TW202339782A - Method for preparing ferment of chinese herbal medicine and skin complexion improved composition - Google Patents

Abstract

The present disclosure provides a method for preparing a ferment of Chinese herbal medicine including the following steps. A fermenting substrate is provided, wherein the fermenting substrate includes a Chinese herbal material and water, and the Chinese herbal material is composed of Astragalus membranaceus, Atractylodes macrocephala, Ampelopsis japonica, Polygonatum sibiricum, Ligusticum striatum, Prunus Amygdalusand Saposhnikovia divaricata. A fermenting fungi solution is provided, wherein the fermenting fungi solution includes a yeast. A fermentation step is performed, wherein the fermenting material and the fermenting fungi solution are mixed so as to obtain a mixture, and the mixture is cultured so as to obtain a fermented product. A solid component of the fermented product is removed so as to obtain the ferment of Chinese herbal medicine. Therefore, the active ingredients of the Chinese herbal material can be effectively extracted and the activities thereof can be retained. In addition, the present disclosure also provides a skin complexion improved composition including the ferment of Chinese herbal medicine.

Description

Preparation method of Chinese herbal fermentation product and composition for improving skin condition

The present invention relates to a method for preparing a fermented product, in particular to a method for preparing a Chinese herbal fermented product and a composition containing the Chinese herbal fermented product for improving skin condition.

According to ancient pharmacopoeia records, mixing a variety of different Chinese medicinal materials and applying them on the face can have the effect of removing acne, dark spots, blisters and fine hairs. However, traditional Chinese medicine is more difficult to obtain, the cost is considerable, and the ingredients in the medicinal materials It is also difficult to obtain active ingredients. Furthermore, traditional Chinese medicine extracts its active ingredients by decoction. The decoction of medicinal materials is darker in color and has a poor texture. In addition, a lot of resources are consumed to extract the active ingredients in the medicinal materials, and the extraction efficiency is also low. This is so ideal that beauty products and cosmetics using Chinese medicinal materials are not popular. Currently, there are few studies on the efficacy of ancient Chinese medicine prescriptions in cosmetics.

Therefore, how to provide a strategy that can effectively apply Chinese medicinal materials in related fields of beauty and skin care is actually the development goal of the relevant academic circles and industry.

One aspect of the present invention is to provide a preparation method of Chinese herbal medicine fermentation product, which includes the following steps. A fermentation substrate is provided, wherein the fermentation substrate includes a Chinese herbal medicine raw material and water, and the Chinese herbal medicine raw material is composed of Astragalus baicalensis, Atractylodes rhizome, Atractylodes baicalensis, Polygonatum sibiricum, Ligusticum chuanxiong, almond, and Fangfeng. A fermentation bacteria liquid is provided, wherein the fermentation bacteria liquid contains a yeast. A fermentation step is performed, which involves mixing the fermentation substrate and the fermentation bacterial liquid to obtain a mixture, and culturing the mixture at 20°C to 50°C for 10 hours to 50 hours to obtain a fermentation product. One solid component of the fermentation product is removed to obtain a Chinese herbal medicine fermentation product.

According to the preparation method of Chinese herbal medicine fermentation product in the aforementioned embodiment, the Chinese herbal medicine raw material can be in the form of a dry powder, and based on 100 wt% of the Chinese herbal medicine raw material, the content of Scutellaria baicalensis can be 10~20 wt%, and the content of Atractylodes rhizome can be 10~20 wt%. 10~20 wt%, the content of Baiji can be 10~20 wt%, the content of Polygonatum can be 15~30 wt%, the content of Ligusticum Chuanxiong can be 15~30 wt%, and the content of almond can be 8~15 wt %, the content of windproof can be 8~15 wt%.

According to the preparation method of Chinese herbal medicine fermentation product in the foregoing embodiment, the yeast may be Saccharomyces cerevisiae .

According to the preparation method of Chinese herbal medicine fermentation product in the aforementioned embodiment, the fermentation matrix may contain 1.15 to 20 g of Chinese herbal medicine raw materials per 100 mL of water.

According to the preparation method of Chinese herbal medicine fermentation product in the above embodiment, the concentration of the fermentation bacterial liquid can be 10 ⁵ CFU/mL to 10 ⁸ CFU/mL, and in the mixture, every 10 g of Chinese herbal medicine raw materials can be Contains 5~32 mL of fermentation bacteria liquid.

According to the preparation method of the Chinese herbal medicine fermentation product in the aforementioned embodiment, the pH value of the Chinese herbal medicine fermentation product may be from 4.5 to 6.8.

According to the above embodiment, the preparation method of Chinese herbal medicine fermentation product may further include a sterilization step, which is to perform a sterilization treatment on the fermentation product, and the solid component may be centrifuged, filtered or a combination of the above methods And remove.

Another aspect of the present invention is to provide a composition for improving skin condition, including an effective dose of a Chinese herbal medicine fermentation product, wherein the Chinese herbal medicine fermentation product is prepared by using the preparation method of a Chinese herbal medicine fermentation product as described in the previous paragraph. have to.

According to the composition for improving skin condition according to the aforementioned embodiment, the composition for improving skin condition may have an antioxidant activity.

According to the composition for improving skin condition according to the aforementioned embodiment, the composition for improving skin condition can be used to improve the moisturizing ability of the skin and/or regulate the sebum secretion ability of the skin.

Thereby, the preparation method of the Chinese herbal medicine fermentation product of the present invention uses yeast to ferment the Chinese herbal medicine raw materials composed of Astragalus, Atractylodes, Bletilla, Polygonatum, Chuanxiong, Almond, and Fangfeng, which can effectively extract Astragalus, Atractylodes, The active ingredients of Bletilla chinensis, Polygonatum sibiricum, Ligusticum chuanxiong, almond and Fangfeng retain their activity, thereby avoiding the loss of active ingredients and the harm of organic solvents caused by traditional extraction methods, thereby making the preparation method of the Chinese herbal medicine fermentation product of the present invention Chinese herbal fermentation can be used in compositions or other cosmetics to improve skin condition, and has the effects of antioxidant, moisturizing and regulating sebum secretion of the skin. The preparation method of Chinese herbal fermentation and the combination of improving skin condition of the present invention The material has excellent market application potential.

Various embodiments of the invention are discussed in greater detail below. However, the embodiments are applicable to various inventive concepts and may be embodied in various specific scopes. The specific embodiments are provided for illustrative purposes only and do not limit the scope of the disclosure.

[Preparation method of Chinese herbal medicine fermentation product of the present invention]

Please refer to Figure 1, which is a step flow chart of the preparation method 100 of the Chinese herbal fermentation product of the present invention. The preparation method 100 of Chinese herbal medicine fermentation product includes step 110, step 120, step 130 and step 140.

