Galikalii-euphorbia tripsacus composite extract as well as preparation method and vagina repair application thereof

Abstract

The invention discloses a galiaceae-euphorbia tripsacus compound extract as well as a preparation method and vagina repair application thereof. The compound extract is prepared from galiaceae herbs and euphorbia tripsacus according to a scientific ratio through compound enzymolysis pretreatment, gradient ethanol solvent step-by-step extraction and double-resin series purification. According to the preparation method, the dissolution rate of the active components is remarkably increased by utilizing an enzymolysis wall breaking technology, the problem that the purity of the multi-polar components (tannin, flavone and polysaccharide) is difficult to increase and extract at the same time is solved through gradient extraction and series purification, and the purity of the total active components is more than or equal to 60%. The obtained composite extract has multiple effects of astringing and tightening, promoting regeneration of elastic fibers and improving contractility of smooth muscles, and is safe and non-irritant. The invention further discloses targeted sustained-release gel containing the extract, and the targeted sustained-release gel can be used for improving vaginal relaxation and deeply repairing private parts.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of biological medicine and female private care, and particularly relates to a Manjakani-Sanmiao composite extract and a preparation method and vaginal repair application thereof. BACKGROUND

[0002] Vaginal relaxation is a common physiological problem for women after childbirth, age growth or hormone level fluctuation. The core pathological mechanism mainly involves two aspects: one is the decrease of vaginal wall smooth muscle contractility; the other is the rupture and degradation of elastic fibers in the lamina propria of vaginal mucosa. This not only reduces the quality of sexual life, but also may lead to incomplete closure of the vagina, thereby increasing the risk of bacterial invasion and causing gynecological inflammation.

[0003] At present, the solutions for vaginal relaxation mainly include surgical treatment, laser treatment and the use of topical products. Among them, surgery and laser treatment have the problems of large trauma, long recovery period and high cost, although they have quick effects. The topical products are concerned due to their convenience, but the existing technology has obvious defects, such as: Many synthetic astringents or hormones are added, and long-term use can easily destroy the vaginal microecological balance, even cause mucosal irritation and allergic reaction, and the safety is questionable; Most of the existing products use single plant raw materials. For example, only Manjakani extract is used, which is rich in tannins and can provide short-term astringency, but cannot repair damaged elastic fibers, and the effect is difficult to last, or only herbal ingredients with repair function are used, which have slow effects and cannot meet the needs of consumers for rapid tightening.

[0004] In addition, the traditional extraction process mostly uses simple water extraction or single concentration alcohol extraction, which is difficult to consider the dissolution of different polar active ingredients in plants. For example, tannins (strong polarity) in Manjakani and flavonoids (weak polarity) in Sanmiao are difficult to achieve high extraction rate at the same time in a single solvent, resulting in low purity (usually ≤30%) of total active ingredients, in addition, the hindrance of plant cell wall makes the dissolution of intracellular active substances insufficient, and the utilization rate of raw materials is low, therefore, a Manjakani-Sanmiao composite extract and a preparation method and vaginal repair application thereof are urgently needed. SUMMARY

[0005] The technical problem to be solved by the present application is that the existing vaginal private care products have the defects of single active ingredient, low purity caused by extensive extraction process, low utilization rate of raw materials and inability to consider the dual effects of rapid astringency and deep repair, and a Manjakani-Sanmiao composite extract and a preparation method and application thereof are provided.

[0006] To solve the above technical problems, the technical scheme adopted by the present application is as follows: In a first aspect, the present application provides a composite extract of Mallotus philippinensis and Smilax china, which is an active composition prepared by mixing Mallotus philippinensis and Smilax china at a mass ratio of 1:1 to 2:1, pre-treating by composite enzymolysis, extracting by gradient ethanol solvent, and purifying by double-resin series connection. In the composite extract, synergistically matched tannin, flavonoid compound and polysaccharide are contained, the content of the tannin is greater than or equal to 15%, the content of the flavonoid compound is greater than or equal to 25%, the content of the polysaccharide is greater than or equal to 20%, and the total active ingredient purity of the composite extract is greater than or equal to 60%.

