Preparation method and application of sapindus mukorossi fermentation product with effects of controlling oil and removing dandruff

Abstract

The invention relates to the technical field of daily chemicals, in particular to a preparation method and application of a sapindus mukorossi fermentation product with oil-controlling and dandruff-removing effects. The preparation method comprises the following steps: (1) drying soapberry in the air, separately powdering peel and seeds, adding 75-80 parts of water, performing ultrasonic extraction at 35 DEG C for 2 hours, then adding 0.5-0.9 part of dipotassium phosphate, sterilizing, and inoculating 3-5% of lactic acid bacteria liquid for fermentation; (2) after the fermentation is finished, adding 3-7 parts of sterilized aloe extract, 2-5 parts of mulberry leaf extract, 5-10 parts of tea bran extract and 6-9 parts of rose extract, adjusting the pH value to 5-7, inoculating 3-5% of schizosaccharomyces pombe, culturing for 24-36 hours at 28-30 DEG C, and introducing oxygen at 0.3-0.5 L / min; and (3) centrifuging at 10000r, and filtering through a sieve plate to obtain clear fermentation liquor, namely the sapindus mukorossi fermented product. The soapberry peel and the soapberry seeds are separately crushed, and the peel powder and the seed powder are mixed according to a ratio of 90%: 10% for use, so that the saponin content and the irritation are favorably controlled, and the use feeling is improved.

Description

Technical Field

[0001] The present invention relates to the field of daily chemical technology, and particularly relates to a preparation method and application of a sapindus ferment with oil control and anti-dandruff effects. Background Art

[0002] In daily life, scalp health problems have attracted more and more attention, among which the problem of dandruff is particularly prominent. The generation of dandruff not only affects personal image, but may also reflect health hazards such as scalp microecological imbalance. The causes of dandruff problems are complex and diverse. In addition to internal factors such as individual genetic factors and diet and work and rest habits, the excessive reproduction of malassezia furfur on the scalp is also a key external factor. When the scalp environment is suitable, malassezia furfur multiplies in large numbers, which will stimulate the excessive proliferation and abnormal shedding of scalp keratinocytes, thus leading to the generation of dandruff.

[0003] To solve the problem of dandruff, in the field of washing and care products, adding anti-dandruff agents has become a common means. At present, the mainstream anti-dandruff agents on the market are mostly chemical substances, such as zinc pyrithione (ZPT), octopirox ethanolamine salt (OCT), selenium disulfide, etc. Although these chemical anti-dandruff agents have strong bactericidal ability and can quickly show anti-dandruff effects in the initial stage of use, they have obvious disadvantages in the long term. They are generally highly irritating, easily damage the scalp, and disrupt the normal metabolic balance of the scalp. More seriously, long-term and frequent use of such chemical anti-dandruff agents will make scalp microorganisms develop drug resistance, and then cause a series of problems such as scalp inflammation and colony imbalance, making scalp problems more frequent and serious.

[0004] With the continuous improvement of consumers' awareness of health and environmental protection, the requirements for washing and care products are also increasing day by day, and natural plant anti-dandruff agents are gradually favored. Natural plant anti-dandruff agents are more suitable for long-term use by consumers because of their advantages such as safety and no toxic side effects. As a potential natural plant resource, sapindus shows unique value in the field of washing and care. Sapindus is a wild deciduous tree widely distributed in tropical and subtropical regions. Its roots and fruits are used as precious traditional Chinese medicines in traditional medicine, with the effects of relieving cough by eliminating phlegm, clearing heat and detoxifying, reducing accumulation, and killing insects. In terms of washing and care applications, sapindus peel extract - sapindus saponin, as a natural non-ionic surfactant, contains components such as triterpenoid saponins, sesquiterpene glycosides, fatty oils, and proteins, has a strong effect of reducing surface tension, has good detergency, can effectively remove dirt, and has no peculiar smell, and can be used for skin cleaning and clothing washing. Sapindus seeds contain a large amount of oil components mainly composed of unsaturated fatty acids (with a mass fraction of more than 80%, including palmitic acid, palmitoleic acid, stearic acid, oleic acid, linoleic acid, linolenic acid, arachidic acid, etc.) and flavonoid components such as kaempferol, quercetin, and apigenin.