Step 110 is to provide a fermentation matrix, wherein the fermentation matrix includes a Chinese herbal medicine raw material and water. The Chinese herbal medicine raw material is composed of Astragalus membranaceus , Atractylodes macrocephala , Ampelopsis japonica , and Polygonatum sibiricum . , Ligusticum striatum , Prunus Amygdalus , Saposhnikovia divaricata . In detail, the ingredients of the Chinese herbal medicine raw materials of the present invention are inspired by the prescription "Zerunfang" in "Puji Prescription" in Sikuquanshu. Among them, Astragalus, also known as Astragalus, has the functions of nourishing qi and raising yang, nourishing the body and strengthening the exterior. It has the functions of diuresis and swelling, treating sores and promoting muscle growth; Atractylodes has the functions of nourishing qi and strengthening the spleen, drying dampness and diuresis, antiperspirant, anti-fetus, etc.; Atractylodes atractylodes has the functions of clearing away heat and detoxification, eliminating carbuncle and dissipating stagnation, promoting muscle production and relieving pain; Polygonatum has the functions of Nourishes the kidneys and lungs, nourishes the spleen and replenishes qi; Ligusticum chuanxiong has the functions of promoting blood circulation and promoting qi, dispelling wind and relieving pain; almond, also known as bitter almond seed, has the functions of relieving cough and asthma, moisturizing the intestines and laxative; and Fangfeng has the functions of dispersing It has the functions of relieving wind and dampness, relieving pain, relieving spasm, and stopping diarrhea.

Furthermore, the Chinese herbal medicine raw material of the present invention can be in the form of a dry powder, and based on 100 wt% of the Chinese herbal medicine raw material, the content of Scutellaria baicalensis can be 10~20 wt%, the content of Atractylodes rhizome can be 10~20 wt%, and the content of Atractylodes atractylodes can be 10~20 wt%. The content of polygonatum can be 10~20 wt%, the content of Polygonatum can be 15~30 wt%, the content of Ligusticum chuanxiong can be 15~30 wt%, the content of almond can be 8~15 wt%, and the content of Fangfeng can be 8~15 wt%. In detail, the Chinese herbal medicine raw materials of the present invention can be dried and ground before subsequent steps, so that they can be in the form of dry powder to increase the reaction area. The drying method can be sun drying, shade drying, oven drying, Vacuum freeze-drying method, far-infrared light drying method or microwave drying method, and the grinding method can be carried out by using grinding equipment such as mortar, pestle or pulverizer, but the present invention is not limited thereto. Specifically, the drying method of the present invention can dry at 70°C to 80°C to obtain dry powder with a particle size of 30 mesh to 80 mesh. Preferably, the particle size of the dry powder can be 70 mesh to facilitate subsequent operations.

Furthermore, the fermentation matrix may contain 1.15 to 20 g of Chinese herbal medicine raw materials per 100 mL of water. Alternatively, the fermentation matrix may contain 5 g of Chinese herbal medicine raw materials per 100 mL of water. Alternatively, the fermentation matrix may contain 3 to 20 g of Chinese herbal medicine raw materials per 300 mL of water.

Step 120 is to provide a fermentation bacteria liquid, wherein the fermentation bacteria liquid includes a yeast. Specifically, yeast can metabolize proteins, lipids and starches in the fermentation matrix to obtain fermentation products rich in polysaccharides, amino acids or other specific components and in specific proportions. Furthermore, the yeast used in the present invention can be, but is not limited to, any strain of Saccharomyces cerevisiae or other species of yeast. Preferably, the yeast used in the present invention can be a brewing yeast deposited at the Taiwan Bioresource Collection and Research Center (BCRC) (Address: No. 331 Food Road, Hsinchu, Taiwan) with the deposit number BCRC 21815 Yeast (hereinafter referred to as "Saccharomyces cerevisiae 21815"). The colonies of Saccharomyces cerevisiae 21815 are dot-shaped, with smooth edges on the surface of the colony. The color of the colony is opaque and milky white, and the yeast exists in the form of single cells. Preferably, the concentration of the fermentation bacteria liquid can be 10 ⁵ CFU/mL to 10 ⁸ CFU/mL, but the present invention is not limited thereto.

Step 130 is to perform a fermentation step, which involves mixing the fermentation substrate and the fermentation bacterial liquid to obtain a mixture, and cultivating the mixture at 20°C to 50°C for 10 hours to 50 hours to obtain a mixture. Products after fermentation. In detail, yeast has the best fermentation efficiency in the above temperature range. If the fermentation time is too short, the active ingredients of the Chinese herbal medicine raw materials will not be fully released; conversely, if the fermentation time is too long, other bacteria may breed and affect the quality of the fermentation. Preferably, the mixture can be cultured at 25°C for 40 hours, but the present invention is not limited thereto.

Furthermore, in the mixture, every 10 g of Chinese herbal medicine raw materials may contain 5 to 32 mL of fermentation bacterial liquid, but the present invention is not limited to this. Alternatively, in the mixture, every 10 g of Chinese herbal medicine raw materials may contain 28 mL of fermentation bacteria liquid. Alternatively, in the mixture, the mixing ratio of fermentation bacterial liquid and Chinese herbal medicine raw materials can be 5~10 mL:2~20 g.

Step 140 is to remove a solid component of the fermentation product to obtain a Chinese herbal medicine fermentation product. Specifically, after the processing from steps 110 to 130, the active ingredients of astragalus, Atractylodes, Bletilla, Polygonatum, Ligusticum chuanxiong, almond and saposhnikovia have all entered into the fermented product. After the solid components are removed, the remaining liquid component is the Chinese herbal medicine fermentation product of the present invention, in which the pH value of the Chinese herbal medicine fermentation product can be 4.5 to 6.8, the viscosity can be 100 cP to 800 cP, and can contain 1.0 wt% to 5.0 wt% of soluble solids, and each milliliter of Chinese herbal medicine fermentation can contain 1.0~5.0 mg of protein, 5.0~10.0 mg of total polysaccharides, 1.0~1.8 mg of total flavonoids, and 0.1~0.6 mg of total phenols.

In addition, the method 100 for preparing fermented Chinese herbal medicine products of the present invention may further include a sterilization step, which is to perform a sterilization treatment on the fermented product. Specifically, the solid components can be removed by centrifugation, filtration, or a combination of the above methods, and the sterilization step can be high-pressure steam treatment, dry heat treatment, or ultraviolet treatment, but the invention is not limited thereto. Specifically, the fermented product can be centrifuged at a centrifugal speed of 4,000 rpm to 12,000 rpm for 15 to 30 minutes and then the supernatant is collected, and the supernatant is the Chinese herbal medicine fermentation product of the present invention. Alternatively, the fermentation product can be centrifuged at 4500 rpm for 20 minutes.

Thereby, the preparation method 100 of the Chinese herbal medicine fermentation product of the present invention uses yeast to ferment the Chinese herbal medicine raw materials of the present invention, which can not only effectively extract the active ingredients of Astragalus root, Atractylodes rhizome, Atractylodes atractylodes, Polygonatum japonica, Chuanxiong rhizome, almond and Fangfeng but also retain Its activity can shorten the fermentation time and avoid bacterial contamination caused by long-term fermentation. Furthermore, the preparation method 100 of the Chinese herbal medicine fermentation product of the present invention does not add any organic reagents during the extraction process, which can avoid the loss of active ingredients and the harm of organic solvent residues caused by traditional extraction methods, and extract the Chinese herbal medicine fermentation product. There is no problem that existing Chinese herbal medicine extracts are too dark in color, so that the Chinese herbal medicine fermented product produced by the preparation method 100 of the Chinese herbal medicine fermented product of the present invention has good biological safety and market aesthetics, and has excellent market application potential.

[Composition for improving skin condition of the present invention]

The composition for improving skin condition of the present invention contains an effective dose of the fermented Chinese herbal medicine of the present invention. The fermented Chinese herbal medicine is prepared by the preparation method of the fermented Chinese herbal medicine of the present invention and has antioxidant activity and the effect of improving the moisturizing ability of the skin and regulating the sebum secretion of the skin. Therefore, the composition for improving the skin condition of the present invention has It has excellent antioxidant, moisturizing, oil control and other functions and has excellent market application potential.

