A peony fermentation liquor, use and skin care product

Abstract

The application belongs to the technical field of skin care, and discloses a peony fermentation liquor, application and skin care product, which is obtained by sequentially fermenting peony roots and peony flowers by Raen's bacteria and Bifidobacterium after mixing the peony roots and the peony flowers; the weight ratio of the peony roots to the peony flowers is 1-2:1-2. The application provides a peony fermentation liquor, which is obtained by sequentially fermenting the roots and flowers of peony by Raen's bacteria and Bifidobacterium so as to obtain the peony fermentation liquor with high soothing effect.

Description

Technical Field

[0001] This application relates to the field of skin care technology, specifically to a peony fermentation liquid, its uses, and skin care products. Background Technology

[0002] With the advancement of skincare technology, plant-based ingredients have become a hot topic in skincare research due to their natural safety and multiple skincare benefits. Among them, peony, as a plant, contains some natural active ingredients that have antibacterial, anti-inflammatory, whitening, moisturizing, antioxidant, and blood circulation-promoting effects, making it an excellent plant with multiple skincare benefits.

[0003] Currently, there are two main directions for the extraction and utilization of plant active ingredients: (1) Extracting active ingredients from plants through extraction methods to obtain plant extracts; for example, prior art 1: Chinese Patent 202411142226.5 discloses an extraction method, anti-hair loss composition and its use. The extraction method of the anti-hair loss composition includes the following steps: Step 1: Adding arborvitae leaves to water for ultrasonic extraction to obtain a mixture; Step 2: Adding Ganoderma lucidum, ginseng root and water to the mixture for compound extraction, and then separating to obtain a filtrate; Step 3: Adding the filtrate to water for ultrasonic treatment to obtain a treated liquid; Step 4: Precipitating the treated liquid with ethanol for 12-14 hours to obtain an ethanol precipitate, and then concentrating to obtain an alcohol-free concentrate; Step 5: Adding an extractant to the alcohol-free concentrate for extraction to obtain an extract; Step 6: Ultrafiltration of the extract to obtain a permeate to obtain the anti-hair loss composition.

[0004] The existing technology involves extracting plant active ingredients through plant extraction methods. The current technology is relatively mature, and it generally uses water extraction, alcohol extraction, and other methods to obtain multiple functional ingredients based on their solubility. However, it is difficult to obtain specific functional ingredients in a targeted manner.

[0005] (2) Through bio-fermentation technology, plant active ingredients are extracted by microorganisms and further converted into other metabolites or active ingredients through the microbial fermentation process; among them, microbial fermentation technology has become a new skin care hotspot due to its milder products, fewer side effects, and the ability to obtain specific functional ingredients through microbial screening and process optimization.

[0006] Currently, there is limited research on the microbial fermentation of peony, especially regarding its skin-soothing effects. Few existing technologies utilize microbial fermentation to specifically enhance the soothing effects of peony components. Summary of the Invention

[0007] One of the purposes of this application is to provide a peony fermentation liquid, which, through microbial fermentation technology, yields a peony fermentation liquid with high soothing effects.

[0008] Another object of this application is to provide a use for peony fermentation liquid.

[0009] Another objective of this application is to provide a soothing skincare product containing the peony fermented liquid of this application, which has a skin-soothing effect.

[0010] To achieve the above objectives, this application provides a peony fermentation broth, which is obtained by mixing peony roots and peony flowers and then fermenting them sequentially with Laenella and Bifidobacterium; the weight ratio of the peony roots to the peony flowers is 1-2:1-2.

[0011] Furthermore, the specific preparation method of the peony fermentation broth includes the following steps:

[0012] Step 1: Crush peony roots and peony flowers and mix them to obtain a mixture, then use the mixture as a fermentation substrate to prepare a fermentation culture medium;

[0013] Step 2: The mixture was inoculated into the fermentation medium with Laenella seed culture and then centrifuged to obtain the primary fermentation broth;

[0014] Step 3: Ferment the primary fermentation broth using Bifidobacterium seed culture, and then centrifuge to obtain the peony fermentation broth.

