Composition for promoting skin microcirculation and preparation method and application thereof

Abstract

The invention relates to the technical field of cosmetics, in particular to a composition for promoting skin microcirculation as well as a preparation method and application thereof, and the key point of the technical scheme is that the composition consists of the following components: rhizoma bletillae, motherwort, poria cocos, moutan bark, ginseng, peach blossom and lotus. According to the composition, the deep-eutectic solvent is combined with a micro-jet extraction technology, so that the extraction efficiency of active ingredients such as flavonoids is remarkably improved, and functional ingredients for resisting oxidation, resisting aging and promoting blood circulation are effectively reserved. According to the composition disclosed by the invention, the comprehensive effects of improving skin microcirculation, relieving dark skin color and delaying senescence are systematically realized by efficiently extracting active substances, activating a microcirculation core pathway and cooperating with coherent effects of a whitening and repairing mechanism.

Description

Technical Field

[0001] This invention relates to the technical field of cosmetics, and more specifically, to a composition that promotes skin microcirculation, its preparation method, and its application. Background Technology

[0002] As the ancients said, "skin like solidified cream" and "fair with a rosy glow" indicate a healthy complexion. A healthy skin condition is one where the skin is not only fair and radiant but also has a healthy, rosy glow. Good skin microcirculation ensures the supply of nutrients and the metabolism of waste products, thereby promoting cell metabolism and increasing skin's radiance and elasticity. When skin microcirculation is impaired, the skin appears dull and lackluster, accelerating skin aging.

[0003] Currently, most skin-brightening products on the market brighten skin tone by inhibiting melanin production or its transfer to the epidermis. However, these methods only temporarily improve skin tone and fail to fundamentally address skin microcirculation issues. With discontinuation of use, melanin may re-accumulate, causing the skin tone to rebound and become even darker. Furthermore, in terms of obtaining active ingredients, traditional extraction processes such as high-temperature boiling and pressure differentials can easily damage heat-sensitive and bioactive components, resulting in insufficient stability and limited efficacy of the final extract. Summary of the Invention

[0004] In view of the shortcomings of the existing technology, the purpose of this invention is to provide a composition that promotes skin microcirculation, which has the advantages of improving microcirculation, alleviating dull skin tone and delaying aging.

[0005] The above-mentioned technical objective of the present invention is achieved through the following technical solution: a composition for promoting skin microcirculation, comprising the following components: Bletilla striata, Leonurus japonicus, Poria cocos, Paeonia suffruticosa, Ginseng, Peach blossom, and Lotus.

[0006] In one embodiment, the composition comprises the following components in parts by weight: 20-60 parts of Bletilla striata, 20-60 parts of Leonurus japonicus, 20-50 parts of Poria cocos, 20-50 parts of Paeonia suffruticosa, 5-30 parts of Panax ginseng, 10-30 parts of Prunus persica, and 10-30 parts of Nelumbo nucifera.

[0007] The method for preparing the above-mentioned composition for promoting skin microcirculation includes the following steps:

[0008] S1. Weigh out Bletilla striata, Leonurus japonicus, Poria cocos, Paeonia suffruticosa, Panax ginseng, Prunus persica and Nelumbo nucifera according to the above weight ratio, mix them, pulverize them, and sieve them through a sieve to obtain medicinal powder.

[0009] S2. Mix the medicinal powder with a eutectic solvent until homogeneous to obtain a mixture;

[0010] S3. The mixture is extracted using microfluidic technology to obtain a crude extract;

[0011] S4. The crude extract is subjected to centrifugation and membrane filtration purification to obtain the target product.

[0012] In one embodiment, the eutectic solvent includes betaine, a hydrogen bond donor, and water.

[0013] In one embodiment, the molar ratio of betaine to hydrogen bond donor is 1:1 to 1:5; and the water mass percentage is 10% to 80%.

[0014] In one embodiment, the hydrogen bond donor includes one or more of propylene glycol, glycerol, butanediol, and urea.

[0015] In one embodiment, in step S2, the mass ratio of the medicinal powder to the eutectic solvent is 1:5 to 1:15.

[0016] In one embodiment, in step S3, the extraction temperature of the extraction process is 15-30°C.

[0017] In one embodiment, in step S4, the centrifugation speed is 4000-8000 rpm and the centrifugation time is 20-40 min; the membrane filtration purification process is ceramic membrane separation and purification, and the pore size of the ceramic membrane is 200-600 μm.

[0018] The above-mentioned composition for promoting skin microcirculation is used in cosmetics for promoting skin microcirculation, improving dull skin tone, or delaying skin aging.

[0019] The above-mentioned composition for promoting skin microcirculation has the following beneficial effects:

[0020] Firstly, this invention significantly enhances the bioactivity and stability of the composition through the synergistic effect of plant components such as Bletilla striata, Leonurus japonicus, Poria cocos, Paeonia suffruticosa, ginseng, lotus, and peach blossom.

[0021] Secondly, the composition of this invention improves microcirculation, alleviates dull skin tone, and delays aging by activating the PI3K / Akt signaling pathway. On one hand, activation of this pathway significantly upregulates the expression of vascular endothelial growth factor and neurogenic nitric oxide synthase, thereby effectively promoting angiogenesis, dilating blood vessels, improving blood flow and skin microcirculation, and fundamentally solving the problem of dull skin tone. On the other hand, through the regulation of downstream nodes such as GSK3β, it can also promote the proliferation and migration of skin cells, accelerate tissue repair, and delay the aging process.

[0022] Thirdly, the composition of this invention significantly reduces oxidative stress by inhibiting the excessive production of oxygen free radicals and enhancing the activity of antioxidant enzymes. This mechanism not only protects cells, promotes vasodilation, and improves blood flow, but also improves problems such as rough skin, disordered collagen fiber arrangement, and pigmentation, achieving whitening and brightening effects.

[0023] Fourth, this invention employs a eutectic solvent system composed of betaine, a hydrogen bond donor, and water, combined with gentle microfluidic extraction technology. This process overcomes the problem that active ingredients such as glycosides, flavonoids, and phenolic compounds are easily destroyed in traditional high-temperature decoction or organic solvent extraction. Under low-temperature conditions, the eutectic solvent can efficiently disrupt plant cell walls, selectively extract target components, and maximize the retention of heat-sensitive and bioactive substances in plant components.

[0024] Fifth, the composition of the present invention can be directly used as a base in cosmetics such as lotions, emulsions, creams, and masks, and has broad market application prospects. Attached Figure Description

[0025] Figure 1 This is a flowchart of the steps of the present invention;

[0026] Figure 2 This is a graph showing the relative angiogenesis length at different concentrations in Example 1;

[0027] Figure 3 This is a diagram showing the cell scratch results from Example 1. Detailed Implementation

[0028] The present invention will now be described in detail with reference to the accompanying drawings and embodiments.

