A yin cleaning and bacterium inhibiting essential oil and a preparation method thereof

Abstract

This invention belongs to the field of pharmaceutical technology, specifically relating to a vaginal cleansing and antibacterial essential oil and its preparation method. This invention utilizes the synergistic effect of three essential oils—oregano, thyme, and lemon balm—to achieve multi-target attack: α-pinene and other components in lemon balm essential oil first disrupt the Candida albicans biofilm, exposing the bacteria; oregano and thyme essential oils further disrupt the cell membranes of the exposed bacteria, and the three synergistically enhance the antibacterial activity of the vaginal cleansing and antibacterial essential oil. A dodecyl-chitosan-β-cyclodextrin ternary graft compound is prepared to encapsulate the composite essential oil into microcapsules, with the dodecyl group inserted into the lipid layer of the mucosal epithelium to provide physical anchoring; in the acidic environment of the vagina, the amino groups in chitosan are protonated and electrostatically bind to the negatively charged mucosa, and the two synergistically reduce drug loss; the hydrophobic cavity of β-cyclodextrin reversibly encapsulates the essential oil, achieving slow release and prolonging the duration of action of the vaginal cleansing and antibacterial essential oil.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically relating to a feminine cleansing and antibacterial essential oil and its preparation method. Background Technology

[0002] Vaginitis is a common gynecological disease that threatens women's lives and mental health. Common pathogens causing vaginitis include Candida, Trichomonas vaginalis, and bacteria, which are mainly transmitted through sexual intercourse or indirect means. Among them, vulvovaginal candidiasis is a common gynecological condition, and the causative agent has been identified as Candida albicans. Candida is widely distributed in nature and also exists in warm and moist areas of the human body, such as the mouth, intestines, and vagina.

[0003] Candida albicans infection accounts for 85-95% of candidal vaginitis. Currently, the main clinically used drugs are azole drugs such as fluconazole, miconazole nitrate, and ketoconazole. However, these antifungal drugs have limitations, including a limited selection range, adverse reactions such as gastrointestinal side effects, and drug resistance due to long-term treatment and repeated administration. Furthermore, the vaginal mucosa's numerous transverse folds and high extensibility facilitate Candida adhesion, preventing the drugs from achieving their full efficacy. Summary of the Invention

[0004] The purpose of this invention is to provide a vaginal cleansing and antibacterial essential oil and its preparation method, in order to solve the technical problem that the vaginal mucosa has many transverse folds and good extensibility, which prevents clinical drugs from exerting their full efficacy.

[0005] To achieve the above-mentioned technical objectives, the technical solution of the present invention is as follows:

[0006] A method for preparing a feminine cleansing and antibacterial essential oil includes the following steps:

[0007] S11. By weight, take 60-80 parts of pharmaceutical grade glycerin, 15-25 parts of modified chitosan-essential oil microcapsules, 5-8 parts of glyceryl monostearate and 3-6 parts of Span80.

[0008] S12. Mix the modified chitosan-essential oil microcapsules with Span80 and grind them to obtain pre-dispersed microcapsules; heat pharmaceutical grade glycerin to 50~60℃, add glyceryl monostearate, and stir until completely dissolved to obtain a glycerin matrix;

[0009] S13. Slowly add the obtained pre-dispersed microcapsules into the glycerol matrix, shear at high speed for 10-15 min, and degas at 0.08-0.095 MPa to obtain the feminine hygiene and antibacterial essential oil;

[0010] The modified chitosan is a ternary graft compound of dodecanal-chitosan-β-cyclodextrin.

[0011] As a further improvement, the modified chitosan-essential oil microcapsules are prepared by the following method:

[0012] S21. By weight, take 20-25 parts of oregano essential oil, 20-25 parts of thyme essential oil and 15-20 parts of sage essential oil, stir and mix evenly at room temperature, add 10-15 parts of Tween-80 to obtain a compound essential oil; slowly drip the obtained compound essential oil into the modified chitosan solution, and shear at high speed for 5-10 minutes to obtain an O / W type emulsion;

[0013] S22. Add 0.5wt% TPP solution dropwise to the O / W type emulsion while stirring. After the addition is complete, continue stirring for 30-60 min to obtain a microcapsule suspension. After centrifugation, discard the supernatant. Wash the obtained precipitate 3-4 times with deionized water, pre-freeze at -40℃ for 4 h, then gradually increase the temperature from -20℃ to 0℃ and dry at 10 Pa for 20 h, and finally dry at 25℃ and 5 Pa for 6 h to obtain modified chitosan-essential oil microcapsules.

