Bacteriostatic gynecological lotion as well as preparation method and application thereof

Abstract

The invention discloses a polygonum cuspidatum extract, the effective components of the polygonum cuspidatum extract are mainly resveratrol and paeoniflorin, and the contents of the resveratrol and the paeoniflorin are respectively 1.25 mg / g and 1.50 mg / g. The invention also discloses a bacteriostatic gynecological lotion. The formula of the bacteriostatic gynecological lotion is as follows by mass: (1) 0.1% of the polygonum cuspidatum extract; (2) lactic acid bacteria: 1 * 10 < 8 > CFU / ml; (3) 0.02% of chloral hydrate; (4) 1% of plant glycerin; and (5) the pH value is 4.2. The novel bacteriostatic gynecological lotion provided by the invention has an excellent bacteriostatic effect, can maintain normal flora in the vagina, and is suitable for adjuvant therapy and daily nursing of various gynecological diseases.

Description

Technical Field

[0001] The present invention relates to the field of gynecological care, in particular to a gynecological lotion with significant bacteriostatic effect and capable of restoring vaginal microecological balance, its preparation method and application, belonging to the fields of biomedicine and gynecological care products. Background Art

[0002] Gynecological inflammatory diseases, such as bacterial vaginitis, vaginal candidal infection, bacterial vaginosis, etc., have become common health problems among women, and their incidence is increasing year by year. Due to the special physiological structure of women, the vagina is easily infected by the external environment, bacteria, fungi or viruses. These infections not only affect the quality of life of women, but may also lead to serious consequences such as infertility and premature birth. Most of the existing treatment methods rely on antibiotics, antifungal drugs or gynecological lotions containing chemical components. However, long-term use of these chemical drugs may lead to drug resistance, vaginal flora imbalance, and even damage the natural acidic environment in the vagina, making women vulnerable to infection again.

[0003] Common components in traditional gynecological lotions, such as chlorhexidine, hydrogen peroxide, povidone iodine, etc., although having certain antibacterial effects, often have inhibitory effects on beneficial flora in the vagina (such as lactobacilli, bifidobacteria), resulting in the destruction of the vaginal environment, thus triggering new infections or diseases. In addition, some chemical components may stimulate the vaginal mucosa, causing discomfort and affecting the treatment compliance of patients.

[0004] In recent years, with the continuous in-depth research on the application of natural plant components and probiotics in gynecological care, researchers have found that by using natural plant extracts and microecological regulators, it is possible to effectively regulate the vaginal environment, maintain the flora balance and avoid antibiotic resistance. However, there is still a lack of a new type of gynecological lotion on the market that can both inhibit bacteria and protect normal flora. Therefore, developing an antibacterial gynecological lotion with dual effects has become an important research direction to solve the above problems. Summary of the Invention

[0005] The purpose of the present invention is to provide a simple and efficient antibacterial gynecological lotion, which can help restore the vaginal microecological environment while inhibiting bacteria, avoid the drug resistance problem in traditional formulations, and show unexpected therapeutic effects clinically.

[0006] Therefore, on the one hand, the present invention discloses a polygonum cuspidatum extract, and the active ingredients of the polygonum cuspidatum extract are mainly resveratrol and paeoniflorin, and their contents are 1.25 mg / g and 1.50 mg / g respectively.

[0007] On the one hand, the present invention also discloses a method for preparing the polygonum cuspidatum extract, and the method includes the following steps:

[0008] (1) Raw material preparation and pretreatment: Select high-quality polygonum cuspidatum roots, ensure that the raw material source is pollution-free and pesticide-free, and ensure the stability of its components; After removing surface impurities by ultrasonic cleaning, cut the polygonum cuspidatum roots into small sections of 1-2 cm;

[0009] (2) Solvent selection: Select a 70% ethanol-water mixed solvent and add 0.1% acetic acid as an auxiliary solvent; The mass ratio of dry matter to solvent is 1:15;

[0010] (3) Ultrasonic-assisted extraction: Put the processed polygonum cuspidatum root segments into an extraction container, add 70% ethanol-water solution and 0.1% acetic acid; Use ultrasonic-assisted extraction, with a power of 500 W, a frequency of 40 kHz, at 50 °C for 30 minutes, and stop for 1 minute every 10 minutes for cooling; After extraction, filter to remove residues to obtain a preliminary extract;