[0029] A composition for promoting skin microcirculation comprises the following components: Bletilla striata, Leonurus japonicus, Poria cocos, Paeonia suffruticosa, Ginseng, Peach blossom, and Lotus. To maximize the content of active ingredients, this invention preferably uses specific parts of each raw material known to be rich in active ingredients: Bletilla striata is extracted from the root; Leonurus japonicus from the stem and leaves; Poria cocos from the fruiting body; Paeonia suffruticosa from the root bark; Ginseng from the root; Peach blossom from the petals; and Lotus from the petals, ensuring maximum preservation of their bioactivity.

[0030] Furthermore, the composition for promoting skin microcirculation comprises the following ingredients in parts by weight: 20-60 parts Bletilla striata, 20-60 parts Leonurus japonicus, 20-50 parts Poria cocos, 20-50 parts Paeonia suffruticosa, 5-30 parts Panax ginseng, 10-30 parts Prunus persica, and 10-30 parts Nelumbo nucifera. A preferred ratio is: 30-50 parts Bletilla striata, 30-50 parts Leonurus japonicus, 25-40 parts Poria cocos, 25-45 parts Paeonia suffruticosa, 5-15 parts Panax ginseng, 10-20 parts Prunus persica, and 10-20 parts Nelumbo nucifera.

[0031] Through the specific ratios mentioned above, the components complement each other, working together to achieve a multi-dimensional skin health promotion effect, from improving microcirculation and regulating metabolism to anti-oxidation and delaying aging.

[0032] like Figure 1 As shown, the preparation method of the above-mentioned composition for promoting skin microcirculation includes the following steps:

[0033] S1. Weigh out Bletilla striata, Leonurus japonicus, Poria cocos, Paeonia suffruticosa, Panax ginseng, Prunus persica and Nelumbo nucifera according to the above weight ratio, mix them, pulverize them, and sieve them through a sieve to obtain medicinal powder.

[0034] This step aims to disrupt the plant tissue structure and increase the specific surface area, creating favorable conditions for subsequent solvent penetration and component dissolution. In this embodiment, the sieve mesh size can be adjusted according to specific process requirements, typically a 30-mesh sieve, to ensure the uniformity of the raw material powder.

[0035] S2. Mix the medicinal powder with a eutectic solvent until homogeneous to obtain a mixture;

[0036] In practical applications, the mass ratio of medicinal powder to eutectic solvent is 1:5-1:15. This solvent system can efficiently break down plant cell walls, promote the dissolution of plant components, and extract the effective components from plants under mild conditions.

[0037] Specifically, the eutectic solvent includes betaine, hydrogen bond donors, and water. Hydrogen bond donors include one or more of propylene glycol, glycerol, butylene glycol, and urea. The molar ratio of betaine to hydrogen bond donor is 1:1 to 1:5, and the mass percentage of water is 10% to 80%. Betaine, as a hydrogen bond acceptor, can work synergistically with hydrogen bond donors to enhance the interaction between the solvent and plant components, ensuring efficient extraction of the active ingredients.

[0038] S3. The mixture is extracted using microfluidic technology to obtain a crude extract;

[0039] This step involves placing the mixture from S2 into a microfluidic extraction device for extraction. Microfluidic extraction utilizes efficient fluid dynamics principles to extract plant components at lower temperatures, maximizing the retention of heat-sensitive components such as glycosides, flavonoids, and phenolic substances, while avoiding the damage caused by traditional high-temperature extraction methods.

[0040] Specifically, the microjets extraction temperature is 15-30℃, the micropore size is 400-800μm, the feed rate is 15-35L / min, and the extraction time is 15-30min. This ensures that the active ingredients in the plant are fully dissolved and prevents over-extraction. Microjets extraction not only improves extraction efficiency but also ensures selective and highly permeable extraction of the components.

[0041] S4. The crude extract is subjected to centrifugation and membrane filtration purification to obtain the target product.

[0042] In practical applications, insoluble impurities are first removed by centrifugation at 4000–8000 rpm for 20–40 minutes to ensure effective separation of liquid and solid components. Then, further filtration is performed using ceramic membrane separation technology with a pore size of 200–600 μm. Ceramic membrane separation efficiently removes macromolecular impurities, ensuring the purity and stability of the target product.

[0043] The above preparation method combines eutectic solvent extraction, microfluidic extraction, and ceramic membrane separation and purification techniques. The resulting composition not only improves the skin microcirculation but also ensures the stability and bioavailability of the active ingredients.

[0044] The above composition is used in cosmetics for promoting skin microcirculation, improving dull skin tone, or delaying skin aging. In this embodiment, there are no special limitations on the cosmetics used, including but not limited to toners, lotions, creams, masks, etc. Depending on different product types and usage needs, this composition can be flexibly added to the formulation, preferably at a concentration of 0.05-5%. The specific amount added can be adjusted according to the formulation requirements, effects, and skin type of different products.

[0045] Example 1

[0046] This embodiment provides a composition for promoting skin microcirculation, which consists of the following components in parts by weight: 50 parts of Bletilla striata, 40 parts of Leonurus japonicus, 30 parts of Poria cocos, 30 parts of Paeonia suffruticosa, 5 parts of Panax ginseng, 15 parts of Prunus persica, and 15 parts of Nelumbo nucifera.

[0047] The specific preparation steps in this embodiment are as follows:

[0048] S1. Preparation of eutectic solvent: weigh betaine and butanediol in a molar ratio of 1:3, add water to prepare a eutectic solvent with a water content of 50 wt%.

[0049] S2. Raw material pretreatment: Weigh out Bletilla striata, Leonurus japonicus, Poria cocos, Paeonia suffruticosa, Ginseng, Peach blossom and Lotus according to the formula ratio, mix them and then pulverize them. Pass them through a 30-mesh sieve to obtain uniform medicinal powder.

[0050] S3. Mixing and extraction: Mix the medicinal powder with the eutectic solvent at a mass ratio of 1:10, place the mixture in a microfluidic extractor, set the extraction temperature to 20℃, the micropore diameter to 600μm, and the feed rate to 25L / min, and extract continuously for 15min to obtain the crude extract.

[0051] S4. Centrifugation and membrane purification: Pour the crude extract into a horizontal spiral centrifuge and centrifuge at 4000 rpm for 30 min. Remove the filter residue and collect the supernatant. Then filter the supernatant through a ceramic membrane system with a pore size of 400 μm, collect the filtrate, and obtain the target product.

