 2 that the median water content of both the sample and HA reached a maximum at 5min after use, and both the sample and HA showed a decreasing trend with increasing time after use, with a percentage change in water content of less than 0 after 20min after use, indicating that the water content at this time point after use was below background data, but the sample and HA data trended similarly.

FIG. 3 shows the trend of percentage change in transdermal water loss for the 10% sample and 0.1% HA, and B shows the trend of median change in transdermal water loss for the 10% sample and 0.1% HA, it can be seen from FIG. 3 that transdermal water loss for both the sample and HA reached a maximum at 5min after application because of the higher water content resulting in greater transdermal water loss, and gradually decreased transdermal water loss with increasing time after application, and the percentage change in TWE L for the sample at 60min after application was less than 0, indicating that TWE L at this time was below the skin background.

Therefore, the rice composite fermentation raw pulp prepared in the example 1 HAs good water replenishing performance, and the water replenishing performance of 10% of samples is equivalent to that of 0.1% of HA.

Third, percutaneous absorption test

1. Preparation of rat skin

The nude mice are killed by taking off the cervical vertebrae, the back hairs are quickly shaved off by a shaver, the back skin is peeled off, subcutaneous fat and blood vessels are removed, the nude mice are repeatedly washed to be clean by distilled water, washed by physiological saline for a plurality of times and stored in a refrigerator at the temperature of 80 ℃ below zero for standby (used up within 5 days).

2. In vitro transdermal absorption experiment of Franz diffusion cell

The experimental procedure was carried out by adding a suitable amount of water to the thermostatic bath of the in vitro permeation diffusion apparatus, turning on the power supply and the thermostatic bath for magnetic stirring, setting the water temperature in the thermostatic bath at 37 + -0.1 deg.C, fixing the prepared rat skin between two diffusion cells with an iron clamp, adding 5m L receiving solution to the receiving cell of the vertical diffusion cell, preheating in the thermostatic bath of the in vitro permeation diffusion apparatus, setting the stirring speed of the receiving cell at 400r/min, adding the feed solution (10% sample, i.e., the large sample prepared in example 1) to the feed cell separatelyPreparing the rice composite fermentation raw stock into 10 percent aqueous solution by volume percentage), and sealing the upper opening by a preservative film. At the beginning of the test, when the samples (i.e. the feeding liquid) permeate for 0, 2, 4, 6, 8, 12 and 24 hours (specific time intervals are determined according to actual samples), 500 mul of samples are respectively taken and placed in a centrifuge tube with a plug, and each sampling is carried out while the same amount of receiving liquid is supplemented into the receiving pool and air bubbles in the pool are removed. The sample content will be determined. Then, the cumulative permeation quantity Q (mg/cm) was calculated according to the following formula²). Wherein the diameter of the bottom of the diffusion cell is 1.50cm, and the contact area of the sample is 1.77cm²。

Wherein Q is the cumulative permeation, S is the transdermal diffusion area, V is the volume of the receiving chamber of the modified Franz diffusion cell, Cn is the concentration of the receiving solution at the nth sampling, Ci is the concentration of the receiving solution at the ith sampling, and 0.5 is the sampling amount. After calculation by the above formula, a plot of accumulated amount versus time is made. A NaCl solution (physiological saline) having a receiving solution concentration of 0.9%; the feed solution was a 10% sample, i.e., a 10% volume aqueous solution prepared from the rice composite fermentation broth prepared in example 1.

3. Results of the experiment

The results of the experiment are shown in FIG. 4. It can be seen from the graph that the transmittance of the sample rapidly increases to 0.40mg/cm from 0 to 2 hours³As described above, the transmittance gradually increased with time, indicating that the sample smoothly permeated through the skin.