[0007] In a second aspect, the present application provides a preparation method of the composite extract of Mallotus philippinensis and Smilax china, which comprises the following steps. In step S1, the raw materials are pre-treated by crushing and mixing Mallotus philippinensis and Smilax china to obtain mixed raw materials. In step S2, the mixed raw materials obtained in step S1 are pre-treated by composite enzymolysis by adding acetic acid-sodium acetate buffer solution with a pH value of 4.5 to 5.5 and adding a composite enzyme solution of cellulase and pectinase, and then performing constant-temperature enzymolysis at 45 to 55 DEG C, and after the enzymolysis is completed, the enzyme is inactivated to obtain an enzymolysis solution. In step S3, the enzymolysis solution obtained in step S2 is extracted by gradient solvent step-by-step. In S301, the enzymolysis solution obtained in step S2 is added with ethanol to adjust the ethanol volume fraction to 50%, and then extracted at 45 to 55 DEG C, and the solid-liquid separation is performed to collect the first filtrate and the filter residue. In S302, the filter residue is added with ethanol solution with a volume fraction of 80%, and then extracted at 55 to 65 DEG C, and the solid-liquid separation is performed to collect the second filtrate. In S303, the first filtrate and the second filtrate are combined to obtain a mixed extract. In step S4, the mixed extract obtained in step S3 is concentrated and passed through AB-8 macroporous resin and polyamide resin in sequence, wherein the AB-8 macroporous resin is used for adsorption and elution to obtain a first eluate, and then the first eluate is passed through the polyamide resin for adsorption and elution to obtain a second eluate. In step S5, the second eluate obtained in step S4 is concentrated and dried to obtain the composite extract.

[0008] In step S2, preferably, the mass ratio of cellulase to pectinase in the composite enzyme solution is 1:1, the addition amount of the composite enzyme solution is 0.3% to 0.8% of the mass of the mixed raw materials, and the constant-temperature enzymolysis time is 1.5 to 2.5 hours.

[0009] Preferably, the double-resin series purification in step S4 is specifically as follows: the mixed extract is loaded onto the AB-8 macroporous resin, impurities are removed by washing with water, and then the first eluent is collected by elution with an ethanol solution with a volume fraction of 60%; The first eluent is directly loaded onto the polyamide resin, and the second eluent is collected by elution with an ethanol solution with a volume fraction of 70%.

[0010] In a third aspect, the present application provides use of the composite extract in the preparation of a female private care product.

[0011] Further, the female private care product is used for improving vaginal relaxation, promoting regeneration of vaginal elastic fibers, or improving contraction of vaginal smooth muscle.

[0012] Preferably, the female private care product is a targeted slow-release gel, and the raw materials of the targeted slow-release gel include the composite extract with a mass percentage of 8-20%, a slow-release carrier, and a penetration enhancer. The slow-release carrier is a chitosan-sodium alginate composite microsphere. The penetration enhancer is a compound of laurocapram and propylene glycol with a volume ratio of 1:2-1:4.

[0013] Compared with the prior art, the present application has the following advantages: The combination of the herb of maringa and the herb of three xiaomao can quickly enhance the contraction of smooth muscle to achieve instant tightening, and the flavonoids and polysaccharides in the herb of three xiaomao can promote the proliferation of elastic fiber cells and inhibit the activity of degrading enzymes, so that the two components form a closed loop of convergence and repair, which is superior to a single raw material product.

[0014] The composite enzymolysis of cellulase and pectinase can effectively destroy the plant cell wall, so that the intracellular active ingredients are fully released. The gradient extraction strategy of 50% ethanol for flavonoids / polysaccharides and 80% ethanol for tannins can solve the problem of co-solubility of different polar components, thereby improving the utilization rate of raw materials. The AB-8 macroporous resin and the polyamide resin are used in series purification, the AB-8 resin is mainly used to remove water-soluble impurities such as sugars and pigments, and the polyamide resin is used to specifically adsorb and enrich flavonoids and tannins. Through the fine purification process, more than 90% of the invalid impurities are removed. The obtained extract has no irritation and no sensitization to vaginal mucosa, thereby improving the safety of the product in private care applications.