[0005] At present, the utilization of Sapindus mukorossi mainly remains in the conventional extraction stage. Common extraction methods include solvent method, ultrasonic method, enzymatic method, microwave extraction method, macroporous resin adsorption method, etc., and often multiple methods are combined for extraction, such as solvent-ultrasonic extraction, solvent-enzymatic assisted extraction, etc. Among them, water extraction method and alcohol extraction method are relatively mature in industrial production, with the advantages of simple operation, controllable quality and low cost; while ultrasonic assisted extraction and microwave method, etc. have relatively high costs and have certain limitations in actual production applications. However, traditional extraction technologies are difficult to fully extract the active ingredients in Sapindus mukorossi, and the content of active ingredients in the extract is limited, resulting in the failure to fully exert the efficacy of Sapindus mukorossi in washing and care products.

[0006] Sapindus mukorossi has a wide range of applications in the daily chemical field. Due to the rich content of saponins in the pericarp, it can be used as a natural surfactant. However, the saponin content of the Sapindus mukorossi extract on the market is low, the extraction cost is high, and the operation is relatively complex. With the rise of fermentation technology, the microbial fermentation method can remove macromolecular substances that are difficult to remove by some physical and chemical methods through the metabolism of microorganisms to achieve the purpose of concentrating active ingredients. At the same time, some proteins and sugars are decomposed into small molecules by microorganisms, such as polypeptides and monosaccharides that are easily absorbed and utilized. At present, it is of great significance to develop a fermentation product of Sapindus mukorossi to enhance its application in washing and care. Summary of the Invention

[0007] To solve the above technical problems, the present invention provides a preparation method of a Sapindus mukorossi fermented product with anti-dandruff efficacy. The steps of the preparation method are as follows: (1) After drying Sapindus mukorossi, the pericarp and seeds are separately ground into powder, 75 - 80 parts of water is added, ultrasonic extraction is carried out at 35°C for 2 h, then 0.5 - 0.9 part of dipotassium hydrogen phosphate is added, after sterilization, 3% - 5% of lactic acid bacteria liquid is inoculated for fermentation; the mass ratio of the pericarp to the seeds is (7 - 10):1; the lactic acid bacteria include Lactobacillus plantarum and Streptococcus thermophilus with a mass ratio of (2 - 4):(1 - 2); (2) After the above fermentation is completed, 3 - 7 parts of sterilized aloe extract, 2 - 5 parts of mulberry leaf extract, 5 - 10 parts of tea bran leaching solution, and 6 - 9 parts of rose extract are added, the pH is adjusted to 5 - 7, 3% - 5% of Schizosaccharomyces pombe is inoculated, and cultured at 28 - 30°C for 24 - 36 h, with oxygen supply of 0.3 - 0.5 L / min; (3) After centrifugation at 10000 r, the clarified fermentation broth obtained by filtration through a sieve plate is the Sapindus mukorossi fermented product; The preparation method of the tea bran leaching solution is: S01: Tea bran pretreatment: The dry tea bran is crushed and sieved to 60 - 80 meshes; then according to the solid-liquid ratio of 50 g:100 mL, 25 g of tea bran powder is added to 50 mL of 0.5% chitosan lactate solution, and left standing at room temperature for 2 hours; then the surface liquid is blotted dry and reserved; S02: Add the pretreated tea bran into 370 mL of acid water extract and control the temperature at 35°C. Stir magnetically for 30 minutes. Collect the supernatant by suction filtration and retain the filter residue. The acid water extract comprises: 0.5% citric acid, 0.3% lactic acid, 10% ethanol, 0.01% sodium ascorbate, 0.05% EDTA-2Na, and the balance water; S03: Add 250 mL of organic acid solution to the residue, control the temperature at 45°C, and extract again for 20 minutes with gentle stirring; filter and combine the supernatant; The composition of the organic acid solution includes: malic acid 0.5%, acetic acid 0.2%, and the balance is deionized water, with a pH of 3.5 to 3.8; S04: Add 0.5% activated carbon powder to the mixed extract, stir thoroughly for 10 minutes, refrigerate and let stand for 24 hours, then filter and take the supernatant; finally, use a 0.45μm filter membrane to filter and sterilize / remove impurities to obtain the finished product.

[0008] As an embodiment of the present invention, the number of lactic acid bacteria is 10 7 ~10 8 CFU; fermentation temperature 35~37℃, fermentation time 36~48h.

[0009] As an embodiment of the present invention, the seed powder is extracted twice with 95% ethanol at a mass ratio of 1:8, filtered and dried.

