Skin care material containing peony root bark extract and method for preparing the same

Abstract

This invention belongs to the field of cosmetic raw material technology, specifically relating to a skincare product raw material containing peony root bark extract and its preparation method. In the skincare product raw material containing peony root bark extract provided by this invention, the volume ratio of peony root bark concentrate: 1,3-butanediol:water is 0.9:6:3.1. The peony root bark concentrate is first prepared using a full-process membrane separation technology. Specifically, the aqueous extract of peony root bark is first clarified and filtered using a ceramic membrane with a pore size of 1μm. Then, it undergoes three-stage retention separation using ultrafiltration membranes with molecular weight cutoffs of 10000 Da, 5000 Da, and 3000 Da. Finally, a fourth-stage retention separation is performed using a nanofiltration membrane with a molecular weight cutoff of 500 Da, followed by concentration via reverse osmosis. The cosmetic raw material prepared by this invention is rich in membrane-purified active ingredients of peony root bark, is green, safe, and non-irritating, and has multiple effects such as moisturizing, whitening, anti-inflammatory, antibacterial, acne-reducing, and oil-controlling properties.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of cosmetic raw materials, and particularly relates to a skin care product raw material containing peony root bark extract and a preparation method thereof. BACKGROUND

[0002] At present, there are various cosmetics with whitening function, but many of them achieve the whitening effect by adding chemical additives, such as common hydroquinone, cysteine, glutathione, vitamin A, glucocorticoid, etc. These compounds have good whitening effect, but the long-term use of chemical reagents will produce certain toxic and side effects on the skin, causing skin sensitivity.

[0003] In addition, in order to inhibit the activity of harmful microorganisms, most cosmetics need to add artificially synthesized organic or inorganic antibacterial agents and preservatives, but these synthetic antibacterial agents and preservatives may cause strong irritation to the eyes, skin or sense of smell to some extent, and show weak antibacterial activity to gram-negative bacteria over a long period of time.

[0004] Research has found that active substances extracted from natural plants have less toxicity and much fewer side effects, therefore, in order to meet the growing demand of skin care effect of cosmetic formula, the application of natural plant extract as raw material in cosmetics has gradually become a global research hotspot in the development of skin care products.

[0005] Peony root bark is rich in paeonol, paeoniflorin, paeonol polysaccharide, gallic acid, volatile oil, phytosterol and other bioactive components. Research shows that these active ingredients have bacteriostatic and bactericidal physiological activity, and paeonol can also inhibit the generation of melanin and prevent the formation of freckles, and has a certain whitening effect. Therefore, peony root bark has become one of the main sources of skin care product raw materials.

[0006] The method of extracting and purifying active ingredients from peony root bark directly determines the types and contents of active ingredients in the extract, so the peony root bark extract obtained by different methods may have different effects when used as a skin care product raw material due to the difference in active ingredients.

[0007] Membrane separation technology is a technology that uses the selectivity of membrane materials to separate, purify and concentrate different components in the feed liquid. Since this process is a physical process that does not require phase change and the addition of additives, the biological activity of the separated components is maintained at a high level. Therefore, membrane separation technology has been widely used in the processing of active ingredients in various natural plants. For example, patent CN115177558A discloses using a membrane with a molecular weight cut-off of 5000-10000 Da to ultrafilter the acetone extract of Sophora flavescens; patent CN110354029A uses an ultrafiltration membrane with a molecular weight cut-off of 600-100000 combined with an adsorption resin to extract soybean isoflavones; and patent CN114259440A discloses using a ceramic membrane to purify a skin care product mixture prepared by the inventors.

[0008] In addition, patents CN104031157A, CN103087128A, CN111363619A, etc. disclose the use of ultrafiltration technology to extract active ingredients such as polysaccharides, paeonol, and peony seed protein from peony seed meal.

[0009] However, there are few reports on the use of ultrafiltration membrane technology to intercept active ingredients from peony root bark.

[0010] After extensive searching, it was found that patent CN101390962A discloses a method of using a membrane to ultrafilter the extract of peony root bark, and then using a macroporous resin to separate paeonol. However, the extraction rate of paeonol obtained by this method is only 2.47%, and the content of paeonol is 47.5%. Therefore, the method results in a large loss of active ingredients in the extract, leading to a low extraction rate of active ingredients. SUMMARY

[0011] To solve the above technical problems, the present application provides a skin care product raw material containing a peony root bark extract and a preparation method thereof.

