A peony root bark extract, a product containing the same, and a preparation method and application thereof

Abstract

The application discloses a peony root bark extract, a product containing the peony root bark extract, and a preparation method and application thereof. The preparation method of the peony root bark extract comprises the following steps: mixing peony root bark powder with a natural eutectic solvent, extracting to obtain an extract; sterilizing the extract to obtain the peony root bark extract; the natural eutectic solvent comprises a hydrogen bond donor, a hydrogen bond acceptor and water; wherein the molar ratio of the hydrogen bond donor and the hydrogen bond acceptor is 1:(2-5); the total mass ratio of the hydrogen bond donor and the hydrogen bond acceptor to the mass of water is 1:(0.8-1.5); the hydrogen bond donor comprises monosaccharide, and the hydrogen bond acceptor comprises polyhydric alcohol. The natural eutectic solvent selected by the application can effectively extract active ingredients in the peony root bark, and after extraction, the extraction solvent does not need to be removed, so that the extraction efficiency is ensured, the pollution of external solvents is reduced, resource waste is reduced, the operation is simple, the safety is high, and the application can be widely applied to the field of skin external agents.

Description

Technical Field

[0001] This application belongs to the field of extraction technology, and in particular relates to a peony root bark extract, products containing the same, preparation methods and applications thereof. Background Technology

[0002] With improved living standards, unhealthy diets, and environmental pollution, inflammatory skin conditions are becoming increasingly common. Currently, there is a wide variety of anti-inflammatory products on the market, with varying effects. However, the amount of truly effective anti-inflammatory ingredients added is very small, resulting in only moderate anti-inflammatory effects. Furthermore, the components in these formulas can burden the skin. Therefore, there is an urgent need in this field to develop a product with better anti-inflammatory effects that will not burden the skin.

[0003] Modern research has found that peony root bark contains various antioxidant active ingredients such as volatile oils, alkaloids, phytosterols, and paeonol, which have anti-inflammatory, antioxidant, and cell-repair-promoting effects. Therefore, peony root extract can be considered a product with good anti-inflammatory properties.

[0004] However, current methods for extracting active ingredients from peony root bark mainly include organic solvent extraction. For example, Chinese patent application "A peony ferment with soothing effects and its preparation method (CN115282103A)" uses anhydrous diethyl ether to extract active ingredients from peony roots. The diethyl ether needs to be removed subsequently, which is cumbersome, energy-intensive, and prone to solvent residue, reducing product safety. Furthermore, the diethyl ether is highly toxic during the preparation process, increasing operational risks.

[0005] Therefore, there is an urgent need in this field to develop a simple and safe method for extracting peony root bark, ensuring that the resulting peony root bark extract has low levels of harmful solvent residues that can irritate the skin, high product safety, and multiple excellent cosmetic effects. Summary of the Invention

[0006] The technical problem this application aims to solve is to overcome the shortcomings of existing technologies, such as cumbersome operation, high risk, poor safety due to residual organic solvents in peony root bark extract, and complex organic solvent removal processes that pollute the environment and waste resources. This application provides a peony root bark extract, products containing it, its preparation method, and its applications. The peony root bark extract obtained using the extraction method of this application has ideal antioxidant, anti-allergic, and cell repair-promoting effects. Using natural organic matter as the extraction agent, there is no need to remove the extraction solvent after extraction, the extraction process is simple, energy-saving and environmentally friendly, reduces costs, and requires a low extraction temperature.

[0007] This application adopts the following technical solution to solve the above-mentioned technical problems:

[0008] This application provides a method for preparing peony root bark extract, which includes the following steps:

[0009] Peony root bark powder was mixed with a natural eutectic solvent and extracted to obtain an extract; the extract was then sterilized to obtain the peony root bark extract.

[0010] The natural eutectic solvent comprises a hydrogen bond donor, a hydrogen bond acceptor, and water; wherein the molar ratio of the hydrogen bond donor to the hydrogen bond acceptor is 1:(2~5); the mass ratio of the total mass of the hydrogen bond donor and the hydrogen bond acceptor to the mass of water is 1:(0.8~1.5); the hydrogen bond donor comprises a monosaccharide, and the hydrogen bond acceptor comprises a polyol.

[0011] In some embodiments, the monosaccharide may include glucose and / or fructose.

[0012] In some embodiments, the polyol may include glycerol and / or 1,3-propanediol.

[0013] In a preferred embodiment, the hydrogen bond donor comprises fructose, and the hydrogen bond acceptor comprises glycerol.

[0014] In some embodiments, the molar ratio of the hydrogen bond donor to the hydrogen bond acceptor is preferably 1:(2~4.5), more preferably 1:(2~4), and even more preferably 1:(2~3).

[0015] In some embodiments, the total mass of the hydrogen bond donor and the hydrogen bond acceptor is preferably compared to the mass of the water in a ratio of 1:(1~1.3), more preferably 1:(1.1~1.3), for example 1:1.2.

[0016] In some embodiments, the water may include deionized water and / or distilled water.

[0017] In some embodiments, the preparation method of the natural eutectic solvent includes the following steps: mixing the hydrogen bond donor, the hydrogen bond acceptor, and the water.

[0018] The mixing temperature can be 70~85℃, preferably 70~80℃.

[0019] The mixing time can be determined according to conventional methods in the art, until a transparent and uniform natural eutectic solvent is formed. The mixing time is 60-120 min, preferably 100-120 min, for example 110 min.

[0020] The mixing can be carried out in a magnetic stirrer as is customary in the art.

[0021] The mixing can be carried out under stirring conditions in accordance with the conventional methods in the art, and the stirring speed can be 150~350 rpm, preferably 150~300 rpm.

[0022] In a preferred embodiment, the method for preparing the natural eutectic solvent includes the following steps: mixing a hydrogen bond donor and a hydrogen bond acceptor once to obtain solution A; and mixing solution A with water a second time to obtain the natural eutectic solvent.

[0023] The initial mixing can be carried out in a magnetic stirrer as is customary in the art.

[0024] The temperature of the first mixing step can be 70~90℃, preferably 75~85℃, for example 80℃.

[0025] The mixing time for one mixing cycle can be 60-100 min, preferably 80-100 min.

[0026] The first mixing can be carried out under stirring conditions in accordance with the conventional practice in the art, and the stirring speed can be 200~500 rpm, for example 300 rpm.

[0027] The secondary mixing can be carried out in a magnetic stirrer in accordance with conventional practices in the art.

[0028] The temperature of the secondary mixing can be 75~85℃, preferably 78~83℃, for example 80℃.

[0029] The secondary mixing time can be 30-60 minutes, preferably 40-60 minutes, for example 50 minutes.

[0030] The secondary mixing can be carried out under stirring conditions in accordance with conventional practices in the art, and the stirring speed can be 150~350 rpm, preferably 150~300 rpm.

[0031] The purpose of the first mixing and the second mixing is to form a transparent and uniform natural eutectic solvent, and the natural eutectic solvent does not precipitate crystals at room temperature.

[0032] In some embodiments, the preparation method of the peony root bark powder includes the following steps: fresh peony root bark is dried, pulverized, and sieved.