[0010] As an embodiment of the present invention, the sterilization step in step (1) is gentle heating at 65-70°C for 30 minutes.

[0011] As an embodiment of the present invention, the preparation method of the mulberry leaf extract in step (2) is as follows: (1) Raw material pretreatment: Select low-temperature dried mulberry leaves, crush them through an 80-mesh sieve and store them; (2) Preparation of acidified solution: Mix citric acid, malic acid, and lactic acid in a mass ratio of 2:1:1, add 0.05% EDTA-2Na and 0.01% sodium ascorbate, and then add solvent: a mixture of deionized water and ethanol in a volume ratio of 8:2; adjust the pH to 3.5-4.0; (3) Extraction: The pretreated mulberry leaf powder and the acidified solution were mixed at a solid-liquid ratio of 1:15 to 1:25 g / mL, and the mixture was subjected to constant temperature magnetic stirring extraction at 40 °C for 30 minutes; the temperature was then lowered to 30 °C, and a second stage of ultrasonic-assisted extraction was performed for 10 minutes; the extract was allowed to stand and then filtered to obtain the first stage extract; (4) Re-extraction: Add the residue to 50% ethanol solution again and re-extract in a 45°C warm water bath for 20 minutes; combine the two extracts, let stand, degas, and filter; (5) Concentrate under reduced pressure at low temperature to 1 / 3 of the original volume; and filter with a 0.45 μm membrane to obtain mulberry leaf extract.

[0012] The second aspect of the present invention provides the preparation method for application in the field of daily chemicals.

[0013] By adopting the above technical solution, the present invention has the following beneficial effects: The soapberry peel and seeds are pulverized separately, and the peel powder and seed powder are mixed in a 90%:10% ratio to control saponin content and irritation, enhancing user experience. Ultrasonic extraction at 35°C for two hours, followed by sterilization with the addition of dipotassium hydrogen phosphate, not only extracts active ingredients but also enhances nutrient release from the post-sterilization fermentation matrix. A two-stage fermentation (primary fermentation with lactic acid bacteria, secondary fermentation with yeast) with rigorous temperature, pH, and aeration control ensures the stable production of active ingredients such as surfactants and antimicrobial peptides, making it suitable for shampooing and conditioning applications (such as anti-dandruff, anti-itching, oil control, and balancing the scalp microbiome). The addition of aloe vera, mulberry leaf, tea bran, and rose flower enhances the product's functional positioning by providing moisturizing, anti-inflammatory, soothing, and oil-control benefits. BRIEF DESCRIPTION OF THE DRAWINGS In order to more clearly illustrate the specific embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the specific embodiments or the description of the prior art. Obviously, the drawings described below are some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0014] Figure 1 This is the product picture of Example 1. DETAILED DESCRIPTION

[0015] The present invention will be further explained below with reference to specific embodiments.

[0016] Example 1 This embodiment provides a method for preparing a fermented soapberry with anti-dandruff effect, and the steps of the preparation method are as follows: (1) After drying 10 parts of soapberry, the peel and seeds were powdered separately, added with 75 parts of water, and ultrasonically extracted at 35°C for 2 hours. Then, 0.6 parts of potassium dihydrogen phosphate were added, and after sterilization (the sterilization step was gentle heating at 65°C for 30 minutes), 3% lactic acid bacteria liquid was added for fermentation; the mass ratio of the peel to the seeds was 9:1; the seed powder was extracted twice with 95% ethanol at a mass ratio of 1:8 (each time for 30 minutes, stirring at 40°C), and filtered and dried.

[0017] The lactic acid bacteria are Lactobacillus plantarum and Streptococcus thermophilus (mass ratio 1:2), and the number of strains is 5*10 7CFU. The fermentation temperature is 36°C and the fermentation time is 36 h.

[0018] Lactobacillus plantarum: China Center of Industrial Culture Collection, CICC®20279.

[0019] Streptococcus thermophilus: China Center of Industrial Culture Collection, CICC®20367.

[0020] (2) After the above fermentation is completed, add 4 parts of sterilized aloe extract, 3 parts of mulberry leaf extract, 8 parts of tea bran leaching solution, and 7 parts of rose extract, adjust the pH to 5 - 7, inoculate 4% of Schizosaccharomyces pombe, culture at 30°C for 24 h, and aerate at 0.4 L / min; Schizosaccharomyces pombe: China Center of Industrial Culture Collection, CICC®31963.