[0014] As a further improvement, the modified chitosan solution is prepared by the following method:

[0015] Take 100-120 parts by weight of dodecaldehyde-chitosan-β-cyclodextrin ternary graft compound, dissolve it in 2000-2500 parts of 1% glacial acetic acid solution, stir at room temperature until completely dissolved, and adjust the pH to 4.0-5.0 to obtain modified chitosan solution.

[0016] As a further improvement, the dodecanoic acid-chitosan-β-cyclodextrin ternary graft compound was prepared by the following method:

[0017] S41. By weight, take 100 parts of N-dodecyl chitosan and dissolve it in 2000~3000 parts of 1% glacial acetic acid solution. Stir at room temperature until completely dissolved, then add 6-Ns-β-CD activating solution and stir at 55~65℃ for 20~28h.

[0018] S42. After the reaction is complete, adjust the pH to 9-10 to precipitate the product. Dissolve the precipitate again in dilute acetic acid and then transfer it to a dialysis bag with a molecular weight cutoff of 8000 Da. Dialyze the product 5-6 times with deionized water and freeze-dry to obtain the dodecaldehyde-chitosan-β-cyclodextrin ternary graft compound.

[0019] As a further improvement, in step S41, the 6-Ns-β-CD activating solution is prepared by the following method:

[0020] S51. By weight, take 80-120 parts of β-cyclodextrin, 100-150 parts of p-nitrobenzenesulfonyl chloride, 150-250 parts of anhydrous DMSO and 60-100 parts of triethylamine;

[0021] S52. Dissolve β-cyclodextrin and p-nitrobenzenesulfonyl chloride together in anhydrous DMSO, add triethylamine, and react at 55~65℃ for 1~2h to obtain 6-Ns-β-CD activated solution.

[0022] As a further improvement, in step S41, the N-dodecyl chitosan is prepared by the following method:

[0023] S61. By weight, take 40-60 parts of dodecyl aldehyde and 2-5 parts of sodium dodecyl sulfonate; add dodecyl aldehyde to chitosan solution, stir for 20-25 min, add sodium dodecyl sulfonate, and reflux at 95-100℃ for 8-12 h.

[0024] S62. After the reflux reaction is completed, the mixture is naturally cooled to room temperature. Under an ice-water bath, 20-30 parts of sodium borohydride are added, and the mixture is stirred at room temperature for 2-4 hours. After the reaction is completed, the pH is adjusted to 9-10. After the precipitate is formed, it is filtered. The precipitate is washed with deionized water until the washing solution is neutral. After vacuum drying, N-dodecyl chitosan is obtained.

[0025] As a further improvement, in step S61, the chitosan solution is prepared by the following method:

[0026] Take 100 parts by weight of chitosan and dissolve it in 2000-2500 parts of 1-2% glacial acetic acid solution. Stir at room temperature for 2-4 hours and adjust the pH to 4-6 to obtain a chitosan solution.

[0027] As a further improvement, the chitosan has a molecular weight of 50~200kDa and a degree of deacetylation ≥90%.

[0028] As a further improvement, the mass ratio of modified chitosan in the modified chitosan solution to TPP in the 0.5wt% TPP solution is 6~10:1.

[0029] The present invention also provides a feminine cleansing and antibacterial essential oil.

[0030] Due to the adoption of the above technical solution, the beneficial effects of the present invention are as follows:

[0031] 1. Utilizing the synergistic effect of oregano essential oil, thyme essential oil, and lemon balm essential oil, this method disrupts and disintegrates the cell membrane and biofilm of Candida albicans: α-pinene and other components in lemon balm essential oil can attack the Candida albicans biofilm, exposing the fungus; the core components of oregano essential oil are carvacrol and thymol, and the core component of thyme essential oil is thymol. Both further disrupt the exposed cell membrane of Candida albicans, enhancing the antibacterial effect. The three components work synergistically to form a multi-target attack on Candida albicans, resisting the development of drug resistance in Candida albicans.