[0033] The drying method can be conventional in the art, and can generally be drying in an oven.

[0034] The drying temperature can be 50~65℃, preferably 60℃.

[0035] The drying time can be 24-60 hours, preferably 45-50 hours, for example 48 hours.

[0036] The mesh size of the sieve used for sieving can be 80 to 200 mesh, for example, 100 mesh.

[0037] In some embodiments, the mass-to-volume ratio of the peony root bark powder to the natural eutectic solvent may be 0.016~0.04 g / mL, preferably 0.018~0.03 g / mL, for example 0.02 g / mL.

[0038] In some embodiments, the extraction includes ultrasonic extraction or heated and stirred extraction.

[0039] The frequency of the ultrasonic extraction can be 30~45kHz, preferably 35~45kHz, for example 40kHz.

[0040] The power of the ultrasonic extraction can be 250~350W, preferably 280~320W, for example 300W.

[0041] The temperature of the ultrasonic extraction can be 40~60℃, preferably 45~55℃, for example 50℃.

[0042] The ultrasonic extraction time can be 90-120 min, preferably 90-110 min, for example 105 min.

[0043] The heating and stirring extraction can be carried out in a magnetic stirrer in accordance with conventional practices in the art.

[0044] The temperature of the heating and stirring extraction can be 40~80℃, preferably 50~70℃.

[0045] The heating and stirring extraction time can be 90-120 min, preferably 100-110 min.

[0046] The rotation speed of the heating and stirring extraction can be 300~800 rpm, preferably 400~600 rpm, for example 500 rpm.

[0047] In some embodiments, the extraction operation may be further followed by a centrifugation operation.

[0048] The centrifugation speed can be 3000~9000 rpm, preferably 4000~6000 rpm, for example 4800 rpm.

[0049] The radius of the centrifuge can be 8 to 15 cm, for example, 12 cm.

[0050] The centrifugation time can be 10-40 min, preferably 20-40 min, for example 30 min.

[0051] In some embodiments, the sterilization method may be high-temperature sterilization.

[0052] When the high-temperature sterilization method is used for sterilization, the sterilization temperature can be 95~110℃, preferably 95~100℃.

[0053] When the high-temperature sterilization method is used for sterilization, the sterilization time can be 20-40 minutes, preferably 25-35 minutes, for example 30 minutes.

[0054] In some embodiments, after sterilization is completed, a further step of mixing with a preservative may be included.

[0055] The preservative may include p-hydroxyacetophenone and / or 1,2-hexanediol.

[0056] When the preservative includes p-hydroxyacetophenone, the p-hydroxyacetophenone accounts for 0.5% to 1% of the mass of the material obtained after sterilization.

[0057] When the preservative includes 1,2-hexanediol, the 1,2-hexanediol accounts for 0.5% to 1% of the mass of the material obtained after sterilization.

[0058] This application also provides a peony root bark extract, which is prepared by the method described above for preparing peony root bark extract.

[0059] This application also provides the use of the peony root bark extract as described above, either directly as a product, as an additive, or as a base in the preparation of topical skin care agents.

[0060] In some embodiments, the peony root bark extract can be used as an antioxidant active ingredient in the topical skin care agent. Preferably, the antioxidant active ingredient is one with DPPH free radical scavenging activity.

[0061] In some embodiments, the peony root bark extract can be used as an anti-allergic active ingredient in the topical skin care agent. Preferably, the anti-allergic active ingredient is one that inhibits hyaluronidase.

[0062] In some embodiments, the peony root bark extract may be used as a cell-repairing component in the topical skin care agent.

[0063] This application also provides a skin care topical agent comprising the peony root bark extract as described above.

[0064] In some embodiments, the skin care topical agent may further include at least one of the following: antioxidant active ingredient, anti-allergic active ingredient, moisturizing active ingredient, anti-aging active ingredient, and anti-inflammatory active ingredient.

[0065] In some embodiments, the skin care topical agent includes a face mask, serum, or toner.

[0066] In some embodiments, the peony root bark extract accounts for 0.5% to 90% of the mass percentage of the skin care topical agent, preferably 0.5% to 70%, more preferably 1% to 10%, and even more preferably 1% to 5%.

[0067] Based on common knowledge in the field, the above-mentioned preferred conditions can be combined arbitrarily to obtain various preferred embodiments of this application.

[0068] All reagents and raw materials used in this application are commercially available.

[0069] The significant advantages of this application are as follows: It utilizes a natural eutectic solvent to effectively extract the active ingredients from peony root bark. After extraction, there is no need to remove the natural eutectic solvent, ensuring extraction efficiency. The resulting product will not burden the skin, while reducing resource waste. The process is simple and highly safe. The obtained peony root bark extract can be used directly in cosmetics, or as a cosmetic additive or substrate, possessing ideal antioxidant, anti-allergic, and anti-aging effects. Attached Figure Description

[0070] This application can be better understood by referring to the description given below in conjunction with the accompanying drawings. These drawings, together with the detailed description below, are included in and form part of this specification, and are used to further illustrate preferred embodiments of the application and explain the principles and advantages of the application. Wherein:

[0071] Figure 1 This is a comparison chart showing the DPPH free radical scavenging capabilities of the products obtained in Examples 1-8 and Comparative Examples 1-4;

[0072] Figure 2 The diagram shows a comparison of cell viability after the products prepared in Examples 1-8 and Comparative Examples 1-4 were treated with cells.

[0073] Figure 3 This is a comparison chart showing the inhibitory ability of the products prepared in Examples 1-8 and Comparative Examples 1-4 on hyaluronidase.

[0074] Figure 4 This is a comparison chart of the total phenol content in the products obtained in Examples 1-8 and Comparative Examples 1-4;

[0075] Figure 5 The chart shows a comparison of the ability of the products prepared in Examples 1-8 and Comparative Examples 1-4 to repair cells after sun exposure. Detailed Implementation

[0076] The present application is further illustrated below by way of embodiments, but these embodiments are not intended to limit the scope of the present application. Experimental methods not specifically described in the following embodiments are performed according to conventional methods and conditions, or as selected in accordance with the product instructions.

[0077] Unless otherwise specified, the experimental methods used in the following examples are conventional methods.

[0078] The peony roots used in the following examples and comparative examples were sourced from Heze City, Shandong Province.

[0079] The glucose used in the following examples and comparative examples was purchased from Beijing Bairui Biotechnology Co., Ltd.

[0080] The 1,3-propanediol used in the following examples and comparative examples was obtained from Shanghai Aladdin Biochemical Technology Co., Ltd.

[0081] The fructose in the following comparative examples is from Shanghai Aladdin Biochemical Technology Co., Ltd.

[0082] The choline chloride in the following comparative examples is from Shanghai Aladdin Biochemical Technology Co., Ltd.

[0083] The betaine in the following comparative examples is from Shanghai Aladdin Biochemical Technology Co., Ltd.

[0084] The glycerol used in the following examples and comparative examples was purchased from Heze City Mudan District Tengfei Chemical Co., Ltd.