[0021] (3) After centrifugation at 10000 r, the clarified fermentation broth obtained by filtration through a sieve plate is the described sapindus mukorossi ferment.

[0022] The preparation method of the tea bran leaching solution is as follows: S01: Tea bran pretreatment: Crush and sieve the dry tea bran to 60 meshes; then, according to the material - liquid ratio of 50 g:100 mL, add 25 g of tea bran powder to 50 mL of 0.5 wt% chitosan lactate solution, and let it stand at room temperature for 2 hours; then suck dry the surface liquid and set aside.

[0023] S02: Put the pretreated tea bran into 370 mL of acid - water extraction solution, control the temperature at 35°C, and stir magnetically for 30 minutes; filter by suction to collect the supernatant, and retain the filter residue.

[0024] The components of the acid - water extraction solution include: citric acid 0.5% (w / v), lactic acid 0.3% (v / v), ethanol 10% (v / v), sodium ascorbate 0.01%, EDTA - 2Na 0.05%, and the balance is water.

[0025] S03: Add 250 mL of organic acid solution to the filter residue, control the temperature at 45°C, extract again for 20 minutes, and stir gently; filter by suction and combine the supernatants.

[0026] The components of the organic acid solution include: malic acid 0.5% (w / v), acetic acid 0.2% (v / v), the balance is deionized water, and the pH is 3.5 - 3.8.

[0027] S04: Add 0.5% activated carbon powder to the mixed extraction solution, stir well for 10 minutes, refrigerate and let it stand for 24 hours, then filter, take the supernatant; finally, filter and sterilize / remove impurities with a 0.45 - μm filter membrane to obtain the finished product.

[0028] The preparation method of the mulberry leaf extract in step (2) is as follows: (1)Raw material pretreatment: Select low-temperature dried mulberry leaves, crush them through an 80-mesh sieve and store them; (2)Preparation of acidified solution: Mix citric acid, malic acid, and lactic acid according to a mass ratio of 2:1:1, add 0.05% EDTA-2Na and 0.01% sodium ascorbate, and then add a solvent: a mixed solution of deionized water and ethanol with a volume ratio of 8:2; Adjust the pH to 3.5 - 4.0; (3)Extraction: According to a solid-liquid ratio of 1:15 g / mL, mix the pretreated mulberry leaf powder and the acidified solution, and carry out constant-temperature magnetic stirring extraction at 40 °C for 30 minutes; Then cool to 30 °C and perform ultrasonic-assisted extraction in the second stage for 10 minutes (frequency 40 kHz, power 250 W); After the extract is allowed to stand, filter it by suction to obtain the first-stage extract; (4)Re-extraction: Add the residue to a 50% ethanol solution (containing 0.2% citric acid) again, and perform re-extraction in a 45 °C water bath for 20 minutes; Combine the two extracts, allow them to stand for degassing and filter; (5)Concentrate under reduced pressure at low temperature to 1 / 3 of the original volume; And filter with a 0.45 μm membrane to obtain the mulberry leaf extract.

[0029] Figure 1 It is the product diagram of Example 1.

[0030] Example 2 The difference between this example and Example 1 is that rose extract and aloe extract are not added in the preparation method of the sapindus ferment.

[0031] Example 3 The difference between this example and Example 1 is that the mulberry leaf extract and the tea bran extract in the preparation method of the sapindus ferment are water extracts, and the specific preparation method is as follows: Put 40 tea bran (or mulberry leaves) in 1 L of water and boil for 20 min, and then concentrate to the same amount as in Example 1.

[0032] Example 4 The difference between this example and Example 1 is that the following is substituted: "(1) After drying the sapindus mukorossi, the pericarp and seeds are separately pulverized, 75 parts of water are added, ultrasonic extraction is carried out at 35°C for 2 h, 0.6 part of dipotassium hydrogen phosphate is added, and after sterilization (the sterilization step is gentle heating at 65°C for 30 min), 3% of lactic acid bacteria liquid is inoculated for fermentation; the mass ratio of the pericarp to the seeds is 9:1; the seed powder is extracted twice with 95% ethanol according to a mass ratio of 1:8 (stirring at 40°C for 30 minutes each time), and then filtered and dried." with "(1) After drying the sapindus mukorossi, the pericarp and seeds are mixed (i.e., the whole sapindus mukorossi) and pulverized, 75 parts of water are added, ultrasonic extraction is carried out at 35°C for 2 h, 0.6 part of dipotassium hydrogen phosphate is added, and after sterilization (the sterilization step is gentle heating at 65°C for 30 min), 3% of lactic acid bacteria liquid is inoculated for fermentation."