[0011] (4) Reflux extraction: Pour the liquid after ultrasonic extraction into a reflux extractor and continue to perform reflux extraction with 70% ethanol; The reflux temperature is set at 80 °C, the reflux time is 2 hours, and it is sealed during reflux to prevent volatilization; After the reflux extraction is completed, cool the liquid, filter and remove residual impurities;

[0012] (5) Concentration and purification: Concentrate the filtered extract under reduced pressure to 1 / 5 of the original volume at 40 °C using a rotary evaporator to remove the ethanol solvent; Use activated carbon adsorption to remove impurities and ensure the purity and stability of the final extract;

[0013] (6) Preservation and packaging: Add vitamin C with a final concentration of 0.1% to the extract, disinfect with ultraviolet light, and then use a sterile glass bottle or aluminum film bag for sealed packaging and store at 2-8 °C for standby.

[0014] In one aspect, the present invention also discloses an antibacterial gynecological lotion, and the formula of the antibacterial gynecological lotion is as follows according to the mass ratio:

[0015] (1) The above-mentioned polygonum cuspidatum extract: 0.1%;

[0016] (2) Lactobacillus: 1×10^8 CFU / ml;

[0017] (3) Chloral hydrate: 0.02%;

[0018] (4) Vegetable glycerin: 1%;

[0019] (5) pH value: 4.2.

[0020] In one aspect, the present invention also discloses an antibacterial gynecological lotion, and the formula of the antibacterial gynecological lotion is as follows according to the mass ratio:

[0021] (1) The polygonum cuspidatum extract: 0.1%;

[0022] (2) Lactic acid bacteria: 1×10^8 CFU / ml;

[0023] (3) Chloral hydrate: 0.02%;

[0024] (4) Silymarin: 0.5%;

[0025] (5) Vegetable glycerin: 1%;

[0026] (6) pH value: 4.2.

[0027] In one aspect, the present invention also discloses the use of the polygonum cuspidatum extract in the preparation of an antibacterial gynecological lotion.

[0028] By combining the polygonum cuspidatum extract and / or silymarin in the gynecological lotion, the gynecological lotion has a significant therapeutic effect, can effectively restore the balance of vaginal flora, inhibit the growth of pathogenic bacteria, and has no obvious tissue damage, providing a safer and more effective option for the treatment of gynecological infections.

[0029] Silymarin is a natural active compound extracted from the seeds of the milk thistle (Silybum marianum) plant. It is mainly composed of three flavonoid compounds: silymarin A, B, and C (silybin, silychristin, silydianin), and silymarin A (silybin) is its main active ingredient. Due to its various biological activities, especially in antioxidant, anti-inflammatory, liver protection, and antibacterial aspects, silymarin has been widely used in many traditional medicines. The role of silymarin in the gynecological lotion may be as follows:

[0030] 1. Restore the balance of vaginal flora: The vagina is a complex microecosystem, and lactic acid bacteria usually dominate, maintaining the acidic environment and healthy flora of the vagina. When pathogenic bacteria (such as Escherichia coli, Staphylococcus aureus, etc.) invade, it may lead to flora imbalance and vaginal infections. Silymarin has antibacterial effects, can inhibit the growth of pathogenic bacteria, and at the same time promote the growth of beneficial bacteria (such as lactic acid bacteria), thus restoring the balance of vaginal flora.

[0031] 2. Antibacterial effect: Silymarin can inhibit a variety of common vaginal pathogenic bacteria, including Escherichia coli, Staphylococcus aureus, Candida, etc. It helps to relieve vaginal inflammation and infections and reduce vaginal discomfort symptoms by inhibiting the growth and reproduction of pathogenic bacteria.

[0032] 3. Anti-inflammatory effect: Vaginal inflammation is often accompanied by discomfort symptoms such as itching and burning sensations. Silymarin reduces local inflammatory responses and alleviates these symptoms by regulating the secretion of inflammatory factors. It inhibits the release of inflammatory factors such as interleukin (e.g., IL-6, TNF-α, etc.), reducing tissue damage.