In addition, the dosage form of the composition for improving skin condition of the present invention can be liquid, emulsion, cream, ampoule, capsule, pill, lozenge, powder, granule or gel, and can be applied to lotion, lotion, essence. , isolation cream, facial cleanser, facial mask or sunscreen, but the invention is not limited thereto.

[Examples and Comparative Examples]

1. Chinese herbal fermentation product of the present invention

The properties of the Chinese herbal ferment obtained by the preparation method of the Chinese herbal ferment of the present invention will be described below with reference to the Chinese herbal ferment from Examples 1 to 4, and its antioxidant activity and application in skin moisturizing and skin conditioning will be further analyzed. sebum secretion ability and other effects. Please refer to Table 1 for the raw materials for preparing the Chinese herbal fermentation products of Examples 1 to 4. Table I Example 1 Example 2 Example 3 Example 4 Chinese herbal medicine raw materials Astragalus (wt%) 15 13 18 16 Atractylodes macrocephala(wt%) 15 13 18 16 White tangerine (wt%) 15 13 18 16 Polygonatum (wt%) 25 twenty two 28 30 Ligusticum chuanxiong(wt%) 25 twenty two 28 30 Almond(wt%) 12 11 15 10 Windproof(wt%) 12 11 15 10 Fermentation bacteria liquid concentration (CFU/mL) 10 ⁶ 10 ⁶ 10 ⁷ 10 ⁷

The Chinese herbal medicine fermentation products of Examples 1 to 4 were prepared by the following steps. First, astragalus, Atractylodes, Atractylodes, Polygonatum, Chuanxiong, almonds and Fangfeng purchased from a Chinese medicine pharmacy were dried in an oven at 75±5°C, and then ground using a grinder to obtain a size of 50-mesh Chinese herbal medicine dry powder, and 15 g of Chinese herbal medicine dry powder and 300 mL of water are mixed into a suspension as the fermentation substrate of the present invention, and mixed with a fermentation bacterial liquid containing yeast for fermentation.

A pretreatment step is performed before the yeast is used for fermentation. In terms of pretreatment steps, Saccharomyces cerevisiae 21815 was first inoculated into potato dextrose agar (PDA) and activated in an incubator at 30°C. Then a single colony was selected for subsequent amplification, and potato was used Liquid culture the glucose agar culture solution (potato dextrose water, PDW) at 20°C to 35°C until the OD value = 0.5~1.0. At this time, the yeast will be in the logarithmic phase and have better activity, and it can be used for subsequent fermentation.

Next, 5 mL of the amplified fermentation culture was inoculated into the fermentation matrix and mixed evenly, then cultured in an incubator at 25°C for 40 hours, and then the obtained fermentation product was heated at 121°C, 1.1 kg/ cm ² of high-pressure steam for sterilization, and then the sterilized fermentation product was centrifuged for 20 minutes at 4500 rpm and a centrifugal radius of 9 cm, and then the supernatant was collected. This supernatant is the Chinese herbal medicine fermentation product of the present invention.

The Chinese herbal medicine fermentation products of Examples 1 to 4 appear as colorless to brown liquids with good touch, and their pH is measured using conventional pH or viscometer methods (such as pH meter and rotational viscometer). The value is 4.5 to 6.8, the viscosity is 100 cP to 800 cP and contains 0.9~2.0 wt% soluble solids, and no pathogenic bacteria are detected. It has a high degree of biological safety and meets the quality requirements of cosmetics.

2. Component analysis of the fermented Chinese herbal medicine of the present invention

This test uses high performance liquid chromatography (HPLC) and refers to the test method GB/T 30483-2013 to analyze the components of the Chinese herbal medicine fermentation products of Examples 1 to 4. The results of high performance liquid chromatography show that each milliliter of the Chinese herbal medicine fermentation products of Examples 1 to 4 contains 1.0~5.0 mg of protein, 5.0~10.0 mg of total polysaccharides, 1.0~1.8 mg of total flavonoids and 0.1 ~0.6 mg of total phenols. Furthermore, since the Chinese herbal medicine fermentation products of Examples 1 to 4 are produced by the same manufacturing method, the only difference lies in the proportion of each component in the Chinese herbal medicine raw materials and the concentration of the fermentation bacteria liquid, and the fermentation products of Examples 1 to 4 are different. The Chinese herbal medicine fermentation product of Example 4 has similar protein, total polysaccharide, total flavonoids and total phenolic contents. Therefore, the following only uses the Chinese herbal medicine fermentation product of Example 1 to perform subsequent analysis of the contents of different active ingredients and their effects to illustrate the present invention. The effect of using fermented Chinese herbal medicine in a composition for improving skin condition is hereby described.

In addition, the following tests may include Comparative Example 1 and Comparative Example 2 at the same time. In Comparative Example 1, the fermentation product was prepared using the method for preparing the Chinese herbal medicine fermentation product of the present invention (hereinafter referred to as the "Chinese herbal medicine fermentation product of Comparative Example 1"). The difference lies only in the ratio of each component in the Chinese herbal medicine raw material of Comparative Example 1. Example 1 is not the same, and the ratio of each component in the Chinese herbal medicine raw materials of Comparative Example 1 is shown in Table 2. Comparative Example 2 uses a traditional decoction method to process the Chinese herbal medicine raw materials of the present invention without any fermentation step (hereinafter referred to as the "Chinese herbal medicine aqueous extract of Comparative Example 2"), and the ratio of the Chinese herbal medicine raw materials in Comparative Example 2 is the same as that in Examples 1, to further analyze the active ingredient content of the Chinese herbal medicine fermentation product prepared by the preparation method of the Chinese herbal medicine fermentation product of the present invention. Table II Comparative example 1 Chinese herbal medicine raw materials Astragalus (wt%) 0.1~1 Atractylodes macrocephala(wt%) 0.1~1 White tangerine (wt%) 0.1~1 Polygonatum (wt%) 1~2 Ligusticum chuanxiong(wt%) 1~2 Almond(wt%) 0.1~1 Windproof(wt%) 0.1~1 Fermentation bacteria liquid concentration (CFU/mL) 10 ⁵ ~10 ⁸

1. Analysis of protein content of Chinese herbal medicine fermentation products of the present invention

After analyzing the components of the Chinese herbal medicine fermentation products of Examples 1 to 4 through high-performance liquid chromatography, this test further analyzed the difference in protein content between Example 1 and Comparative Example 1.

In terms of experiments, first accurately measure 0.0 mL, 0.2 mL, 0.4 mL, 0.6 mL, 0.8 mL, and 1.0 mL of bovine serum albumin standard solutions into 10 mL test tubes, add water to make up to 1.0 mL, and place in the test tubes. Add 1.0 mL of alkaline copper test solution to the solution, shake well, and place at room temperature for 10 minutes. Then, add 4.0 mL of folinol test solution (2 mol/L acid concentration), mix well and let it stand at room temperature for 30 minutes. Use a spectrophotometer to measure the absorbance value at a wavelength of 650 nm, and draw a standard curve based on the obtained value. Next, measure 1 mL of the Chinese herbal medicine fermentation product of Example 1 and the Chinese herbal medicine fermentation product of Comparative Example 1 and place them in 10 mL test tubes respectively. Conduct the experiment according to the experimental method for preparing the standard curve and measure the absorbance value at a wavelength of 650 nm. And converted to the concentration of protein in the sample according to the standard curve.