[0032] 2. Preparation of a ternary graft compound of dodecyl aldehyde-chitosan-β-cyclodextrin, which is then mixed and emulsified with a complex essential oil to form microcapsules: The dodecyl group in dodecyl aldehyde can insert into the lipid bilayer gap of the fold mucosal epithelial cells, providing physical anchoring force; in the acidic environment of the vagina (pH 3.5~4.5), the amino groups on chitosan are highly protonated and electrostatically bind to the negatively charged vaginal mucosa, further preventing the drug from being lost with secretions; the hydrophobic cavity of β-cyclodextrin can hydrophobically bind with the complex essential oil. As the microcapsules open in the acidic environment, the released essential oil is absorbed or metabolized, resulting in the slow release of the remaining essential oil and prolonging the effect time of the vaginal cleansing and antibacterial essential oil. Detailed Implementation

[0033] The technical solution of the present invention will be clearly and completely described below with reference to specific embodiments. However, those skilled in the art will understand that the embodiments described below are some embodiments of the present invention, but not all embodiments, and are only used to illustrate the present invention, and should not be regarded as limiting the scope of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention. Where specific conditions are not specified in the embodiments, conventional conditions or manufacturer's conditions shall be followed. Where the manufacturers of reagents or instruments are not specified, they are all conventional products that can be purchased commercially.

[0034] The oregano oil, thyme oil and sage oil used in the examples and comparative examples were all prepared by steam distillation, a well-known method.

[0035] The Candida albicans ATCC 10231, Sabouraud dextrose agar, and yeast extract peptone glucose liquid broth used in the test can all be purchased through conventional channels.

[0036] Example 1: A method for preparing a feminine cleansing and antibacterial essential oil, comprising the following steps:

[0037] S1. Take 100 parts by weight of chitosan (molecular weight of 50kDa, degree of deacetylation ≥90%), dissolve it in 2000 parts by volume of 1% glacial acetic acid solution, stir at 300 rpm for 2 hours at room temperature, and adjust the pH to 4 with 0.1 mol / L NaOH solution to obtain chitosan solution.

[0038] S2. By weight, take 40 parts of dodecyl aldehyde and 2 parts of sodium dodecyl sulfonate; add dodecyl aldehyde to the chitosan solution obtained in S1, stir and emulsify for 20 min, add sodium dodecyl sulfonate, and stir and reflux at 95℃ for 12 h to generate C=N bonds from the aldehyde group (-CHO) of dodecyl aldehyde and the amino group (-NH2) of chitosan through a Schiff base reaction; the specific reaction mechanism is as follows:

[0039] ;

[0040] Where n and p are both integers greater than 0;

[0041] S3. After the reflux reaction of S2 is completed, the mixture is naturally cooled to room temperature. Under an ice-water bath, 20 parts of sodium borohydride are added, and the mixture is stirred at room temperature for 2 hours to reduce the Schiff base (C=N) to a more stable secondary amine (CN). After the reaction is completed, the pH is adjusted to 9 with 0.1 mol / L NaOH solution until a precipitate is formed. The precipitate is then filtered, and the precipitate is repeatedly washed with deionized water until the washing solution is neutral. The precipitate is then vacuum dried at 60°C for 12 hours to obtain N-dodecyl chitosan.

[0042] S4, p-nitrobenzenesulfonation of β-cyclodextrin

[0043] By weight, 80 parts of β-cyclodextrin and 100 parts of p-nitrobenzenesulfonyl chloride were dissolved in 150 parts of anhydrous dimethyl sulfoxide (DMSO), and 60 parts of triethylamine were added. The mixture was reacted at 55°C for 1 hour to obtain an activated solution of 6-p-nitrobenzenesulfonyl-β-cyclodextrin (6-Ns-β-CD). The reaction mechanism is as follows:

[0044] ;

[0045] S5. By weight, 100 parts of N-dodecyl chitosan were dissolved in 2000 parts of 1% (v / v) glacial acetic acid solution. After complete dissolution at room temperature, 6-Ns-β-CD activating solution was added, and the mixture was stirred at 55°C for 20 h. After the reaction was completed, the pH was adjusted to 9 with 0.1 mol / L NaOH, and a precipitate was formed. After filtration, the precipitate was redissolved in 6 wt% dilute acetic acid and transferred to a dialysis bag with a molecular weight cutoff of 8000 Da. After repeated dialyzing with deionized water 5 times to remove unreacted β-CD derivatives and impurities, the product was freeze-dried at -55°C to obtain the dodecylaldehyde-chitosan-β-cyclodextrin ternary graft compound. The reaction mechanism is as follows:

[0046] ;

[0047] Where m = npz, and m, n, p, and z are all integers greater than 0;

[0048] S6. By weight, take 100 parts of dodecaldehyde-chitosan-β-cyclodextrin ternary graft compound and dissolve it in 2000 parts of 1% glacial acetic acid solution. Stir at 300 rpm for 30 min at room temperature. Adjust the pH to 4.0 with 0.1 mol / L NaOH solution to obtain modified chitosan solution for later use.