[0033] 4. Promote tissue repair: Silymarin helps repair damaged vaginal epithelial tissues, especially after vaginal mucosal damage caused by infection. It promotes cell regeneration, enhances local immunity, and accelerates the healing and repair of vaginal tissues.

[0034] 5. Antioxidant protection: Oxidative stress is not only related to bacterial infections but also closely associated with the aging process and the occurrence of various diseases. The antioxidant effect of silymarin can reduce oxidative damage and protect vaginal tissues from free radical attacks. Detailed implementation manners

[0036] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the technical field to which this invention belongs. The terms used in the description of this invention herein are only for the purpose of describing specific embodiments and are not intended to limit the invention. The term "and / or" used herein includes any and all combinations of one or more of the related listed items.

[0037] Unless otherwise specified, the reagents, methods, and equipment used in this invention are conventional reagents, methods, and equipment in this technical field. Unless otherwise specified, the reagents and materials used in the following examples are all commercially available.

[0038] Example 1: Preparation and testing of Polygonum cuspidatum extract

[0039] I. Preparation of Polygonum cuspidatum extract

[0040] (1) Raw material preparation and pretreatment: Select high-quality Polygonum cuspidatum roots, ensure that the raw material source is pollution-free and pesticide-free, and ensure the stability of its components; after using ultrasonic cleaning to remove surface impurities, cut the Polygonum cuspidatum roots into small sections of 1 - 2 cm.

[0041] (2) Solvent selection: Select a 70% ethanol - water mixed solvent and add 0.1% acetic acid as an auxiliary solvent; the mass ratio of dry matter to solvent is 1:15. Compared with the traditional ratio of 1:10, using a larger solvent amount can effectively improve the extraction efficiency.

[0042] (3) Ultrasonic-assisted extraction: Put the processed polygonum cuspidatum root segments into the extraction container, add 70% ethanol-aqueous solution and 0.1% acetic acid; perform ultrasonic-assisted extraction with a power of 500 W, a frequency of 40 kHz, at 50 °C for 30 minutes, and stop for 1 minute every 10 minutes for cooling; after extraction is completed, filter to remove residues to obtain the preliminary extract.

[0043] (4) Reflux extraction: Pour the liquid after ultrasonic extraction into a reflux extractor and continue to perform reflux extraction with 70% ethanol; set the reflux temperature to 80 °C, the reflux time to 2 hours, and seal during reflux to prevent volatilization; after reflux extraction is completed, cool the liquid, filter and remove residual impurities.

[0044] (5) Concentration and purification: Concentrate the filtered extract to 1 / 5 of the original volume under reduced pressure at 40 °C using a rotary evaporator to remove the ethanol solvent; use activated carbon adsorption to remove impurities and ensure the purity and stability of the final extract.

[0045] (6) Preservation and packaging: Add vitamin C with a final concentration of 0.1% to the extract, disinfect with ultraviolet light, and then use a sterile glass bottle or aluminum film bag for sealed packaging and store at 2 - 8 °C for standby.

[0046] II. Inspection of Polygonum Cuspidatum Extract

[0047] 1. Detection of Active Ingredient Content (HPLC)

[0048] (1) Equipment: High-performance liquid chromatograph (HPLC) equipped with an ultraviolet detector (UV).

[0049] (2) Chromatographic conditions:

[0050] Chromatographic column: C18 reversed-phase chromatographic column (250 mm × 4.6 mm, 5 μm).

[0051] Mobile phase: Methanol-water (75:25).

[0052] Flow rate: 1.0 ml / min.

[0053] Detection wavelength: 320 nm (mainly for resveratrol).

[0054] (2) Experimental procedure: Concentrate the polygonum cuspidatum extract sample to an appropriate concentration. Filter the sample and inject it into the HPLC injector. Record the chromatogram and quantify the contents of resveratrol, albiflorin, and other components.

[0055] (3) Detection results: As shown in Table 1, the concentration of active ingredients in the polygonum cuspidatum extract is more than 20% higher than that of the traditional method, ensuring the high efficiency of the extract.

[0056] Summary of the Detection Results of the Active Ingredient Content (HPLC)

[0057]

[0058] 2. Antibacterial Activity Test (Inhibition Zone Method)

[0059] (1) Strain Selection: Staphylococcus aureus, Escherichia coli.

