Preparation method of a composition containing asiaticoside-paeonoside and application thereof

Abstract

The application belongs to the field of natural product extraction and cosmetic raw materials, and discloses a preparation method and application of a composition containing asiaticoside-paeoniflorin, which comprises two key and functionally different stages: ① extraction stage: using asiaticoside as an extraction aid and using propylene glycol as a complex medium, efficiently dissolving and forming an "asiaticoside-paeoniflorin" supramolecular complex; ② complexing and stabilizing stage: using glycerol as a stabilizer to maintain the long-term kinetic stability of the formed supramolecular complex and prevent its dissociation and aggregation. The method solves the technical problem that the extraction rate and stability of paeoniflorin are difficult to be considered simultaneously in the field by functional division of extraction and stabilization, and the composition containing asiaticoside-paeoniflorin obtained finally has excellent soothing performance, not only rapid effect, but also persistent and stable soothing protection effect, which can effectively resist and repair the influence of external stimulation on the skin.

Description

Technical Field

[0001] This invention belongs to the field of natural product extraction and cosmetic raw materials, and mainly relates to a method for preparing a composition containing asiaticoside-paeoniflorin and its application. Background Technology

[0002] Peony ( Paeonia lactiflora Pall. ) is a commonly used Chinese medicinal herb of the Ranunculaceae family, and its hallmark active ingredient is paeoniflorin ( Paeoniflorin Paeoniflorin possesses a variety of confirmed pharmacological activities, including anti-inflammatory, analgesic, hepatoprotective, and neuroprotective effects, and is a core quality marker for many traditional Chinese medicine compound preparations. However, in existing industrial extraction and preparation processes, the inherent physicochemical properties of paeoniflorin have led to the long-standing inability to effectively overcome the following technical challenges: First, when using conventional solvents such as water or lower alcohols for extraction, the transfer rate (i.e., yield) of the target component is not ideal, resulting in a waste of medicinal resources; second, and more critically, the obtained extracts or preparations are prone to chemical degradation and physical precipitation during storage, manifested as decreased solution clarity and precipitate formation, thereby reducing the effective content and unstable product quality, severely limiting the clinical efficacy and commercial shelf life of related products.

[0003] Existing technologies have made various attempts to address the above problems, such as optimizing extraction temperature and time or using different proportions of mixed solvents, but all have had limited effects and have failed to fundamentally improve the long-term stability of paeoniflorin in the final product. Therefore, developing a paeoniflorin extraction method that can simultaneously achieve high extraction rates and high stability, and obtaining paeoniflorin compositions that meet specific applications, has become an urgent technical challenge to be solved in this field. Summary of the Invention

[0004] During in-depth research, the inventors, for the first time, creatively conceived and verified a technical approach to simultaneously improve the dissolution efficiency and stability of paeoniflorin by introducing specific auxiliary components to form a molecular complex. Specifically, this invention proposes using asiaticoside as an extraction aid, with the following steps: ① In the extraction stage, asiaticoside is used as an extraction aid and propylene glycol is used as a complex medium to efficiently dissolve and form a "asiaticoside-paeoniflorin" supramolecular complex; ② In the compound stabilization stage, glycerol is used as a stabilizer to maintain the long-term kinetic stability of the formed asiaticoside-paeoniflorin supramolecular complex and prevent its dissociation and aggregation.

[0005] The inventors first used density functional theory (DFT) to perform calculations and simulations. The results clearly showed that there is a significant negative binding energy between asiaticoside and paeoniflorin molecules. This indicates that the two can spontaneously form a stable complex through intermolecular interactions such as hydrogen bonds and van der Waals forces, and do not form a stable covalent bond between them. Therefore, asiaticoside and paeoniflorin can be detected separately by HPLC. Asiaticoside, also known as Asian saponin or asiaticoside glycoside, is a pentacyclic triterpenoid saponin extracted from the whole herb of Centella asiatica, a plant in the Apiaceae family. Its molecular formula is C0. 48 H 78 O 19 Molecular weight 959.12, CAS Registry Number 16830-15-2.