Fourth, antioxidant free radical scavenging performance test

1. Experimental methods

Preparing ABTS +. stock solution, namely preparing 2.45 mmol/L potassium persulfate, dissolving ABTS by using potassium persulfate to prepare 7 mmol/L ABTS stock solution, and standing for 12-16 h at room temperature under the condition of keeping out of the sun, wherein the stock solution can be stable for 3-4 d.

ABTS working solution is prepared by mixing 5m L7 mmol/L ABTS and 88 mu L140 mmol/L potassium persulfate solution, standing overnight at room temperature in a dark condition to form the ABTS working solution, diluting the ABTS working solution with absolute ethyl alcohol before use to ensure that the absorbance at 734nm is 0.70 +/-0.02.

Experiment operation, adding 40 mu L sample solution to be detected (namely the rice composite fermentation raw stock aqueous solution with the volume concentration of 5%, 10%, 20%, 50% and 100% prepared in advance) into 4m L ABTS working solution, accurately oscillating for 30s, and measuring an absorbance A sample at 734nm wavelength after reacting for 6 min.

Calculated according to the following formula:

ABTS + clearance/% (1-A sample/0.700) × 100

2. Results of the experiment

The results of the experiment are shown in FIG. 5. It can be seen from the figure that samples of different concentrations have different ABTS free radical scavenging abilities and exhibit dose dependence, with 100% of the samples having about 30% ability to scavenge ABTS free radicals. The rice composite fermentation raw stock prepared in example 1 has certain antioxidant performance.

Fifth, human skin wrinkle and roughness test

1. Test method

The Antera 3D camera can emit parallel light with various wavelengths, a skin image is constructed by collecting the wavelength of reflected light on the surface of the skin in a detected area, and parameters such as skin wrinkles, textures, pigmentation degrees and the like can be accurately measured according to a built-in complex algorithm.

10 volunteers meeting the conditions are selected to participate in the test, the test place has no direct light and no wind, the room temperature is 22-24 ℃, the humidity is 40% -60%, the face is cleaned by facial cleanser before the test, the face is rested for 30min, the canthus position of a subject is taken as a test part, the face mask soaked with a sample (100% of rice composite fermentation raw stock prepared in example 1) is respectively cut into the size of 3 × 3cm, the face mask is attached to the canthus, the face is taken down after 15min, the non-dried essence of the test part is lightly stained by cosmetic cotton, the timing is started, the canthus skin pores, fine lines and wrinkles are analyzed by Antera 3D face imaging before (background), 5min, 20min and 60min before and after the use, and the changes of the canthus skin pores, fine lines and wrinkles are measured.

2. Test results

The test results are shown in fig. 6-9.

FIG. 6 is a picture of the change analysis of the canthus skin of one volunteer during the sample application, wherein the pictures are from top to bottom for analyzing pores, fine lines and wrinkles, and from left to right for analyzing the skin before application (background) and 5min, 20min and 60min after application of the sample; it can be seen from the figure that pores, fine lines and wrinkles of the volunteers before use are obvious, and the rice composite fermentation raw stock prepared in example 1 is obviously improved.

Further, data were captured by software for trend analysis, resulting in fig. 8 to 9.

Fig. 7 shows the variation trend of the number of pores in the canthus area analyzed by the volunteers, and it can be seen from the graph that the background number of pores in the same area is about 160, and the number of pores scanned by the instrument 5min, 20min and 60min after the application is significantly less than the background number of pores, indicating that the sample has the potential effect of reducing the pores.

FIG. 8 shows the variation trend of average depth of fine lines in the canthus area analyzed by volunteers, and it can be seen that the average depth of fine lines on the skin is 0.035mm in the same area, the depth of fine lines is slightly reduced after 5min of use, and the skin fine lines 20min and 60min after use are not scanned by the instrument, indicating that the sample has fine line resistance.

FIG. 9 shows the variation trend of the average fine line depth in the canthus area analyzed by the volunteers, and it can be seen that the background skin fine line depth in the same area is 0.06-0.07 mm, the average skin wrinkle depth 5min and 20min after application is lower than the background, and the wrinkle depth 60min after application is increased, probably due to the water loss of the sample after application, which indicates that the sample has anti-wrinkle capability.