[0052] Example 2

[0053] This embodiment provides a composition for promoting skin microcirculation, which consists of the following components in parts by weight: 45 parts of Bletilla striata, 50 parts of Leonurus japonicus, 25 parts of Poria cocos, 30 parts of Paeonia suffruticosa, 20 parts of Panax ginseng, 10 parts of Prunus persica, and 10 parts of Nelumbo nucifera.

[0054] The specific preparation steps in this embodiment are as follows:

[0055] S1. Preparation of eutectic solvent: weigh betaine and butanediol in a molar ratio of 1:3, add water to prepare a eutectic solvent with a water content of 40wt%.

[0056] S2. Raw material pretreatment: Weigh out Bletilla striata, Leonurus japonicus, Poria cocos, Paeonia suffruticosa, Ginseng, Peach blossom and Lotus according to the formula ratio, mix them and then pulverize them. Pass them through a 30-mesh sieve to obtain uniform medicinal powder.

[0057] S3. Mixing and extraction: Mix the medicinal powder with the eutectic solvent at a mass ratio of 1:15, place the mixture in a microfluidic extractor, set the extraction temperature to 25℃, the micropore diameter to 800μm, and the feed rate to 20L / min, and extract continuously for 20min to obtain the crude extract.

[0058] S4. Centrifugation and membrane purification: Pour the crude extract into a horizontal spiral centrifuge and centrifuge at 6000 rpm for 20 min. Remove the filter residue and collect the supernatant. Then filter the supernatant through a ceramic membrane system with a pore size of 600 μm, collect the filtrate, and obtain the target product.

[0059] Example 3

[0060] This embodiment provides a composition for promoting skin microcirculation, which consists of the following components in parts by weight: 35 parts of Bletilla striata, 45 parts of Leonurus japonicus, 35 parts of Poria cocos, 40 parts of Paeonia suffruticosa, 10 parts of Panax ginseng, 15 parts of Prunus persica, and 10 parts of Nelumbo nucifera.

[0061] The specific preparation steps in this embodiment are as follows:

[0062] S1. Preparation of eutectic solvent: Weigh betaine and butanediol in a molar ratio of 1:2.5, add water to prepare a eutectic solvent with a water content of 30wt%.

[0063] S2. Raw material pretreatment: Weigh out Bletilla striata, Leonurus japonicus, Poria cocos, Paeonia suffruticosa, Ginseng, Peach blossom and Lotus according to the formula ratio, mix them and then pulverize them. Pass them through a 30-mesh sieve to obtain uniform medicinal powder.

[0064] S3. Mixing and extraction: Mix the medicinal powder with the eutectic solvent at a mass ratio of 1:15, place the mixture in a microfluidic extractor, set the extraction temperature to 20℃, the micropore diameter to 400μm, and the feed rate to 30L / min, and extract continuously for 15min to obtain the crude extract.

[0065] S4. Centrifugation and membrane purification: Pour the crude extract into a horizontal spiral centrifuge and centrifuge at 4000 rpm for 30 min. Remove the filter residue and collect the supernatant. Then filter the supernatant through a ceramic membrane system with a pore size of 200 μm, collect the filtrate, and obtain the target product.

[0066] Example 4

[0067] This embodiment provides a composition for promoting skin microcirculation, which consists of the following components in parts by weight: 40 parts of Bletilla striata, 40 parts of Leonurus japonicus, 35 parts of Poria cocos, 35 parts of Paeonia suffruticosa, 15 parts of Panax ginseng, 10 parts of Prunus persica, and 15 parts of Nelumbo nucifera.

[0068] The specific preparation steps in this embodiment are as follows:

[0069] S1. Preparation of eutectic solvent: Weigh betaine and butanediol in a molar ratio of 1:2.5, add water to prepare a eutectic solvent with a water content of 40wt%.

[0070] S2. Raw material pretreatment: Weigh out Bletilla striata, Leonurus japonicus, Poria cocos, Paeonia suffruticosa, Ginseng, Peach blossom and Lotus according to the formula ratio, mix them and then pulverize them. Pass them through a 30-mesh sieve to obtain uniform medicinal powder.

[0071] S3. Mixing and extraction: Mix the medicinal powder with the eutectic solvent at a mass ratio of 1:10, place the mixture in a microfluidic extractor, set the extraction temperature to 15℃, the micropore diameter to 600μm, and the feed rate to 15L / min, and extract continuously for 30min to obtain the crude extract.

[0072] S4. Centrifugation and membrane purification: Pour the crude extract into a horizontal spiral centrifuge and centrifuge at 8000 rpm for 10 min. Remove the filter residue and collect the supernatant. Then filter the supernatant through a ceramic membrane system with a pore size of 300 μm, collect the filtrate, and obtain the target product.

[0073] Example 5

[0074] This embodiment provides a composition for promoting skin microcirculation, which consists of the following components in parts by weight: 30 parts of Bletilla striata, 55 parts of Leonurus japonicus, 30 parts of Poria cocos, 45 parts of Paeonia suffruticosa, 10 parts of Panax ginseng, 10 parts of Prunus persica, and 10 parts of Nelumbo nucifera.

[0075] The specific preparation steps in this embodiment are as follows:

[0076] S1. Preparation of eutectic solvent: Weigh betaine and butanediol in a molar ratio of 1:2.5, add water to prepare a eutectic solvent with a water content of 40wt%.

[0077] S2. Raw material pretreatment: Weigh out Bletilla striata, Leonurus japonicus, Poria cocos, Paeonia suffruticosa, Ginseng, Peach blossom and Lotus according to the formula ratio, mix them and then pulverize them. Pass them through a 30-mesh sieve to obtain uniform medicinal powder.

[0078] S3. Mixing and extraction: Mix the medicinal powder with the eutectic solvent at a mass ratio of 1:10, place the mixture in a microfluidic extractor, set the extraction temperature to 25℃, the micropore diameter to 500μm, and the feed rate to 25L / min, and extract continuously for 15min to obtain the crude extract.

[0079] S4. Centrifugation and membrane purification: Pour the crude extract into a horizontal spiral centrifuge and centrifuge at 6000 rpm for 20 min. Remove the filter residue and collect the supernatant. Then filter the supernatant through a ceramic membrane system with a pore size of 200 μm, collect the filtrate, and obtain the target product.

[0080] Comparative Example 1

[0081] This comparative example provides a composition for promoting skin microcirculation, comprising the following components in parts by weight: 15 parts Bletilla striata, 15 parts Leonurus japonicus, 10 parts Poria cocos, 10 parts Paeonia suffruticosa, 60 parts Panax ginseng, 35 parts Prunus persica, and 35 parts Nelumbo nucifera.

[0082] The specific preparation steps for this comparative example are the same as those for Example 1.

[0083] Comparative Example 2

[0084] This comparative example provides a composition that promotes skin microcirculation, and its raw material composition is exactly the same as that of Example 1.