[0015] Furthermore, the specific operation of step 1 is as follows:

[0016] The peony root and peony flower were crushed and mixed to obtain a mixture. Then, the mixture was prepared by mixing the peony root and peony flower with 20g of glucose, 75g of the mixture, and the remainder water per kg of peony root and adjusting the pH of the solution to 7.0-7.2.

[0017] Furthermore, the specific operation of step 2 is as follows:

[0018] Inoculate the seed culture of *Laenia* into the fermentation medium at an inoculation rate of 2-5% (v / v), control the dissolved oxygen at 20-40%, and ferment at a constant temperature of 30℃ for 24 hours to obtain the primary fermentation broth.

[0019] Preferably, the viable count of the Larynella seed culture is 2.6 × 10⁻⁶. 9 cfu / g.

[0020] It should be noted that the Larnella seed solution is obtained by Larnella through activation culture and expansion culture in sequence. Those skilled in the art can obtain the Larnella seed solution of this application using conventional technical means.

[0021] More preferably, the specific preparation steps of the Larynella seed solution are as follows:

[0022] Prepare seed culture medium by mixing 20g of glucose and water per 1kg of solution and adjusting the pH to 7.0-7.2, then allow it to cool naturally; then prepare the frozen-preserved *Lahn's bacterium* solution (concentration: 0.3 × 10⁻⁶). 9 CFU / g was added to activation medium for primary culture, and the culture temperature was set at 30℃. After 24 hours of culture, *Rahn's bacillus* seed culture was obtained. The viable cell concentration in the *Rahn's bacillus* seed culture was detected to be 2.6 × 10⁻⁶. 9 cfu / g; the weight of Raenella bacteria solution added to each 1 kg of seed culture medium is 5 g.

[0023] Furthermore, the specific operation of step 3 is as follows:

[0024] Fermentation medium was prepared by mixing 20g of glucose, 50g of primary fermentation broth, and the remainder water per kg of culture medium and adjusting the pH of the solution to 6.0. Bifidobacterium seed solution was inoculated into the fermentation medium and fermented at 37℃ for 24 hours to obtain fermentation mixture A. 3g of Bifidobacterium seed solution was inoculated into each kg of fermentation medium.

[0025] Fermentation mixture A was separated by centrifugation at 14,000 rpm. The supernatant A of the centrifuged fermentation mixture was collected and sterilized at 121°C and 0.11 MPa for 20 minutes. Then, it was clarified and purified to obtain peony fermentation broth.

[0026] Preferably, the viable bacterial concentration in the Bifidobacterium seed culture is 5.0 x 10⁻⁶. 9 cfu / g

[0027] This application also provides the use of the above-mentioned peony fermentation liquid in the preparation of skin care products.

[0028] This application further provides a soothing skin care product containing 0.5-50 wt% of the above-mentioned peony fermentation liquid.

[0029] Beneficial effects

[0030] Compared with the prior art, this application provides a peony fermentation liquid, which obtains a peony fermentation liquid with high soothing effect by sequentially fermenting the roots and flowers of peony with Laenella and Bifidobacterium. Detailed Implementation

[0031] The present application will be further described below with reference to embodiments, but this does not constitute any limitation on the present application. Any limited modifications made within the scope of the claims of the present application shall still be within the scope of the claims of the present application.

[0032] To illustrate the technical content of this application in detail, the following description is provided in conjunction with the embodiments.

[0033] In the following examples and comparative examples, the *Laenia lanuginosa* was purchased from the Guangdong Provincial Microbial Culture Collection Center (GDMCC), product serial number GDMCC NO: 1.998, product name *Laenia lanuginosa*; the *Bifidobacterium* was purchased from Minsheng Zhongke Jiayi (Shandong) Biotechnology Co., Ltd.; and the peony root and peony flower were both purchased from Shandong Pain Biotechnology Co., Ltd.

[0034] Unless otherwise specified, in the following examples and comparative examples, the parts and % refer to parts by weight and weight percentage, respectively.

[0035] In the following examples and comparative examples, the Larynella seed culture was prepared using the following steps:

[0036] Prepare seed culture medium by mixing 20g of glucose and water per 1kg of solution and adjusting the pH to 7.0-7.2, then allow it to cool naturally; then prepare the frozen-preserved *Lahn's bacterium* solution (concentration: 0.3 × 10⁻⁶). 9 CFU / g was added to activation medium for primary culture, and the culture temperature was set at 30℃. After 24 hours of culture, *Rahn's bacillus* seed culture was obtained. The viable cell concentration in the *Rahn's bacillus* seed culture was detected to be 2.6 × 10⁻⁶. 9 cfu / g; the weight of Raenella bacteria solution added to each 1 kg of seed culture medium is 5 g.