Please refer to Table 3, which is the analysis result of protein content of the Chinese herbal medicine fermentation product of Example 1 and the Chinese herbal medicine fermentation product of Comparative Example 1. It can be seen from the contents of Table 3 that the Chinese herbal medicine fermentation product of Example 1 contains 1.706 mg of protein per milliliter, which is better than the protein content of the Chinese herbal medicine fermentation product of Comparative Example 1 which is 1.404 mg. Table 3 Protein content (mg/mL) Example 1 1.706±0.17 Comparative example 1 1.404±0.14

2. Analysis of polysaccharide content of Chinese herbal medicine fermentation products of the present invention

After analyzing the components of the Chinese herbal medicine fermentation products of Examples 1 to 4 through high-performance liquid chromatography, this test further analyzed the difference in polysaccharide content between Example 1 and Comparative Example 1.

In terms of experiments, first accurately measure 0.2 mL, 0.4 mL, 0.6 mL, 0.8 mL, and 1.0 mL of glucose standard solutions (90 μg/mL) into 10 mL test tubes, add water to make up to 1.0 mL, and place in the test tubes. Add 1 mL of 5% phenol solution and shake well, then add 5 mL of concentrated sulfuric acid, shake well, and then bathe in boiling water for 20 minutes. Next, cool the sample in an ice bath for 5 minutes and then use a spectrophotometer to measure the absorbance value at a wavelength of 488 nm, and draw a standard curve based on the obtained value. Next, 1 mL of the Chinese herbal medicine fermentation product of Example 1 and the Chinese herbal medicine fermentation product of Comparative Example 1 were measured and placed in 10 mL test tubes. Further experiments were conducted according to the experimental method for preparing the standard curve and the absorbance value at a wavelength of 488 nm was measured. , and converted to the polysaccharide content in the sample according to the aforementioned standard curve and formula (I), the formula (I) is as follows. W = Formula (I); Among them, m ₁ is the sugar content (μg) in the sample measurement solution corresponding to the standard curve, V ₁ is the constant volume of the sample (mL), and V ₂ is the volume of the sample measurement solution removed during colorimetric measurement (mL). ), and m ₂ is the sampling amount of the sample (g).

Please refer to Table 4, which is the analysis result of the polysaccharide content of the Chinese herbal medicine fermentation product of Example 1 and the Chinese herbal medicine fermentation product of Comparative Example 1. It can be seen from the contents of Table 4 that the Chinese herbal medicine fermentation product of Example 1 contains 7.069 mg of polysaccharides per milliliter, which is better than the polysaccharide content of the Chinese herbal medicine fermentation product of Comparative Example 1 which is 4.629 mg. Table 4 Polysaccharide content (mg/mL) Example 1 7.069±0.707 Comparative example 1 4.629±0.463

3. Analysis of total phenolic content of Chinese herbal medicine fermentation products of the present invention

After analyzing the components of the Chinese herbal medicine fermentation products of Examples 1 to 4 through high-performance liquid chromatography, this test further analyzed the difference in total phenolic content between Example 1 and Comparative Example 1.

In terms of experiments, first dilute with deionized water to prepare gallic acid solutions with concentrations of 10 μg/mL, 20 μg/mL, 30 μg/mL, 40 μg/mL, and 50 μg/mL as a standard curve. For this purpose, a 7.5% Na ₂ CO ₃ solution is also prepared as an experimental reagent. Next, take 1 mL of the Chinese herbal medicine fermentation product of Example 1 and the Chinese herbal medicine fermentation product of Comparative Example 1 and mix them evenly with 5 mL of 10 mol/L% folinol reagent, and then add 4 mL of 7.5% Na ₂ CO _3. Mix evenly and let it stand for 1 hour in the dark at room temperature. Use a spectrophotometer to measure the absorbance value at a wavelength of 765 nm. Convert the obtained data into the equivalent of gallic acid according to the standard curve, and then use the formula ( II) Calculate the total phenolic content, the formula (II) is as follows: X = C × 10 × n Formula (II); where The total phenol content in the test solution (mg/L), and n is the dilution factor of the sample.

Please refer to Table 5, which is the analysis result of the total phenolic content of the Chinese herbal medicine fermentation product of Example 1 and the Chinese herbal medicine fermentation product of Comparative Example 1. It can be seen from the contents of Table 5 that the Chinese herbal medicine fermentation product of Example 1 contains 0.422 mg of total phenols per milliliter, which is better than the total phenol content of the Chinese herbal medicine fermentation product of Comparative Example 1 which is 0.235 mg. Table 5 Total phenolic content (mg/mL) Example 1 0.422±0.042 Comparative example 1 0.235±0.024

4. Analysis of amino acid content of Chinese herbal medicine fermentation products of the present invention

After analyzing the components of the Chinese herbal medicine fermentation products of Examples 1 to 4 through high-performance liquid chromatography, this test further analyzed the difference in amino acid content between Example 1 and Comparative Example 1.

In terms of experiments, first accurately measure 0.0 mL, 0.2 mL, 0.4 mL, 0.6 mL, 0.8 mL, and 1.0 mL of amino acid standard solutions (equivalent to 0 μg, 40 μg, 80 μg, 120 μg, 160 μg, 200 μg amino acids) were placed in 25 mL test tubes and added water to make up to 4 mL, then added 1 mL of 2% ninhydrin solution and phosphate buffer (pH = 8.04), shaken well, and placed in a boiling water bath for 15 minute. Next, cool the sample to room temperature in an ice bath and let it stand for 15 minutes. Then use a spectrophotometer to measure the absorbance value at a wavelength of 570 nm, and draw a standard curve based on the obtained value. Next, 1 mL of the Chinese herbal medicine fermentation product of Example 1 and the Chinese herbal medicine fermentation product of Comparative Example 1 were measured and placed in 25 mL test tubes respectively. The experiment was carried out according to the experimental method for preparing the standard curve and the absorbance value at the wavelength of 570 nm was measured. And according to the standard curve and formula (III), it is converted into the content of amino acids in the sample. Formula (III) is as follows. X = Formula (III); Among them, C ₁ is the amount of amino acid in the sample obtained from the standard curve (μg), C ₀ is the value of the blank sample obtained from the standard curve (μg), V is the amount of sample solution removed, and m is the sample. Amount (g).

Please refer to Table 6, which is the analysis result of the amino acid content of the Chinese herbal medicine fermentation product of Example 1 and the Chinese herbal medicine fermentation product of Comparative Example 1. It can be seen from the contents of Table 6 that the Chinese herbal medicine fermentation product of Example 1 contains 0.671 mg of amino acids per milliliter, which is better than the amino acid content of the Chinese herbal medicine fermentation product of Comparative Example 1 which is 0.293 mg. Table 6 Amino acid content (mg/mL) Example 1 0.671±0.07 Comparative example 1 0.293±0.03

5. Analysis of total flavonoid content of Chinese herbal medicine fermentation products of the present invention

After analyzing the components of the Chinese herbal medicine fermentation products of Examples 1 to 4 through high-performance liquid chromatography, this test further analyzed the difference in total flavonoid content between Example 1 and Comparative Example 1.