[0085] The specific preparation steps of this comparative example differ from those of Example 1 in that the extraction solvent used is changed from a eutectic solvent to water, while the remaining steps are the same as those of Example 1.

[0086] Comparative Example 3

[0087] This comparative example provides a composition that promotes skin microcirculation, and its raw material composition is exactly the same as that of Example 1.

[0088] The specific preparation steps of this comparative example differ from those of Example 1 in that the extraction process is changed from microfluidic extractor to 80°C water bath for 2 hours, while the remaining steps are the same as those of Example 1.

[0089] Comparative Example 4

[0090] This comparative example provides a composition that promotes skin microcirculation, and its raw material composition is exactly the same as that of Example 1.

[0091] The specific preparation steps of this comparative example differ from those of Example 1 in that the extraction process was changed from microfluidic extraction to room temperature ultrasonication for 30 min (50 Hz), while the remaining steps are the same as those of Example 1.

[0092] Comparative Example 5

[0093] This comparative example provides a composition that promotes skin microcirculation, and its raw material composition is exactly the same as that of Example 1.

[0094] The specific preparation steps of this comparative example differ from those of Example 1 in that the raw material for preparing the eutectic solvent is changed to an aqueous solution of betaine and 1,3-propanediol (molar ratio of 3:1) (water content 50wt%), while the remaining steps are the same as those in Example 1.

[0095] Test 1: Determination of total flavonoid content

[0096] 1. Principle

[0097] This method employs the NaNO2-Al(NO3)3-NaOH colorimetric method. Flavonoids with ortho-dihydroxyl structures, after reduction with NaNO2, react with Al... 3+ The complex breaks down under alkaline conditions (NaOH) to form a chalcone structure, resulting in a reddish-orange color. Within a certain range, the intensity of the color is directly proportional to the total flavonoid content, allowing for colorimetric determination to calculate the total flavonoid content.

[0098] 2. Instruments: UV-Vis spectrophotometer, analytical balance, pipette.

[0099] 3. Reagents: The water used in this standard is pure water or water of equivalent purity. Unless otherwise specified, all reagents used are of analytical grade. Analytical grade reagents include 0.2 mg / mL rutin standard solution, 5% sodium nitrite solution, 10% aluminum nitrate solution, and 1% sodium hydroxide solution.

[0100] 4. Analysis Steps

[0101] 4.1 Construction of standard curve: Accurately pipette rutin standard solutions of different concentrations, add reagents according to standard procedures, and finally use a spectrophotometer to measure the absorbance at a wavelength of 510 nm to construct a standard curve.

[0102] 4.2 Sample Determination: Filter or vacuum filter the sample until it is clear, and dilute it to an appropriate concentration. Add reagents according to the standard curve method, and finally measure the absorbance of the sample. Calculate the total flavonoid concentration using the regression equation of the standard curve.

[0103] 5. Test Results

[0104] Table 1. Results of Total Flavonoid Content Test

[0105]

[0106] As shown in Table 1, in the total flavonoid content test, the total flavonoid content of Examples 1-5 was significantly higher than that of Comparative Examples 1-5, with Example 1 reaching 4.05 mg / mL, far exceeding the 1.39 mg / mL of Comparative Example 1. This indicates that the formulation and extraction process of the present invention can significantly improve the extraction efficiency of flavonoid active ingredients. Flavonoids have significant antioxidant, anti-inflammatory, and vasodilatory effects, and their high content directly supports the efficacy of the composition in improving microcirculation, anti-aging, and brightening skin tone. In addition, the total flavonoid content obtained by the water extraction method of Comparative Example 2 and the high-temperature water bath extraction method of Comparative Example 3 was significantly lower than that of the Example group, indicating that the low-temperature eutectic solvent combined with microfluidic extraction technology can effectively avoid the destruction of heat-sensitive flavonoids, thereby improving the retention rate and stability of active ingredients.

[0107] Test 2: DPPH free radical scavenging experiment

[0108] 1. Testing Principles

[0109] DPPH (2,2-diphenyl-1-picrylhydrazyl) is a stable free radical that appears purple in organic solvents and possesses a single-electron structure. DPPH can accept an electron or a hydrogen ion, causing its color to lighten, with a maximum absorption wavelength of 517 nm. In the presence of free radical scavengers, the single electron of DPPH is captured, leading to a decrease in absorbance. The change in absorbance is linearly related to the activity of the antioxidant; therefore, the degree of decrease in absorbance reflects the strength of the antioxidant activity. Antioxidant capacity is expressed as a scavenging rate (inhibition rate); the higher the inhibition rate, the stronger the antioxidant capacity.

[0110] 2. Reagents and Samples

[0111] 1) DPPH working solution: Accurately weigh 0.0100g of DPPH powder, dissolve it in 95% ethanol and dilute to 250mL to prepare a 0.1mmol / L solution.

[0112] 2) Test solutions: The extracts from each example and the comparative example were prepared into 1.0% and 2.0% test solutions using 95% ethanol aqueous solution. A 1 mg / mL vitamin C ethyl ether solution was also prepared as a control.

[0113] 3. Antioxidant capacity test

[0114] a. Add 4.0 mL of DPPH solution and 1.0 mL of 95 wt% ethanol to a 10 mL test tube, mix and shake well, react in the dark for 30 min, and after stabilization, measure the absorbance at 517 nm using 95 wt% ethanol as a reference, and record it as A0.

[0115] b. Add 4.0 mL of DPPH solution and 1.0 mL of the test sample solution to a 10 mL test tube, mix and shake well, react in the dark for 30 min, and after stabilization, measure the absorbance at 517 nm using 95 wt% ethanol as a reference, and record it as Ar.

[0116] c. Add 4.0 mL of 95 wt% ethanol solution and 1.0 mL of the test sample solution to a 10 mL test tube, mix and shake well, react in the dark for 30 min, and after stabilization, measure the absorbance at 517 nm using 95 wt% ethanol as a reference, and record it as As.

[0117] d. Calculation formula: Scavenging rate of DPPH free radicals by the sample:

[0118]

[0119] 5. Test Results

[0120] Table 2 DPPH Removal Results

[0121]

[0122] As shown in Table 2, in the DPPH free radical scavenging ability test, Example 1 exhibited a free radical scavenging rate of 77.71% at a concentration of 2%, significantly higher than the 51.47% of Comparative Example 1 and other comparative examples. This indicates that the composition of the present invention has strong antioxidant capacity, effectively neutralizing free radicals and mitigating the damage of oxidative stress to skin cells. The increased DPPH free radical scavenging rate indicates that the composition of the present invention has strong anti-aging and skin brightening effects, and has potential application value in resisting skin oxidation and maintaining skin health. The low DPPH free radical scavenging rates of the water extraction method in Comparative Example 2 and the high-temperature water bath extraction method in Comparative Example 3 further confirm that traditional extraction methods easily lead to the loss of antioxidant components, while the extraction system used in the present invention can better retain active ingredients.