[0015] The composite extract is prepared into a targeted slow-release gel, and the chitosan-sodium alginate composite microsphere is used as a carrier to effectively avoid the rapid loss of traditional gels due to the flushing of secretions, thereby realizing the long-term residence and continuous effect of the drug. BRIEF DESCRIPTION OF DRAWINGS

[0016] The accompanying drawings are provided to further illustrate the invention and form part of the specification. They are used together with the embodiments of the invention to explain the invention and do not constitute a limitation thereof.

[0017] Figure 1 This is a flow chart of the preparation process of the Manjacaranda-Trifolium repens compound extract provided in the embodiments of the present invention; Figure 2 This is a bar chart comparing the improvement rate of vaginal contractility between the compound extract and the single-raw-ingredient extract of this invention; Figure 3 This is a schematic diagram of the microstructure and release principle of the targeted sustained-release gel provided in the embodiments of the present invention. Detailed Implementation

[0018] The technical solutions of the present invention will be clearly and completely described below with reference to the accompanying drawings of the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments.

[0019] Unless otherwise specified, the raw materials and reagents used in the embodiments and comparative examples of this invention are all commercially available products.

[0020] Manjakani (Quercus infectoria): Dried insect galls, medicinal grade, purchased from Bozhou Traditional Chinese Medicine Trading Center, Anhui Province.

[0021] Three-leafed herb (Curculigo orchioides): Dried rhizome, medicinal grade, purchased from Bozhou Traditional Chinese Medicine Trading Center, Anhui Province. Enzyme preparation: Cellulase: Enzyme activity ≥10,000 U / g.

[0022] Pectinase: CAS No.: 9032-75-1, enzyme activity ≥30,000 U / g, purchased from Sigma-Aldrich.

[0023] Cellulase: CAS No.: 9012-54-8, enzyme activity ≥10,000 U / g, purchased from Shanghai Yuanye Biotechnology Co., Ltd.

[0024] Polyamide resin: CAS No.: 63428-84-2, chromatography grade (14-30 mesh), purchased from Nanjing Dulai Biotechnology Co., Ltd.

[0025] Resin: AB-8 macroporous adsorption resin: weakly polar copolymer styrene type.

[0026] Reference standards: gallic acid (for tannin determination), rutin (for flavonoid determination), and anhydrous glucose (for polysaccharide determination), all with a purity ≥98%.

[0027] Among them, gallic acid (CAS No.: 149-91-7, purity ≥99%) was purchased from Zhangjiajie Jiurui Biotechnology Co., Ltd.

[0028] Rutin: CAS No.: 153-18-4, purity ≥98%, purchased from Wuhan Yuqing Jiaheng Pharmaceutical Co., Ltd.

[0029] Anhydrous glucose: CAS No.: 50-99-7, purity ≥99% (biotechnology grade), purchased from Shanghai Maclean Biochemical Technology Co., Ltd.

[0030] Example 1 like Figure 1 As shown, a method for preparing a compound extract of Manjacaranda and Trifolium repens is described, with the following specific steps: Step S1: Select 150g of dried Manjakali herbs that are free of impurities and pollution and 100g of fresh Sanxianmao grass (mass ratio 1.5:1), crush them separately and pass them through an 80-mesh sieve, mix them evenly, and obtain 250g of mixed raw material powder.

[0031] Step S2: Add 3750 mL of acetate-sodium acetate buffer solution (solid-to-liquid ratio 1:15 g / mL) with a pH of 5.0 to the mixed raw materials obtained in step S1. Add a compound enzyme solution of cellulase and pectinase (mass ratio of 1:1), with a total enzyme addition of 0.5% (i.e. 1.25g) of the mass of the mixed raw materials. The enzyme was stirred and hydrolyzed at a constant temperature of 50℃ for 2 hours. After the hydrolysis was completed, the enzyme was heated to 90℃ and held for 10 minutes to inactivate it. The solution was then cooled to room temperature to obtain the hydrolysate.