[0006] Based on the above theoretical predictions, the inventors further systematically optimized the extraction process. The study found that a phased solvent system is key to achieving high extraction rates and high stability. First, in the extraction stage, when propylene glycol is chosen as the solvent, it exhibits a significant synergistic effect with asiaticoside in promoting the dissolution of paeoniflorin, making it the preferred medium for achieving high extraction rates. More importantly, in the subsequent compound stabilization stage, we unexpectedly discovered that the introduction of glycerol is crucial for maintaining the long-term stability of paeoniflorin. Compared to control examples that do not use asiaticoside or use other glycol solvents, the specific two-stage solvent system of "propylene glycol (extraction) - glycerol (stabilization)" constructed in this invention, acting synergistically with asiaticoside, not only significantly improves the extraction rate of paeoniflorin but also achieves unexpected technical effects: paeoniflorin in the final product can remain dissolved for a long time, greatly inhibiting precipitation, thereby fundamentally improving product stability and extending its shelf life.

[0007] This invention provides a method for preparing a composition containing asiaticoside and paeoniflorin based on a novel mechanism of action. The core of this method lies in the first-time application of asiaticoside as an extraction aid and its combination with a specific two-stage solvent system of propylene glycol and glycerol. This combination, through the functional division of labor in extraction and stabilization, synergistically solves the long-standing technical problem in the field of balancing extraction rate and stability.

[0008] The specific technical solution of this application is as follows:

[0009] A method for preparing a composition containing asiaticoside-paeoniflorin, the specific steps of which are as follows:

[0010] Peony root is pulverized using a pulverizer and passed through a 20-40 mesh sieve. The peony powder that passes through the sieve is added to a compound solvent at a weight ratio of 1:15-25. The compound solvent consists of asiaticoside and propylene glycol, with a ratio of 1-5 mmol:11-45 g. After soaking at room temperature for 2 hours, the temperature is raised to 40-75℃ and extracted by stirring for 3 hours. After extraction, the liquid is filtered through a 200-mesh gauze and then subjected to fine filtration. The turbidity of the filtrate is controlled to be <10.0 FNU to obtain the fine filtrate S1.

[0011] The peony mentioned therein is selected from white peony or red peony, both of which contain paeoniflorin and can be used as extraction raw materials.

[0012] Then, add glycerol of the same mass as the filtrate S1 to the filtrate S1, stir at room temperature for 15-30 minutes to compound, and finally sterilize the compound solution at 80-85℃ for 30 minutes to obtain a composition containing asiaticoside-paeoniflorin.

[0013] Preferably, the temperature for heat preservation and stirring extraction is 60℃.

[0014] The ratio of asiaticoside and propylene glycol in the composite solvent is 5 mmol: 22.5 g.

[0015] The fine filtration mentioned above is achieved by using filter paperboard or filter membrane with a pore size of less than 0.45 micrometers.

[0016] To verify the effectiveness of the above technical solution, the inventors compared it with the traditional water extraction method and found that the method provided in this application can improve the yield by up to 56.1%.

[0017] The beneficial effects achieved by this invention are as follows:

[0018] 1. This study is the first to use asiaticoside as an extraction aid for paeoniflorin, and uses propylene glycol as a composite medium to achieve efficient dissolution and the formation of a supramolecular complex of asiaticoside-paeoniflorin, thereby improving the extraction efficiency of paeoniflorin.

[0019] 2. Glycerin was used as a compounding solvent in the compounding stage, which enabled paeoniflorin in the final product to remain in a dissolved state for a long time, greatly inhibiting precipitation and thus fundamentally improving the stability of the product and extending its shelf life.

[0020] 3. The provided composite solvent extraction method, while significantly improving the extraction efficiency of paeoniflorin, unexpectedly produced a synergistic enhancement effect on the dissolution and enrichment of flavonoids in the extract and their in vitro antioxidant activity;

[0021] 4. The composition containing asiaticoside-paeoniflorin provided by the present invention has excellent soothing properties. It not only takes effect quickly, but also provides a long-lasting and stable soothing and protective effect, effectively combating and repairing the effects of external stimuli on the skin. Attached Figure Description

[0022] Figure 1 This is a schematic diagram of the preparation process of the asiaticoside-paeoniflorin complex in this invention;

[0023] Figure 2 A schematic diagram showing the surface electrostatic trends of paeoniflorin and asiaticoside molecules;

[0024] Figure 3 The standard curve for the determination of paeoniflorin by HPLC method;

[0025] Figure 4 This is a schematic diagram of HPLC detection of paeoniflorin content in Comparative Example 1.