[0033] Example 5 This example provides a method for preparing a sapindus mukorossi ferment with an anti-dandruff effect. The steps of the preparation method are as follows: (1) After drying 10 parts of sapindus mukorossi, the pericarp and seeds are separately pulverized. The mass ratio of the pericarp to the seeds is 9:1; the seed powder is extracted twice with 95% ethanol according to a mass ratio of 1:8 (stirring at 40°C for 30 minutes each time), and then filtered and dried.

[0034] Add 4 parts of sterilized aloe vera extract, 3 parts of mulberry leaf extract, 8 parts of tea bran leaching solution, and 7 parts of rose extract to the pericarp powder and seed powder processed in step (1), adjust the pH to 5 - 7, add 75 parts of water, carry out ultrasonic extraction at 35°C for 2 h, then add 0.6 part of dipotassium hydrogen phosphate, and after sterilization (the sterilization step is gentle heating at 65°C for 30 min), inoculate 7% of the strain and culture at 30°C for 24 h, with oxygen supply at 0.4 L / min; The strain is Lactobacillus plantarum, Streptococcus thermophilus, Schizosaccharomyces pombe (1:2:4).

[0035] (3) After centrifugation at 10000 r, the clarified fermentation broth obtained by filtration through a sieve plate is the sapindus mukorossi ferment.

[0036] The preparation method of the tea bran leaching solution is as follows: S01: Tea bran pretreatment: Crush and sieve the dry tea bran to 60 meshes; then, according to a solid-liquid ratio of 50 g:100 mL, add 25 g of tea bran powder to 50 mL of 0.5 wt% chitosan lactate solution, and let it stand at room temperature for 2 hours; then suck dry the surface liquid and set aside.

[0037] S02: Put the pretreated tea bran into 370 mL of acid water extraction solution, control the temperature at 35°C, and stir magnetically for 30 minutes; filter and collect the supernatant, and retain the filter residue.

[0038] The components of the acidified water extract include: citric acid 0.5% (w / v), lactic acid 0.3% (v / v), ethanol 10% (v / v), sodium ascorbate 0.01%, EDTA-2Na 0.05%, and the balance is water.

[0039] S03: Add 250 mL of organic acid solution to the filter residue, control the temperature at 45 °C, extract again for 20 minutes with gentle stirring; filter with suction and combine the supernatant.

[0040] The components of the organic acid solution include: malic acid 0.5% (w / v), acetic acid 0.2% (v / v), the balance is deionized water, and the pH is 3.5 - 3.8.

[0041] S04: Add 0.5% activated carbon powder to the mixed extract, stir well for 10 minutes, refrigerate and let stand for 24 hours, then filter, take the supernatant; finally, filter and sterilize / remove impurities with a 0.45 μm filter membrane to obtain the finished product.

[0042] The preparation method of the mulberry leaf extract in step (2) is as follows: (1) Raw material pretreatment: Select low-temperature dried mulberry leaves, pulverize them and pass through an 80-mesh sieve for storage; (2) Preparation of acidified solution: Mix citric acid, malic acid, and lactic acid in a mass ratio of 2:1:1, then add 0.05% EDTA-2Na and 0.01% sodium ascorbate, and then add the solvent: a mixed solution of deionized water and ethanol with a volume ratio of 8:2; adjust the pH to 3.5 - 4.0; (3) Extraction: According to a solid-liquid ratio of 1:15 g / mL, mix the pretreated mulberry leaf powder and the acidified solution, and perform constant-temperature magnetic stirring extraction at 40 °C for 30 minutes; then cool to 30 °C and perform ultrasonic-assisted extraction in the second stage for 10 minutes (frequency 40 kHz, power 250 W); let the extract stand and then filter with suction to obtain the first-stage extract; (4) Re-extraction: Add the residue to a 50% ethanol solution (containing 0.2% citric acid) again, and perform re-extraction in a 45 °C water bath for 20 minutes; combine the two extracts, let stand, degas, and filter; (5) Concentrate under reduced pressure at low temperature to 1 / 3 of the original volume; and filter with a 0.45 μm membrane to obtain the mulberry leaf extract.