[0085] The ultrasonic device KQ-300E ultrasonic cleaner used in the following examples and comparative examples was purchased from Kunshan Ultrasonic Instrument Co., Ltd.

[0086] Example 1

[0087] (1) Preparation of natural eutectic solvent: The molar ratio of glucose to glycerol is 1:4. 45.04g of glucose and 92.09g of glycerol are mixed once at a temperature of 80℃ for 100min. The first mixing is carried out in a magnetic stirrer at a speed of 300rpm. Solution A is obtained after the first mixing. The mass ratio of solution A to deionized water is 1:1. Solution A is mixed with 137.13g of deionized water a second time using a magnetic stirrer. The mixture is heated to dissolve. The temperature of the second mixing is 80℃ for 50min. The stirring speed of the second mixing is 300rpm. After the second mixing, a transparent and uniform natural eutectic solvent is formed, and no crystals precipitate in the natural eutectic solvent at room temperature.

[0088] (2) Fresh peony root bark was dried in an oven at 60°C for 48 hours. After drying, it was pulverized and passed through a 100-mesh sieve to obtain peony root bark powder. 4g of peony root bark powder was subjected to ultrasonic extraction with 200g of the natural eutectic solvent prepared in step (1). The ultrasonic extraction frequency was 40kHz, the ultrasonic extraction power was 300W, the ultrasonic extraction temperature was 50°C, and the ultrasonic extraction time was 105min. After ultrasonic extraction, centrifugation was performed at 4800rpm, with a centrifugation radius of 12cm and a centrifugation time of 30min. The supernatant was collected after centrifugation to obtain the extract. The extract was sterilized at 100°C for 30min to obtain peony root bark extract.

[0089] Example 2

[0090] Compared with Example 1, the only difference is that the molar ratio of glucose to glycerol in step (1) is 1:2, and the other conditions and parameters are the same as in Example 1, specifically including the following steps:

[0091] (1) Preparation of natural eutectic solvent: 90g of glucose and 92g of glycerol were mixed once at a temperature of 80℃ for 100min. The first mixing was carried out in a magnetic stirrer at a speed of 300rpm. Solution A was obtained after the first mixing. The mass ratio of solution A to deionized water was 1:1. Solution A was mixed with 182g of deionized water a second time using a magnetic stirrer. The mixture was heated to dissolve at a temperature of 80℃ for 50min. The stirring speed for the second mixing was 300rpm. After the second mixing, a transparent and uniform natural eutectic solvent was formed, and no crystals precipitated in the natural eutectic solvent at room temperature.

[0092] (2) Fresh peony root bark was dried in an oven at 60°C for 48 hours. After drying, it was pulverized and passed through a 100-mesh sieve to obtain peony root bark powder. 4g of peony root bark powder was subjected to ultrasonic extraction with 200g of the natural eutectic solvent prepared in step (1). The ultrasonic extraction frequency was 40kHz, the ultrasonic extraction power was 300W, the ultrasonic extraction temperature was 50°C, and the ultrasonic extraction time was 105min. After ultrasonic extraction, centrifugation was performed at 4800rpm, with a centrifugation radius of 12cm and a centrifugation time of 30min. The supernatant was collected after centrifugation to obtain the extract. The extract was sterilized at 100°C for 30min to obtain peony root bark extract.

[0093] Example 3

[0094] The preparation method is the same as in Example 1, with a molar ratio of glucose to glycerol of 1:4.5, and other conditions and parameters are the same as in Example 1. Specifically, the steps include:

[0095] (1) Preparation of natural eutectic solvent: 36g of glucose and 82.8g of glycerol were mixed once at a temperature of 80℃ for 100min. The first mixing was carried out in a magnetic stirrer at a speed of 300rpm. Solution A was obtained after the first mixing. The mass ratio of solution A to deionized water was 1:1. Solution A was mixed with 118.8g of deionized water a second time using a magnetic stirrer and heated to dissolve. The temperature of the second mixing was 80℃ for 50min and the stirring speed was 300rpm. After the second mixing, a transparent and uniform natural eutectic solvent was formed, and no crystals precipitated in the natural eutectic solvent at room temperature.

[0096] (2) Fresh peony root bark was dried in an oven at 60°C for 48 hours. After drying, it was pulverized and passed through a 100-mesh sieve to obtain peony root bark powder. 4g of peony root bark powder was subjected to ultrasonic extraction with 200g of the natural eutectic solvent prepared in step (1). The ultrasonic extraction frequency was 40kHz, the ultrasonic extraction power was 300W, the ultrasonic extraction temperature was 50°C, and the ultrasonic extraction time was 105min. After ultrasonic extraction, centrifugation was performed at 4800rpm, with a centrifugation radius of 12cm and a centrifugation time of 30min. The supernatant was collected after centrifugation to obtain the extract. The extract was sterilized at 100°C for 30min to obtain peony root bark extract.

[0097] Example 4

[0098] Compared with Example 1, the only difference is that in step (1), the ratio of solution A to water is 1:1.2, and other conditions and parameters are the same as in Example 1, specifically including the following steps:

[0099] (1) Preparation of natural eutectic solvent: 45.04g of glucose and 92.09g of glycerol were mixed once at a temperature of 80℃ for 100min. The first mixing was carried out in a magnetic stirrer at a speed of 300rpm. Solution A was obtained after the first mixing. The mass ratio of solution A to deionized water was 1:1.2. Solution A was mixed with 164.56g of deionized water a second time using a magnetic stirrer. The mixture was heated to dissolve. The temperature of the second mixing was 80℃ for 50min. The stirring speed of the second mixing was 300rpm. After the second mixing, a transparent and uniform natural eutectic solvent was formed, and no crystals precipitated in the natural eutectic solvent at room temperature.

[0100] (2) Fresh peony root bark was dried in an oven at 60°C for 48 hours. After drying, it was pulverized and passed through a 100-mesh sieve to obtain peony root bark powder. 4g of peony root bark powder was ultrasonically extracted with 200g of the natural eutectic solvent prepared in step (1). The ultrasonic extraction frequency was 40kHz, the ultrasonic extraction power was 300W, the ultrasonic extraction temperature was 50°C, and the ultrasonic extraction time was 105min. After ultrasonic extraction, centrifugation was performed at 4800rpm, with a centrifugation radius of 12cm and a centrifugation time of 30min. The supernatant was collected after centrifugation to obtain extract B. Extract B was sterilized at 100°C for 30min to obtain peony root bark extract.

[0101] Example 5

[0102] Compared with Example 1, the only difference is that the ultrasonic extraction time is 90 min, while other conditions and parameters are the same as in Example 1, specifically including the following steps:

[0103] (1) Preparation of natural eutectic solvent: The molar ratio of glucose to glycerol is 1:4. 45.04g of glucose and 92.09g of glycerol are mixed once at a temperature of 80℃ for 100min. The first mixing is carried out in a magnetic stirrer at a speed of 300rpm. Solution A is obtained after the first mixing. The mass ratio of solution A to deionized water is 1:1. Solution A is mixed with 137.13g of deionized water a second time using a magnetic stirrer. The mixture is heated to dissolve. The temperature of the second mixing is 80℃ for 50min. The stirring speed of the second mixing is 300rpm. After the second mixing, a transparent and uniform natural eutectic solvent is formed, and no crystals precipitate in the natural eutectic solvent at room temperature.