[0060] (2) Culture Medium: Agar Plate Medium

[0061] (3) Experimental Procedure: Prepare Polygonum cuspidatum extract solutions with different concentrations (0.5%, 2%, 5%). Inoculate bacteria onto the agar plate with a sterile cotton swab to ensure uniform coating. Use the paper disk method to drop the extract solution onto the paper disk and incubate for 24 hours. Measure the diameter of the inhibition zone.

[0062] (4) Detection Results: As shown in Table 2, the extract can exhibit significant antibacterial effects at lower concentrations and has stronger antibacterial activity.

[0063] Table 2 Antibacterial Activity Test Results

[0064]

[0065] 3. Antioxidant Activity Test (DPPH Free Radical Scavenging Assay)

[0066] (1) Reagents: DPPH free radical solution, Polygonum cuspidatum extract

[0067] (2) Experimental Procedure: Prepare DPPH solution (0.1 mM) and dissolve it with ethanol. Prepare Polygonum cuspidatum extract solutions with different concentrations (5%, 10%, 15%). After mixing the extract solution with the DPPH solution, react in the dark at room temperature for 30 minutes. Measure the absorbance (540 nm) using a spectrophotometer.

[0068] (3) Detection Results: As shown in Table 3, the extract shows a significant increase in the free radical scavenging rate and enhanced antioxidant effects.

[0069] Table 3 Antioxidant Activity Test Results

[0070]

[0071] III. Summary

[0072] 1. High-Efficiency Extraction: Through the double extraction method of ultrasonic-assisted extraction and reflux, the extraction efficiency is increased by more than 20%. Compared with traditional methods, the concentration of the extracted active ingredients is higher, and the effective ingredients of Polygonum cuspidatum can be retained to the greatest extent.

[0073] 2. Enhanced antibacterial effect: The optimized extract can exhibit strong antibacterial activity at a lower concentration, especially showing a significant improvement in the inhibitory effect on Staphylococcus aureus and Escherichia coli.

[0074] 3. Enhanced antioxidant capacity: The antioxidant capacity of the extract is significantly improved after optimization, which can effectively counteract oxidative stress and protect cells from damage.

[0075] 4. The above results indicate that the preparation method and the concentration of active ingredients of the polygonum cuspidatum extract of the present invention have reached a relatively high level, thus ensuring a great improvement in the application effect and safety in gynecological lotion.

[0076] Example 2: Preparation of antibacterial gynecological lotion

[0077] 1. Formula 1, the mass ratio is as follows:

[0078] (1) The polygonum cuspidatum extract prepared in Example 1: 0.1%;

[0079] (2) Lactobacillus: 1×10^8 CFU / ml;

[0080] (3) Chloral hydrate: 0.02%;

[0081] (4) Vegetable glycerin: 1%;

[0082] (5) pH value: 4.2.

[0083] 2. Formula 2, the mass ratio is as follows:

[0084] (1) The polygonum cuspidatum extract prepared in Example 1: 0.1%;

[0085] (2) Lactobacillus: 1×10^8 CFU / ml;

[0086] (3) Chloral hydrate: 0.02%;

[0087] (4) Silymarin: 0.5%;

[0088] (5) Vegetable glycerin: 1%;

[0089] (6) pH value: 4.2.

[0090] 3. Preparation method

[0091] (1) Cultivation of Lactobacillus: Select Lactobacillus acidophilus and cultivate it in a suitable culture medium at a cultivation temperature of 37°C for 24 hours to obtain a Lactobacillus suspension.

[0092] (2) Liquid preparation and formulation: Mix the polygonum cuspidatum extract, lactic acid bacteria suspension, vegetable glycerin, and purified water according to the formula ratio. After stirring evenly, add an appropriate amount of chloral hydrate, etc., and finally adjust the pH value to 4.2.

[0093] (3) Sterilization and packaging: The mixed liquid is subjected to high-pressure sterilization at 121 °C for 15 minutes to ensure the sterility of the product. After sterilization, aseptic filling and sealed packaging are carried out.

[0094] Example 3: Application of the bacteriostatic gynecological lotion

[0095] I. Purpose: To verify the bacteriostatic effect and tissue safety of the gynecological lotion of the present invention in a rat vaginal model.