[0043] Application examples Add the sapindus extracts prepared in Examples 1 - 5 to the following formulation in Table 1 for subsequent tests to obtain Application Examples 1 - 5.

[0044] Table 1 Shampoo formulation

[0045] The preparation method of the said application examples is as follows: 1. Weigh the above components by weight parts. When the temperature of deionized water is about 65°C, slowly add sodium lauroamphoacetate, lauryl hydroxysulfobetaine, coconut oil amide DEA, TEA salt of coconut oil acyl glutamate, potassium cocoyl glycinate, and zinc PCA, and mix well.

[0046] 2. Continue to cool down. Add sodium chloride and betaine at about 45°C and stir well. Then add the remaining components, stir well, and discharge the product.

[0047] Performance Test Test 1: Evaluation of Oil Control Efficacy 1 Test Purpose and Principle The oil control efficacy is mainly characterized by evaluating the inhibition rate of the test sample on 5α-reductase. 5α-Reductase (5α-R) is an important androgen metabolic enzyme in the skin, which can irreversibly convert testosterone (T) into dihydrotestosterone (DHT). DHT is the most active androgen, which can induce the sebaceous glands to secrete excessive lipids. By inhibiting the metabolism of 5α-reductase, the secretion of oil can be controlled, the formation of dihydrotestosterone can be controlled, and sebum secretion can be reduced.

[0048] 2 Scope of Application and Relevance This test method is an in vitro method, which is applicable to evaluate cosmetic products and raw materials that claim to achieve oil control effects by inhibiting 5α-reductase. As a supplement to the subjective and objective evaluation methods of the human body, the in vitro method is one of the in vivo sebum secretion pathways and has a certain correlation with the results of testing the oil control efficacy by the in vivo method.

[0049] Through experimental design, the inhibition rate results of 5α-reductase of the test sample group and the blank group are compared. Repeat three times, calculate the mean value and standard deviation, and use mean comparison and variance analysis between the in vitro oil control test groups. If the inhibition rate of 5α-reductase in the test group is better than that in the blank group and the significance analysis P < 0.05, it can be considered that the test sample has a certain oil control efficacy. (Note: P < 0.05 is recorded as: *; P < 0.01 is recorded as: **; P < 0.001 is recorded as: ***) By measuring the inhibition rate of the sample on 5α-reductase, it can be reflected whether the sample has oil control efficacy; the higher the inhibition rate of 5α-reductase, the better the oil control efficacy of the test substance.

[0050] 3. Test Setup: Sample Group and Blank Control Group; Concentration Setting: The products of Examples 1 to 5 are diluted to 1% concentration with purified water; Detection Method: The sample reacts with 5α-reductase and its substrate, and the absorbance value is measured at a wavelength of 340 nm; Result calculation: Calculate the 5α-reductase inhibition rate based on the absorbance value.

[0051] All samples were tested in triplicate, and the mean and standard deviation were calculated. Mean comparison and one-way analysis of variance (ANOVA) were used for statistical tests.

[0052] If the enzyme inhibition rate of the sample group is higher than that of the blank group and the statistical difference is significant (P < 0.05), it is determined that the sample has a certain oil control effect.

[0053] Significance markers are as follows: P < 0.05: Significant difference (*) *P < 0.01: Highly significant difference (**) P < 0.001: Extremely significant difference (***) Table 2 Test results of Test 1

[0054] As can be seen from Table 2, Example 1 had the highest inhibition rate (57.41%), indicating that it had a good blocking effect on the oil secretion-related pathway and a significant oil control effect. Example 4 (whole fruit fermentation) and Example 5 (combination of three strains) also had strong inhibitory effects. Example 2 without aloe and rose, and Example 3 with water extraction instead of organic extraction both showed a decrease.

[0055] Test 2: 1 Test purpose and principle Malassezia is a dimorphic conditional pathogenic fungus that parasitizes the normal skin surface of humans and animals, and has two phenotypes: the spore phase (yeast form) and the hyphal phase (mold form). The dominant strains often detected in patients with dandruff or seborrheic dermatitis are generally Malassezia restricta, Malassezia globosa, Malassezia furfur, and Malassezia sympodialis, etc. The presence of Malassezia in dandruff, the species composition, and Malassezia infection are closely related to dandruff. The inhibitory effect of anti-dandruff products on Malassezia can effectively improve the dandruff condition.