[0012] The skin care product raw material prepared in the present application comprises a peony root bark concentrate, a glycol solvent, and water, and the volume ratio of the peony root bark concentrate, the glycol solvent, and water is 0.5-2:6:3-4.

[0013] Preferably, the amount of the skin care product raw material containing the peony root bark extract added to the skin care product is 0.5-2.0% (v / v).

[0014] Preferably, the amount of the skin care product raw material containing the peony root bark extract added to the skin care product is 1.0-2.0% (v / v).

[0015] Preferably, the diol solvent is selected from at least one of 1,2-propanediol, 1,3-propanediol, 1,2-butanediol, 1,3-butanediol, 1,4-butanediol, 1,2-hexanediol, and 1,6-hexanediol.

[0016] Preferably, the skin care raw material containing the peony root bark extract includes the following components: peony root bark concentrate, 1,3-butanediol, and water, and the volume ratio of the peony root bark concentrate, 1,3-butanediol, and water is 0.5-2:6:3-4.

[0017] More preferably, in the skin care raw material, the volume ratio of the peony root bark concentrate, 1,3-butanediol, and water is 0.9:6:3.1.

[0018] The peony root bark concentrate is compounded with a diol reagent, and preferably the diol reagent is 1,3-butanediol, because it has good moisturizing and water-locking effects and can also achieve a certain degree of preservation effect, which is conducive to the storage and effect of the peony root bark concentrate. As shown in the experimental results of the present application, the skin care raw material composition obtained by compounding the peony root bark concentrate with the diol reagent can inhibit melanin synthesis when the volume ratio is greater than 0.5%, and the raw material after compounding can achieve the effect of inhibiting melanin production when applied to cosmetics, thereby achieving good whitening effect.

[0019] In addition, the content of the diol reagent needs to be strictly controlled. If the content is too high, it will irritate the skin, but if the content is too low, it will affect the moisturizing and water-locking effects and the skin feel.

[0020] The preparation method of the skin care raw material containing the peony root bark extract provided by the present application includes the following steps:

[0021] (1) The dried peony root bark is crushed and sieved, and then water extraction is performed at 40-60 DEG C to obtain a clear filtrate of the peony root bark water extract;

[0022] (2) The clear filtrate of the peony root bark water extract obtained in (1) is subjected to a molecular weight cut-off of 500-10000 Da, and finally concentrated by a reverse osmosis membrane to obtain a peony root bark concentrate;

[0023] (3) The peony root bark concentrate obtained in (2) is mixed with 1,3-butanediol and water at a volume ratio of 0.5-2:6:3-4, and the pH of the mixed solution is adjusted to 4.5-6.0, and then filtered through a polypropylene membrane to obtain a skin care raw material containing the peony root bark extract.

[0024] In the above steps, preferably, in (1), a ceramic membrane with a pore size of 1 mu m is used to clarify and filter the water extract of the peony root bark.

[0025] Preferably, the ceramic membrane is a tubular SiC ceramic membrane, which adopts a fluid separation process in the form of "cross-flow filtration". The raw liquid flows at high speed in the membrane tube, and the clear permeate containing small molecular components is driven by pressure to permeate the membrane outward in the vertical direction, and the turbid concentrate containing large molecular components is intercepted by the membrane, so as to separate, concentrate and purify the fluid. Compared with other ceramic membranes made of metal and Al2O3, the tubular SiC ceramic membrane has the characteristics of large membrane flux and high temperature resistance. The filter cake layer is not easy to accumulate, and the high-efficiency and continuous operation of filtration can be maintained. In addition, the membrane has the characteristics of long flow channel, low pretreatment precision requirement for the liquid, easy cleaning and high filtration efficiency.

[0026] Preferably, in (2), three-stage interception separation is carried out by using ultrafiltration membranes with molecular weight cut-off of 10000 Da, 5000 Da and 3000 Da in sequence, and then fourth-stage interception is carried out by using a nanofiltration membrane with molecular weight cut-off of 500 Da.