[0104] (2) Fresh peony root bark was dried in an oven at 60°C for 48 hours. After drying, it was pulverized and passed through a 100-mesh sieve to obtain peony root bark powder. 4g of peony root bark powder was subjected to ultrasonic extraction with 200g of the natural eutectic solvent prepared in step (1). The ultrasonic extraction frequency was 40kHz, the ultrasonic extraction power was 300W, the ultrasonic extraction temperature was 50°C, and the ultrasonic extraction time was 90min. After ultrasonic extraction, centrifugation was performed at 4800rpm, with a centrifugation radius of 12cm and a centrifugation time of 30min. The supernatant was collected after centrifugation to obtain the extract. The extract was sterilized at 100°C for 30min to obtain peony root bark extract.

[0105] Example 6

[0106] Compared with Example 1, the only difference is that the extraction method is heating and stirring extraction, and the operation steps of heating extraction are as follows:

[0107] (1) Preparation of natural eutectic solvent: The molar ratio of glucose to glycerol is 1:4. 45.04g of glucose and 92.09g of glycerol are mixed once at a temperature of 80℃ for 100min. The first mixing is carried out in a magnetic stirrer at a speed of 300rpm. Solution A is obtained after the first mixing. The mass ratio of solution A to deionized water is 1:1. Solution A is mixed with 137.13g of deionized water a second time using a magnetic stirrer. The mixture is heated to dissolve. The temperature of the second mixing is 80℃ for 50min. The stirring speed of the second mixing is 300rpm. After the second mixing, a transparent and uniform natural eutectic solvent is formed, and no crystals precipitate in the natural eutectic solvent at room temperature.

[0108] (2) Fresh peony root bark was dried in an oven at 60°C for 48 hours. After drying, it was pulverized and passed through a 100-mesh sieve to obtain peony root bark powder. 4g of peony root bark powder and 200g of the natural eutectic solvent prepared in step (1) were heated and stirred at 50°C for 100 minutes and 500 rpm for extraction. After extraction, centrifugation was performed at 4800 rpm with a centrifugation radius of 12 cm for 30 minutes. The supernatant was collected after centrifugation to obtain the extract. The extract was sterilized at 100°C for 30 minutes to obtain peony root bark extract.

[0109] Example 7

[0110] Compared with Example 1, the difference is that glucose, glycerol and water are mixed simultaneously, while other conditions and parameters are the same as in Example 1. The specific operation is as follows:

[0111] (1) Mix 45.04g glucose, 92.09g glycerol and 137.13g deionized water at a temperature of 80℃ for 100min. Mix in a magnetic stirrer at a speed of 300rpm to form a transparent and uniform natural eutectic solvent. No crystals precipitate in the natural eutectic solvent at room temperature.

[0112] (2) Fresh peony root bark was dried in an oven at 60°C for 48 hours. After drying, it was pulverized and passed through a 100-mesh sieve to obtain peony root bark powder. 4g of peony root bark powder was subjected to ultrasonic extraction with 200g of the natural eutectic solvent prepared in step (1). The ultrasonic extraction frequency was 40kHz, the ultrasonic extraction power was 300W, the ultrasonic extraction temperature was 50°C, and the ultrasonic extraction time was 105min. After ultrasonic extraction, centrifugation was performed at 4800rpm, with a centrifugation radius of 12cm and a centrifugation time of 30min. The supernatant was collected after centrifugation to obtain the extract. The extract was sterilized at 100°C for 30min to obtain peony root bark extract.

[0113] Example 8

[0114] Compared with Example 1, the only difference is the type of natural eutectic solvent; all other conditions and parameters are the same as in Example 1, specifically including the following steps:

[0115] (1) Preparation of natural eutectic solvent: The molar ratio of fructose and 1,3-propanediol is 1:4. 60.05g of fructose and 101.45g of 1,3-propanediol are mixed once at a temperature of 80℃ for 100min. The first mixing is carried out in a magnetic stirrer at a speed of 300rpm. Solution A is obtained after the first mixing. The mass ratio of solution A to deionized water is 1:1. Solution A is mixed with 161.5g of deionized water a second time using a magnetic stirrer and heated to dissolve. The temperature of the second mixing is 80℃ for 50min and the stirring speed is 300rpm. After the second mixing, a transparent and uniform natural eutectic solvent is formed, and no crystals precipitate in the natural eutectic solvent at room temperature.

[0116] (2) Fresh peony root bark was dried in an oven at 60°C for 48 hours. After drying, it was pulverized and passed through a 100-mesh sieve to obtain peony root bark powder. 4g of peony root bark powder was subjected to ultrasonic extraction with 200g of the natural eutectic solvent prepared in step (1). The ultrasonic extraction frequency was 40kHz, the ultrasonic extraction power was 300W, the ultrasonic extraction temperature was 50°C, and the ultrasonic extraction time was 105min. After ultrasonic extraction, centrifugation was performed at 4800rpm, with a centrifugation radius of 12cm and a centrifugation time of 30min. The supernatant was collected after centrifugation to obtain the extract. The extract was sterilized at 100°C for 30min to obtain peony root bark extract.

[0117] Comparative Example 1

[0118] Extraction was performed by replacing 200g of natural eutectic solvent with 200g of deionized water, with other conditions and parameters the same as in Example 1.

[0119] Comparative Example 2

[0120] Compared with Example 1, the only difference is the type of natural eutectic solvent; all other conditions and parameters are the same as in Example 1, specifically including the following steps:

[0121] (1) Preparation of natural eutectic solvent: The molar ratio of choline chloride to glycerol is 1:4. 69.81g of choline chloride and 184.18g of glycerol are mixed once at a temperature of 80℃ for 100min. The first mixing is carried out in a magnetic stirrer at a speed of 300rpm. Solution A is obtained after the first mixing. The mass ratio of solution A to deionized water is 1:1. Solution A and 254g of deionized water are mixed a second time using a magnetic stirrer. The mixture is heated to dissolve. The temperature of the second mixing is 80℃ for 50min. The stirring speed of the second mixing is 300rpm. After the second mixing, a transparent and uniform natural eutectic solvent is formed, and no crystals precipitate in the natural eutectic solvent at room temperature.

[0122] (2) Fresh peony root bark was dried in an oven at 60°C for 48 hours. After drying, it was pulverized and passed through a 100-mesh sieve to obtain peony root bark powder. 4g of peony root bark powder was ultrasonically extracted with 200g of the natural eutectic solvent prepared in step (1). The ultrasonic extraction frequency was 40kHz, the ultrasonic extraction power was 300W, the ultrasonic extraction temperature was 50°C, and the ultrasonic extraction time was 105min. After ultrasonic extraction, centrifugation was performed at 4800rpm, with a centrifugation radius of 12cm and a centrifugation time of 30min. The supernatant was collected after centrifugation to obtain extract B. Extract B was sterilized at 100°C for 30min to obtain peony root bark extract.