[0056] 2 Scope of application and relevance This test method is an in vitro method and is applicable to evaluating cosmetic products and raw materials that claim to achieve anti-dandruff efficacy by inhibiting Malassezia. As a supplement to the subjective and objective evaluation methods of the human body, the in vitro method has economy and convenience. Currently, there is research evidence showing the relevance between the inhibition of Malassezia by the in vitro method and the test results of the in vivo anti-dandruff efficacy.

[0057] Through experimental design, the inhibition rate of Malassezia of the test sample group was compared with that of the blank group. This was repeated three times, and the mean value and standard deviation were calculated. For the in vitro anti-dandruff test, mean comparison and analysis of variance were used between groups. If the Malassezia inhibition rate of the test group was better than that of the control group and the P value of the significance analysis was less than 0.05, it could be considered that the test sample had a certain anti-dandruff effect. (Note: P < 0.05 is denoted as *; P < 0.01 is denoted as **; P < 0.001 is denoted as ***) By measuring the inhibition rate of the sample against Malassezia, it was reflected whether the sample had an anti-dandruff effect; the higher the inhibition rate of Malassezia, the better the anti-dandruff effect of the test substance. An inhibition zone diameter of not less than 7 mm indicated that the sample had an antibacterial effect.

[0058] Sample treatment: The test sample was prepared into a solution to be tested according to the required concentration. Cultivation conditions: The sample and Malassezia were inoculated on a culture medium plate and co-cultured for 48 hours; Concentration setting: The products of Examples 1 to 5 were diluted to a concentration of 0.5% with purified water. Detection index: Observe and measure the diameter of the inhibition zone (unit: mm). Result determination: An inhibition zone diameter ≥ 7 mm indicated that the sample had antibacterial activity. Calculate the inhibition rate of Malassezia for quantitative analysis of the anti-dandruff effect. Each experiment was repeated three times, and the results were expressed as mean ± standard deviation.

[0059] Inter-group mean comparison and one-way analysis of variance (ANOVA) were used for statistical tests: If there was a significant difference between the test group and the blank control group (P < 0.05), it could be determined that the sample had an anti-dandruff effect.

[0060] Table 3 Test results of Test 2

[0061] The test results showed that Example 1 also had the most significant anti-dandruff effect, indicating that the selected composite plant components and the two-stage fermentation system had an obvious synergistic effect. Example 5 also had a good effect, corroborating the enhancing effect of the co-fermentation of lactic acid bacteria and yeast on antifungal activity. In Examples 2 and 3, removing some auxiliary extraction components or changing to water extraction led to a decrease in activity, reflecting the influence of the extraction process in the formula on the efficacy.

[0062] Test 3: Store the shampoos of Application Examples 1 to 5 and observe their stability changes at different temperatures over a certain period of time. Test at 4 °C, 25 °C, and 45 °C for 30 days respectively; for the thermal cycling test, it was 4 °C (24 h) - 45 °C (24 h), 1 cycle = 48 h, and 15 cycles were carried out.

[0063] Test Results of Test 3 in Table 4

[0064] Example 1: Under various temperature and hot-cold cycle conditions, the appearance, odor, color, and foaming property remained relatively stable. Only slight delamination and odor change occurred at 45°C and during the hot-cold cycle, and the overall stability was the best. Examples 2 and 3: Odor fluctuations and a decrease in foaming property occurred under high temperature or hot-cold cycle, indicating that removing some functional components or replacing water extraction may affect the system balance.