[0027] Preferably, in (3), citric acid solution and NaOH solution are used to adjust the pH of the mixed solution.

[0028] In the present application, water is used to extract the active ingredients in the root bark of tree peony. Since water has a large polarity, the water-soluble polysaccharides and other components contained in the root bark of tree peony will inevitably be dissolved therein. Therefore, the water extract of the root bark of tree peony contains a large amount of solid impurities and components with relatively large molecular weight, such as polysaccharides and starch. If a membrane with a molecular weight cut-off of 3000 Da is directly used for ultrafiltration interception, most of the high molecular weight impurities and ineffective components will cause membrane pollution, reduce the membrane flux and shorten the service life of the membrane. Therefore, by gradually reducing the molecular weight cut-off of the membrane, the loss of the filter membrane can be reduced, the utilization rate of the membrane can be improved, and the filtration effect can be improved.

[0029] The present application has the following advantages:

[0030] Firstly, the whole-process membrane separation technology significantly improves the purity of the active ingredients in the water extract of the root bark of tree peony.

[0031] The molecular weight of the effective components in the root bark of Paeonia suffruticosa is mostly not more than 1000 Da, wherein the molecular weight of paeonol is 166.174 Da, the molecular weight of paeoniflorin is 480.462 Da, and the ineffective components are above 50000 Da, therefore, in the application, the water extract of the root bark of Paeonia suffruticosa is pretreated by using a 1 mu m SiC ceramic membrane to remove the large solid impurities and macromolecular substances with relatively large molecular weight in the water extract, and then the water extract of the root bark of Paeonia suffruticosa is separated and purified by using the four-stage membrane cutting technology, so that the impurities and ineffective components in the filtrate are gradually separated and removed from large to small, the ultrafiltration membrane with a certain cutting molecular weight is selected to perform the grading cutting, which not only can reduce the influence of the extract on the membrane flux, membrane pollution and membrane service life in the membrane separation process, but also can retain the main effective components in the root bark of Paeonia suffruticosa, and finally the filtrate is concentrated by using a reverse osmosis membrane, and the effective component purity of the concentrated solution of the root bark of Paeonia suffruticosa obtained is higher.

[0032] Secondly, a green and safe skin care product raw material containing the root bark of Paeonia suffruticosa extract is provided.

[0033] By mixing and compounding the root bark of Paeonia suffruticosa concentrate, 1,3-butanediol and water at a volume ratio of 0.9:6:3.1, without adding any preservative, only by means of the active components contained in the root bark of Paeonia suffruticosa concentrate, a good bacteriostatic effect can be achieved, in addition, the 1,3-butanediol added in the raw material is a small molecule alcohol, which is beneficial to the dissolution and absorption of the root bark of Paeonia suffruticosa cut-off liquid after being mixed with water, and has a certain preservative effect, and each raw material is green and safe.

[0034] Thirdly, the skin care product raw material containing the root bark of Paeonia suffruticosa extract provided by the application has an inhibition rate of tyrosinase activity as high as 86.43% when the addition amount is 2.0%, so as to inhibit the synthesis of cell melanin and achieve a good whitening effect, in addition, when the extract is treated at a concentration of 2.0%, the average concentration of inflammatory factor IL-1 alpha is 4.94 pg / mL, and the concentration level of IL-6 is 6.42 pg / mL, which shows a stronger inhibition activity of inflammatory factor than the blank group, and the average bacteriostatic rate of the skin care product raw material is 78.1% after acting on Malassezia furfur for 20 min, the bacteriostatic rate of Propionibacterium acnes is 78.5%, and in addition, the composition also shows a good oil control effect. BRIEF DESCRIPTION OF DRAWINGS

[0035] Figure 1 It is the preparation process flow chart of the skin care product raw material containing the root bark of Paeonia suffruticosa extract in embodiment 1 of the application;

[0036] Figure 2 It is the in vitro whitening evaluation result chart of the root bark of Paeonia suffruticosa extract in test example 1 of the application;

[0037] Figure 3Inhibition result of the root-bark of Paeonia suffruticosa extract on inflammatory factor IL-1α in the test example 2 of the present application;

[0038] Figure 4 Inhibition result of the root-bark of Paeonia suffruticosa extract on inflammatory factor IL-6 in the test example 2 of the present application;

[0039] Figure 5 Sebaceous gland cell lipid droplet staining result of each group in the test example 4 of the present application;

[0040] Figure 6 Oil control efficacy evaluation result chart of the root-bark of Paeonia suffruticosa extract in the test example 4 of the present application. DETAILED DESCRIPTION

[0041] In order to enable those skilled in the art to better understand the present application, the present application will be further described in conjunction with specific embodiments.