[0032] Step S3: Perform gradient solvent stepwise extraction, specifically as follows: S301: Add anhydrous ethanol to the enzymatic hydrolysate obtained in step S2, adjust the ethanol volume fraction of the system to 50%, and extract by stirring at a constant temperature of 50°C for 1.5 hours. After filtration by a plate and frame filter press, the first filtrate and the filter residue are collected separately. S302: Add 3000 mL (approximately 12 times the volume) of 80% ethanol solution to the filter residue, and extract by stirring at 60°C for 2 hours; filter and collect the second filtrate. S303: Combine the first and second filtrates, concentrate under reduced pressure to recover the ethanol, and obtain a mixed extract (concentrated to a relative density of 1.08, measured at 60°C).

[0033] Step S4: Dual Resin Tandem Purification After cooling the mixed extract obtained in step S3, it is loaded onto a pretreated AB-8 macroporous resin column (diameter-to-height ratio 1:9) at a flow rate of 2.5 BV / h. First, rinse with 4 BV of purified water to remove sugars and water-soluble pigment impurities; then elute with 60% ethanol solution at a flow rate of 1.5 BV / h, and collect the first eluent (approximately 3.5 BV). The first eluent was directly loaded onto a polyamide resin column (diameter-to-height ratio 1:7) at a flow rate of 1.5 BV / h. After adsorption equilibrium was reached, the column was eluted with a 70% ethanol solution at a flow rate of 1 BV / h, and the second eluent (approximately 2.5 BV) was collected.

[0034] Step S5: Concentrate the second eluent under reduced pressure at a vacuum of 0.08 MPa and 50°C until no alcohol odor remains, to obtain a thick paste with a relative density of 1.23; The mixture was transferred to a freeze dryer and dried at -45°C and 15 Pa for 30 hours. After being pulverized and passed through a 120-mesh sieve, the compound extract powder was obtained (weighed at 51.3g, with an extraction rate of 20.5%).

[0035] Example 2 This embodiment provides another method for preparing a compound extract, which differs from Example 1 only in the adjustment of the enzymatic hydrolysis parameters in step S2; the remaining steps are the same. Step S2: Adjust the pH of the buffer solution to 4.5, the amount of enzyme added to 0.5%, the enzymatic hydrolysis temperature to 50℃, and extend the enzymatic hydrolysis time to 2.5 hours. Under these conditions, it is beneficial to further break down the plant cell wall and promote the release of deep polysaccharides.

[0036] Example 3 This embodiment provides another method for preparing a compound extract, using a raw material mass ratio of 1:1.

[0037] Step S1: Mix 100g of *Manjacali* herb and 100g of *Trifolium repens* herb evenly. Step S2: Buffer pH 5.5, enzyme addition 0.3%, enzymatic hydrolysis temperature 45℃, enzymatic hydrolysis time 1.5 hours. Step S3: Extraction temperature 45℃ for S301; extraction temperature 55℃ for S302, ethanol concentration 80%. Step S4: Elute with AB-8 resin at 60% ethanol concentration; elute with polyamide resin at 70% ethanol concentration.

[0038] The remaining operations are the same as in Example 1.

[0039] To verify the inventiveness of the process of this invention, the following comparative examples are set up: Comparative Example 1: Compared with Example 1, the enzyme addition and enzymatic hydrolysis process in step S2 was omitted, and the sample was directly soaked in buffer solution for 2 hours before extraction in step S3. The remaining steps were the same.

[0040] Comparative Example 2: Compared with Example 1, step S3 was changed from gradient extraction to: ethanol was directly added to the enzymatic hydrolysate to adjust the concentration to 70%, and extraction was performed for 3.5 hours in a single step. The remaining steps were the same.

[0041] Comparative Example 3: Compared with Example 1, step S4 only used AB-8 macroporous resin for purification, and the 60% ethanol eluent was collected and then directly dried.