In terms of experiments, firstly, accurately draw 0.0 mL, 1.0 mL, 2.0 mL, 3.0 mL, 4.0 mL, 5.0 mL, and 6.0 mL of rutin standard stock solutions (purity ≥ 90.0%) into 25 mL volumetric flasks and add water to 6 mL, then add 1 mL of 5% sodium nitrite solution (let it stand for 6 minutes), 1 mL of 10% aluminum nitrate solution (let it stand for 6 minutes), and 10 mL of 4.3% sodium hydroxide test solution (let it stand for 6 minutes). After standing for 15 minutes), prepare rutin solutions with concentrations of 0.0 μg/mL, 8.0 μg/mL, 16 μg/mL, 24 μg/mL, 32 μg/mL, 40 μg/mL, and 48 μg/mL. , used as a standard curve, and prepare a 10% aluminum nitrate solution as an experimental reagent. Next, take 2 mL of the Chinese herbal medicine fermentation product of Example 1 and the Chinese herbal medicine fermentation product of Comparative Example 1 and add water to 6 mL, and then add 1 mL of 5% sodium nitrite solution in sequence (let it stand for 6 minutes to react), 1 After mL of 10% aluminum nitrate solution (let to react for 6 minutes) and 10 mL of 4.3% sodium hydroxide test solution (let to react for 15 minutes), use a spectrophotometer to measure the absorbance value at a wavelength of 510 nm, and compare the obtained The data were converted into rutin equivalents according to the standard curve, and then the total flavonoid content was calculated according to formula (IV). Formula (IV) is as follows: X = Formula (IV); Among them _, Volume (mL), V ₂ is the volume of the sample solution (mL), V ₃ is the color development volume (mL), and M is the sample volume (g or mL).

Please refer to Table 7, which is the analysis result of the total flavonoid content of the Chinese herbal medicine fermentation product of Example 1 and the Chinese herbal medicine fermentation product of Comparative Example 1. It can be seen from the contents of Table 7 that the Chinese herbal medicine fermentation product of Example 1 contains 0.197 mg of total flavonoids per milliliter, which is better than the total flavonoid content of the Chinese herbal medicine fermentation product of Comparative Example 1 which is 0.104 mg. Table 7 Total flavonoid content (mg/mL) Example 1 0.197±0.02 Comparative example 1 0.104±0.01

6. Analysis of glutathione content of the Chinese herbal medicine fermentation product of the present invention

Glutathione, also known as glutathione or glutamine sulfide, is a small molecule protein composed of three amino acids, including glutamic acid, cysteine and glycine. An important component of the antioxidant enzyme Glutathione Peroxidase (GPx), it can help human cells fight free radicals and reduce the occurrence of diseases. Therefore, this test further used high-performance liquid chromatography to analyze the glutathione content of the Chinese herbal medicine fermentation product of Example 1 and the Chinese herbal medicine aqueous extract of Comparative Example 2, and the detection of glutathione content was analyzed by high-performance liquid chromatography. The conditions are listed in Table 8. Table 8 separation column Shim-pack GIST C18 (4.6mm×250 mm, 5 μm) mobile phase Mobile phase A: 2 g sodium octane sulfonate 5.4 g potassium dihydrogen phosphate 1000 mL water Mobile phase B: methanol The mixing ratio is A:B = 90:10 Detection wavelength (nm) 210 Temperature (°C) room temperature Flow rate (mL/min) 1 Sample processing Take 25 mL of the sample and perform ultrasonic extraction with 25 mL of methanol, then centrifuge to remove the protein. Filter it through a 0.45 μm filter membrane and use it as the test solution. Injection volume 10 μL

Please refer to Table 9, which is the analysis result of glutathione content of the Chinese herbal medicine fermentation product of Example 1 and the Chinese herbal medicine aqueous extract of Comparative Example 2. It can be seen from the contents of Table 9 that the Chinese herbal medicine fermentation product of Example 1 contains 10.94 μg of glutathione per milliliter, which is a 30% increase in the glutathione content compared to the Chinese herbal medicine aqueous extract of Comparative Example 2. Table 9 Glutathione content (μg/mL) Peak time (minutes) Example 1 10.94 2.125 Comparative example 2 8.37 1.978

7. Analysis of ellagic acid content of the Chinese herbal medicine fermentation product of the present invention

Ellagic acid is a natural phenolic antioxidant found in many fruits and vegetables, especially in the fruits of blackberries, strawberries, pomegranates, wolfberries, raspberries, white acorns, cranberries, pecans and other plants. The most abundant content, and the anti-cancer cell proliferation and antioxidant activity of ellagic acid make it widely used in various fields. Therefore, this experiment further used high-performance liquid chromatography to analyze the ellagic acid content of the Chinese herbal medicine fermentation product of Example 1 and the Chinese herbal medicine aqueous extract of Comparative Example 2, and the detection conditions of high-performance liquid chromatography for analyzing the ellagic acid content are as follows: Shown in Table 10. Table 10 separation column Shim-pack GIST C18 (4.6mm×250 mm, 5 μm) mobile phase Mobile phase A: acetonitrile Mobile phase B: 0.2% phosphoric acid aqueous solution The mixing ratio is A:B = 20:80 Detection wavelength (nm) 254 Temperature (°C) 35 Sample processing Dilute the sample with ethanol to a concentration of 10%, filter it through a 0.45 μm filter membrane, and use it as the test solution.

Please refer to Table 11, which is the analysis result of the ellagic acid content of the Chinese herbal medicine fermentation product of Example 1 and the Chinese herbal medicine aqueous extract of Comparative Example 2. It can be seen from the contents of Table 11 that the Chinese herbal medicine fermentation product of Example 1 contains 245.281 μg of ellagic acid per milliliter, which is 86.19% higher than the ellagic acid content of the Chinese herbal medicine aqueous extract of Comparative Example 2. Table 11 Ellagic acid content (μg/mL) Peak time (minutes) Example 1 245.281 12.067 Comparative example 2 131.732 12.033

8. Analysis of ferulic acid content of Chinese herbal medicine fermentation products of the present invention

Ferulic acid is one of the derivatives of cinnamic acid. It is an abundant phenolic compound present in plant cell walls. It has the functions of scavenging free radicals and inhibiting tyrosinase activity. Therefore, this experiment further used high-performance liquid chromatography to analyze the ferulic acid content of the Chinese herbal medicine fermentation product of Example 1 and the Chinese herbal medicine aqueous extract of Comparative Example 2. The detection conditions of high-performance liquid chromatography for analyzing the ferulic acid content are as follows: Shown in Table 12. Table 12 separation column Shim-pack GIST C18 (4.6mm×250 mm, 5 μm) mobile phase Mobile phase A: acetonitrile Mobile phase B: 0.1% phosphoric acid aqueous solution The mixing ratio is A:B = 45:55 Detection wavelength (nm) 325 Temperature (°C) 35 Sample processing Dilute the sample with ethanol to a concentration of 10%, filter it through a 0.45 μm filter membrane, and use it as the test solution.