[0037] The Bifidobacterium seed solution was prepared using the following steps:

[0038] Step a: Prepare activation medium and seed culture medium by mixing 25g of glucose per kg of glucose solution with the remainder being water, and adjusting the pH of the solution to 6.0. Sterilize both the activation medium and seed culture medium at 121℃ and 0.11 MPa for 20 minutes. After the activation medium cools, add 1g of a 1.0 x 10⁻⁶ solution to each kg of activation medium. 7 A CFU / g Bifidobacterium bacterial suspension was incubated at 37°C for 24 hours in activation medium to obtain a viable bacterial concentration of 3.0 x 10⁻⁶. 8 Bifidobacterium activating solution with cfu / g;

[0039] Step b: Inoculate the cooled seed culture medium with Bifidobacterium activation solution and incubate at 37°C for 24 hours to obtain a viable bacterial concentration of 5.0 x 10⁻⁶. 9 Bifidobacterium seed culture with cfu / g; the weight of Bifidobacterium activation solution added to each 1 kg seed culture medium is 2 g.

[0040] In the following examples and comparative examples, the specific process for obtaining primary fermentation broth or fermentation mixture A by constant temperature fermentation at 30℃ for 24 hours is as follows: During fermentation 0-12 hours, the rotation speed is set to 0 rpm; during fermentation 12-14 hours, the rotation speed is set to 60 rpm, and the dissolved oxygen content in the fermentation tank is set to 40%; during fermentation 14-18 hours, the rotation speed is set to 150 rpm, and the dissolved oxygen content in the fermentation tank is set to 40%; during fermentation 18-24 hours, the dissolved oxygen content in the fermentation tank is set to 20%, and the stirring speed is automatically adjusted in accordance with the dissolved oxygen content; stirring is stopped after 24 hours of fermentation.

[0041] In the following examples and comparative examples, the specific process for obtaining primary fermentation broth or fermentation mixture A by constant temperature fermentation at 37℃ for 24 hours is as follows: during fermentation 0-12 hours, the rotation speed is set to 0 rpm; during fermentation 12-16 hours, the rotation speed is set to 180 rpm; during fermentation 16-19 hours, the rotation speed is set to 120 rpm; during fermentation 19-24 hours, the rotation speed is set to 50 rpm; and stirring is stopped after 24 hours of fermentation.

[0042] In the following examples and comparative examples, the specific operation of the clarification and purification is as follows: clarification filtration is performed using a 0.22µm filter membrane.

[0043] Example 1

[0044] A peony fermentation broth is prepared by the following steps:

[0045] Step 1: Crush peony roots and peony flowers at a mass ratio of 1:1 and mix them to obtain a mixture. Then, mix the mixture according to the ratio of 20g glucose, 75g of the mixture, and the remainder water per kg and adjust the pH of the solution to 7.2 to prepare a fermentation culture medium.

[0046] Step 2: Inoculate the Laenella seed culture into the fermentation medium at an inoculation rate of 3% (v / v) and ferment at a constant temperature of 30℃ for 24 hours to obtain the primary fermentation broth;

[0047] Fermentation medium was prepared by mixing 20g of glucose, 50g of primary fermentation broth, and the remainder water per kg of culture medium and adjusting the pH of the solution to 6.0. Bifidobacterium seed solution was inoculated into the fermentation medium and fermented at 37℃ for 24 hours to obtain fermentation mixture A. 3g of Bifidobacterium seed solution was inoculated into each kg of fermentation medium.

[0048] Fermentation mixture A was separated by centrifugation at 14,000 rpm. The supernatant A of the centrifuged fermentation mixture was collected and sterilized at 121°C and 0.11 MPa for 20 minutes. Then, it was clarified and purified to obtain peony fermentation broth.

[0049] Example 2

[0050] It is largely the same as Example 1, except that the mass ratio of peony root to peony flower in step 1 is 2:1.