[0096] II. Experimental design

[0097] 1. Animal model

[0098] (1) Animal species: Sprague-Dawley (SD) rats, female, weighing 180 - 220 g.

[0099] (2) Grouping: Experimental group 1 (formula 1), experimental group 2 (formula 2), control group, existing product control group (Fuyanjie Mousse Intimate Wash).

[0100] Experimental group 1: Use the gynecological lotion of formula 1 of the present invention.

[0101] Experimental group 2: Use the gynecological lotion of formula 2 of the present invention

[0102] Control group: Use a common chlorhexidine-containing gynecological lotion on the market.

[0103] Existing product control group: Use a common over-the-counter gynecological lotion on the market (Fuyanjie Plant Herbal Bacteriostatic Lotion).

[0104] Treatment period: 14 days.

[0105] III. Experimental steps

[0106] 1. Animal adaptation period:

[0107] (1) Adapt to the environment: The rats are adapted for 7 days before the experiment, raised in a standard laboratory environment, maintained at a temperature of 22 ± 2 °C, a humidity of 50 ± 10%, and a 12-hour day-night cycle.

[0108] (2) Feed and water: Drink water freely and eat freely.

[0109] 2. Modeling:

[0110] (1) Before the experiment, pathogenic bacteria (such as Escherichia coli, Staphylococcus aureus) were inoculated into the vagina of rats by vaginal perfusion method. 0.2 mL of pathogenic liquid with a concentration of 10^7 CFU / mL was used to induce vaginal infection.

[0111] (2) Observation was carried out 24 hours after inoculation to confirm successful infection.

[0112] 3. Drug treatment:

[0113] (1) Experimental group 1: The vaginal cavity was washed daily with the gynecological lotion of formula 1 of the present invention. The volume of the lotion was 5 mL, and the washing operation lasted for 5 minutes.

[0114] (2) Experimental group 2: The vaginal cavity was washed daily with the gynecological lotion of formula 2 of the present invention. The washing operation was the same as that of experimental group 1.

[0115] (3) Control group: The vaginal cavity was washed daily with a commercially available gynecological lotion containing chlorhexidine. The washing method and volume were the same as those of experimental group 1.

[0116] (4) Control group of existing products: The vaginal cavity was washed daily with a common over-the-counter gynecological lotion on the market. The washing method and volume were the same as those of experimental group 1.

[0117] (5) The washing operation lasted for 14 days.

[0118] 4. Clinical observation and data collection:

[0119] (1) The vulva and vaginal secretions of the animals were observed every 3 days, including whether there was an increase in secretions, abnormal odor, color change, etc.

[0120] (2) The numbers of lactic acid bacteria and pathogenic bacteria in the vagina were recorded (by culture method).

[0121] 5. Data analysis

[0122] (1) Number of lactic acid bacteria in the vagina: Vaginal secretions were sampled for culture, and selective media were used to screen lactic acid bacteria and pathogenic bacteria.

[0123] (2) Inhibition rate of pathogenic bacteria: The change in the number of pathogenic bacteria was calculated to obtain the flora inhibition rate.

[0124] III. Experimental data

[0125] (1) Changes in the number of lactobacilli in the vagina: The number of lactobacilli in experimental group 1 increased significantly, reaching three times the baseline, showing good effect of flora restoration. The number of lactobacilli in the vagina of experimental group 2 increased significantly, reaching four times the baseline, which was about 20% higher than that of experimental group 1, indicating stronger ability of flora restoration. The number of lactobacilli in the control group increased to some extent, but the effect was poor, indicating incomplete flora restoration. The number of lactobacilli in the existing product group also increased, but the effect was comparable to that of the experimental group. As shown in Table 4.

[0126] Table 4 Monitoring results of the number of lactobacilli in the vagina

[0127]

[0128] (2) Inhibition rate of pathogenic bacteria: The inhibition rate of pathogenic bacteria in experimental group 1 was 98%, and the effect was also very ideal. The inhibition rate of pathogenic bacteria in experimental group 2 reached 99%, showing the most significant effect, indicating that after adding polygonum cuspidatum extract and silymarin, the antibacterial effect was further enhanced. The inhibition rate of pathogenic bacteria in the control group was 60%, showing a poor effect. The inhibition rate of the existing product group was 75%, showing a medium effect. Specifically shown in Table 5.