[0049] S7. By weight, take 20 parts of oregano essential oil, 20 parts of thyme essential oil and 20 parts of sage essential oil, stir at 200 rpm for 10 minutes at room temperature to mix evenly, add 10 parts of emulsifier Tween-80 to obtain a compound essential oil; under high-speed shearing at 8000 rpm, slowly drip the compound essential oil into the modified chitosan solution, emulsify for 5 minutes to obtain an oil-in-water (O / W) emulsion;

[0050] S8. Take 12 parts by weight of sodium tripolyphosphate (TPP) and dissolve it in 2400 parts by weight of deionized water to prepare a 0.5 wt% TPP solution. Under stirring at 400 rpm, slowly add the TPP solution dropwise to the O / W type emulsion at a rate of 1 mL / s to allow the modified chitosan and TPP to undergo an ionic cross-linking reaction. After the addition is complete, continue stirring for 30 min to obtain a microcapsule suspension.

[0051] S9. Centrifuge the obtained microcapsule suspension at 3000 rpm for 10 min, discard the supernatant, wash the obtained precipitate three times with deionized water, pre-freeze at -40℃ for 4 h, then gradually increase the temperature from -20℃ to 0℃ and dry at a vacuum of 10 Pa for 20 h, and finally dry it again at 25℃ and a vacuum of 5 Pa for 6 h to obtain modified chitosan-essential oil microcapsules.

[0052] S10. By weight, take 60 parts of pharmaceutical grade glycerin, 15 parts of modified chitosan-essential oil microcapsules, 5 parts of glyceryl monostearate and 3 parts of Span-80.

[0053] S11. Mix the modified chitosan-essential oil microcapsules with Span-80, grind them in a mortar for 10 minutes to uniformly coat the surface of the microcapsules with wetting agent, and obtain pre-dispersed microcapsules for later use; heat pharmaceutical grade glycerol to 50°C, add glyceryl monostearate, stir until completely dissolved, and obtain glycerol matrix for later use.

[0054] S12. Slowly add the pre-dispersed microcapsules to the glycerol matrix, shear at 5000 rpm for 10 min, then place in a vacuum drying oven and degas at 0.08 MPa for 60 min to obtain the feminine hygiene and antibacterial essential oil.

[0055] Example 2: A method for preparing a feminine cleansing and antibacterial essential oil, comprising the following steps:

[0056] S1. Take 100 parts by weight of chitosan (molecular weight of 150kDa, degree of deacetylation ≥90%), dissolve it in 2250 parts by volume of 1.5% glacial acetic acid solution, stir at 400 rpm for 3 hours at room temperature, and adjust the pH to 5 with 0.1 mol / L NaOH solution to obtain chitosan solution.

[0057] S2. Take 50 parts by weight of dodecyl aldehyde and 3 parts by weight of sodium dodecyl sulfonate; add dodecyl aldehyde to the chitosan solution obtained in S1, stir and emulsify for 23 min, add sodium dodecyl sulfonate, and stir and reflux at 100℃ for 10 h.

[0058] S3. After the reflux reaction of S2 is completed, cool naturally to room temperature, add 25 parts of sodium borohydride under an ice-water bath, and continue to stir the reaction at room temperature for 3 hours. After the reaction is completed, adjust the pH to 10 with 0.1 mol / L NaOH solution until a precipitate is formed. Filter the precipitate and wash it repeatedly with deionized water until the washing solution is neutral. Dry it under vacuum at 70°C for 12 hours to obtain N-dodecyl chitosan.

[0059] S4, p-nitrobenzenesulfonation of β-cyclodextrin

[0060] Take 105 parts by weight of β-cyclodextrin and 125 parts by weight of p-nitrobenzenesulfonyl chloride, dissolve them in 200 parts of anhydrous DMSO, add 80 parts of triethylamine, and react at 60℃ for 2 hours to obtain 6-Ns-β-CD activated solution.