[0026] Figure 5 This is a schematic diagram of the HPLC detection of paeoniflorin content in Example 3;

[0027] Figure 6 This is a comparison chart of the effects of lactic acid stinging in Experiment Example 5. Detailed Implementation

[0028] The present invention will be further described below with reference to specific embodiments, which will enable those skilled in the art to more fully understand the invention, but will not limit the invention in any way. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without inventive effort are within the scope of protection of the present invention.

[0029] Example 1: A method for preparing a composition containing asiaticoside-paeoniflorin, the specific steps of which are as follows:

[0030] Take the medicinal material, white peony root, pulverize it with a pulverizer and pass it through a 40-mesh sieve. Take 20g of the peony powder that passes through the sieve and add it to 400g of a compound solvent. The composition ratio of the compound solvent is 1 mmol of asiaticoside and 22.5g of propylene glycol.

[0031] After soaking at room temperature (25℃) for 2 hours, the temperature is raised to 60℃ and stirred for 3 hours for extraction. After the liquid is cooled to room temperature, it is then filtered through 200-mesh gauze and a filter membrane with a pore size of less than 0.45 micrometers to obtain fine filtrate S1 by controlling the turbidity of the filtrate to <10.0 FNU.

[0032] Subsequently, an equal mass of glycerol was added to the filtrate S1, and the mixture was stirred at room temperature for 15-30 minutes to prepare it. Finally, the prepared solution was sterilized at 80-85℃ for 30 minutes to complete the preparation and obtain a composition containing asiaticoside-paeoniflorin.

[0033] Example 2: A method for preparing a composition containing asiaticoside-paeoniflorin, the specific steps of which are the same as in Example 1, except that the composition ratio of the composite solvent is 2 mmol asiaticoside: 22.5 g propylene glycol.

[0034] Example 3: A method for preparing a composition containing asiaticoside-paeoniflorin, the specific steps of which are the same as in Example 1, except that the composition ratio of the composite solvent is 5 mmol asiaticoside: 22.5 g propylene glycol.

[0035] Example 4: A method for preparing a composition containing asiaticoside-paeoniflorin, the specific steps of which are the same as in Example 1, except that the composition ratio of the composite solvent is 1 mmol asiaticoside: 22.5 g glycerol.

[0036] Example 5: A method for preparing a composition containing asiaticoside-paeoniflorin, the specific steps of which are the same as in Example 1, except that the composition ratio of the composite solvent is 1 mmol asiaticoside: 22.5 g 1,3-butanediol.

[0037] Example 6: A method for preparing a composition containing asiaticoside-paeoniflorin, the specific steps of which are the same as in Example 1, except that the composition ratio of the composite solvent is 1 mmol asiaticoside: 11.25 g propylene glycol.

[0038] Example 7 A method for preparing a composition containing asiaticoside-paeoniflorin, the specific steps are the same as in Example 1, except that the composition ratio of the composite solvent is 1 mmol asiaticoside: 45.00g propylene glycol.

[0039] Example 8: A method for preparing a composition containing asiaticoside-paeoniflorin, the specific steps are the same as in Example 1, except that the temperature of the heat preservation and stirring extraction is adjusted from 60°C to 40-45°C.

[0040] Example 9: A method for preparing a composition containing asiaticoside-paeoniflorin, the specific steps are the same as in Example 1, except that the temperature of the heat preservation and stirring extraction is adjusted from 60°C to 70-75°C.

[0041] Example 10: A method for preparing a composition containing asiaticoside-paeoniflorin, the specific steps of which are the same as in Example 1, except that propylene glycol of equal mass is added to the filtrate S1.

[0042] Example 11 A method for preparing a composition containing asiaticoside-paeoniflorin, the specific steps are the same as in Example 1, except that 1,3-butanediol of equal mass is added to the filtrate S1.

[0043] Comparative Example 1: Obtaining paeoniflorin by conventional water extraction method

[0044] The extraction process of paeoniflorin is as follows: Take peony powder (white peony root) that has been pulverized and passed through a 20-40 mesh sieve. Add 400g of pure water at a material-to-liquid ratio of 1:20. Soak at room temperature for 2 hours, then heat to 60-65℃ and maintain the temperature with stirring for 3 hours. After the liquid cools to room temperature, filter it through 200-mesh gauze and then through a fine filter, controlling the turbidity to ≤10.0 FNU, to obtain fine filtrate S1. Then, add an equal mass of glycerol to fine filtrate S1 and stir at room temperature for 15-30 minutes to reconstitute. Finally, sterilize the reconstituted solution at 80-85℃ for 30 minutes to obtain a mixture containing paeoniflorin.