[0042] Example 1

[0043] The skin care product raw material containing the root-bark of Paeonia suffruticosa extract is prepared by the following steps:

[0044] (1) The dried root-bark of Paeonia suffruticosa is crushed, sieved, and water extracted at 60℃. The water extract of the root-bark of Paeonia suffruticosa is filtered by using a ceramic membrane with a pore size of 1 μm to obtain a clear filtrate of the water extract of the root-bark of Paeonia suffruticosa;

[0045] (2) The clear filtrate of the water extract of the root-bark of Paeonia suffruticosa obtained in (1) is subjected to three-stage ultrafiltration separation by using ultrafiltration membranes with a molecular weight cut-off of 10000 Da, 5000 Da, and 3000 Da, respectively, and then subjected to fourth-stage cut-off separation by using a nanofiltration membrane with a molecular weight cut-off of 500 Da. Finally, reverse osmosis membrane is used for concentration to obtain a root-bark of Paeonia suffruticosa concentrate;

[0046] (3) The concentrate in (2) is mixed with 1,3-butanediol and water at a volume ratio of 0.9:6:3.1, and the pH of the mixture is adjusted to 5.0 by using citric acid and NaOH solution. Finally, the mixture is filtered by using a polypropylene membrane to obtain a skin care product raw material containing the root-bark of Paeonia suffruticosa extract.

[0047] In this embodiment, the preparation process of the root-bark of Paeonia suffruticosa concentrate is shown in the attached Figure 1

[0048] In the cosmetic raw material composition prepared in this embodiment, 1,3-butanediol is added, so that water molecules can be adsorbed to a certain extent, and the product has a strong water supplementing and moisturizing effect, and the texture is relatively refreshing without stickiness.

[0049] Example 2

[0050] ​Different from Example 1, the obtained Paeonia suffruticosa Andr. root-bark concentrated solution and 1,2-propanediol were compounded in a volume ratio of 0.9:6:3.1 in (3).

[0051] Compared with the cosmetic raw material prepared in Example 1, the cosmetic raw material composition obtained by compounding 1,2-propanediol and Paeonia suffruticosa Andr. root-bark concentrated solution in this example has better viscosity and hygroscopicity, but the moisturizing effect and refreshing property are not as good as those of the raw material prepared in Example 1, and the raw material shows greater irritation to the skin due to the addition of 1,2-propanediol.

[0052] Example 3

[0053] Different from Example 1, the volume ratio of Paeonia suffruticosa Andr. root-bark concentrated solution, 1,3-butanediol and water in (3) is 2:4:4.

[0054] The cosmetic raw material composition obtained by using the combination ratio in this example has a good moisturizing effect, but the clarity and skin feel are slightly worse than those of the product in Example 1.

[0055] The reason for this phenomenon may be that the main active ingredient in the Paeonia suffruticosa Andr. root-bark concentrated solution is paeonol, which is insoluble in cold water and easily soluble in alcohol. Therefore, in this example, the content of the concentrated solution is increased, and the content of 1,3-butanediol is greatly reduced, so that the clarity of the obtained composition after mixing of the components is affected, which may further affect the gloss of the prepared cosmetic.

[0056] In addition, a high content of 1,3-butanediol is beneficial to the improvement of the moisturizing properties of the raw material and the cosmetic, but it also easily makes the skin feel heavy and the skin burden heavier.

[0057] Example 4

[0058] Different from Example 1, the volume ratio of Paeonia suffruticosa Andr. root-bark concentrated solution, 1,3-butanediol and water in (3) is 0.5:6.5:3.