[0061] S5. By weight, 100 parts of N-dodecyl chitosan were dissolved in 2500 parts of 1% glacial acetic acid solution. After stirring at room temperature until completely dissolved, 6-Ns-β-CD activating solution was added, and the mixture was stirred at 60℃ for 24 hours. After the reaction was completed, the pH was adjusted to 10 with 0.1 mol / L NaOH, and a precipitate was formed. After filtration, the precipitate was redissolved in 6 wt% dilute acetic acid and transferred to a dialysis bag with a molecular weight cutoff of 8000 Da. After repeated dialyzing with deionized water 5-6 times to remove unreacted β-CD derivatives and impurities, the mixture was freeze-dried at -55℃ to obtain dodecyl-chitosan-β-cyclodextrin ternary graft compound.

[0062] S6. By weight, take 110 parts of dodecaldehyde-chitosan-β-cyclodextrin ternary graft compound and dissolve it in 2250 parts of 1% glacial acetic acid solution. Stir at 400 rpm for 30 min at room temperature. Adjust the pH to 5.0 with 0.1 mol / L NaOH solution to obtain modified chitosan solution for later use.

[0063] S7. By weight, take 23 parts of oregano essential oil, 23 parts of thyme essential oil and 18 parts of sage essential oil, stir at 200 rpm for 10 minutes at room temperature to mix evenly, add 13 parts of emulsifier Tween-80 to obtain a compound essential oil; under high-speed shearing at 9000 rpm, slowly drip the compound essential oil into the modified chitosan solution, emulsify for 8 minutes to obtain an O / W type emulsion;

[0064] S8. Take 13 parts by weight of TPP and dissolve them in 2800 parts of deionized water to prepare a 0.5wt% TPP solution. Under stirring at 500 rpm, slowly add the TPP solution dropwise into the O / W type emulsion at a rate of 2 mL / s to allow the modified chitosan and TPP to undergo an ionic cross-linking reaction. After the addition is complete, continue stirring for 45 min to obtain a microcapsule suspension.

[0065] S9. Centrifuge the obtained microcapsule suspension at 3500 rpm for 13 min, discard the supernatant, wash the obtained precipitate with deionized water 3-4 times, pre-freeze at -40℃ for 4 h, then heat it at -20℃ to 0℃ under vacuum of 10 Pa for 20 h, and finally dry it again at 25℃ under vacuum of 5 Pa for 6 h to obtain modified chitosan-essential oil microcapsules.

[0066] S10. By weight, take 70 parts of pharmaceutical grade glycerin, 20 parts of modified chitosan-essential oil microcapsules, 7 parts of glyceryl monostearate and 4 parts of Span-80.

[0067] S11. Mix the modified chitosan-essential oil microcapsules with Span-80, grind them in a mortar for 13 minutes to uniformly coat the surface of the microcapsules with wetting agent, and obtain pre-dispersed microcapsules for later use; heat pharmaceutical grade glycerol to 55°C, add glyceryl monostearate, stir until completely dissolved, and obtain glycerol matrix for later use.

[0068] S12. The pre-dispersed microcapsules were slowly added to the glycerol matrix and sheared at 7000 rpm for 13 min. Then, the mixture was placed in a vacuum drying oven and degassed at 0.085 MPa for 45 min to obtain the feminine hygiene and antibacterial essential oil.

[0069] Example 3: A method for preparing a feminine cleansing and antibacterial essential oil, comprising the following steps:

[0070] S1. Take 100 parts by weight of chitosan (molecular weight of 200kDa, degree of deacetylation ≥90%), dissolve it in 2500 parts by volume of 2% glacial acetic acid solution, stir at 500rpm for 4h at room temperature, and adjust the pH to 6 with 0.1mol / L NaOH solution to obtain chitosan solution.

[0071] S2. Take 60 parts by weight of dodecaldehyde and 5 parts by weight of sodium dodecyl sulfonate; add dodecaldehyde to the chitosan solution obtained in S1, stir and emulsify for 25 min, add sodium dodecyl sulfonate, and stir and reflux at 110℃ for 12 h.