[0051] Example 3

[0052] It is largely the same as Example 1, except that the mass ratio of peony root to peony flower in step 1 is 1:2.

[0053] Comparative Example 1

[0054] A peony fermentation broth is prepared by the following steps:

[0055] Step 1: Crush peony roots and peony flowers at a mass ratio of 1:1 and mix them to obtain a mixture. Then, mix the mixture according to the ratio of 20g glucose, 75g of the mixture, and the remainder water per kg and adjust the pH of the solution to 7.0 to prepare a fermentation culture medium.

[0056] Step 2: Inoculate the Laenella seed culture into the fermentation medium at an inoculation rate of 3% (v / v) and ferment at a constant temperature of 30℃ for 24 hours to obtain the primary fermentation broth;

[0057] Step 3: Prepare fermentation medium by mixing 20g of glucose, 50g of primary fermentation broth, and the remainder water per kg of culture medium and adjusting the pH of the solution to 7.0. Inoculate the *Laenia* seed culture medium at an inoculation rate of 3% (v / v) and ferment at 30℃ for 24 hours to obtain fermentation mixture A. Separate fermentation mixture A using a centrifuge at a speed of 14000 rpm and collect the supernatant A. Sterilize the supernatant A at 121℃ and 0.11 MPa for 20 minutes, and then clarify and purify it.

[0058] Comparative Example 2

[0059] A peony fermentation liquid, which is largely the same as that in Example 1, except that in step 1 the mixture is replaced with peony flowers.

[0060] Comparative Example 3

[0061] A peony fermentation liquid, largely the same as in Example 1, except that in step 1 the mixture is replaced with peony roots.

[0062] Comparative Example 4

[0063] A peony fermentation broth is prepared by the following steps:

[0064] Step 1: Prepare a fermentation medium by mixing 20g of glucose, 75g of the mixture, and the remainder being water per 1kg of water, and adjusting the pH of the solution to 6.0.

[0065] Step 2: Inoculate the fermentation medium with Bifidobacterium seed liquid and ferment at 37℃ for 24 hours to obtain the primary fermentation broth;

[0066] Inoculate 3g of Bifidobacterium seed solution into each 1kg of fermentation medium;

[0067] Fermentation medium was prepared by mixing 20g of glucose, 50g of primary fermentation broth, and the remainder water per kg of culture medium and adjusting the pH of the solution to 6.0. Bifidobacterium seed solution was inoculated into the fermentation medium and fermented at 37℃ for 24 hours to obtain fermentation mixture A. 3g of Bifidobacterium seed solution was inoculated into each kg of fermentation medium.

[0068] Fermentation mixture A was separated by centrifugation at 14,000 rpm. The supernatant A of the centrifuged fermentation mixture was collected and sterilized at 121°C and 0.11 MPa for 20 minutes. Then, it was clarified and purified to obtain peony fermentation broth.

[0069] Comparative Example 5

[0070] A peony fermentation broth is prepared by the following steps:

[0071] Step 1: Crush peony roots and peony flowers at a mass ratio of 1:1 and mix them to obtain a mixture. Then, mix the mixture according to the ratio of 20g glucose, 75g of the mixture, and the remainder water per kg and adjust the pH of the solution to 6.0-7.2 to prepare a fermentation culture medium.

[0072] Step 2: Inoculate the fermentation medium with Bifidobacterium seed liquid and ferment at 37℃ for 24 hours to obtain the primary fermentation broth;

[0073] Fermentation medium was prepared by mixing 20g of glucose, 50g of primary fermentation broth, and the remainder of water per kg of culture medium and adjusting the pH of the solution to 7.2. The seed culture of *Laenia* was inoculated into the fermentation medium at an inoculation rate of 3% (v / v) and fermented at a constant temperature of 30℃ for 24 hours to obtain fermentation mixture A.

[0074] Fermentation mixture A was separated by centrifugation at 14,000 rpm. The supernatant A of the centrifuged fermentation mixture was collected and sterilized at 121°C and 0.11 MPa for 20 minutes. Then, it was clarified and purified to obtain peony fermentation broth.