[0059] The prepared Paeonia suffruticosa Andr. root-bark extract has a significantly increased content of 1,3-butanediol compared with Example 1, and shows stronger irritation to the skin. At the same time, the concentration of the Paeonia suffruticosa Andr. root-bark concentrated solution is too low, which inevitably leads to weak whitening and bacteriostatic effects of the skin care product raw material.

[0060] Comparative Examples 1-3

[0061] Different from Example 1, the water extract of Paeonia suffruticosa Andr. is filtered by using SiC ceramic membranes with pore sizes of 0.1 μm, 0.5 μm and 3 μm, respectively, in (1) to obtain the clarified filtrate of Paeonia suffruticosa Andr. water extract.

[0062] Comparative Example 4

[0063] Different from Example 1, the water extract of the root bark of Paeonia suffruticosa was filtered by using an Al2O3 ceramic membrane with a pore size of 1 μm in (1) to obtain a clear filtrate of the water extract of the root bark of Paeonia suffruticosa.

[0064] The turbidity removal rate and the recovery rate of the membrane flux of the crude water extract of the root bark of Paeonia suffruticosa filtered by using ceramic membranes with different pore sizes and types in Example 1 and Comparative Examples 1-4 are shown in Table 1.

[0065] Table 1 Filtration by using ceramic membranes with different types

[0066] Group Turbidity removal rate (%) Membrane flux recovery rate (%) Example 1 98.8 98.7 Comparative Example 1 98.9 97.2 Comparative Example 2 98.7 97.8 Comparative Example 3 97.3 98.7 Comparative Example 4 97.8 97.8

[0067] The data in the table show that the turbidity removal rate and the recovery rate of the membrane flux of the crude water extract of the root bark of Paeonia suffruticosa filtered by using SiC ceramic membranes with different pore sizes in Comparative Examples 1-3 are both higher than 97%, but compared with Example 1, the use of ceramic membranes with smaller pore sizes can reduce the membrane flux and increase the possibility of membrane pollution, and the use of ceramic membranes with larger pore sizes can affect the clarity and impurity removal effect of the filtrate, which is not conducive to subsequent membrane separation operations.

[0068] Although the Al2O3 ceramic membrane used in Comparative Example 4 can also be used for the clarification filtration of the water extract of the root bark of Paeonia suffruticosa, the turbidity removal rate and the recovery rate of the membrane flux are not as good as those of the SiC ceramic membrane.

[0069] In order to verify the performance (whitening, anti-inflammatory, antibacterial, acne-removing, and oil-controlling) of the skin care product raw material containing the extract of the root bark of Paeonia suffruticosa provided in the present application, the skin care product raw material containing the extract of the root bark of Paeonia suffruticosa prepared in Example 1 (hereinafter referred to as the extract of the root bark of Paeonia suffruticosa) was subjected to the following test verification.

[0070] Test Example 1

[0071] The skin care product raw material prepared in Example 1 was subjected to in vitro whitening evaluation, and the whitening effect of the product in Example 1 on the melanin synthesis of human melanoma cells was determined by cell experiments to determine the whitening effect of the extract of the root bark of Paeonia suffruticosa. The specific operation is as follows:

[0072] (1) Collect G361 human melanoma cells in the logarithmic growth phase and inoculate cells into a 24-well plate at a seeding density of 2×10 5 cells / well, and incubate in an incubator (37℃, 5% CO2) for 24 h. According to the cell toxicity results, add the drugs according to the experimental grouping in Table 2 below, and use untreated cells as a blank control (BC). At the same time, use an experiment group to which kojic acid is added as a positive control (PC), and set 3 parallels for each group.

[0073] Table 2 Experimental grouping

[0074]

[0075] (2) After adding the drug, continue to culture in the incubator (37℃, 5% CO2) for 24h, discard the supernatant, add 0.5mL of 10% DMSO-containing 1M NaOH, and incubate in a 80℃ constant temperature oven for 1h, after recovering to room temperature, transfer 200μL per well to a 96-well plate, take 10% DMSO-containing 1M NaOH as a blank control, read the absorbance value at 405nm, and calculate the relative inhibition rate of cell melanin synthesis.

[0076] The calculation formula of the inhibition rate (%) of cell melanin synthesis is as follows:

[0077]

[0078] The inhibition test results of each experimental group on cell melanin synthesis are shown in the following Figure 2 and Table 3.