Please refer to Table 13, which is the analysis result of ferulic acid content of the Chinese herbal medicine fermentation product of Example 1 and the Chinese herbal medicine aqueous extract of Comparative Example 2. It can be seen from the contents of Table 13 that the ferulic acid content of the Chinese herbal medicine fermentation product of Example 1 contains 2.667 μg of ferulic acid per milliliter, which is a 71.62% increase in the ferulic acid content compared to the Chinese herbal medicine aqueous extract of Comparative Example 2. Table 13 Ferulic acid content (μg/mL) Peak time (minutes) Example 1 2.667 8.784 Comparative example 2 1.554 8.776

9. Analysis of Cimicin Glycoside Content of Chinese Herbal Medicine Fermentation Products of the Invention

Studies have shown that prim-O-glucosylcimifugin in sapsroot and other medicinal materials has antipyretic, analgesic, anti-inflammatory, swelling, and anti-platelet coagulation effects. Therefore, this experiment further analyzed the examples using high-performance liquid chromatography. The content of cimicin glycosides in the Chinese herbal medicine fermentation product of 1 and the aqueous extract of Chinese herbal medicine in Comparative Example 2, and the detection conditions of high-performance liquid chromatography for analyzing the content of cimicin glycosides are listed in Table 14. Table 14 separation column Shim-pack GIST C18 (4.6mm×250 mm, 5 μm) mobile phase Mobile phase A: methanol Mobile phase B: 0.05% phosphoric acid aqueous solution The mixing ratio is A:B = 45:55 Detection wavelength (nm) 254 Temperature (°C) 35 Sample processing Dilute the sample with ethanol to a concentration of 50%, filter it through a 0.45 μm filter membrane, and use it as the test solution.

Please refer to Table 15, which is the analysis result of the cimicin glycoside content of the Chinese herbal medicine fermentation product of Example 1 and the Chinese herbal medicine aqueous extract of Comparative Example 2. From the contents of Table 15, it can be seen that the Chinese herbal medicine fermentation product of Example 1 contains 6.289 μg of cimicin glycosides per milliliter, which is 73.44% higher than the content of cimicin glycosides of the Chinese herbal medicine aqueous extract of Comparative Example 2. Table 15 Cimicifugarin content (μg/mL) Peak time (minutes) Example 1 6.289 5.960 Comparative example 2 3.626 5.944

10. Analysis of the callisoflavone glucoside content of the Chinese herbal medicine fermentation product of the present invention

Scutellaria baicalensis contains the active ingredient calycosin-7-glucoside, which has the effects of replenishing qi, detoxifying, promoting muscle growth, and diluting water. Therefore, this experiment further analyzed Example 1 by high-performance liquid chromatography. The fermentation product of Chinese herbal medicine and the aqueous extract of Chinese herbal medicine in Comparative Example 2 have the content of calycose isoflavone glucoside, and the detection conditions of high performance liquid chromatography for analyzing the content of calebasin isoflavone glucoside are listed in Table 16. Table 16 separation column Shim-pack GIST C18 (4.6mm×250 mm, 5 μm) mobile phase Mobile phase A: acetonitrile Mobile phase B: 0.2% formic acid aqueous solution The mixing ratio is A:B = 20:80 Detection wavelength (nm) 260 Temperature (°C) 35 Sample processing Dilute the sample with ethanol to a concentration of 20%, filter it through a 0.45 μm filter membrane, and use it as the test solution. Injection volume 10 μL

Please refer to Table 17, which is the analysis result of the callisoflavone glucoside content of the Chinese herbal medicine fermentation product of Example 1 and the Chinese herbal medicine aqueous extract of Comparative Example 2. It can be seen from the contents of Table 17 that the Chinese herbal medicine fermentation product of Example 1 contains 131.254 μg of ellagic acid per milliliter, which is a 56.92% increase in the callisoflavone glucoside content compared to the Chinese herbal medicine aqueous extract of Comparative Example 2. . Table 17 Content of mullein isoflavone glucoside (μg/mL) Peak time (minutes) Example 1 131.254 9.949 Comparative example 2 83.642 9.926

It can be seen from the above results that the preparation method of the Chinese herbal medicine fermentation product of the present invention can more effectively extract glutathione, ellagic acid, ferulic acid, and cohosh contained in the Chinese herbal medicine raw materials of the present invention than the traditional extraction method. The fermented Chinese herbal medicine fermented product prepared by the preparation method of the Chinese herbal medicine fermented product of the present invention has excellent market application potential.

3. Analysis of antioxidant activity of Chinese herbal medicine fermentation products of the present invention

This test utilizes the hydroxyl radicals generated by the reaction of hydrogen peroxide and ferrous ions in the Fenton reaction, and uses salicylic acid to capture the hydroxyl radicals generated by the reaction to produce a purple product to measure the characteristics of the present invention. The ability of Chinese herbal fermentation products to scavenge hydroxyl radicals. In detail, since the aforementioned purple product has a large absorbance value for light with a wavelength of 510 nm, if the Chinese herbal fermentation product of the present invention has antioxidant effects, it is added with hydrogen peroxide, ferrous ions and salicylic acid. After using the reaction solution, the absorbance value of the reaction solution for light with a wavelength of 510 nm can be reduced, and the antioxidant activity of the Chinese herbal medicine fermentation product of the present invention can be evaluated.

In terms of experiments, the Chinese herbal medicine fermentation product of Example 1 and the Chinese herbal medicine aqueous extract of Comparative Example 2 and 9 mmol/L FeSO ₄ were taken with volume percentage concentrations of 10%, 15%, 20%, 25% and 30% respectively. After mixing 8.8 mmol/L H ₂ O ₂ and 9 mmol/L salicylic acid ethanol solution, place in a water bath at 37°C for 15 minutes and measure the absorbance value of each sample at a wavelength of 510 nm with a spectrophotometer, and Calculate the hydroxyl radical scavenging rate according to the following formula (V): X% = [1-(AU/AU ₀ )] × 100% Formula (V); wherein The relative absorbance value of AU ₀ represents the relative absorbance value of Comparative Example 2.

Furthermore, this test also includes Comparative Example 1 and Comparative Example 2, in which the absorbance values of Comparative Example 1 and Comparative Example 2 are used as the blank background values of Example 1 and Comparative Example 2 respectively to exclude the presence of The change in absorbance value caused by factors other than color reaction is used to obtain the standardized relative absorbance value. Please refer to Table 18 for the reagent ratios used for testing in Example 1, Comparative Example 2, Comparative Example 1 and Comparative Example 2. Table 18 Reagent(mL) Example 1 Comparative example 1 Comparative example 2 Comparative example 2 FeSO ₄ 3 3 3 3 H ₂ O ₂ 3 0 3 0 salicylic acid 3 3 3 3 Chinese herbal fermentation products 1 1 0 0 Chinese herbal water extract 0 0 1 1 distilled water 0 3 1 4 Number of measurements 3 1 3 1

Please refer to Figure 2, which is an analysis chart of the hydroxyl radical scavenging ability of the Chinese herbal medicine fermentation product of the present invention. As shown in Figure 2, as the volume percentage concentration increases, the hydroxyl radical scavenging rate of the Chinese herbal medicine fermentation product of Example 1 gradually increases, and when the volume percentage concentration of the Chinese herbal medicine fermentation product of Example 1 is 30% When, its hydroxyl radical scavenging rate is 18.60%, which is better than the Chinese herbal medicine aqueous extract of Comparative Example 2 with the same concentration. It can be seen that the Chinese herbal medicine fermentation product prepared by the preparation method of the Chinese herbal medicine fermentation product of the present invention is relatively good. Compared with the extract obtained by traditional extraction methods, it has better antioxidant effect and has the potential to be used in compositions for improving skin condition or other cosmetics and has application potential in related markets.