[0123] Comparative Example 3

[0124] Compared with Example 1, the only difference is the type of natural eutectic solvent; all other conditions and parameters are the same as in Example 1, specifically including the following steps:

[0125] (1) Preparation of natural eutectic solvent: The molar ratio of betaine to citric acid is 1:1:. 58.58 g of betaine and 96.07 g of citric acid are mixed once at a temperature of 80 °C for 100 min. The first mixing is carried out in a magnetic stirrer at a speed of 300 rpm. Solution A is obtained after the first mixing. The mass ratio of solution A to deionized water is 1:1. Solution A is mixed with 154.65 g of deionized water a second time using a magnetic stirrer. The mixture is heated to dissolve. The temperature of the second mixing is 80 °C for 50 min. The stirring speed of the second mixing is 300 rpm. After the second mixing, a transparent and uniform natural eutectic solvent is formed, and no crystals precipitate in the natural eutectic solvent at room temperature.

[0126] (2) Fresh peony root bark was dried in an oven at 60°C for 48 hours. After drying, it was pulverized and passed through a 100-mesh sieve to obtain peony root bark powder. 4g of peony root bark powder was ultrasonically extracted with 200g of the natural eutectic solvent prepared in step (1). The ultrasonic extraction frequency was 40kHz, the ultrasonic extraction power was 300W, the ultrasonic extraction temperature was 50°C, and the ultrasonic extraction time was 105min. After ultrasonic extraction, centrifugation was performed at 4800rpm, with a centrifugation radius of 12cm and a centrifugation time of 30min. The supernatant was collected after centrifugation to obtain extract B. Extract B was sterilized at 100°C for 30min to obtain peony root bark extract.

[0127] Comparative Example 4

[0128] The preparation method is the same as in Example 1, except that the mass ratio of solution A to water is 1:4. All other conditions and parameters are the same as in Example 1, and the specific steps include:

[0129] 45.04g of glucose and 92.09g of glycerol were mixed once at 80℃ for 100 minutes using a magnetic stirrer at 300 rpm. Solution A was obtained after the first mixing. The mass ratio of solution A to deionized water was 1:4. 40g of solution A was then mixed a second time with 160g of deionized water using a magnetic stirrer, heated to dissolve, at 80℃ for 50 minutes at 300 rpm. The second mixing resulted in a transparent, homogeneous natural eutectic solvent, and no crystals precipitated at room temperature.

[0130] (2) Fresh peony root bark was dried in an oven at 60°C for 48 hours. After drying, it was pulverized and passed through a 100-mesh sieve to obtain peony root bark powder. 4g of peony root bark powder was subjected to ultrasonic extraction with 200g of the natural eutectic solvent prepared in step (1). The ultrasonic extraction frequency was 40kHz, the ultrasonic extraction power was 300W, the ultrasonic extraction temperature was 50°C, and the ultrasonic extraction time was 105min. After ultrasonic extraction, centrifugation was performed at 4800rpm, with a centrifugation radius of 12cm and a centrifugation time of 30min. The supernatant was collected after centrifugation to obtain the extract. The extract was sterilized at 100°C for 30min to obtain peony root bark extract.

[0131] Example 1: DPPH free radical scavenging experiment

[0132] DPPH is an early synthesized organic free radical often used to evaluate the hydrogen-donating capacity of antioxidants. It is very stable in organic solvents, exhibits a purple color, and has a characteristic absorption peak at 517 nm. When it encounters a free radical scavenger, the lone pair electrons of DPPH are paired, causing it to fade, meaning the absorbance at the maximum absorption wavelength decreases. Therefore, the scavenging effect of a sample on DPPH free radicals can be evaluated by measuring the change in absorbance.

[0133] The preparation steps of the test solution are as follows: dilute the peony root bark extracts prepared in Examples 1-8 and Comparative Examples 1-4 by 10 times with deionized water to obtain the test solution.

[0134] The specific experimental steps of the DPPH free radical scavenging experiment are as follows:

[0135] (1) Take an equal volume (1 mL) of the test solution and 2 × 10 -4 Mix the mol / L DPPH solution thoroughly (tube A1).

[0136] (2) Take an equal volume (1 mL) of anhydrous ethanol and 2 × 10 -4 Mix the mol / L DPPH solution thoroughly (tube A2).

[0137] (3) Take an equal volume (1 mL) of anhydrous ethanol and mix it with the test solution (A3 tube).

[0138] (4) After reacting in the dark for 30 minutes, the absorbance values ​​of tubes A1, A2, and A3 were measured at 517 nm (the testing equipment was a Sunrise microplate reader, manufactured by Diken Trading Co., Ltd.). Each group was tested in triplicate, and the average value was taken. The DPPH free radical scavenging rate was calculated using the following formula. The results are shown in Table 1 and Figure 1 .

[0139] DPPH free radical scavenging rate = [(A2+A3)-A1] / A2×100%.

[0140] Table 1

[0141]

[0142] from Figure 1 As can be seen from the results in Table 1, the DPPH free radical scavenging ability of the peony root bark extracts prepared in Examples 1-8 of this application is significantly higher than that of the products prepared in Comparative Examples 1-4. Figure 1 In this context, ns indicates no statistically significant difference compared to Example 1; p < 0.05 indicates a statistically significant difference compared to Example 1; p < 0.01 indicates a statistically significant difference compared to Example 1, with a significant reduction; p < 0.001 indicates a highly significant statistical difference compared to Example 1, representing a highly significant reduction.

[0143] Example 2: Human skin fibroblast toxicity experiment

[0144] This experiment used human skin fibroblasts from the Chinese Scientific Cell Bank to verify the cytotoxicity of the products prepared in the above examples and comparative examples.

[0145] Reagents: 0.25% (EDTA-containing) trypsin was manufactured by GIBCO, USA; DMEM medium was manufactured by GIBCO, USA; penicillin-dextrose antibody was manufactured by Corning, USA; CCK-8 was manufactured by Beijing Bairui Biotechnology Co., Ltd.; fetal bovine serum was manufactured by GIBCO, USA; phosphate buffer was manufactured by Beijing Bairui Biotechnology Co., Ltd.

[0146] Equipment: The WJ-80A-Ⅱ CO2 constant temperature incubator is manufactured by Shanghai Shengke Instrument Equipment Co., Ltd.; the Sunrise microplate reader is manufactured by Diken Trading Co., Ltd.; the TL80-2 medical centrifuge is manufactured by Jiangsu Tianli Medical Equipment Co., Ltd.; and the NUNC 96-well cell culture plate is manufactured by Thermo Fisher Scientific.

[0147] 1. Experimental steps:

[0148] The products obtained in Examples 1-8 and Comparative Examples 1-4 were respectively prepared into test solutions with a volume percentage of 1% using serum-free DMEM medium.