[0075] Efficacy testing

[0076] I. Hyaluronidase Inhibition Rate

[0077] 1. Experimental Objective and Principle

[0078] Hyaluronidase is a specific enzyme that cleaves hyaluronic acid and is involved in allergic reactions, showing a strong correlation with histamine release from mast cells. The soothing effect of a test sample can be determined using the hyaluronidase inhibition rate; a higher inhibition rate indicates a stronger soothing effect, and vice versa.

[0079] This experiment referenced the laboratory method (HMC-WI-029 hyaluronidase inhibition rate) and compared the results of the hyaluronidase inhibition rate test between the test samples and the negative control.

[0080] 2 Test Indicators

[0081] Criteria for determining hyaluronidase inhibition rate: If the hyaluronidase inhibition rate of the sample is higher than that of the negative control and there is a significant difference, the test sample can be considered to have a certain soothing effect.

[0082] 3. Experimental Materials and Methods

[0083] 3.1 Instruments and Equipment

[0084] BSA224S analytical balance;

[0085] L6s UV spectrophotometer.

[0086] 3.2 Reagents

[0087] Hyaluronidase, BR;

[0088] Sodium hyaluronate, BR.

[0089] 3.3 Test Methods

[0090] (1) Treatment of control materials and test samples

[0091] Sample group: 50% of Example 1 and Comparative Examples 1-5;

[0092] Positive control (dipotassium glycyrrhizate, purity ≥98%): diluted with water to a concentration of 3%;

[0093] Negative control: pure water.

[0094] (2) Experimental operation procedures

[0095] Set up a sample group, a sample background group, a solvent group, and a solvent background group. Each group should have 3 replicates. Add different reagent solutions to each of the four groups, shake well, and let stand at room temperature for 30 minutes to develop color. Measure the absorbance value at a wavelength of 528 nm using a UV spectrophotometer.

[0096] (3) Calculation formula

[0097]

[0098] In the formula: A—is the absorbance of the reaction solution without sample and the reaction solution containing enzyme;

[0099] B—The absorbance of the reaction solution without the sample and enzyme;

[0100] C—is the absorbance of the reaction solution containing the sample and enzyme;

[0101] D—The absorbance of the reaction solution containing the sample and the reaction solution without the enzyme.

[0102] The above testing methods were used to test Example 1 and Comparative Examples 1-5, and the results are shown in Table 1. Since the data comparison in Table 1 is mainly based on Example 1 and Comparative Examples 1-5, the data analysis only focuses on Example 1 and Comparative Examples 1-5.

[0103] Table 1. Results of hyaluronidase inhibition rate tests at 50% concentration in Example 1 and Comparative Examples 1-5

[0104] Example 1 48.208 Comparative Example 1 25.980 Comparative Example 2 33.824 Comparative Example 3 37.255 Comparative Example 4 27.206 Comparative Example 5 14.216 Positive control 62.334 negative control -2.653

[0105] According to the results in Table 1:

[0106] According to the data comparison of Example 1 and Comparative Examples 1-4, the fermentation scheme adopted in this application can significantly improve the hyaluronidase inhibition rate of peony fermentation broth.

[0107] According to the data comparison between Example 1 and Comparative Example 1, the technical solution of fermenting peony roots and peony flowers sequentially with Larynella and Bifidobacterium in this application, compared with the technical solution of simply using Larynella to ferment peony roots and peony flowers twice, can significantly improve the hyaluronidase inhibition rate of the product. It is speculated that Larynella fully decomposes peony roots and peony flowers, and the resulting primary fermentation broth has significantly improved bioavailability, thereby enhancing the subsequent fermentation effect of Bifidobacterium, thus significantly improving the hyaluronidase inhibition rate of the peony fermentation broth.

[0108] Based on the data comparison of Example 1 and Comparative Examples 2 and 3, it can be seen that when Laryn's bacterium and Bifidobacterium are fermented sequentially, the hyaluronidase inhibition rate is significantly weaker than that of Example 1 when the fermentation substrate is only peony root or peony flower. It is speculated that the fermentation products obtained by fermenting peony root and peony flower in sequence with Laryn's bacterium and Bifidobacterium produce a synergistic effect, which can synergistically improve the hyaluronidase inhibition rate of peony fermentation broth.