[0042] The content and extraction rate of active ingredients were determined using the methods described in the Pharmacopoeia of the People's Republic of China and the instructions for this application (HPLC for tannins, UPLC for flavonoids, and phenol-sulfuric acid method for polysaccharides) to detect the extracts obtained in the above examples and comparative examples.

[0043] Table 1 compares the content and extraction rate of active ingredients in each group of extracts.

[0044] Group Tannin content (%) Flavonoid content (%) Polysaccharide content (%) Total purity (%) Extraction rate (%) Note Example 1 15.8 26.3 21.5 63.6 20.5 Overall optimal Example 2 15.2 27.1 22.8 65.1 21.2 Long enzyme digestion time, slightly high polysaccharide Example 3 14.5 24.8 19.6 58.9 19.1 Raw material ratio adjustment impact Comparative Example 1 15.5 18.2 12.5 46.2 13.4 No enzyme digestion, poor dissolution of intracellular components Comparative Example 2 16.1 20.4 14.2 50.7 10.5 Single solvent, weakly polar substances are not extracted Comparative Example 3 11.2 19.5 13.8 44.5 16.8 Incomplete impurity removal, low purity As can be seen from the above, compared with Comparative Example 1, the compound enzymatic hydrolysis pretreatment significantly improved the dissolution of flavonoids and polysaccharides (the extraction rate increased by about 53%), proving that enzymatic hydrolysis can effectively break the cell wall.

[0045] Compared with Comparative Example 1 and Comparative Example 2, gradient extraction (50% first, then 80%) can balance the dissolution of flavonoids / polysaccharides (weakly polar / polar) and tannins (strongly polar), and the total extraction rate is nearly doubled.

[0046] Compared with Comparative Example 1 and Comparative Example 3, the total purity was increased from 44.5% to 63.6% by dual-resin tandem purification, demonstrating that AB-8 impurity removal and polyamide adsorption have a significant synergistic purification effect.

[0047] In addition, for the verification of vaginal repair efficacy, the compound extract prepared in Example 1 was selected and prepared into a targeted sustained-release gel with a drug content of 12% (as described in Example 4).

[0048] Fifty subjects with grade I-II vaginal laxity were selected and treated once daily for 8 consecutive weeks.

[0049] Vaginal contractility improved: an average increase of 78.3%.

[0050] Elastic fiber integrity rate: 87.2% was observed through pathological sections.

[0051] Safety: No redness, stinging, or allergic reactions occurred during the trial, and the mucosal irritation score was 0. Example 4 This embodiment provides a feminine intimate care product, specifically a targeted sustained-release gel with deep repair and long-lasting firming effects. This targeted sustained-release gel uses the Manjacaranda-Trifolium repens compound extract prepared in Example 1 as its core active ingredient, combined with a specific sustained-release carrier and penetration-enhancing system. The microstructure of this targeted sustained-release gel is as follows: Figure 3 As shown, the chitosan-sodium alginate composite microspheres form a sustained-release shell with a mesh structure, encapsulating the core active ingredient (Manjacali-Trifolium repens composite extract). During use, the active ingredient is slowly released through the pores of the microspheres, thereby maintaining an effective drug concentration on the vaginal mucosa surface for a long time, achieving long-lasting repair.

[0052] Formula composition (total 100g): Example 1: Compound extract: 12.0 g Mucosal repair factors: Ceramide NP 2.0 g and Sodium Hyaluronate 1.0 g Penetration enhancer: 0.5 mL laurocapram and 1.5 mL propylene glycol Sustained-release carrier: 6.0 g of chitosan-sodium alginate composite microspheres. The microspheres can be purchased from Suzhou Nanomicro Technology Co., Ltd. or prepared in-house. The in-house preparation process is as follows: Prepared using the conventional ionic crosslinking method: Sodium alginate was dissolved in distilled water to prepare a 1.5% (w / v) solution; chitosan was dissolved in 1% acetic acid solution to prepare a 0.5% (w / v) solution, and 1% calcium chloride was added; under magnetic stirring, the sodium alginate solution was dripped into the chitosan-calcium chloride mixed solution through a syringe needle, solidified for 30 minutes, filtered, washed, and dried to obtain composite microspheres with a particle size of 10-50 μm.