[0123] Test 3: ABTS Free Radical Scavenging Ability Test

[0124] 1. Testing Principles

[0125] Using ABTS (2,2'-Azino-bis (3-ethylbenzothiazoline-6-sulfonic acid)diammonium salt) as a water-soluble free radical initiator, stable blue-green cationic free radical ABTS+· is generated by reactive oxygen species oxidation. After the addition of an antioxidant, the antioxidant reacts with ABTS+, causing the solution to decolorize. After 6 minutes of reaction, the change in absorbance (A734) is detected at 734 nm.

[0126] 2. Reagent preparation

[0127] a. PBS solution: Weigh 8g NaCl, 0.2g KCl, 1.44g Na2HPO4 and 0.24g KH2PO4, adjust pH to 7.2 and bring the volume to 1000mL.

[0128] b. 2.45 mmol / L potassium persulfate: To prepare 100 mL, take 0.0662 g of potassium persulfate and dilute to 100 mL with water.

[0129] c. 7 mmol / L ABTS stock solution: Weigh 0.0384 g ABTS, dissolve in 2.45 mmol / L potassium persulfate, and make up to 10 mL. The solution is dark green.

[0130] d. Preparation of ABTS+ assay solution: Dilute the ABTS+ stock solution with phosphate buffer (pH=7.2) until the absorbance reaches 0.700±0.020 at 734 nm. When preparing approximately 0.12 mmol / L ABTS+, the measured absorbance is 0.716 Å (preparing 4 mL ABTS+ in 1 mL PBS) – approximately 1.7 mL of mmol / L ABTS stock solution is diluted to 100 mL.

[0131] 3. Testing Methods

[0132] a. Take 4 mL of ABTS+ assay solution → 1 mL of sample → mix for 10 s → let stand at 25℃ in the dark for 6 min → measure the absorbance at 734 nm. Use 4 mL of PBS + 1 mL of sample as a reference, and record it as sample A;

[0133] b. Take 4 mL of ABTS+ assay solution → 1 mL of PBS → mix for 10 s → incubate at 25°C in the dark for 6 min → measure the absorbance at 734 nm. Use PBS as a reference and record A0.

[0134] Inhibition rate = (A0 - A sample) / A0 × 100%

[0135] 4. Test Results

[0136] Table 3 Results of ABTS free radical scavenging rate

[0137]

[0138] As shown in Table 3, in the ABTS free radical scavenging ability test, Example 1 achieved a free radical scavenging rate of 100% at both 1% and 2% concentrations, significantly higher than all comparative examples. The ABTS free radical scavenging ability reflects the comprehensive antioxidant efficacy of the composition; its high scavenging capacity indicates that this composition can comprehensively combat multiple free radicals and protect the skin from oxidative damage. Compared to Comparative Example 1, the scavenging rate of Example 1 was nearly doubled, demonstrating that the optimization of the formulation design and extraction process in this invention significantly enhanced its antioxidant performance.

[0139] Test 4: Relative angiogenesis length

[0140] 1. Detection Principle

[0141] Zebrafish embryos, due to their transparent bodies and clearly visible blood vessel growth, have become a classic model for studying angiogenesis and microcirculation improvement. Microscopic observation of angiogenesis between the segments of zebrafish embryos allows for the evaluation of the sample's role in promoting blood circulation.

[0142] 2. Conditions for raising zebrafish

[0143] In the breeding and specialized aquaculture system for adult wild zebrafish, the pH range is 7.0-8.0, the conductivity range is 500-800 μS / cm, the room temperature range is 26℃±1℃, the water temperature range is 28℃±1℃, and the photoperiod is 14h light and 10h darkness.

[0144] 3. Preparation of experimental embryos

[0145] Healthy zebrafish embryos were collected and placed in a constant temperature incubator at (28±0.5℃) with embryo buffer added (see Table 4 for embryo buffer preparation method). When the embryos reached 24 hpf, the normally developing embryos were selected for the experiment.

[0146] Table 4 Preparation method of embryo buffer solution

[0147]

[0148] Add the ingredients to the measuring cup according to Table 4, and bring the volume to 4.5L with ultrapure water. Filter using a vacuum filter (0.2μm pore size), sterilize using an autoclave, and store at room temperature.

[0149] 4. Test substance pretreatment methods

[0150] The samples from Example 1 and Comparative Example 1 were prepared into 40 g / L stock solutions at a ratio of 0.3811 g sample to 9.5275 mL embryo buffer, and then diluted to the required final concentration for testing.

[0151] Table 5 Specific information of experimental samples

[0152]

[0153] 5. Test Methods

[0154] 5.1 Experimental Grouping

[0155] This experiment consisted of 1+1+1+3 groups, namely, blank control group, model control group, positive control group, and sample concentration groups*3.

[0156] 5.2 Group Design

[0157] Use 24-well cell culture plates, with 3 wells per group and 10 embryos per well. See Table 6 for details.

[0158] Table 6 Dosage Design Table

[0159]

[0160] 5.3 Experimental Procedure

[0161] 1) Collect wild-type zebrafish embryos at 24 hpf and assign them to groups of 30 embryos each, according to the group design in 5.2;

[0162] 2) Add 2 mL of the corresponding treatment culture medium to each well: the blank control group was given embryo buffer without any drugs; the model control group was given embryo culture medium containing 2 μmol / L sorafenib; the positive control group was given embryo culture medium containing both 2 μmol / L sorafenib and 5 g / L asiaticoside; each group in Example 1 was given embryo culture medium containing both 2 μmol / L sorafenib and the corresponding concentration of the sample in Example 1; each group in Comparative Example 1 was given embryo culture medium containing both 2 μmol / L sorafenib and the corresponding concentration of the sample in Comparative Example 1.

[0163] 3) Incubate at 28℃ for 6 hours, then change the solution and continue incubation until 72 hpf;

[0164] 4) After the embryo is removed, it is fixed and angiogenesis is observed using a fluorescence microscope.

[0165] 5.4 Data Analysis and Processing

[0166] The length of intersegmental angiogenesis in zebrafish was statistically analyzed using Photoshop. The results were imported into Excel for data processing, and the mean and standard error (SEM) for each group were calculated. All results are expressed as mean ± standard error (Mean ± SEM). GraphPad Prism 8 was used for statistical analysis, and differences between groups were analyzed using a t-test. Generally, P < 0.05 was considered statistically significant.