[0109] Based on the data comparison of Example 1 and Comparative Example 4, it can be seen that the technical solution of using Laenella and Bifidobacterium to ferment peony roots and peony flowers sequentially, compared with the technical solution of simply using Bifidobacterium to ferment peony roots and peony flowers twice, can significantly improve the hyaluronidase inhibition rate of the product. It is speculated that this may be because Laenella fully decomposes peony roots and peony flowers, and the resulting primary fermentation broth has significantly improved bioavailability, thereby enhancing the subsequent fermentation effect of Bifidobacterium, thus significantly improving the hyaluronidase inhibition rate of the peony fermentation broth.

[0110] According to the data comparison between Example 1 and Comparative Example 5, if Bifidobacterium and Laenella are used sequentially to ferment peony roots and peony flowers, the hyaluronidase inhibition rate of the resulting peony fermentation broth is significantly reduced.

[0111] In summary, the fermentation sequence and selection of fermentation strains in this application are key to improving the hyaluronidase inhibition rate.

[0112] II. Elastase Inhibition Rate

[0113] 1. Experimental Objective and Principle

[0114] Anti-wrinkle efficacy is primarily characterized by evaluating the inhibition rate of elastase in the test samples. Elastase is mainly synthesized and secreted by fibroblasts and can degrade elastin in the skin, leading to skin aging. The experimental principle of elastase inhibition is that porcine pancreatic elastase undergoes a catalytic reaction with its substrate. After adding the active substance, the absorbance changes, and the magnitude of the absorbance change reflects the inhibition rate of the elastase inhibitor.

[0115] This experiment referenced the laboratory method (HMC-WI-028 elastase inhibition rate) and compared the results of the elastase inhibition rate test between the test samples and the negative control.

[0116] 2. Test Indicators

[0117] Elastase inhibition rate: If the elastase inhibition rate of the sample is significantly higher than that of the negative control, the test sample can be considered to have anti-wrinkle effect.

[0118] 3. Experimental Materials and Methods

[0119] 3.1 Instruments and Equipment

[0120] BSA224S Analytical Balance;

[0121] RT-6100 ELISA reader.

[0122] 3.2 Reagents

[0123] elastase (porcine pancreas), BR;

[0124] N-succinyl-L-alanyl-L-alanyl-L-alanine, 98%;

[0125] Epigallocatechin gallate (EGCG), 98%.

[0126] 3.3 Test Methods

[0127] (1) Treatment of control materials and test samples

[0128] Sample group: Example 1 with a 50% concentration;

[0129] Positive control (EGCG): Dilute with pure water to a concentration of 0.1%;

[0130] Negative control: pure water.

[0131] (2) Experimental operation procedures

[0132] Set up a sample group, a sample background group, a solvent group, and a solvent background group. Each group should have 3 replicates. Add different reagent solutions to 96-well plates, shake gently, incubate at 25°C for 15 min, and then place them in an ELISA reader to measure the absorbance at 410 nm.

[0133] (3) Calculation formula

[0134]

[0135] In the formula: A — is the absorbance of the reaction solution containing enzymes but without the sample;

[0136] B—The absorbance of the reaction solution without the sample and enzyme;

[0137] C—is the absorbance of the reaction solution containing the sample and enzyme;

[0138] D—The absorbance of the reaction solution containing the sample and the reaction solution without the enzyme.

[0139] Example 1 was tested according to the above test method, and the results are shown in Table 2.

[0140] Table 2 Results of 50% elastase inhibition rate in Example 1

[0141] Example 1 31.250 Positive control 75.980 negative control -1.472

[0142] According to the results in Table 2:

[0143] The technical solution of this application has a high elastase inhibition rate and excellent anti-wrinkle effect on the skin.

[0144] III. Free radical scavenging rate

[0145] 1. Experimental Objective and Principle

[0146] Excessive production of free radicals can lead to natural skin aging and photoaging, resulting in wrinkles.

[0147] This experiment referenced the laboratory method (HMC-WI-030 DPPH free radical scavenging rate) and compared the DPPH free radical scavenging rate test results between the test samples and the negative control.

[0148] 2. Test Indicators

[0149] DPPH free radical scavenging rate: If the DPPH scavenging rate of the sample is significantly higher than that of the negative control, the test sample can be considered to have antioxidant effect.