[0045] Experiment Example 1: DFT Calculation Simulation

[0046] 2D-to-3D modeling and MM2 pre-optimization of paeoniflorin and asiaticoside were performed using ChemDraw or GaussView, and calculations were performed using Gauss software. The ground state method was used to calculate the ground state, and DFT was used to represent the density functional theory. The B3LYP-D3 exchange-correlated functional basis set was used with 6-311 G. ** The basis set underwent structural optimization and frequency analysis, and was applied to B3LYP-D3 / 6-311+G. ** The energy at a single point is calculated horizontally, and the structure of the molecule is considered to be optimized when the frequency calculation has no imaginary frequency.

[0047] DFT calculations revealed that asiaticoside and paeoniflorin molecules can form supramolecular structures. Supramolecular asiaticoside-paeoniflorin combinations in different molar ratios are prepared via ion exchange reactions between the two molecules, arising through ionic or hydrogen bonding interactions rather than covalent bonds. Figure 1 Therefore, asiaticoside and paeoniflorin can be detected separately by HPLC. Furthermore, the surface electrostatic potential (ESP) of paeoniflorin and asiaticoside molecules can be analyzed. Figure 2The study found that van der Waals surface penetration is significant in the hydrogen bond formation regions, with red and blue surfaces interpenetrating, demonstrating the complementary characteristics of electrostatic potentials and reflecting the electrostatic attraction of hydrogen bonds. Paeoniflorin molecules contain hydroxyl groups (negatively charged, red ESP region) and hydrophobic regions (positively charged, blue region), providing hydrogen bond donor / acceptor and hydrophobic binding sites. Centella asiatica molecules contain sugar chains (negatively charged) and a terpenoid skeleton (positively charged), with a complex distribution of polar and hydrophobic regions, enhancing multimodal interactions. The two molecules bind through hydroxyl-hydroxyl hydrogen bonds (polar region) and methyl-terpenoid ring hydrophobic stacking (hydrophobic region), with complementary negative (red) and positive (blue) ESP charges, stabilizing the supramolecular structure. Therefore, the inventors ultimately confirmed that the intermolecular forces between Centella asiatica and paeoniflorin can be utilized to improve the extraction efficiency of paeoniflorin.

[0048] Experimental Example 2: Liquid Chromatography Detection of Paeoniflorin Content

[0049] Based on the standard methods in existing pharmacopoeias, a C18 column (150 mm × 4.6 mm, 5 μm) was selected. The mobile phase A was set to 0.1% phosphoric acid (v / v), and the mobile phase B to acetonitrile, with a v / v ratio of A:B = 86:14. Isocratic elution was used; the flow rate was 1 ml / min; the injection volume was 10 μL; and the detection wavelength was 230 nm. R... 2 The standard curve with a value of 0.9998 is: y = 14161.0750x + 10920.6500. The graph of the above standard curve can be found in [link to graph]. Figure 3 .

[0050] Figure 4 This is a schematic diagram of HPLC detection of paeoniflorin content in Comparative Example 1. Figure 5 This is a schematic diagram of the HPLC detection of paeoniflorin content in Example 3.

[0051] The area of ​​commercially available paeoniflorin standard is shown in Table 1 below; the paeoniflorin content in the comparative example, the paeoniflorin content in the experimental example, and the improvement rate compared to the comparative example are shown in Table 2 below.

[0052] Table 1. Area of ​​paeoniflorin standard at different concentrations

[0053]

[0054] Table 2. Content of paeoniflorin in comparative examples and embodiments

[0055]

[0056] As shown in Table 2 above, the preparation method of the composition containing asiaticoside-paeoniflorin provided by the present invention, compared with the traditional water extraction process (Comparative Example 1), achieved a significant improvement in the extraction efficiency of paeoniflorin in all embodiments. More specifically, in the preferred technical solutions, namely Examples 1, 2, 3, 6, 7, 8, and 9 using the "asiaticoside-propylene glycol" composite solvent system, the improvement in paeoniflorin extraction efficiency was particularly outstanding, with an improvement rate ranging from 31.71% to 56.10%. Among them, in the most preferred Example 3, the paeoniflorin content reached 1.28 mg / mL, and the extraction efficiency was improved by 56.10% compared with Comparative Example 1, achieving an unexpected technical effect. Through a systematic investigation of the influence of different process parameters, the present invention further reveals the following key findings:

[0057] Dose-dependent effect of auxiliary solvent: In the composite solvent system used, as the amount of asiaticoside adjuvant increased from 1 mmol (Example 1) to 5 mmol (Example 3), the extraction efficiency of paeoniflorin showed a clear dose-dependent increase, which proved the core role of asiaticoside in promoting the dissolution of the target component.