[0079] Table 3 Inhibition test results of each experimental group on cell melanin synthesis

[0080]

[0081] Kojic acid, also known as kojic acid, has the chemical name of 5-hydroxy-2-hydroxymethyl-1, 4-pyrone, which is a melanin-specific inhibitor. After entering the skin cells, it can complex with copper ions in the cells, change the structure of tyrosinase, prevent the activation of tyrosinase, and thus inhibit the formation of melanin. It has been formulated into various cosmetics.

[0082] From the data in the following Figure 2 and Table 3, it can be seen that the inhibition rate of kojic acid with a concentration of 0.3% on cell melanin synthesis is relatively high, which is 85.75%, and different concentrations of peony root bark extract also show certain inhibition effect on cell melanin synthesis. The data shows that with the gradual increase of the concentration of peony root bark extract from 0.5% to 1.0% to 2.0% (V / V), the inhibition ability on cell melanin synthesis also gradually increases, and when the content of peony root bark extract is 0.5%, the inhibition effect of the extract is poor, and when the content is 2.0%, it shows better effect than kojic acid. This not only shows that the prepared peony root bark extract has the ability to inhibit cell melanin synthesis, but also when the concentration is 2.0%, the inhibition effect on cell melanin synthesis is higher than that of kojic acid, and the inhibition rate reaches 86.43%.

[0083] Test Example 2

[0084] The Paeonia suffruticosa Andr. root bark extract prepared in Example 1 was subjected to in vitro anti-inflammatory evaluation, and the average concentration of IL-6 and IL-1α was determined by ELISA kit to determine the in vitro anti-inflammatory efficacy of the Paeonia suffruticosa Andr. root bark extract.

[0085] The specific method is as follows:

[0086] (1) Collecting RAW264.7 cells in logarithmic growth phase, inoculating cells into a 24-well plate at a seeding density of 1×10 5 cells per well, and placing into an incubator (37℃, 5% CO2) for culture. According to the cytotoxicity result, preparing the test work solution as the sample group according to Table 4, using the cells without any treatment as blank control (BC), using dexamethasone as positive control (PC), using the cells only stimulated by LPS without other treatment as negative control (NC), and setting 3 parallel holes for each group;

[0087] Table 4 Experimental grouping

[0088]

[0089] (2) Dosing: according to the experimental design table of Table 4, when the plating rate of cells in the 24-well plate reaches 40% to 60%, grouping and dosing, and setting 3 parallel holes for each group;

[0090] (3) Stimulation: according to the experimental design table of Table 4, adding 100 μL 10×LPS work solution into the dosed hole plate, shaking the hole plate left and right to mix the drug in the hole plate, and the final concentration of LPS is 1 μg / mL, and continuing to culture in the incubator (37℃, 5% CO2) for 24 h;

[0091] (4) Sampling: after incubation, collecting the cell culture supernatant in an EP tube (remark: according to the detection index to determine the amount of collected sample), and after collection, storing the sample in a-80℃ refrigerator for frozen preservation;

[0092] (5) Detection of inflammatory factor content

[0093] IL-1α content detection: according to the operation instruction of Mouse IL-1α ELISA kit for detection, and IL-6 content detection: according to the operation instruction of Mouse IL-6 ELISA kit for detection.

[0094] The detection results of IL-1α content are shown in Table 5, and the detection results of IL-6 content are shown in Table 6, and the corresponding detection results are shown in the accompanying Figures 3-4 .

[0095] Table 5 Detection results of IL-1α content

[0096] Sample name / group Average concentration (pg / mL) SD p-value Blank control (BC) 5.27 0.80 / Negative control (NC) 8.27 0.42 0.008 ## ]] Positive control (PC) 5.14 0.39 0.030* Peony root bark extract (0.5%) 5.86 0.36 0.047* Peony root bark extract (1.0%) 5.60 0.51 0.038* Peony root bark extract (2.0%) 4.94 0.44 0.028*

[0097] Note: When using t-test method for statistical analysis, the significance of comparison between NC group and BC group is indicated by #, p-value < 0.05 is indicated by # , and p-value < 0.01 is indicated by ## . The significance of comparison between sample group and PC group and NC group is indicated by *, p-value < 0.05 is indicated by *, and p-value < 0.01 is indicated by **. The same applies hereinafter.