[0150] 3. Experimental Materials and Methods

[0151] 3.1 Instruments and Equipment

[0152] BSA224S Analytical Balance;

[0153] L6s UV spectrophotometer.

[0154] 3.2 Reagents

[0155] DPPH (1,1-diphenyl-2-picrylhydrazine), 98%.

[0156] 3.3 Test Methods

[0157] (1) Treatment of control materials and test samples

[0158] Sample group: Example 1 with a 50% concentration;

[0159] Positive control (vitamin E, purity ≥96%): diluted with 95% ethanol to a concentration of 0.1%.

[0160] Negative control: pure water.

[0161] (2) Experimental operation procedures

[0162] Set up sample tubes, sample background tubes, DPPH tubes, and solvent background tubes, with three parallel tubes for each group. Add different reagent solutions to each of the four groups, gently shake to mix, and let stand at room temperature for 5 minutes. Transfer the reaction solutions of each group into a 1 cm cuvette and measure the absorbance at 517 nm.

[0163] (3) Calculation formula

[0164]

[0165] Where: T—absorbance of the sample tube, i.e., absorbance of the solution after the sample reacts with DPPH;

[0166] T0—Sample background absorbance;

[0167] The C-DPPH tube absorbance is the average of three measurements, which is the absorbance of the DPPH solution without any sample added.

[0168] C0 — Solvent background absorbance.

[0169] Example 1 was tested according to the above test method, and the results are shown in Table 3.

[0170] Table 3. Results of DPPH free radical scavenging rate in Example 1 (50%)

[0171] Example 1 50.672 Positive control 94.924 negative control -3.889

[0172] According to the results in Table 3:

[0173] The technical solution of this application has a high DPPH free radical scavenging rate and excellent skin antioxidant effect.

[0174] The embodiments presented herein are merely selected implementations based on combinations of all possible embodiments. The appended claims should not be limited to the embodiments described herein. Some numerical ranges used in the claims include sub-ranges within them, and variations within these ranges should also be covered by the appended claims.

Claims

1. A peony fermentation liquid, characterized in that, It is obtained by fermenting a mixture of peony root and peony flower with Laenella and Bifidobacterium in sequence; the weight ratio of peony root to peony flower is 1-2:1-2. The specific preparation method of the peony fermentation broth includes the following steps: Step 1: Crush peony roots and peony flowers and mix them to obtain a mixture, then use the mixture as a fermentation substrate to prepare a fermentation culture medium; Step 2: Inoculate the *Laenia* seed culture into the fermentation medium at an inoculation rate of 3% (v / v) and ferment at a constant temperature of 30℃ for 24 hours to obtain the primary fermentation broth; the *Laenia* strain was purchased from the Guangdong Provincial Microbial Culture Collection Center (GDMCC), product serial number GDMCC NO: 1.998, and the viable count of the *Laenia* seed culture was 2.6 × 10⁻⁶. 9 cfu / g; Step 3: Prepare fermentation medium by mixing 20g of glucose, 50g of primary fermentation broth, and the remainder water per kg of culture medium and adjusting the pH to 6.0; inoculate the fermentation medium with Bifidobacterium seed solution and ferment at 37℃ for 24 hours to obtain fermentation mixture A; inoculate 3g of Bifidobacterium seed solution per kg of fermentation medium; separate fermentation mixture A using a centrifuge at 14000 rpm, collect the supernatant A after centrifugation, sterilize the supernatant A at 121℃ and 0.11 MPa for 20 minutes, and then clarify and refine to obtain peony fermentation broth; The viable cell concentration in the Bifidobacterium seed culture was 5.0 x 10⁻⁶. 9 cfu / g.

2. The peony fermentation broth according to claim 1, characterized in that, The specific operation of step 1 is as follows: The peony root and peony flower were crushed and mixed to obtain a mixture. Then, the mixture was prepared by mixing the peony root and peony flower with 20g of glucose, 75g of the mixture, and the remainder water per kg of peony root and adjusting the pH of the solution to 7.0-7.

2.

3. Use of peony fermentation liquid as described in claim 1 or 2 to prepare skin care products.

4. A soothing skincare product, characterized in that, Contains 0.5-50 wt% of the peony fermentation broth as described in claim 1 or 2.

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