[0058] Specificity and synergistic effect of solvent system: Examples using propylene glycol as the extraction solvent generally showed superior extraction results. In contrast, the control examples using the glycerol system (Example 4) or the 1,3-butanediol system (Example 5) showed significantly lower extraction efficiency improvements (15.85% and 4.88%, respectively) than the propylene glycol system. This demonstrates the outstanding synergistic effect and irreplaceability of the specific combination of "asiaticoside-propylene glycol" in this invention.

[0059] Wide adaptability to process parameters: The technical solution of this invention exhibits good adaptability to changes in key process parameters. For example, under the conditions of Example 9 (high-temperature extraction, 70-75℃) and Example 8 (low-temperature extraction, 40-45℃), the extraction efficiency improvement rate of paeoniflorin remained stable at a relatively high level of 52.44% and 34.15%, respectively. Similarly, under different propylene glycol concentrations (Examples 6 and 7), the extraction efficiency was also significantly better than that of the comparative examples.

[0060] The data from the above embodiments fully demonstrate that the present invention, through the creative application of the specific composite solvent system of "asiaticoside-propylene glycol", has successfully solved the technical bottleneck of low extraction efficiency of paeoniflorin by traditional water extraction method, and obtained a composition containing asiaticoside-paeoniflorin.

[0061] Experimental Example 3: Detection of Flavonoid Content and DPPH Free Radical Scavenging Rate

[0062] The total antioxidant capacity and 1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging ability of the obtained compositions containing asiaticoside-paeoniflorin were detected. The DPPH scavenging ability at a sample concentration of 2.5% was tested. The DPPH scavenging test was performed in accordance with the instructions of the DPPH scavenging ability kit (Nanjing Jiancheng).

[0063] Total flavonoids determination method: The yield of flavonoids in plant extracts was determined by the NaNO2-Al(NO3)3 colorimetric method (GB / T20574-2006 Determination of Total Flavonoids in Propolis - Spectrophotometric Colorimetric Method). The detection results of the corresponding examples and comparative examples are as follows:

[0064] Table 3. Study on flavonoid content and DPPH free radical scavenging effect of different samples

[0065]

[0066] As shown in Table 3 above, the composition containing asiaticoside-paeoniflorin provided by the present invention, while significantly improving the extraction efficiency of paeoniflorin, unexpectedly exhibits a synergistic enhancement effect on the dissolution and enrichment of flavonoids in the composition and their in vitro antioxidant activity. Specifically, compared with the traditional water extraction method (Comparative Example 1), all embodiments of the present invention achieve a simultaneous and significant increase in flavonoid content and DPPH free radical scavenging rate. In the most preferred embodiment 3, the flavonoid content reaches 0.28 mg / mL, with an increase rate as high as 75.00%, and the corresponding DPPH free radical scavenging rate also reaches 82.06%, with an activity increase of over 60%, achieving unexpected technical effects. Different examples demonstrate that the examples using the "asiaticoside-propylene glycol" composite solvent system (such as Examples 1, 2, 3, 6, 7, 8, and 9) exhibit significantly and consistently superior flavonoid content (enhancement rate of 31.25%–75.00%) and antioxidant activity (DPPH scavenging rate of 72.08%–82.06%) compared to the examples using the glycerol system (Example 4) or the 1,3-butanediol system (Example 5). These results strongly confirm the significant synergistic effect and irreplaceable nature of the "asiaticoside-propylene glycol" combination in the efficient extraction and enrichment of flavonoid active ingredients.

[0067] Furthermore, the experimental data clearly demonstrate that the increase in flavonoid content in the composition is significantly positively correlated with the enhancement of DPPH free radical scavenging rate. This proves that the present invention, by optimizing the extraction solvent system, not only achieves efficient transfer of active ingredients in quantity, but also completely preserves and even synergistically enhances their inherent biological activity in quality, overcoming the technical defects of traditional extraction methods that may lead to damage to the function of active ingredients due to process limitations.