[0098] Attached Figure 3 The experimental results in Table 5 show that the IL-1α content in the cells of the blank control group was 5.27 pg / mL, and the IL-1α content in the cells of the positive control group to which dexamethasone was added was 5.14 pg / mL. It can be seen that dexamethasone showed a certain inhibitory effect on IL-1α, but the inhibition rate of dexamethasone on IL-1α was only 2.47%. Dexamethasone is a long-acting adrenal glucocorticoid that can inhibit the level of inflammatory factors, and is therefore widely used in clinical practice for anti-inflammatory and anti-allergic purposes.

[0099] In addition, the skin care product raw material prepared by the present application containing the peony root bark extract showed almost no inhibition of IL-1α when the addition amount was 0.5% to 1.0%. Only when the addition amount was as high as 2.0%, did the product show a high inhibitory effect on the IL-1α level, with an average concentration of IL-1α of 4.94 pg / mL. After calculation, the inhibition rate of the product on IL-1α was 6.26%, i.e., at a high concentration, the product had a high inhibitory efficiency on the inflammatory factor IL-1α.

[0100] Table 6: Results of detection of IL-6 content

[0101] Sample name / group Average concentration (pg / mL) SD p-value Blank control (BC) 8.23 1.87 / Negative control (NC) 27.76 1.97 0.017 # ]] Positive control (PC) 9.42 1.00 0.009** Peony root bark extract (0.5%) 9.63 1.67 0.015* Peony root bark extract (1.0%) 7.91 2.02 0.001** Peony root bark extract (2.0%) 6.42 0.76 0.003**

[0102] The results in Table 6 show that the inhibitory effect of the product in Example 1 on the inflammatory factor IL-6 was much higher than that of dexamethasone. When the concentration of the extract was 2.0%, the concentration level of the inflammatory factor IL-6 was 6.42 pg / mL, and the inhibition rate was 21.99% relative to the IL-6 concentration level of the blank control.

[0103] Test Example 3: Anti-dandruff and Malassezia inhibition rate

[0104] I. According to T / GDCA 010-2022 "Anti-dandruff product anti-dandruff effect test method" article 7, under the test temperature 20℃±1℃ conditions, through 5 repeated tests, the product prepared in Example 1 has an average inhibition rate of 78.1%, 64.4%, and 57.1% on Pityrosporum ovale at concentrations of 2.0%, 1.0%, and 0.5% for 20 minutes, respectively, which has a significant difference (p<0.05) compared with the control group, as shown in Table 7.

[0105] Table 7 Inhibition effect of samples with different concentrations on Pityrosporum ovale

[0106]

[0107] Note: Negative control group: PBS, no bacterial growth in the medium.

[0108] After inspection, the product of Example 1 showed an inhibition effect at different concentrations of 2.0%, 1.0%, and 0.5% on Pityrosporum ovale after 20 minutes, with an average inhibition rate of 78.1%, 64.4%, and 57.1%, respectively.

[0109] II. Anti-acne-P. acnes inhibition rate

[0110] Test basis: QB / T 2738-2012 "Evaluation method for antibacterial and bacteriostatic effect of daily chemical products" 7.3.

[0111] The specific operation of the bacteriostatic test is as follows:

[0112] (1) Preparation of bacterial suspension: Dilute the test bacterial suspension with PBS solution as required, with a concentration of 1×10 4 ~ 9×10 4 CFU / mL;

[0113] (2) According to the requirements, use the sample to be tested (prepared with sterile water at concentrations of 2.0%, 1.0%, and 0.5% (V / V)), and act on P. acnes for 20 minutes at a temperature of 20℃±1℃, with three repeated tests;

[0114] (3) After 12 hours of action to the set time, 0.5mL of the sample and test bacterial mixture is added to a test tube containing 4.5mL of sterilized PBS, and mixed thoroughly;

[0115] (4) Take 1mL of the sample solution and place it on a nutrient agarose culture medium plate, and place the plate in an anaerobic environment in a incubator at 37℃ for 72 hours.

[0116] Table 8 Inhibition effect of samples with different concentrations on P. acnes

[0117]

[0118]

[0119] Note: negative control group: PBS, medium without bacterial growth.