[0053] Gel matrix: Carbomer 940 1.2 g pH adjuster: Triethanolamine (appropriate amount) Solvent: Add purified water to 100 g The preparation process, specifically the steps are as follows: (1) Disperse Carbomer 940 in 80 mL of purified water and let it stand to swell for 24 hours; (2) Add ceramide NP, sodium hyaluronate and chitosan-sodium alginate composite microspheres, and stir at low speed at 50°C for 1 hour to mix evenly; (3) Add the premixed compound extract, lauryl acetone and propylene glycol to the gel matrix and stir at 40°C for 2 hours; (4) Adjust the pH to 4.2 with triethanolamine (to match the vaginal environment), degas under vacuum, and fill into a sterile administration device.

[0054] Release tests of the gel in simulated vaginal fluid in vitro showed a cumulative release rate of 75% after 8 hours and 92% after 12 hours, demonstrating excellent sustained-release performance and preventing rapid loss of active ingredients.

[0055] To further verify the synergistic effect of the combination of Manjakalia and Sanxianmao grass in this invention, as well as the clinical efficacy of the compound extract in practical applications, a single-raw-material control group was set up for comparative testing in this experiment.

[0056] Experimental sample: The present invention group: the targeted sustained-release gel (containing 12% of the compound extract of the present invention) prepared in Example 4.

[0057] Single mangacalis group: The preparation process is the same as in Example 4, but the compound extract is replaced with an equal amount of single mangacalis extract (extraction process is the same as in Example 1).

[0058] Single herb group: The preparation process is the same as in Example 4, but the compound extract is replaced with an equal amount of single herb extract (extraction process is the same as in Example 1).

[0059] Subjects and Methods: 150 subjects diagnosed with grade I-II vaginal laxity (aged 25-45 years) were randomly divided into 3 groups (n=50 per group). Each group used the corresponding gel once daily for 8 weeks.

[0060] The test results are shown in Table 2.

[0061] Table 2. Comparison of the efficacy of the compound extract and single-raw material extract of the present invention Test index Inventive group (Example 4) Single Macropiper methysticum group Single Trifolium repens group Improvement rate (relative to single group mean) 8-week vaginal contractility improvement rate 78.30% 42.00% 38.00% +38.3% Elastic fibroblast proliferation rate 62.50% 25.00% 58.00% +21.0% Elastic fiber degradation enzyme inhibition rate 48.20% 18.00% 40.00% +19.2% 8-week elastic fiber integrity rate 87.20% 62.00% 78.00% +17.2% Mucous membrane irritation score (0-4 points) 0 (no irritation) 1.2 (slight) 0.8 (extremely slight) Safer As shown in Table 2, the single Manjaca lanceolata group performed reasonably well in improving contractility (42%), but had a poor effect on elastic fiber proliferation (25%); the single Trifolium repens group, on the other hand, showed strong repair ability (58%) but limited improvement in contractility (38%). The present invention combines the advantages of both, achieving a dual improvement in "contraction + repair".

[0062] The shrinkage strength improvement rate (78.3%) of the present invention group is not only better than any single group, but also close to the sum of two single groups, which proves that under the specific enzymatic hydrolysis and gradient extraction process, the active ingredients are released more fully and achieve unexpected technical effects.

[0063] Thanks to the removal of most impurities by the dual-resin tandem purification process, the mucosal irritation score of the present invention group was 0, which was significantly better than the single control group that had not undergone fine purification.

[0064] To more intuitively demonstrate the synergistic effect of the present invention, the key data in Table 2 are plotted as a bar chart, such as...Figure 2 As shown. From Figure 2 It can be clearly seen that the vaginal contraction force improvement rate of the present invention group (black column) is higher than that of the single Manjacali group and the single Trichosanthes kirilowii group (gray column), and the improvement is greater than the simple sum of the two, which confirms the unexpected technical effect brought about by the combination of dual raw materials and specific extraction process.