4. Evaluation of the moisturizing ability of the Chinese herbal fermentation product of the present invention

This test uses transepidermal water loss (TEWL) as an indicator of the moisture content of the skin surface to evaluate the moisturizing ability of the Chinese herbal fermentation product in Example 1. In addition, this test also includes the Chinese herbal medicine aqueous extract of the aforementioned comparative example 2 to further evaluate the moisturizing effect of the Chinese herbal medicine fermentation product prepared by the preparation method of the Chinese herbal medicine fermentation product of the present invention.

In terms of experiments, first, a control area and an experimental area (each 3 cm × 3 cm) were set on the arm skin of three subjects, and the control area was detected using a commercially available transepidermal water loss meter TM300 (Courage + Khazaka Electronic). Transepidermal water loss from the skin of the regional and experimental areas. Next, the Chinese herbal fermentation product of Example 1 and the Chinese herbal aqueous extract of Comparative Example 2 were applied to the skin of the experimental area in a single dose at a dosage of 2.0 ± 1.0 mg/cm ² , but no treatment was performed on the skin of the control area. The transepidermal water loss was measured before the test, on the 1st day after use, on the 5th day after use, on the 7th day after use and on the 14th day after use, and the measured data were used as the transepidermal water loss in the control area. The skin moisture content assessment results are obtained by correcting the volume data.

Please refer to Figure 3, which is an analysis result of the improvement of skin moisture content by the Chinese herbal medicine fermentation product of the present invention. As shown in Figure 3, on the 1st day after use, the 5th day after use, the 7th day after use, and the 14th day after use, the skin moisture content of the subjects after using the Chinese herbal fermentation product of Example 1 was all obvious. The skin moisture content is higher than that of Comparative Example 2, indicating that the amount of transcutaneous water loss after using the Chinese herbal fermentation product of Example 1 is less. It can be seen from this that the Chinese herbal fermentation product prepared by the preparation method of the Chinese herbal fermentation product of the present invention has excellent moisturizing effect and can improve the moisturizing ability of the skin, and has the potential to be used in compositions or other cosmetics to improve skin condition. Application potential in relevant markets.

5. Analysis of the sebum conditioning effect of the Chinese herbal fermentation product of the present invention

This test is to test the oil content of the skin after applying the Chinese herbal fermentation product of Example 1 to evaluate its effect on regulating the sebum secretion of the skin. In addition, this test also includes the aqueous Chinese herbal medicine extract of Comparative Example 2 mentioned above to further evaluate the sebum conditioning effect of the Chinese herbal medicine fermentation product prepared by the preparation method of the Chinese herbal medicine fermentation product of the present invention.

In terms of experiments, first, a control area and an experimental area (each 3 cm × 3 cm) were set on the forehead skin of three subjects, and the control area was detected using the commercially available skin oil tester Sebumeter SM300 (Courage + Khazaka Electronic). and the oil content of the skin in the experimental area. Next, the Chinese herbal fermentation product of Example 1 and the Chinese herbal aqueous extract of Comparative Example 2 were applied to the skin of the experimental area in a single dose at a dosage of 2.0 ± 1.0 mg/cm ² , but no treatment was performed on the skin of the control area. The oil content of the skin was measured before the test, on the 1st day after use, on the 5th day after use, on the 7th day after use and on the 14th day after use, and the measured data were compared with the skin oil content data of the control area. The result of correction is the difference in skin oil content.

Please refer to Figure 4, which is an analysis result of the sebum conditioning effect of the Chinese herbal fermentation product of the present invention. As shown in Figure 4, on the 5th day after use, the 7th day after use, and the 14th day after use, the difference in skin oil content of the subjects after using the Chinese herbal fermentation product of Example 1 was significantly lower than that of Comparative Example 2. The difference in skin oil content shows that the Chinese herbal fermentation product of Example 1 has a better sebum conditioning effect after being applied to the skin, and can effectively regulate the sebum secretion ability of the skin, and has the potential to be used in compositions to improve skin condition. or other cosmetics and have application potential in relevant markets.

6. Evaluation of the anti-allergic and soothing effect of the Chinese herbal fermentation product of the present invention

In this test, the Chinese herbal fermentation product of Example 1 was applied to the skin of three subjects to observe the fading of red blood streaks and evaluate the anti-allergic and soothing effect of the Chinese herbal medicine fermentation product of the present invention.

In terms of experiments, first, an experimental area (each 3 cm × 3 cm) was set on the arm skin of three subjects, and the Chinese herbal fermentation product of Example 1 was applied to the experiment in an amount of 2.0 ± 1.0 mg/cm ² Apply to the skin of the area twice a day for 14 days, and take skin images before the test and on the 14th day after use, and calculate the average redness value based on the images obtained.

Please refer to Figure 5A, Figure 5B, Figure 6 and Table 19. Figure 5A is a test image of the anti-allergic and soothing effect of the Chinese herbal ferment of the present invention, and Figure 5B is a test image of the Chinese herbal ferment of the present invention. Another test image of the anti-allergic and soothing effect. Figure 6 is an analysis result chart of the anti-allergic and soothing effect of the Chinese herbal ferment of the present invention. Table 19 records the results of Subject 1 and Subject 2 without using Example 1. The average value of red blood cells of the Chinese herbal medicine fermented product and after 14 days of use, and the average value of red blood cells of subject 3 without using the Chinese herbal medicine fermented product of Example 1 and after using it for 7 days and 14 days. Table 19 red blood filament average Unused After 14 days of use Subject 1 1.28 1.248 Subject 2 1.316 1.211 Unused After 7 days of use After 14 days of use Subject 3 1.471 1.4 1.274

It can be seen from Figure 5A, Figure 5B, Figure 6 and Table 19 that after 14 days of using the Chinese herbal fermentation product of Example 1, the skin redness and swelling of subjects 2 and 3 subsided significantly. The average value of red blood streaks was also significantly reduced, and the skin red blood streaks of the three subjects were reduced by 8.2% on average, indicating that the Chinese herbal fermentation product prepared by the preparation method of the Chinese herbal medicine fermentation product of the present invention has considerable anti-allergic and soothing effects. It has the potential to be used in compositions or other cosmetics to improve skin condition and has application potential in relevant markets.

7. Safety evaluation of the fermented Chinese herbal medicine of the present invention

This test uses a closed patch test to evaluate the potential irritation of the Chinese herbal fermentation product of Example 1 to the skin to confirm the safety of the Chinese herbal fermentation product of the present invention when applied to the human body. In terms of experiment, first, 0.2 mL to 0.25 mL of the Chinese herbal medicine fermentation product of Example 1 was dropped onto the filter paper piece, and then the filter paper piece was placed in a spot tester for later use. Regarding the subjects, a total of 30 subjects ranging in age from 18 to 60 years old were randomly selected, and 2 patch testers were fixed on the back of each subject with non-irritating tape, so that the The filter paper piece was attached to the subject's back skin for 24 hours. The spot tester was not removed during the test, and the spot tester was also tried to avoid being contaminated with sweat or water. In addition, each test group corresponds to a control group, and the control group is tested with 0.20 mL to 0.25 mL of distilled water. Remove the patch tester 24 hours after the test, and observe the skin under the patch tester (hereinafter referred to as the "test area") and the subject 30 minutes, 24 hours and 48 hours after removal of the patch tester. Whether the skin around the area shows symptoms of allergies or inflammation such as erythema, infiltration, edema, papules and/or herpes.