[0167] 6. Test Results

[0168] See zebrafish angiogenesis in the sample Figure 1 , Figure 2 And Table 7.

[0169] Table 7. Statistical results of relative angiogenesis length in zebrafish.

[0170]

[0171] Note: Compared with the blank control group, significance is indicated by #, p < 0.05 is indicated by #, p < 0.01 is indicated by ##, p < 0.001 is indicated by ###; compared with the model control group, significance is indicated by *, p < 0.05 is indicated by *, p < 0.01 is indicated by **, p < 0.001 is indicated by ***.

[0172] like Figure 1 , Figure 2 As shown in Table 7, in the angiogenesis experiment of zebrafish embryos, Example 1 exhibited a significant angiogenesis-promoting effect at different concentrations. At a concentration of 0.5 g / L, the relative angiogenesis length of Example 1 was 0.30 ± 0.03, which was significantly increased compared to 0.28 ± 0.03 in the model control group (P < 0.05). With increasing concentration, the angiogenesis length of Example 1 at concentrations of 1 g / L and 2 g / L was 0.36 ± 0.05 and 0.37 ± 0.04, respectively, which were significantly increased compared to the model control group (P < 0.01 and P < 0.001). Furthermore, the angiogenesis effect of Example 1 at a concentration of 0.5 g / L was significantly better than that of Comparative Example 1. These results indicate that the composition of the present invention can effectively promote blood circulation and microcirculation, contributing to improved skin blood supply and health.

[0173] Test 5: Detection of expression levels of genes related to angiogenesis

[0174] 1. Detection Principle

[0175] Zebrafish angiogenesis after injury is regulated by many related genes. By testing the expression of AKT1 (protein kinase B1), NOS1 (neural nitric oxide synthase), and vegfaa (vascular endothelial growth factor Aa subtype) genes in zebrafish and comparing the relative expression levels of related genes in the model control group and the sample group, the promoting effect of the samples on vascular microcirculation can be evaluated.

[0176] 2. Zebrafish rearing conditions: Same as those in Test 4.

[0177] 3. Preparation of experimental embryos: Same as preparation for test 4.

[0178] 4. Test substance pretreatment method: Same as test 4.

[0179] 5. Test Methods

[0180] 5.1 Experimental Grouping

[0181] This experiment consisted of 1+1+1+3 groups, namely, a blank control group, a model control group, a positive control group, and three groups with different sample concentrations.

[0182] 5.2 Group design: Use 6-well cell culture plates, with 3 wells per group and 30 embryos per well. See Table 8 for details.

[0183] Table 8 Dosage Design Table

[0184]

[0185] 5.3 Experimental Procedure

[0186] 1) Collect wild-type zebrafish embryos at 24 hpf and allocate them according to the 5.2 grouping design, with 90 embryos in each group;

[0187] 2) Add 2 mL of the corresponding treatment culture medium to each well: the blank control group was given embryo buffer without any drugs; the model control group was given embryo culture medium containing 2 μmol / L sorafenib; the positive control group was given embryo culture medium containing both 2 μmol / L sorafenib and 5 g / L asiaticoside; each group in Example 1 was given embryo culture medium containing both 2 μmol / L sorafenib and the corresponding concentration of the sample in Example 1.

[0188] 3) Incubate at 28℃ for 6 hours, then change the solution and continue incubation until 72 hpf;

[0189] 4) Wash the sample three times with PBS, extract RNA, perform reverse transcription, and then prepare a system for real-time quantitative PCR analysis;

[0190] 5) Statistically analyze the changes in the expression levels of AKT1, NOS1, and vegfaa genes.

[0191] 5.4 Data Analysis and Processing

[0192] The statistical results were imported into Excel for data processing. The mean and standard error (SEM) of each group were calculated, and the results were expressed as mean ± standard error (Mean ± SEM). The statistical analysis software GraphPad Prism 8 was used to analyze the data, and the differences between different groups were analyzed using a t-test. Generally, P < 0.05 was considered statistically significant.

[0193] 6. Test Results

[0194] The expression levels of genes related to angiogenesis in zebrafish samples are shown in the figure. Figure 1 And Tables 9, 10, and 11.

[0195] Table 9. Statistical results of expression levels of zebrafish-related genes (AKT1)

[0196]

[0197] Note: Compared with the blank control group, significance is indicated by #, p < 0.05 is indicated by #, p < 0.01 is indicated by ##, p < 0.001 is indicated by ###; compared with the model control group, significance is indicated by *, p < 0.05 is indicated by *, p < 0.01 is indicated by **, p < 0.001 is indicated by ***.

[0198] Table 10. Statistical results of gene expression levels in zebrafish (NOS1)

[0199]

[0200] Note: Compared with the blank control group, significance is indicated by #, p < 0.05 is indicated by #, p < 0.01 is indicated by ##, p < 0.001 is indicated by ###; compared with the model control group, significance is indicated by *, p < 0.05 is indicated by *, p < 0.01 is indicated by **, p < 0.001 is indicated by ***.

[0201] Table 11 Statistical results of expression levels of zebrafish-related genes (vegfaa)

[0202]

[0203] Note: Compared with the blank control group, significance is indicated by #, p < 0.05 is indicated by #, p < 0.01 is indicated by ##, p < 0.001 is indicated by ###; compared with the model control group, significance is indicated by *, p < 0.05 is indicated by *, p < 0.01 is indicated by **, p < 0.001 is indicated by ***.

[0204] As shown in Tables 9-11, the composition of the present invention, namely Example 1, can significantly upregulate the expression of three key genes closely related to angiogenesis in zebrafish embryos: AKT1, NOS1, and vegfaa, thereby revealing its mechanism of action in promoting skin microcirculation at the molecular level.

[0205] Specifically, compared with the model group, the AKT1 gene expression level in the Example 1 group was significantly different at a concentration of 2 g / L (P<0.01), indicating that it promoted AKT1 gene expression. AKT1 is a core node in the PI3K / AKT signaling pathway, and its activation can promote cell survival, proliferation, and metabolism, providing a basis for angiogenesis.

[0206] Meanwhile, compared with the model group, the NOS1 gene expression level in the Example 1 group showed significant differences at concentrations of 0.5 g / L, 1 g / L, and 2 g / L (P < 0.05 or P < 0.01), indicating that Example 1 has the effect of promoting NOS1 gene expression. NOS1 encodes nitric oxide synthase, which participates in the synthesis of nitric oxide. Nitric oxide, as an important vasodilator, can improve blood flow and promote microcirculation function.

[0207] Furthermore, compared with the model group, the vegfaa gene expression level in the Example 1 group showed a highly significant difference at concentrations of 1 g / L and 2 g / L (P < 0.001), indicating that Example 1 can significantly promote the expression of the vegfaa gene. vegfaa is a core factor regulating vascular endothelial cell proliferation, migration, and angiogenesis, and its increased expression level is beneficial to the occurrence of angiogenesis.