[0068] Experiment Example 4: Stability Assessment

[0069] To verify the long-term stability of the compositions obtained in this invention, the samples prepared in Comparative Example 1, Example 3, Example 10, and Example 11 were left to stand at room temperature for 24 hours, and the precipitation was observed. The results are shown in Table 4 below:

[0070] Table 4. Precipitation in Examples 3, 10, 11 and Comparative Example 1

[0071]

[0072] The stability test data clearly revealed an unexpected phenomenon: although the core "Centella asiatica-propylene glycol" solvent system of this invention was used in the extraction stage, the type of subsequent compound solvent had a decisive influence on the stability of the final product.

[0073] Specifically, the example using glycerol for compounding after the extraction stage (Example 3) exhibited excellent stability. However, when the compounding solvent was changed to propylene glycol (Example 10) or 1,3-butanediol (Example 11), precipitation occurred in both cases after standing.

[0074] The above findings demonstrate that the technical solution provided by this invention is a multi-step, multi-component, precisely synergistic systematic solution. The use of propylene glycol as a composite medium in the extraction stage and the use of glycerol as a stabilizer in the compounding stage are two interrelated and indispensable technical features, together constituting a complete technical means to ensure high extraction rates of paeoniflorin and high stability of the composition. Separating these two features or simply replacing them with other glycol solvents cannot achieve the unexpected technical effects obtained by this invention. This highlights the non-obviousness and outstanding substantive characteristics of the technical solution of this invention.

[0075] Based on the mechanism of action of supramolecular complexes, the inventors offer the following explanation for the aforementioned stability:

[0076] The technical solution of this invention comprises two key and functionally distinct stages: ① Extraction stage: The core task of this stage is to efficiently dissolve and form the "asiaticoside-paeoniflorin" supramolecular complex. Propylene glycol, due to its polarity and molecular structure, optimally promotes the intermolecular interactions (such as hydrogen bonds) between asiaticoside and paeoniflorin at this stage, making it an indispensable medium and participant in the formation of this complex. ② Complex stabilization stage: The core task of this stage is to maintain the long-term kinetic stability of the formed supramolecular complex, preventing its dissociation and aggregation. Glycerol plays a unique role in the stabilization stage: Experimental results show that glycerol plays an irreplaceable "stabilizer" role in the complex stabilization stage.

[0077] Compared to straight-chain diols such as propylene glycol or 1,3-butanediol, glycerol molecules contain three hydroxyl groups, resulting in a higher hydrogen bond density and a stronger ability to form a three-dimensional hydrogen bond network. Glycerol molecules, through their multi-hydroxyl structure, can form a more stable and sterically hindered solvation layer or protective hydrogen bond network around the already formed "asiaticoside-paeoniflorin" complex, effectively inhibiting the dissociation of the complex and the re-aggregation and precipitation of paeoniflorin molecules. While propylene glycol or 1,3-butanediol in other solvents are structurally similar to glycerol, they contain only two hydroxyl groups, and their ability to form a stable hydrogen bond network is far inferior to that of glycerol. They cannot provide the same effective stabilizing environment during the compounding stage, leading to the gradual dissociation of the supramolecular complex during long-term static storage, ultimately triggering the precipitation of the target component.

[0078] Experimental Example 5: Lactic Acid Sting Test

[0079] 1. Experimental Objectives and Model Construction

[0080] This invention uses the internationally recognized "lactic acid stinging test" as a human efficacy evaluation model. This model actively induces a controllable and measurable skin stinging and redness reaction by applying a high concentration of lactic acid solution to the nasolabial folds of sensitive skin subjects, thereby accurately simulating the chemical stimulation received by the skin.

[0081] A total of 7 subjects were included, 3 males and 4 females, with 7 ultimately completing the study. The subjects were aged 20-22 years, with an average age of 20.43 years.

[0082] 2. Experimental methods and evaluation indicators

[0083] After baseline data collection was completed in a constant temperature and humidity environment, lactic acid stinging modeling was performed on the selected eligible subjects. Subsequently, the sample of Example 3 (Sample 2) and the sample of Comparative Example 1 (Sample 1) prepared according to this invention were applied to both cheeks of the subjects, respectively; the composition of Sample 1 and Sample 2 is shown in Table 5 below:

[0084] Table 5 Preparation methods of Sample 1 and Sample 2

[0085]

[0086] At multiple key time points after product use (5 min, 15 min, 30 min, 1 h), the skin redness value (a-value) of specific areas of the face was quantitatively recorded using a high-precision Antera 3D imager. This instrument, through 3D reconstruction and multispectral analysis, can objectively and accurately capture minute color changes in the skin. A decrease in the skin a-value directly indicates a reduction in facial redness and is the most crucial objective indicator for evaluating the soothing and redness-reducing efficacy.