Please refer to Table 20, which is the safety test results of the Chinese herbal medicine fermentation product in Example 1. The safety test results are divided into five levels. The picture number "-" represents a negative reaction in the test area without any symptoms of allergy or inflammation. The picture number "±" represents a suspicious reaction in the test area (such as a mild reaction). erythema), the figure number "+" represents a weak positive reaction in the tested area (also known as erythema reaction, that is, erythema, infiltration, edema and/or papules), the figure number "++" represents a strong positive reaction in the tested area Reaction (also known as herpetic reaction, that is, the occurrence of erythema, infiltration, edema, papules, herpes, and the herpetic reaction extends beyond the skin outside the tested area), the picture number "+++" represents a very strong positive in the tested area Reaction (also known as confluent herpetic reaction, that is, obvious erythema, severe infiltration, edema, confluent papules, and confluent herpetic reaction extends beyond the skin outside the tested area). In addition, in Table 20, the number of people whose reaction degree is positive (i.e., the figure numbers are +, ++, and +++) are also calculated to more clearly confirm the Chinese herbal fermentation obtained by the preparation method of the Chinese herbal fermentation product of the present invention. human safety of the object. Table 20 degree of reaction Number of responders (after removing the spot tester) Number of positive reactions 30 minutes 24 hours 48 hours control group - 30 30 30 0 ± 0 0 0 + 0 0 0 ++ 0 0 0 +++ 0 0 0 Example 1 - 30 30 30 0 ± 0 0 0 + 0 0 0 ++ 0 0 0 +++ 0 0 0

As shown in Table 20, 30 minutes, 24 hours or 48 hours after removing the spot tester, the subject's skin in and around the test area showed no erythema, infiltration, edema, papules, herpes or fusion. Herpes and other reactions show that the Chinese herbal fermentation product prepared by the preparation method of the Chinese herbal medicine fermentation product of the present invention has excellent biological safety and has application potential in relevant markets.

It should be added that although the present invention uses specific strains, specific materials, specific ratios, specific processes, specific analysis methods or specific instruments as examples to illustrate that the Chinese herbal fermentation product of the present invention has antioxidant, moisturizing, and However, anyone with ordinary knowledge in the technical field to which the present invention belongs will know that the ingredients and functions of the Chinese herbal fermentation product of the present invention can also be used in other strains without departing from the spirit and scope of the present invention. , other materials, other proportions, other processes, other analysis methods or other instruments for analysis, and the present invention is not limited thereto.

To sum up, the Chinese herbal medicine raw materials of the present invention are composed of Astragalus, Atractylodes, Bletilla, Polygonatum, Ligusticum chuanxiong, almond, and Fangfeng, and the above-mentioned medicinal materials can be fermented by yeast through the preparation method of the Chinese herbal fermentation product of the present invention. A Chinese herbal ferment containing a high concentration of active ingredients is obtained, and the aforementioned active ingredients have excellent antioxidant, moisturizing, and regulating sebum secretion abilities of the skin under the synergistic effect of each other, and have excellent biological safety and can be used It has excellent market application potential in compositions for improving skin condition.

Although the present invention has been disclosed in the above embodiments, it is not intended to limit the present invention. Anyone skilled in the art can make various modifications and modifications without departing from the spirit and scope of the present invention. Therefore, the protection of the present invention is The scope shall be determined by the appended patent application scope.

100: Preparation method of Chinese herbal medicine fermentation products 110,120,130,140: steps

In order to make the above and other objects, features, advantages and embodiments of the present invention more apparent and understandable, the accompanying drawings are described as follows: Figure 1 is a flow chart showing the steps of the preparation method of Chinese herbal medicine fermentation product of the present invention; Figure 2 is an analysis chart of the hydroxyl radical scavenging ability of the Chinese herbal medicine fermentation product of the present invention; Figure 3 is a diagram showing the analysis results of the Chinese herbal medicine fermentation product of the present invention for improving skin moisture content; Figure 4 is a diagram showing the analysis results of the sebum conditioning effect of the Chinese herbal medicine fermentation product of the present invention; Figure 5A is a test image of the anti-allergic and soothing effect of the Chinese herbal fermentation product of the present invention; Figure 5B is another test image of the anti-allergic and soothing effect of the Chinese herbal ferment of the present invention; and Figure 6 is a graph showing the analysis results of the anti-allergic and soothing effect of the Chinese herbal fermentation product of the present invention.

100: Preparation method of Chinese herbal medicine fermentation products

110,120,130,140: steps

Claims (10)

A preparation method of Chinese herbal medicine fermentation product, including: A fermentation matrix is provided, wherein the fermentation matrix contains a Chinese herbal medicine raw material and water, and the Chinese herbal medicine raw material is composed of Astragalus baicalensis, Atractylodes rhizome, Bletilla chinensis, Polygonatum japonica, Ligusticum chuanxiong, almond, and sapsroot; Provide a fermentation bacteria liquid, wherein the fermentation bacteria liquid contains a yeast; Perform a fermentation step, which is to mix the fermentation substrate and the fermentation bacterial liquid to obtain a mixture, and culture the mixture at 20°C to 50°C for 10 hours to 50 hours to obtain a fermentation product; and One solid component of the fermentation product is removed to obtain a Chinese herbal medicine fermentation product. The preparation method of Chinese herbal medicine fermentation product as described in claim 1, wherein the Chinese herbal medicine raw material is in the form of dry powder, and based on 100 wt% of the Chinese herbal medicine raw material, the content of the Astragalus is 10~20 wt%, and the content of the Atractylodes is 10~20 wt%. 10~20 wt%, the content of Baiji is 10~20 wt%, the content of Polygonatum is 15~30 wt%, the content of Ligusticum Chuanxiong is 15~30 wt%, the content of almond is 8~15 wt %, the content of the windproof is 8~15 wt%. The preparation method of Chinese herbal medicine fermentation product as described in claim 1, wherein the yeast is Saccharomyces cerevisiae . The preparation method of Chinese herbal medicine fermentation product as described in claim 1, wherein in the fermentation matrix, every 100 mL of the water contains 1.15~20 g of the Chinese herbal medicine raw material. The preparation method of Chinese herbal medicine fermentation product as described in claim 1, wherein the concentration of the fermentation bacterial liquid is 10 ⁵ CFU/mL to 10 ⁸ CFU/mL, and in the mixture, every 10 g of the Chinese herbal medicine raw material contains 5~ 32 mL of this fermentation broth. The preparation method of Chinese herbal medicine fermentation product as described in claim 1, wherein the pH value of the Chinese herbal medicine fermentation product is 4.5 to 6.8. The preparation method of Chinese herbal medicine fermentation product as described in claim 1 further includes: A sterilization step is to perform a sterilization treatment on the fermentation product, and the solid components are removed using a centrifugal method, a filtration method, or a combination of the above methods. A composition for improving the condition of the skin, comprising: An effective dose of a Chinese herbal medicine fermentation product, wherein the Chinese herbal medicine fermentation product is prepared by using any one of the preparation methods of claim 1 to claim 7. The composition for improving skin condition as claimed in claim 8, wherein the composition for improving skin condition has an antioxidant activity. The composition for improving skin condition as claimed in claim 8, wherein the composition for improving skin condition is used to improve the moisturizing ability of the skin and/or to regulate the sebum secretion ability of the skin.

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