In summary, in the embodiments according to the present disclosure, the present disclosure provides the following technical solutions, but is not limited thereto:

scheme 1, a preparation method of a rice composite fermentation product with an anti-aging effect is characterized by comprising the following steps: inoculating the zymophyte into a fermentation substrate consisting of rice, burdock root, silybum marianum and water for fermentation culture, and then carrying out sterilization treatment to obtain the rice composite fermentation product with the anti-aging effect.

The method according to claim 2 or 1, wherein the fermentation tubes are lactic acid bacteria.

The method according to claim 3 or 2, wherein the lactic acid bacteria are selected from any one or two or more of the following:

lactobacillus delbrueckii subsp. Bulgaricus (L actinobacillus delbruuchii) with a collection number of CGMCC 1.16075;

lactobacillus buchneri (L actinobacillus buchneri) with the preservation number of CGMCC 1.15607;

bifidobacterium bifidum (Bifidobacterium bifidum) with the preservation number of CGMCC 1.5029.

The manufacturing method according to claim 4 or any one of claims 1 to 3, wherein the rice is rice grains or rice flour, and the amount of the rice in the fermentation substrate is 0.5 to 3% by weight based on the weight of water.

The preparation method according to scheme 5 or any one of schemes 1 to 4, wherein the burdock root is a dry product in a sheet or a ground state, and the burdock root accounts for 0.05 to 0.2 percent of the weight of water in the fermentation substrate.

The preparation method according to claim 6 or 5, wherein the burdock root is a pulverized dry product passing through a 20-mesh screen.

Scheme 7, the preparation method according to any one of schemes 1 to 6, characterized in that silybum marianum is a dried lean fruit product, and the amount of silybum marianum in the fermentation substrate is 0.05-0.15% of the weight of water.

The method according to any one of schemes 8 to 7, wherein the fermentation tubes are Lactobacillus delbrueckii subsp.

Scheme 9, the preparation method according to scheme 8, wherein the preparation process of the lactobacillus delbrueckii subspecies bulgaricus seed liquid for inoculation to fermentation substrate is as follows:

and (3) purification: carrying out gradient dilution on the activated liquid strain, inoculating the liquid strain into an MRS solid culture medium plate, and carrying out standing culture at 35-37 ℃ for 40-48 h;

and (3) performing amplification culture, namely measuring the bacterial liquid obtained by liquid culture by using 10% of inoculation amount, inoculating the bacterial liquid into a 100m L MRS liquid culture medium, and culturing for 10-14h at 35-37 ℃ to obtain seed liquid.

Scheme 10, the preparation method according to scheme 9, wherein the MRS liquid medium formulation is: 10.0g of peptone, 10.0g of beef extract, 5.0g of yeast powder, 20.0g of glucose and K₂HPO₄·7H₂O2.0 g, NaO 3H₂O5.0g, triammonium citrate 2.0g, MgSO₄·7H₂O0.05g, Tween 801.0 m L, pH6.2, and water content is supplemented to 1000m L, the MRS solid culture medium is obtained by adding 1.5% agar powder into the MRS liquid culture medium, and the culture medium is sterilized for 15-20min at 115-121 ℃.

The method according to claim 11 or any one of claims 2 to 10, wherein the concentration of the seed liquid of lactic acid bacteria to be inoculated to the fermentation substrate is 10⁷-10¹⁰CFU/m L, wherein the volume ratio of the seed liquid to the fermentation substrate is 1-3%.

Scheme 12, the preparation method according to any of scheme 1-11, characterized in that the temperature of the fermentation culture is 40-50 ℃ and the time is 6-10 h.

Scheme 13, the preparation method according to any one of schemes 1 to 12, wherein the temperature of the sterilization treatment is 115 to 121 ℃ and the time is 15 to 20 min.