[0149] Human skin fibroblasts were cultured in a solution containing 10% fetal bovine serum and 1% penicillin-dextrose antibody (1×10⁻⁶). 5 Cells were cultured in DMEM medium containing 100 mg / L penicillin and 100 mg / L streptomycin. Cells were grown in a 37°C, 5% CO2 saturated humidity incubator. When cell confluence reached 85% or higher, logarithmic growth phase cells were digested with 0.05% trypsin, and the digestion was terminated with serum-containing DMEM. Cells were counted using a cell counting chamber, and the cell suspension concentration was adjusted to 7 × 10⁻⁶ cells / mL. 4 Cells were seeded at a rate of 100 μL / well in 96-well plates and incubated at 37°C and 5% CO2 for 12 h. The old culture medium was removed, and the cells were washed twice with phosphate-buffered saline (PBS). 100 μL of the filtered and sterilized test solution was added to each well of the experimental group, with six replicates per test solution. The control group contained cells and was treated with 100 μL of serum-free DMEM medium. The blank control group contained no cells and was treated with 100 μL of PBS. The cells were then incubated at 37°C and 5% CO2 for 24 h. Then, 10 μL of CCK-8 solution was added to each well, and the cells were incubated for another 3 h. The absorbance was measured at 450 nm, and the cell viability was calculated. The results are shown in Table 2. Figure 2 .

[0150] The formula for calculating cell viability is as follows:

[0151] Cell viability (%) = (A experimental group - A blank control group) / (A control group - A blank control group) × 100%.

[0152] Table 2

[0153]

[0154] from Figure 2 As can be seen from the results in Table 2, the cell viability of the peony root bark extracts prepared in Examples 1-8 of this application is significantly higher than that of the products prepared in Comparative Examples 1-4. Figure 2 In this context, ns indicates no statistically significant difference compared to Example 1; p < 0.05 indicates a statistically significant difference compared to Example 1; p < 0.01 indicates a statistically significant difference compared to Example 1, with a significant reduction; p < 0.001 indicates a highly significant statistical difference compared to Example 1, representing a highly significant reduction.

[0155] Example 3: Hyaluronidase Inhibition Experiment

[0156] Hyaluronidase is a lysosome that breaks down polysaccharides. It hydrolyzes potassium hyaluronate to produce β-N-acetylglucosamine, which condenses with acetylacetone under alkaline conditions to form the chromogen 2-methyl-3-diacetylpyrrole derivative. The chromogen reacts with Ehrlich's reagent in concentrated hydrochloric acid and ethanol to produce a colorimetric reaction. Hyaluronidase is strongly correlated with inflammation and allergies and is involved in type I hypersensitivity reactions. Therefore, in vitro hyaluronidase inhibition assays can serve as a rapid method for assessing anti-allergy levels.

[0157] Reagents: Hyaluronidase, sodium hyaluronate, anhydrous ethanol, sodium hydroxide, anhydrous sodium carbonate, concentrated hydrochloric acid, p-dimethylaminobenzaldehyde, acetylacetone, glacial acetic acid, anhydrous calcium chloride.

[0158] Equipment: The Sunrise microplate reader is manufactured by Diken Trading Co., Ltd.; the digital display constant temperature water bath is from the Medical Equipment Factory of Shanghai Boxun Industrial Co., Ltd.

[0159] The test sample solution was prepared by diluting Examples 1-8 and Comparative Examples 1-4 by 2 times.

[0160] Take 0.1 mL of CaCl2 aqueous solution (0.25 mmol / L) and 0.5 mL of hyaluronidase solution (100 U / mL) and incubate at 37°C in a water bath for 20 min; add 0.5 mL of the above-prepared test sample solution and continue incubation for 20 min; then add 0.5 mL of sodium hyaluronate solution (0.5 mg / mL), incubate at 37°C in a water bath for 30 min, remove and place at room temperature for 5 min; add 0.1 mL of NaOH aqueous solution (0.4 mol / L) and 0.5 mL of acetylacetone solution (3.5 mL acetylacetone dissolved in... The solution was added to 50 mL of 1.0 mol / L sodium carbonate solution and heated in a boiling water bath for 15 min, then immediately transferred to an ice-water bath for cooling for 5 min. 1.0 mL of Ehrlich's reagent (0.8 g p-dimethylaminobenzaldehyde dissolved in 15 mL concentrated hydrochloric acid and 15 mL anhydrous ethanol) was added dropwise, and the solution was diluted with 3.0 mL of anhydrous ethanol. The solution was allowed to stand at room temperature for 20 min for color development. The absorbance was measured at 540 nm using a spectrophotometer. Each group was performed in triplicate, and the average value was taken. The hyaluronidase inhibition rate was calculated using the following formula. The results are shown in Table 3. Figure 3 .

[0161] Hyaluronidase inhibition rate = [(AB) - (CD)] / (AB) × 100%;

[0162] Where: A—Absorbance value of the control solution (using acetate buffer solution instead of the test sample solution); B—Absorbance value of the control blank solution (using acetate buffer solution instead of the test sample solution and enzyme solution); C—Absorbance value of the test sample solution; D—Absorbance value of the test sample blank solution (using acetate buffer solution instead of enzyme solution);

[0163] Table 3

[0164]

[0165] from Figure 3 As can be seen from the results in Table 3, the hyaluronidase inhibition rate of the peony root bark extracts prepared in Examples 1-8 of this application is significantly higher than that of the products prepared in Comparative Examples 1-4. The peony root bark extracts prepared in the examples of this application have ideal anti-allergic effects. Figure 3 In this context, ns indicates no statistically significant difference compared to Example 1; p < 0.05 indicates a statistically significant difference compared to Example 1; p < 0.01 indicates a statistically significant difference compared to Example 1, with a significant reduction; p < 0.001 indicates a highly significant statistical difference compared to Example 1, representing a highly significant reduction.

[0166] Example 4: Determination of total phenol content

[0167] In alkaline solutions, polyphenolic compounds can reduce tungsticolaminosic acid (W... 6+ Become W 3+ It produces a blue compound with maximum absorption at 760 nm, and the intensity of the color is positively correlated with the polyphenol content. Gallic acid (or pyrogallic acid) is generally used as a reference standard, and the total polyphenol content in the extract is expressed as the amount equivalent to gallic acid.

[0168] Experimental drugs:

[0169] 1. Pyrogallic acid standard;

[0170] 2. Folin-phenol test solution;

[0171] 3.26.7% Na2CO3 solution: Weigh 26.7g of Na2CO3 solid powder, add distilled water to 100g, and stir to dissolve.

[0172] Experimental instruments: precision balance, spectrophotometer.

[0173] Experimental procedure:

[0174] 1. Plotting the standard curve

[0175] Accurately weigh 44.3 mg of pyrogallol standard, vacuum dried to constant weight, and dissolve it in distilled water to a final volume of 100 mL. Take 2.0, 4.0, 8.0, 12.0, 16.0, and 20.0 mL of each solution and place them in 100 mL volumetric flasks, respectively, and dilute to the mark with distilled water. Add 1 mL of each of these solutions to a 10 mL colorimetric tube, then add 1 mL of distilled water, 0.5 mL of a 2-fold diluted Folin-phenol solution, and 1.5 mL of a 26.7 wt% Na₂CO₃ aqueous solution. Dilute to a final volume of 10 mL with water and allow to react at room temperature for 2 h. Measure the absorbance at 760 nm. Plot the absorbance against the standard concentration to obtain a typical standard curve. The total phenol standard curve is: y = 6.3036x + 0.0048, R₀ = 0.0048. 2 =0.9997.