[0065] Finally, it should be noted that the above examples are only used to illustrate the technical solutions of the present invention and are not intended to limit them. Although the present invention has been described in detail with reference to the foregoing examples, those of ordinary skill in the art should understand that they can still modify the technical solutions described in the foregoing examples, or perform equivalent replacements for some or all of the technical features; and these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A preparation method of sapindus ferment with the functions of controlling oil and removing dandruff, characterized in that, The steps of the preparation method are as follows: (1) After drying the sapindus mukorossi, the pericarp and seeds are separately powdered, 75 - 80 parts of water are added, ultrasonic extraction is carried out at 35°C for 2 hours, then 0.5 - 0.9 parts of dipotassium hydrogen phosphate are added, after sterilization, 3% - 5% of lactic acid bacteria liquid is inoculated for fermentation; the mass ratio of the pericarp to the seeds is (7 - 10):1; the lactic acid bacteria include Lactobacillus plantarum and Streptococcus thermophilus with a mass ratio of (2 - 4):(1 - 2); (2) After the above fermentation is completed, 3 - 7 parts of sterilized aloe vera extract, 2 - 5 parts of mulberry leaf extract, 5 - 10 parts of tea bran leaching solution, and 6 - 9 parts of rose extract are added, the pH is adjusted to 5 - 7, 3% - 5% of Schizosaccharomyces pombe is inoculated, cultured at 28 - 30°C for 24 - 36 hours, and oxygen is passed at 0.3 - 0.5 L / min; (3) After centrifugation at 10000 r, the clarified fermentation broth obtained by filtration through a sieve plate is the sapindus mukorossi ferment; The preparation method of the tea bran leaching solution is as follows: S01: Tea bran pretreatment: The dry tea bran is crushed and sieved to 60 - 80 meshes; then according to the material - liquid ratio of 50 g:100 mL, 25 g of tea bran powder is added to 50 mL of 0.5% chitosan lactate solution, and left standing at room temperature for 2 hours; then the surface liquid is blotted dry and reserved; S02: The pretreated tea bran is put into 370 mL of acid - water extraction solution, the temperature is controlled at 35°C, and magnetic stirring is carried out for 30 minutes; suction filtration is carried out to collect the supernatant, and the filter residue is reserved; Among them, the components of the acid - water extraction solution include: 0.5% citric acid, 0.3% lactic acid, 10% ethanol, 0.01% sodium ascorbate, 0.05% EDTA - 2Na, and the balance is water; S03: 250 mL of organic acid solution is added to the filter residue, the temperature is controlled at 45°C, and extraction is carried out again for 20 minutes with gentle stirring; suction filtration is carried out to combine the supernatant; The components of the organic acid solution include: 0.5% malic acid, 0.2% acetic acid, the balance is deionized water, and the pH is 3.5 - 3.8; S04: 0.5% activated carbon powder is added to the mixed extraction solution, stirred thoroughly for 10 minutes, refrigerated and left standing for 24 hours, then filtered, and the supernatant is taken; finally, filtration and sterilization / removal of impurities are carried out with a 0.45 μm filter membrane to obtain the finished product.

2. The preparation method according to claim 1, characterized in that, The number of lactic acid bacteria strains is 10 7 ~10 8 CFU; the fermentation temperature is 35~37°C, and the fermentation time is 36~48h.

3. The preparation method according to claim 1, characterized in that, The seed powder is extracted twice with 95% ethanol according to the mass ratio of 1:8, and suction - filtered and dried.

4. The preparation method according to claim 1, characterized in that, The sterilization step in step (1) is gentle heating at 65 - 70°C for 30 minutes.

5. The preparation method according to claim 1, characterized in that, The preparation method of the mulberry leaf extract in step (2) is as follows: (1) Raw material pretreatment: Select low - temperature dried mulberry leaves, crush and sieve through 80 - mesh sieve and store; (2) Acidified solution preparation: Citric acid, malic acid, and lactic acid are mixed according to the mass ratio of 2:1:1, then 0.05% EDTA - 2Na and 0.01% sodium ascorbate are added, and then the solvent: a mixed solution of deionized water and ethanol with a volume ratio of 8:2 is added; the pH is adjusted to 3.5 - 4.0; (3) Extraction: According to the material - liquid ratio of 1:15 - 1:25 g / mL, the pretreated mulberry leaf powder and the acidified solution are mixed, and magnetic stirring extraction is carried out at a constant temperature of 40°C for 30 minutes; then the temperature is reduced to 30°C, and ultrasonic - assisted extraction is carried out in the second stage for 10 minutes; the extraction solution is left standing and then suction - filtered to obtain the first - stage extraction solution; (4) Re-extraction: Add the residue into 50% ethanol solution again, and perform re-extraction at 45 °C in a warm water bath for 20 minutes; Combine the two extraction solutions, let it stand for degassing and filtration; (5) Concentrate at low temperature under reduced pressure to 1 / 3 of the original volume; After filtering with a 0.45 μm membrane, the mulberry leaf extract is obtained.

6. The preparation method according to any one of claims 1 to 5, characterized in that, It is applied to the daily chemical field.

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