[0120] After testing, the product containing the peony root bark extract in Example 1 has bacteriostatic effect on Propionibacterium acnes at 2.0%, 1.0%, and 0.5% concentrations for 20 min, and the bacteriostatic rates are 78.5%, 62.9%, and 54.1% respectively, which has good acne-removing ability.

[0121] Study on the oil control effect of the product of Test Example 4

[0122] Excessive secretion of sebum is the main factor causing various skin problems, and the main factor affecting the secretion of sebum is the disorder of hormone level in the body. Sebum gland cells can synthesize a large amount of oil and secrete it to the outside of the cells in the form of lipid droplets, which affects the function of the cells and causes various skin problems. The amount of lipid droplets is the main indicator for measuring the degree of oil control. In the present application, SZ95 sebaceous gland cells are used as the research object, 5α-DHT is used as the induction condition, and isotretinoin is used as the positive marker substance to evaluate the inhibitory effect of the cosmetic on the secretion of cell sebum and judge the oil control effect.

[0123] (1) Cell inoculation: inoculate cells into a 24-well plate at a seeding density of 1×10 5 cells / well, and incubate in an incubator (37℃, 5% CO2) overnight;

[0124] (2) Liquid preparation: prepare the working solution of the test substance according to Table 9 of the experimental design;

[0125] (3) Drug administration: according to the experimental design table 9, when the cell plating rate in the 24-well plate reaches 40%-60%, group administration is performed, and each group has 3 replicate wells. Continue to culture in an incubator (37℃, 5% CO2) for 24h;

[0126] (4) Discard the culture medium, rinse the cells with PBS, and after fixation, add Nile red for staining for 15min. After rinsing, take photos under a fluorescence microscope;

[0127] (5) Result analysis: quantitatively analyze the fluorescence intensity by using Image Pro Plus software.

[0128] Table 9 Experimental design

[0129]

[0130] The staining results of the sebaceous gland cell lipid droplets in the present example are shown in the accompanying Figure 5 photograph.

[0131] The test results of the lipid droplet content are shown in Table 10 and the following figures. Figure 6

[0132] Table 10 Test results of the lipid droplet content

[0133]

[0134] As shown in the table, the Nile red fluorescence signal of the NC group is significantly higher than that of the BC group (p<0.01), indicating that the 5α-DHT lipid droplet modeling in this experiment is successful.

[0135] Compared with the NC group, the Nile red fluorescence signal of the PC group is significantly lower (p<0.01), indicating that the positive control detection in this experiment is effective.

[0136] Under the conditions of this experiment, compared with the NC group, the skin care product raw material containing the peony root bark extract prepared in the present application has a significantly reduced Nile red fluorescence signal (p<0.05) at the concentrations of 1% and 2% (V / V), indicating that the product has good oil control efficacy.​

Claims

1. A skin care raw material containing an extract of Paeonia suffruticosa root bark, characterized in that, The components include peony root-bark concentrate, 1,3-butanediol, and water, and the volume ratio of peony root-bark concentrate:1,3-butanediol:water is 0.9:6:3.1; The skin care product raw material containing peony root-bark extract is prepared by the following method: (1) dry peony root-bark is ground and sieved, and then water extraction is performed at 60°C; the water extract of peony root-bark is filtered by using a SiC ceramic membrane with a pore size of 1 μm to obtain a clear filtrate of peony root-bark water extract; (2) the clear filtrate of peony root-bark water extract obtained in (1) is subjected to three-stage ultrafiltration separation by using ultrafiltration membranes with a molecular weight cut-off of 10000 Da, 5000 Da, and 3000 Da, respectively, and then subjected to fourth-stage cut-off separation by using a nanofiltration membrane with a molecular weight cut-off of 500 Da, and finally concentrated by using a reverse osmosis membrane to obtain peony root-bark concentrate; (3) the concentrate in (2) is mixed with 1,3-butanediol and water according to a volume ratio of 0.9:6:3.1, and the pH of the mixture is adjusted to 5.0 by using citric acid and NaOH solution, and finally filtered by using a polypropylene membrane to obtain a skin care product raw material containing peony root-bark extract.

Images