[0072] S3. After the reflux reaction of S2 is completed, cool naturally to room temperature, add 30 parts of sodium borohydride under an ice-water bath, and continue to stir the reaction at room temperature for 4 hours. After the reaction is completed, adjust the pH to 10 with 0.1 mol / L NaOH solution until a precipitate is formed. Filter the precipitate and wash it repeatedly with deionized water until the washing solution is neutral. Dry it under vacuum at 80°C for 12 hours to obtain N-dodecyl chitosan.

[0073] S4, p-nitrobenzenesulfonation of β-cyclodextrin

[0074] Take 120 parts by weight of β-cyclodextrin and 150 parts by weight of p-nitrobenzenesulfonyl chloride, dissolve them in 250 parts of anhydrous DMSO, add 100 parts of triethylamine, and react at 65℃ for 2 hours to obtain 6-Ns-β-CD activated solution.

[0075] S5. By weight, 100 parts of N-dodecyl chitosan were dissolved in 3000 parts of 1% glacial acetic acid solution. After stirring at room temperature until completely dissolved, 6-Ns-β-CD activating solution was added, and the mixture was stirred at 65℃ for 28 hours. After the reaction was completed, the pH was adjusted to 10 with 0.1 mol / L NaOH, and a precipitate was formed. After filtration, the precipitate was redissolved in 6 wt% dilute acetic acid and transferred to a dialysis bag with a molecular weight cutoff of 8000 Da. After repeated dialyzing with deionized water 5-6 times to remove unreacted β-CD derivatives and impurities, the mixture was freeze-dried at -55℃ to obtain dodecyl-chitosan-β-cyclodextrin ternary graft compound.

[0076] S6. By weight, take 120 parts of dodecaldehyde-chitosan-β-cyclodextrin ternary graft compound and dissolve it in 2500 parts of 1% glacial acetic acid solution. Stir at 500 rpm for 30 min at room temperature. Adjust the pH to 5.0 with 0.1 mol / L NaOH solution to obtain modified chitosan solution for later use.

[0077] S7. By weight, take 25 parts of oregano essential oil, 25 parts of thyme essential oil and 15 parts of sage essential oil, stir at 200 rpm for 10 minutes at room temperature to mix evenly, add 15 parts of emulsifier Tween-80 to obtain a compound essential oil; under high-speed shearing at 10000 rpm, slowly drip the compound essential oil into the modified chitosan solution, emulsify for 10 minutes to obtain an O / W type emulsion;

[0078] S8. Take 15 parts by weight of TPP and dissolve them in 3000 parts of deionized water to prepare a 0.5wt% TPP solution. Under stirring at 600 rpm, slowly add the TPP solution dropwise to the O / W type emulsion at a rate of 2 mL / s to allow the modified chitosan and TPP to undergo an ionic cross-linking reaction. After the addition is complete, continue stirring for 60 min to obtain a microcapsule suspension.

[0079] S9. Centrifuge the obtained microcapsule suspension at 4000 rpm for 15 min, discard the supernatant, wash the obtained precipitate 4 times with deionized water, pre-freeze at -40℃ for 4 h, then heat it at -20℃ to 0℃ under vacuum of 10 Pa for 20 h, and finally dry it again at 25℃ under vacuum of 5 Pa for 6 h to obtain modified chitosan-essential oil microcapsules.

[0080] S10. By weight, take 80 parts of pharmaceutical grade glycerin, 25 parts of modified chitosan-essential oil microcapsules, 8 parts of glyceryl monostearate and 6 parts of Span-80.

[0081] S11. Mix the modified chitosan-essential oil microcapsules with Span-80, grind them in a mortar for 15 minutes to uniformly coat the surface of the microcapsules with wetting agent, and obtain pre-dispersed microcapsules for later use; heat pharmaceutical grade glycerol to 60°C, add glyceryl monostearate, stir until completely dissolved, and obtain glycerol matrix for later use.

[0082] S12. Slowly add the pre-dispersed microcapsules to the glycerol matrix, shear at 8000 rpm for 15 min, then place in a vacuum drying oven and degas at 0.095 MPa for 30 min to obtain the feminine hygiene and antibacterial essential oil.