[0176] 2. Sample determination

[0177] Take 1 mL of the test solution (the products prepared in Examples 1-8 and Comparative Examples 1-4 diluted 30 times) and add it to a 10 mL test tube. Then add 1 mL of distilled water, 0.5 mL of 2-fold diluted Folin-phenol test solution, and 1.5 mL of 26.7% Na2CO3 solution sequentially. Make up the volume to 10 mL with water and react at room temperature for 2 hours. Measure the absorbance at 760 nm. Perform three replicates for each group, and take the average value. The results are shown in Table 4. Calculate the polyphenol content in the products prepared in Examples 1-8 and Comparative Examples 1-4 based on the standard curve and the dilution factor of the test solution. The results are shown in Table 4. Figure 4 .

[0178] Table 4

[0179]

[0180] from Figure 4 As can be seen from the results in Table 4, the total phenol content of the peony root bark extracts prepared in Examples 1-8 of this application is significantly higher than the total phenol content of the products prepared in Comparative Examples 1-4. Figure 4 In this context, ns indicates no statistically significant difference compared to Example 1; p < 0.05 indicates a statistically significant difference compared to Example 1; p < 0.01 indicates a statistically significant difference compared to Example 1, with a significant reduction; p < 0.001 indicates a highly statistically significant difference compared to Example 1, representing a highly significant reduction.

[0181] Example 5: External Post-Sun Repair

[0182] 1. Experimental steps:

[0183] The products obtained in the above examples and comparative examples were respectively prepared into experimental test solutions with a volume percentage of 1% using serum-free DMEM medium.

[0184] HaCaT cells were cultured in a solution containing 10% fetal bovine serum and 1% penicillin-antibody (1×10⁻⁶ cells / mL). 5 Cells were cultured in DMEM medium containing 100 mg / L penicillin and 100 mg / L streptomycin. Cells were grown in a 37°C, 5% CO2 saturated humidity incubator. When cell confluence reached 85% or higher, logarithmic growth phase cells were digested with 0.05% trypsin, and the digestion was terminated with serum-containing DMEM. Cells were counted using a cell counting chamber, and the cell suspension concentration was adjusted to 7 × 10⁻⁶ cells / mL. 4 Cells were seeded at a rate of 100 μL per well in 96-well plates and incubated at 37°C with 5% CO2 for 12 h. After incubation, the old culture medium was removed, and the cells were washed twice with phosphate-buffered saline (PBS). Then, 100 μL of PBS was added. Both the model and experimental groups were irradiated with UVB at a dose of 15 mJ / cm². 2 The negative control group was not irradiated. PBS was discarded. 100 μL of serum-free DMEM medium was added to the model group and negative control group. 100 μL of the filtered and sterilized experimental test solution prepared above was added to each well of the experimental group, with 6 replicates for each test solution. The blank control group was cell-free and contained 100 μL of PBS. The cells were then incubated at 37℃ and 5% CO2 for 24 h. Then, 10 μL of CCK-8 solution was added to each well, and the cells were incubated for another 3 h. The absorbance was measured at 450 nm, and the cell viability of each group was calculated. The results are shown in Table 5. Figure 5 .

[0185] The formula for calculating cell viability is as follows:

[0186] Cell viability (%) = (A experimental group or model group - A blank control group) / (A negative control group - A blank control group) × 100%.

[0187] Table 5

[0188]

[0189] from Figure 5 As can be seen from the results in Table 5, the cell survival rate of the peony root bark extracts prepared in Examples 1-8 of this application is significantly higher than that of the products prepared in Comparative Examples 1-4, indicating that the peony root bark extracts prepared in this application have ideal post-sun exposure repair effects. Figure 5 In this context, ns indicates no statistically significant difference compared to the model group; p < 0.05 indicates a statistically significant difference compared to the model group; p < 0.01 indicates a statistically significant difference compared to the model group, representing a significant decrease; p < 0.001 indicates a highly statistically significant difference compared to the model group, or a highly significant decrease. ### p < 0.001 indicates a highly significant difference compared to Example 1, a significant increase. Finally, it should be noted that the terms "comprising," "including," or any other variations thereof in this application are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such process, method, article, or apparatus.

[0190] Although this application has been disclosed above through the description of specific embodiments, it should be understood that those skilled in the art can devise various modifications, improvements, or equivalents to this application within the spirit and scope of the appended solutions. Such modifications, improvements, or equivalents should also be considered to be included within the scope of protection claimed in this application.

Claims

1. A method for preparing a peony root bark extract, characterized in that, It includes the following steps: Peony root bark powder is mixed with a natural eutectic solvent and extracted to obtain an extract. The extract is then sterilized to obtain the peony root bark extract. The natural eutectic solvent comprises a hydrogen bond donor, a hydrogen bond acceptor, and water. The molar ratio of the hydrogen bond donor to the hydrogen bond acceptor is 1:(2~5). The mass ratio of the total mass of the hydrogen bond donor and the hydrogen bond acceptor to the mass of water is 1:(0.8~1.5). The hydrogen bond donor comprises a monosaccharide, and the hydrogen bond acceptor comprises a polyol.

2. The method for preparing the peony root bark extract as described in claim 1, characterized in that, The method for preparing the peony root bark extract satisfies at least one of the following conditions: The monosaccharides include glucose and / or fructose; The polyols include glycerol and / or 1,3-propanediol; The molar ratio of the hydrogen bond donor to the hydrogen bond acceptor is 1:(2~4.5). The total mass ratio of the hydrogen bond donor and the hydrogen bond acceptor to the mass of the water is 1:(1~1.3). The water includes deionized water and / or distilled water; The mass-to-volume ratio of the peony root bark powder to the natural eutectic solvent is 0.016~0.04 g / mL.

3. The method for preparing peony root bark extract as described in claim 2, characterized in that, The method for preparing the peony root bark extract satisfies at least one of the following conditions: The molar ratio of the hydrogen bond donor to the hydrogen bond acceptor is 1:(2~4). The total mass ratio of the hydrogen bond donor and the hydrogen bond acceptor to the mass of the water is 1:(1.1~1.3). The mass-to-volume ratio of the peony root bark powder to the natural eutectic solvent is 0.018~0.03 g / mL.

4. The method for preparing the peony root bark extract as described in claim 3, characterized in that, The method for preparing the peony root bark extract satisfies at least one of the following conditions: The molar ratio of the hydrogen bond donor to the hydrogen bond acceptor is 1:(2~3). The total mass ratio of the hydrogen bond donor and the hydrogen bond acceptor to the mass ratio of the water is 1:1.2; The mass-to-volume ratio of the peony root bark powder to the natural eutectic solvent is 0.02 g / mL.

5. The method for preparing the peony root bark extract as described in claim 1, characterized in that, The preparation method of the natural eutectic solvent includes the following steps: mixing the hydrogen bond donor, the hydrogen bond acceptor and the water.

6. The method for preparing the peony root bark extract as described in claim 5, characterized in that, The mixing temperature is 70~85℃; the mixing time is 60~120min.