The method according to claim 14 or any one of claims 1 to 13, further comprising a separation treatment before the sterilization treatment.

The production method according to claim 15 or 14, wherein the separation treatment is a centrifugation treatment.

Scheme 16, the preparation method according to scheme 15, wherein the speed of the centrifugal treatment is 3500-8000r/min, and the time is 20-60 min.

The method according to claim 17 or any one of claims 1 to 16, further comprising a drying step after the sterilization step.

The method according to claim 18 or 17, wherein the drying treatment is spray drying or/and vacuum freeze drying.

Scheme 19, a rice composite fermented dry powder prepared by the preparation method as described in scheme 17 or 18.

Scheme 20, a rice composite fermentation raw pulp prepared by the preparation method according to any one of schemes 1-16.

The application of the rice composite fermented dry powder in the scheme 21 and the scheme 19 in the preparation of cosmetics.

Scheme 22, application of the rice composite fermentation raw pulp in preparation of cosmetics according to scheme 20.

In the use of scheme 23, or scheme 21 or 20, the cosmetic is a mask, a serum, or a toner.

Finally, it is also noted that, in the present disclosure, relational terms such as first and second, and the like are used solely to distinguish one entity or action from another entity or action without necessarily requiring or implying any actual such relationship or order between such entities or actions. Also, the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus. Without further limitation, an element defined by the phrase "comprising an … …" does not exclude the presence of other identical elements in a process, method, article, or apparatus that comprises the element.

While the disclosure has been disclosed above by the description of specific embodiments thereof, it should be understood that various modifications, improvements or equivalents of the disclosure may be devised by those skilled in the art within the spirit and scope of the appended claims. Such modifications, improvements and equivalents are intended to be included within the scope of the present disclosure as claimed.

Claims

1. A preparation method of a rice composite fermentation product with an anti-aging effect is characterized by comprising the following steps: inoculating the zymophyte into a fermentation substrate consisting of rice, burdock root, silybum marianum and water for fermentation culture, and then carrying out sterilization treatment to obtain the rice composite fermentation product with the anti-aging effect.

2. The method according to claim 1, wherein the fermentation tubes are lactic acid bacteria.

3. The method according to claim 2, wherein the lactic acid bacteria are selected from any one or two or more of the following:

4. The method according to any one of claims 1 to 3, wherein the rice is rice grains or rice flour, and the amount of the rice in the fermentation substrate is 0.5 to 3% by weight based on the weight of water.

5. The method according to any one of claims 1 to 4, wherein the burdock root is a dried product in a sheet or a ground state, and the burdock root accounts for 0.05 to 0.2 percent of the weight of water in the fermentation substrate.

6. The method according to claim 5, wherein the burdock root is a pulverized dry product passing through a 20-mesh screen.

7. The preparation method according to any one of claims 1 to 6, wherein the silybum marianum is a dried lean fruit, and the amount of the silybum marianum in the fermentation substrate is 0.05 to 0.15 percent of the weight of water.

8. The method according to any one of claims 3 to 7, wherein the fermentation tubes are Lactobacillus delbrueckii subsp.

9. The method of claim 8, wherein the seed liquid of Lactobacillus delbrueckii subspecies bulgaricus for inoculation into a fermentation substrate is prepared by the following procedure:

10. The preparation method according to claim 9, wherein the MRS liquid medium formula is as follows: 10.0g of peptone, 10.0g of beef extract, 5.0g of yeast powder, 20.0g of glucose and K₂HPO₄·7H₂O2.0 g, NaO 3H₂O5.0g, triammonium citrate 2.0g, MgSO₄·7H₂O0.05g, Tween 801.0 m L, pH6.2, and water content is supplemented to 1000m L, the MRS solid culture medium is obtained by adding 1.5% agar powder into the MRS liquid culture medium, and the culture medium is sterilized for 15-20min at 115-121 ℃.