[0087] 3. Experimental Results and Advantages Analysis

[0088] As shown in Table 6 and Figure 6 As shown, the experimental data clearly demonstrate the superior soothing performance of the sample (sample 2) in Example 3 of this invention:

[0089] Significant immediate soothing effect: Just 5 minutes after application, the skin a* value on the side where sample 2 was applied showed a negative increase of -8.33%, indicating rapid relief of redness. In stark contrast, sample 1 showed a change of -3.43% at the same time point. This data demonstrates that the sample of this invention exhibits a significant advantage in terms of the speed of its effect.

[0090] Long-lasting efficacy stability: Throughout the 1-hour observation period, the rate of change of a* value at each time point of Sample 2 (-8.33% to -5.28%) was consistently superior to the corresponding value of Sample 1 (-3.43% to -2.32%). This demonstrates that the composition provided by this invention not only has a rapid onset of action but also provides a long-lasting and stable soothing and protective effect, effectively combating and repairing the effects of external stimuli on the skin.

[0091] Table 6. Skin a* value test results (sample size = 7)

[0092]

[0093] Where a* change rate = (test value after using the product for a certain period of time - test value before using the product) / test value before using the product × 100%; the above experimental results corroborate the aforementioned in vitro extraction rate, stability, and antioxidant data, together forming a complete chain of technical evidence. The results show that the composition containing asiaticoside-paeoniflorin prepared by this invention through a specific "asiaticoside-propylene glycol" composite solvent system successfully transforms its superior physicochemical properties (high content, high stability, high activity) into a definite and significantly superior clinical soothing effect compared to the control in the final product. This fully demonstrates that the technical solution of this invention not only solves the technical bottleneck in the basic extraction process but also provides solid evidence of human efficacy for its application in high-end functional cosmetics, skincare products, and other fields, generating unexpected commercial value.

[0094] The above description is merely a preferred embodiment of the present invention and is not intended to limit the invention in any other way. Any person skilled in the art can utilize the above technical content to make changes or modifications to create equivalent embodiments. Any simple modifications, equivalent changes, and modifications made to the above embodiments without departing from the technical essence of the present invention shall still fall within the protection scope of the present invention.

Claims

1. A method for preparing a composition containing asiaticoside-paeoniflorin, characterized in that, The specific steps are as follows: Peony root is pulverized using a pulverizer and passed through a 20-40 mesh sieve. The sifted peony powder is added to a composite solvent at a weight ratio of 1:15-25. The composite solvent consists of asiaticoside and propylene glycol, with a ratio of 1-5 mmol:11-45 g. After soaking at room temperature for 2 hours, the temperature is raised to 40-75℃ and stirred for 3 hours. After extraction, the liquid is filtered through 200-mesh gauze and then finely filtered to control the turbidity of the filtrate to <10.0 FNU, thus obtaining fine filtrate S1. Then, an equal mass of glycerol is added to fine filtrate S1, and the mixture is stirred at room temperature for 15-30 minutes for compounding. Finally, the compound solution is sterilized at 80-85℃ for 30 minutes to obtain a composition containing asiaticoside and paeoniflorin.

2. The method for preparing the composition containing asiaticoside-paeoniflorin according to claim 1, characterized in that, The peony mentioned is selected from white peony or red peony.

3. The method for preparing the composition containing asiaticoside-paeoniflorin according to claim 1, characterized in that, The extraction temperature was 60℃ with stirring and heat preservation.

4. The method for preparing the composition containing asiaticoside-paeoniflorin according to claim 1, characterized in that, The ratio of asiaticoside and propylene glycol in the composite solvent is 5 mmol: 22.5 g.

5. The method for preparing the composition containing asiaticoside-paeoniflorin according to claim 1, characterized in that, The fine filtration mentioned above is achieved by using filter paperboard or filter membrane with a pore size of less than 0.45 micrometers.

6. The use of the composition containing asiaticoside-paeoniflorin obtained by the preparation method of claim 1 in the preparation of cosmetics.

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