7. The method for preparing the peony root bark extract as described in claim 6, characterized in that, The mixing temperature is 70~80℃; the mixing time is 100~120min.

8. The method for preparing the peony root bark extract as described in claim 5, characterized in that, The mixing is carried out in a magnetic stirrer; when the magnetic stirrer is used for mixing, the mixing speed is 150~350 rpm.

9. The method for preparing the peony root bark extract as described in claim 8, characterized in that, When using the magnetic stirrer for mixing, the mixing speed is 150~300 rpm.

10. The method for preparing the peony root bark extract according to any one of claims 5-9, characterized in that, The preparation method of the natural eutectic solvent includes the following steps: mixing the hydrogen bond donor and the hydrogen bond acceptor once to obtain solution A; mixing solution A with water a second time to obtain the natural eutectic solvent.

11. The method for preparing the peony root bark extract as described in claim 10, characterized in that, The method for preparing the peony root bark extract must meet at least one of the following conditions: The temperature for the first mixing step is 70~90℃; The mixing time for each mixing cycle is 60-100 minutes. The first mixing is carried out under stirring conditions, and the stirring speed is 200~500 rpm; The temperature for the secondary mixing is 75~85℃; The secondary mixing time is 30-60 minutes; The secondary mixing is carried out under stirring conditions, and the stirring speed is 150~350 rpm.

12. The method for preparing the peony root bark extract as described in claim 11, characterized in that, The method for preparing the peony root bark extract must meet at least one of the following conditions: The temperature for the first mixing step is 75~85℃; The mixing time for one cycle is 80-100 minutes; The first mixing is carried out under stirring conditions, and the stirring speed is 300 rpm; The temperature for the secondary mixing is 78~83℃; The secondary mixing time is 40-60 minutes; The secondary mixing is carried out under stirring conditions, and the stirring speed is 150~300 rpm.

13. The method for preparing the peony root bark extract as described in claim 1, characterized in that, The preparation method of the peony root bark powder includes the following steps: drying, pulverizing and sieving fresh peony root bark.

14. The method for preparing the peony root bark extract as described in claim 13, characterized in that, The method for preparing the peony root bark extract must meet at least one of the following conditions: The drying process involves drying in an oven. The drying temperature is 50~65℃; The drying time is 24-60 hours; The sieve used for sieving has a mesh size of 80 to 200.

15. The method for preparing the peony root bark extract as described in claim 14, characterized in that, The method for preparing the peony root bark extract must meet at least one of the following conditions: The drying temperature is 60°C; The drying time is 45-50 hours; The sieve used for sieving has a mesh size of 100.

16. The method for preparing the peony root bark extract as described in claim 1, characterized in that, The extraction includes ultrasonic extraction or heating and stirring extraction.

17. The method for preparing the peony root bark extract as described in claim 16, characterized in that, The method for preparing the peony root bark extract must meet at least one of the following conditions: The frequency of the ultrasonic extraction is 30~45kHz; The power of the ultrasonic extraction is 250~350W; The temperature for ultrasonic extraction is 40~60℃; The ultrasonic extraction time is 90-120 min; The temperature for heating and stirring extraction is 40~80℃; The heating and stirring extraction time is 90-120 min; The rotation speed for heating and stirring extraction is 300~800 rpm.

18. The method for preparing the peony root bark extract as described in claim 17, characterized in that, The method for preparing the peony root bark extract must meet at least one of the following conditions: The frequency of the ultrasonic extraction is 35~45kHz; The power of the ultrasonic extraction is 280~320W; The temperature for ultrasonic extraction is 45~55℃; The ultrasonic extraction time is 90-110 min; The temperature for the heating and stirring extraction is 50~70℃; The heating and stirring extraction time is 100-110 min; The rotation speed for heating and stirring during extraction is 400~600 rpm.

19. The method for preparing the peony root bark extract as described in claim 18, characterized in that, The method for preparing the peony root bark extract must meet at least one of the following conditions: The frequency of the ultrasonic extraction is 40kHz; The power of the ultrasonic extraction is 300W; The temperature for ultrasonic extraction is 50℃; The ultrasonic extraction time was 105 min; The heating and stirring extraction speed is 500 rpm.

20. The method for preparing the peony root bark extract as described in claim 1, characterized in that, The method for preparing the peony root bark extract satisfies at least one of the following conditions: The extraction operation is followed by a centrifugation operation; The sterilization method is high-temperature sterilization. After sterilization is completed, the process also includes mixing with preservatives.

21. The method for preparing the peony root bark extract as described in claim 20, characterized in that, The method for preparing the peony root bark extract satisfies at least one of the following conditions: The centrifuge speed is 3000~9000 rpm; the centrifuge radius is 8~15 cm; preferably, the centrifuge time is 10~40 min; When the high-temperature sterilization method is used for sterilization, the sterilization temperature is 95~110℃ and the sterilization time is 20~40min. The preservatives include p-hydroxyacetophenone and / or 1,2-hexanediol.

22. The method for preparing the peony root bark extract as described in claim 21, characterized in that, The method for preparing the peony root bark extract satisfies at least one of the following conditions: The centrifuge speed is 4000~6000 rpm; the centrifuge radius is 12 cm; the centrifuge time is 20~40 min; When the high-temperature sterilization method is used for sterilization, the sterilization temperature is 95~100℃ and the sterilization time is 25~35min; The p-hydroxyacetophenone accounts for 0.5% to 1% of the mass of the material obtained after sterilization; the 1,2-hexanediol accounts for 0.5% to 1% of the mass of the material obtained after sterilization.

23. A peony root bark extract, characterized in that, It is prepared by the method for preparing peony root bark extract as described in any one of claims 1 to 22.

24. The use of the peony root bark extract as described in claim 23, either directly as a product, as an additive, or as a base, in the preparation of topical skin care agents.

25. The application as described in claim 24, characterized in that, The application must satisfy at least one of the following conditions: The peony root bark extract is used as the antioxidant active ingredient in the topical skin care agent. The peony root bark extract is used as the anti-allergic active ingredient in the topical skin care agent. The peony root bark extract is used as a cell-repairing component in the topical skincare product.

26. The application as described in claim 25, characterized in that, The antioxidant active ingredient is an antioxidant active ingredient with DPPH free radical scavenging effect; The anti-allergic active ingredient is an anti-allergic active ingredient with hyaluronidase inhibitory activity.

27. A topical skin care agent, characterized in that, Includes the peony root bark extract as described in claim 23.

28. The skin care topical agent as described in claim 27, characterized in that, The topical skin care agent must meet at least one of the following conditions: The skin care topical agent also includes at least one of the following: antioxidant active ingredients, anti-allergic active ingredients, moisturizing active ingredients, anti-aging active ingredients, and anti-inflammatory active ingredients; The skin care topical agents include face masks, serums, or toners; The peony root bark extract accounts for 1% to 10% of the mass of the topical skin care agent.

29. The skin care topical agent as described in claim 28, characterized in that, The peony root bark extract accounts for 1% to 5% of the total mass of the topical skin care agent.

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