[0208] In summary, Example 1 can simultaneously promote the expression of key angiogenesis-related genes such as AKT1, NOS1, and vegfaa, forming a synergistic pro-angiogenic signaling pattern. This molecular-level evidence is completely consistent with the significant angiogenesis phenotype observed in Test 4, together strongly demonstrating that the composition of the present invention can fundamentally promote microcirculation by activating the PI3K / AKT-VEGF signaling pathway.

[0209] Test 6: Melanin Inhibition Test

[0210] 1. Detection Principle

[0211] Melanin production occurs through the conversion of tyrosine into melanin catalyzed by tyrosinase. Pigmentation in zebrafish embryos is similar to that in humans, and the embryos are transparent, facilitating observation. By analyzing changes in melanin area in zebrafish embryos, the melanin inhibition rate was calculated, and the inhibitory effect of the product was evaluated.

[0212] 2. Main reagents and consumables

[0213] 2.1 Water: Complies with GB / T 6682.

[0214] 2.2 Methanol, dimethyl sulfoxide, ethanol and other co-solvents were of analytical grade.

[0215] 2.3 Water for broodstock culture: Prepared by dissolving 4g of sea salt in 10L of water, with a salinity of 0.25-0.50‰, conductivity of 500-800µS / cm, dissolved oxygen ≥80% saturation, pH of 6.5-8.5, and hardness of 30-300mg / L (calculated as calcium carbonate).

[0216] 2.4 Artemia salina eggs: Dormant artemia eggs need to be dried, protected from light, and refrigerated. Weigh out about 15g of sea salt and dissolve it in 1L of water to prepare incubation water for artemia eggs. The density should not exceed 4g / L of artemia eggs.

[0217] 2.5 Fish embryo culture medium: Prepared by dissolving 2940 mg anhydrous calcium chloride, 1233 mg magnesium sulfate heptahydrate, 630 mg sodium bicarbonate, and 55 mg potassium chloride in 10 L of water. pH value: 6.5–8.5. All chemicals are of analytical grade.

[0218] 2.6 Phenylephrine stock solution: Prepared by dissolving 15 mg of phenylthionine in 10 mL of water.

[0219] 2.7 Phenylephrine working solution: Dilute the phenylthiourea stock solution 10 times with fish embryo culture medium to 0.15 g / L, and prepare 5 mL fresh before use.

[0220] 2.8 Methylcellulose solution: Prepared by dissolving 2-4g of methylcellulose in 100mL of water.

[0221] 2.9 Kojic acid working solution: 2.5 g / L

[0222] 3. Experimental Methods

[0223] 3.1 Test organisms: Healthy wild-type AB strain zebrafish were used and domesticated for 14 days.

[0224] 3.2 Spawning and Fertilization: Spawning takes place under appropriate water temperature and light conditions. The eggs are collected and cleaned. Healthy embryos are selected and cultured for 6-8 hours after fertilization.

[0225] 4. Test substance preparation: The test substance is dissolved directly in fish embryo culture medium to prepare the test solution.

[0226] 5. Testing Procedures

[0227] 5.1 Test Grouping

[0228] Blank / solvent control group (fish embryo culture medium / solvent solution), positive control group (kojic acid working solution), and test substance group.

[0229] 1) Blank control group setup: 24 fish embryos were randomly selected and transferred to 24-well plates, with each well containing 6 fish embryos and 1 mL of fish embryo culture medium / solvent solution.

[0230] 2) Setting up a positive control group: 24 fish embryos were randomly selected and transferred to a 24-well plate, with 6 fish embryos and kojic acid working solution in each well.

[0231] 3) Test substance treatment: 20 fish embryos were randomly selected and transferred to a 48-well plate, with each well containing one fish embryo and the test substance solution.

[0232] Incubate in a constant temperature incubator at 28±1℃ for 72±1h after fertilization.

[0233] 5.2 Microscopic Analysis of Samples: At least 8 fish embryos were randomly selected from each test group, covered with 2-4% methylcellulose, and placed back-side up. They were then photographed under an upright microscope. All fish embryos were photographed using standardized parameters.

[0234] 5.3 Data and Result Calculation: The melanin area was measured using image analysis software (such as ImageJ), and the melanin inhibition rate was calculated. The formula for calculating the melanin inhibition rate is as follows:

[0235]

[0236] Where T is the average signal intensity of the fish embryos in the test substance treatment group; C is the average signal intensity of the fish embryos in the blank control group.

[0237] A two-tailed t-test was performed on the melanin area of ​​fish embryos in the test group and the melanin area in the blank control group to obtain the p-value.

[0238] At least 90% of the fish embryos must survive after exposure in each test group; otherwise, the results of the corresponding test group are invalid.

[0239] 6. Test Results

[0240] The results of the inhibition rate of melanin production in zebrafish samples are shown in Table 12.

[0241] Table 12 Effect of samples on inhibition of melanin production in fish embryos (%)

[0242]

[0243] Note: Significance compared to the blank control group is indicated by *, p < 0.05 is indicated by *, p < 0.01 is indicated by **, and p < 0.001 is indicated by ***.

[0244] As shown in Table 12, at a concentration of 12.5 g / L, Example 1 showed a significant inhibitory effect on melanin production in fish embryos, with a melanin inhibition rate of 35.55%, significantly better than the blank control group (P<0.05). Compared with the positive control group, Example 1 showed a similar inhibitory effect (positive control group inhibition rate 42.15%), but its melanin inhibition effect was slightly inferior. Comparative Example 1 showed a melanin inhibition rate of 28.62% at the same concentration, indicating a lower inhibitory effect, which was also significantly different (P<0.05). These results indicate that Example 1 can effectively inhibit melanin production in zebrafish embryos, thereby helping to improve skin pigmentation.

[0245] Test 7: Cell proliferation promotion rate

[0246] 1. Detection Principle

[0247] CCK-8 is a commonly used reagent for detecting cell proliferation and toxicity. Its full name is Cell Counting Kit-8. It assesses cell proliferation and toxicity by detecting metabolic activity in living cells.

[0248] 2. Experimental Materials

[0249] 2.1 Reagents and consumables: DMEM culture medium, fetal bovine serum (FBS), penicillin, Streptomycin (PS), CCK-8 kit, 96-well white plate, 6-well plate

[0250] 2.2 Cell Information: HUVEC cell line; Culture medium: DMEM + 10% FBS + PS

[0251] 3. Test Methods

[0252] 1) Set up 9 experimental groups (0-5%) with different concentrations in a 96-well plate, with 3 replicates for each group;

[0253] 2) After culturing the cells for 24 hours, aspirate the culture medium and add drug-containing medium. The control group is treated with drug-free medium.