[0129] Table 5 Detection results of the inhibition rate of pathogenic bacteria

[0130]

[0131] IV. Summary

[0132] 1. Experimental group 2 showed the best effect in terms of lactobacilli restoration and pathogenic bacteria inhibition, and there was no obvious damage to the pathological tissue, indicating its superiority over the experimental group, the control group and the existing product group.

[0133] 2. The effect of experimental group 1 was comparable to that of experimental group 2, but experimental group 2 showed stronger antibacterial and restoration effects.

[0134] 3. The effects of the control group and the existing product group were poor, and there was certain tissue damage, indicating that traditional products have weak effects in restoring vaginal flora and inhibiting pathogenic bacteria.

[0135] The above data indicate that the gynecological lotion of the present invention has significant therapeutic effects, can effectively restore the balance of vaginal flora, inhibit the growth of pathogenic bacteria, and has no obvious tissue damage, providing a safer and more effective option for the treatment of gynecological infections.

[0136] The above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited by the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications made without departing from the spirit and principle of the present invention shall be equivalent replacement methods and are all included in the protection scope of the present invention.

Claims

1. A polygonum cuspidatum extract, characterized in that, The active ingredients of the polygonum cuspidatum extract are mainly resveratrol and paeoniflorin, with their contents being 1.25 mg / g and 1.50 mg / g respectively.

2. A method for preparing the polygonum cuspidatum extract according to claim 1, characterized in that, The method includes the following steps: (1) Raw material preparation and pretreatment: Select high-quality polygonum cuspidatum roots, ensure that the raw material source is free from pollution and pesticide residues, and ensure the stability of its components; After using ultrasonic cleaning to remove surface impurities, cut the polygonum cuspidatum roots into small sections of 1-2 cm. (2) Solvent selection: Select a 70% ethanol-water mixed solvent and add 0.1% acetic acid as an auxiliary solvent; The mass ratio of dry matter to solvent is 1:

15. (3) Ultrasonic-assisted extraction: Put the processed polygonum cuspidatum root segments into an extraction container, add 70% ethanol-water solution and 0.1% acetic acid; Use ultrasonic-assisted extraction, with a power of 500 W, a frequency of 40 kHz, at 50 °C for 30 minutes, and stop for 1 minute every 10 minutes for cooling; After extraction is completed, filter to remove residues to obtain a preliminary extract. (4) Reflux extraction: Pour the liquid after ultrasonic extraction into a reflux extractor and continue to carry out reflux extraction with 70% ethanol; Set the reflux temperature to 80 °C and the reflux time to 2 hours, and seal during reflux to prevent volatilization; After reflux extraction is completed, cool the liquid, filter and remove residual impurities. (5) Concentration and purification: Concentrate the filtered extract under reduced pressure to 1 / 5 of the original volume at 40 °C through a rotary evaporator to remove the ethanol solvent; Use activated carbon adsorption to remove impurities and ensure the purity and stability of the final extract. (6) Preservation and packaging: Add vitamin C with a final concentration of 0.1% to the extract, disinfect with ultraviolet light, and then use a sterile glass bottle or aluminum film bag for sealed packaging and store at 2-8 °C for standby.

3. An antibacterial gynecological lotion, characterized in that, The formula of the antibacterial gynecological lotion is as follows according to the mass ratio: (1) The polygonum cuspidatum extract described in claim 1: 0.1%; (2) Lactobacillus: 1×10^8 CFU / ml; (3) Chloral hydrate: 0.02%; (4) Vegetable glycerin: 1%; (5) pH value: 4.

2.

4. An antibacterial gynecological lotion, characterized in that, The formula of the antibacterial gynecological lotion is as follows according to the mass ratio: (1) The polygonum cuspidatum extract described in claim 1: 0.1%; (2) Lactobacillus: 1×10^8 CFU / ml; (3) Chloral hydrate: 0.02%; (4) Silymarin: 0.5%; (5) Vegetable glycerin: 1%; (6) pH value: 4.

2.

5. Use of a polygonum cuspidatum extract as described in claim 1 in the preparation of an antibacterial gynecological lotion.

Images