[0065] The above description is only a preferred embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any equivalent substitutions or modifications made by those skilled in the art within the scope of the technology disclosed in the present invention, based on the technical solution and inventive concept of the present invention, should be covered within the scope of protection of the present invention.

Claims

1. A compound extract of Manjacaranda and Trifolium repens, characterized in that, The compound extract is an active composition prepared by mixing Manjakalia herb and Trifolium repens in a mass ratio of 1:1 to 2:1, followed by compound enzymatic hydrolysis pretreatment, stepwise extraction with gradient ethanol solvent, and purification by dual resin tandem. The composite extract contains synergistically proportioned tannins, flavonoids, and polysaccharides, wherein the content of tannins is ≥15%, the content of flavonoids is ≥25%, the content of polysaccharides is ≥20%, and the purity of the total active ingredients in the composite extract is ≥60%.

2. The preparation method of the Manjacaranda-Trifolium repens compound extract according to claim 1, characterized in that, Includes the following steps: Step S1, Raw material pretreatment: The Manjakali herb and the Three-Spear Grass are crushed and mixed separately to obtain mixed raw materials; Step S2, compound enzymatic hydrolysis pretreatment: Add an acetate-sodium acetate buffer solution with a pH of 4.5 to 5.5 to the mixed raw materials obtained in step S1, and add a compound enzyme solution of cellulase and pectinase. Perform enzymatic hydrolysis at a constant temperature of 45 to 55°C. After the enzymatic hydrolysis is completed, inactivate the enzyme to obtain the enzymatic hydrolysate. Step S3: Perform gradient solvent stepwise extraction, wherein: S301: Add ethanol to the enzymatic hydrolysate obtained in step S2 to adjust the ethanol volume fraction of the system to 50%, extract at 45-55°C, separate solid and liquid, and collect the first filtrate and filter residue respectively. S302: Add an 80% ethanol solution to the filter residue, extract at 55-65°C, separate the solid and liquid, and collect the second filtrate; S303: Combine the first filtrate and the second filtrate to obtain a mixed extract; Step S4, Dual Resin Tandem Purification: The mixed extract obtained in step S3 is concentrated and passed sequentially through AB-8 macroporous resin and polyamide resin; wherein, the first eluent is obtained by adsorption and elution through the AB-8 macroporous resin, and then the first eluent is obtained by adsorption and elution through the polyamide resin to obtain the second eluent. Step S5, Post-processing: The second eluent obtained in step S4 is concentrated and dried to obtain the composite extract.

3. The preparation method according to claim 2, characterized in that, In step S2, the mass ratio of cellulase to pectinase in the compound enzyme solution is 1:1, the amount of compound enzyme solution added is 0.3% to 0.8% of the mass of the mixed raw materials, and the isothermal enzymatic hydrolysis time is 1.5 to 2.5 hours.

4. The preparation method according to claim 2, characterized in that, The dual-resin tandem purification in step S4 specifically includes: loading the mixed extract onto the AB-8 macroporous resin, rinsing with water to remove impurities, then eluting with a 60% (v / v) ethanol solution, and collecting the first eluent; The first eluent was directly loaded onto the polyamide resin, eluted with a 70% (v / v) ethanol solution, and the second eluent was collected.

5. The application of the compound extract according to claim 1 in the preparation of feminine intimate care products.

6. The application according to claim 5, characterized in that, The aforementioned feminine hygiene products are used to improve vaginal laxity, promote the regeneration of vaginal elastic fibers, or enhance the contractile force of vaginal smooth muscles.

7. The application according to claim 5, characterized in that, The feminine hygiene products mentioned are gels, suppositories, or creams.

8. The application according to claim 7, characterized in that, The feminine intimate care product is a targeted sustained-release gel, the raw materials of which include 8% to 20% by weight of the aforementioned compound extract, as well as a sustained-release carrier and a penetration enhancer; The sustained-release carrier is a chitosan-sodium alginate composite microsphere.

9. The application according to claim 8, characterized in that, The penetration enhancer is a compound of laurocapram and propylene glycol in a volume ratio of 1:2 to 1:4.

Images