[0254] 3) After adding CCK-8 solution, incubate for 2 hours, measure the OD value at 450nm using a microplate reader, and analyze and process the data.

[0255] 4. Test Results

[0256] Table 13 HUVEC cell growth status

[0257]

[0258] As shown in Table 13, Example 1 had a significant effect on HUVEC cell proliferation. At a concentration of 0.04%, the cell proliferation rate was 100.74%, similar to the 100% in the control group, indicating that this concentration had a relatively small effect on cell proliferation. With increasing concentration, the proliferation rate was 111.14% at 0.16% and further increased to 124.71% at 0.31%, indicating that this concentration significantly promoted cell proliferation. However, when the concentration reached 1.25% and above, the proliferation rate significantly decreased, dropping to a maximum of 36.12%. These results indicate that the cell proliferation-promoting effect of the composition of the present invention is concentration-dependent; moderate concentrations can effectively promote cell proliferation, while excessively high concentrations may inhibit proliferation.

[0259] Test 8: Cellular Scratch Repair

[0260] 1. Experimental Principle

[0261] The cell scratch assay assesses cell migration ability. A straight line is drawn across a fully confluent cell layer to create a blank area, and cell migration to fill the blank is observed, thus assessing the speed and efficiency of cell migration.

[0262] 2. Experimental Materials

[0263] 2.1 Reagents and consumables: DMEM culture medium, fetal bovine serum (FBS), penicillin, Streptomycin (PS), CCK-8 kit, 96-well white plate, 6-well plate

[0264] 2.2 Cell Information: HUVEC cells; Culture Medium: DMEM + 10% FBS + PS

[0265] 3. Test Methods

[0266] 1) Seed the cells in 6-well plates and culture for 24 hours until they are fully confluent.

[0267] 2) Use a sterile scraper or pipette tip to make incisions in the cell layer to create blank areas.

[0268] 3) Add different concentrations (0.075%, 0.15%, 0.3%) of Example 1 or PBS as controls.

[0269] 4) Continue culturing and observe the healing of the scratch at 24 hours and 48 hours respectively.

[0270] 5) Use ImageJ software to analyze the scratch healing rate and calculate the cell migration rate.

[0271] 4. Test Results

[0272] Table 14 Statistical Table of Cell Scratch Migration Rate

[0273]

[0274] like Figure 3 As shown in Table 14, the cell migration rate of Example 1 at a concentration of 0.075% was 48.05% after 24 hours and 68.66% after 48 hours. With increasing concentration, the migration rate at 0.15% concentration was 49.85% after 24 hours and 79.49% after 48 hours; at a concentration of 0.3%, the migration rate was 52.97% after 24 hours and 81.47% after 48 hours. These results indicate that Example 1 significantly promotes HUVEC cell migration, especially at a concentration of 0.3%, further demonstrating that the composition of the present invention can promote cell repair processes.

[0275] In summary, tests 1-3 confirm that by using a eutectic solvent combined with microfluidic extraction technology, this composition significantly increases the content of flavonoid active ingredients and exhibits strong DPPH and ABTS free radical scavenging capabilities, laying the material foundation for its antioxidant and activity protection.

[0276] Tests 4-5 in a zebrafish model revealed that this composition significantly promoted angiogenesis and upregulated the expression of key genes such as AKT1, VEGFA, and NOS1. This reveals its core mechanism of action at both the animal phenotype and molecular levels: directly promoting microcirculation improvement by activating the PI3K / AKT-VEGF signaling pathway.

[0277] Tests 6-8 further demonstrated that the composition not only has the effect of inhibiting melanin production and brightening the skin, but also promotes the proliferation and migration of vascular endothelial cells, thereby enhancing the skin's self-repair ability.

[0278] In summary, the composition of the present invention achieves comprehensive effects of improving skin microcirculation, alleviating dull skin tone, and delaying aging through efficient extraction of active substances, activation of microcirculation core pathways, and synergistic effects of whitening and repair mechanisms.

[0279] The embodiments described above are merely illustrative of several implementations of the present invention, and while the descriptions are specific and detailed, they should not be construed as limiting the scope of the present invention. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these modifications and improvements all fall within the scope of protection of the present invention. Therefore, the scope of protection of this patent should be determined by the appended claims.

Claims

1. A composition for promoting skin microcirculation, characterized in that: It is composed of the following ingredients: Bletilla striata, Leonurus japonicus, Poria cocos, Paeonia suffruticosa, Ginseng, Peach blossom, and Lotus.

2. The composition for promoting skin microcirculation according to claim 1, characterized in that: It is composed of the following components in parts by weight: Bletilla striata 20-60 parts, Leonurus japonicus 20-60 parts, Poria cocos 20-50 parts, Paeonia suffruticosa 20-50 parts, Panax ginseng 5-30 parts, Peach blossom 10-30 parts, and Lotus 10-30 parts.

3. The method for preparing the composition for promoting skin microcirculation according to claim 2, characterized in that: Includes the following steps: S1. Weigh out Bletilla striata, Leonurus japonicus, Poria cocos, Paeonia suffruticosa, Panax ginseng, Peach blossom and Lotus in proportion, mix them, pulverize them, and sieve them through a sieve to obtain medicinal powder. S2. Mix the medicinal powder with a eutectic solvent until homogeneous to obtain a mixture; S3. The mixture is extracted using microfluidic technology to obtain a crude extract; S4. The crude extract is subjected to centrifugation and membrane filtration purification to obtain the target product.

4. The preparation method according to claim 3, characterized in that: The eutectic solvent includes betaine, hydrogen bond donors, and water.

5. The preparation method according to claim 4, characterized in that: The molar ratio of betaine to hydrogen bond donor is 1:1 to 1:5; the mass percentage of water is 10% to 80%.

6. The preparation method according to claim 4, characterized in that: The hydrogen bond donors include one or more of propylene glycol, glycerol, butanediol, and urea.

7. The preparation method according to claim 3, characterized in that: In step S2, the mass ratio of the medicinal powder to the eutectic solvent is 1:5 to 1:

15.

8. The preparation method according to claim 3, characterized in that: In step S3, the extraction temperature for the extraction process is 15-30℃.

9. The preparation method according to claim 3, characterized in that: In step S4, the centrifugation speed is 4000-8000 rpm and the centrifugation time is 20-40 min; the membrane filtration purification process is ceramic membrane separation and purification, and the pore size of the ceramic membrane is 200-600 μm.

10. The use of the composition for promoting skin microcirculation as described in any one of claims 1-2 in cosmetics for promoting skin microcirculation, improving dull skin tone, or delaying skin aging.

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