[0083] Comparative Example 1: A method for preparing a feminine cleansing and antibacterial essential oil, differing from Example 1 in that it does not contain lemon balm essential oil, and specifically includes the following steps:

[0084] S1. Take 100 parts by weight of chitosan (molecular weight of 50kDa, degree of deacetylation ≥90%), dissolve it in 2000 parts by volume of 1% glacial acetic acid solution, stir at 300 rpm for 2 hours at room temperature, and adjust the pH to 4 with 0.1 mol / L NaOH solution to obtain chitosan solution.

[0085] S2. Take 40 parts by weight of dodecyl aldehyde and 2 parts by weight of sodium dodecyl sulfonate; add dodecyl aldehyde to the chitosan solution obtained in S1, stir and emulsify for 20 min, add sodium dodecyl sulfonate, and stir and reflux at 95°C for 12 h.

[0086] S3. After the reflux reaction of S2 is completed, cool naturally to room temperature, add 20 parts of sodium borohydride under an ice-water bath, and continue to stir the reaction at room temperature for 2 hours. After the reaction is completed, adjust the pH to 9 with 0.1 mol / L NaOH solution until a precipitate is formed. Filter the precipitate and wash it repeatedly with deionized water until the washing solution is neutral. Dry it under vacuum at 60°C for 12 hours to obtain N-dodecyl chitosan.

[0087] S4, p-nitrobenzenesulfonation of β-cyclodextrin

[0088] Take 80 parts by weight of β-cyclodextrin and 100 parts by weight of p-nitrobenzenesulfonyl chloride, dissolve them in 150 parts of anhydrous DMSO, add 60 parts of triethylamine, and react at 55℃ for 1 h to obtain 6-Ns-β-CD activated solution.

[0089] S5. By weight, 100 parts of N-dodecyl chitosan were dissolved in 2000 parts of 1% glacial acetic acid solution. After stirring at room temperature until completely dissolved, 6-Ns-β-CD activating solution was added, and the mixture was stirred at 55℃ for 20 h. After the reaction was completed, the pH was adjusted to 9 with 0.1 mol / L NaOH, and a precipitate was formed. After filtration, the precipitate was redissolved in 6 wt% dilute acetic acid and transferred to a dialysis bag with a molecular weight cutoff of 8000 Da. After repeated dialyzing with deionized water 5 times to remove unreacted β-CD derivatives and impurities, the mixture was freeze-dried at -55℃ to obtain dodecyl-chitosan-β-cyclodextrin ternary graft compound.

[0090] S6. By weight, take 100 parts of dodecaldehyde-chitosan-β-cyclodextrin ternary graft compound and dissolve it in 2000 parts of 1% glacial acetic acid solution. Stir at 300 rpm for 30 min at room temperature. Adjust the pH to 4.0 with 0.1 mol / L NaOH solution to obtain modified chitosan solution for later use.

[0091] S7. By weight, take 30 parts of oregano essential oil and 30 parts of thyme essential oil, stir at 200 rpm for 10 minutes at room temperature to mix evenly, add 10 parts of emulsifier Tween-80 to obtain a compound essential oil; under high-speed shearing at 8000 rpm, slowly drip the compound essential oil into the modified chitosan solution, emulsify for 5 minutes to obtain an O / W type emulsion;

[0092] S8. Take 12 parts by weight of TPP and dissolve them in 2400 parts of deionized water to prepare a 0.5wt% TPP solution. Stir at 400 rpm and slowly add the TPP solution dropwise to the O / W type emulsion at a rate of 1 mL / s. After the addition is complete, continue stirring for 30 min to obtain the microcapsule suspension.

[0093] S9. Centrifuge the obtained microcapsule suspension at 3000 rpm for 10 min, discard the supernatant, wash the obtained precipitate three times with deionized water, pre-freeze at -40℃ for 4 h, then heat it at -20℃ to 0℃ under vacuum of 10 Pa for 20 h, and finally dry it again at 25℃ under vacuum of 5 Pa for 6 h to obtain modified chitosan-essential oil microcapsules.

[0094] S10. By weight, take 60 parts of pharmaceutical grade glycerin, 15 parts of modified chitosan-essential oil microcapsules, 5 parts of glyceryl monostearate and 3 parts of Span-80.

[0095] S11. Mix the modified chitosan-essential oil microcapsules with Span-80, grind them in a mortar for 10 minutes to uniformly coat the surface of the microcapsules with wetting agent, and obtain pre-dispersed microcapsules for later use; heat pharmaceutical grade glycerol to 50°C, add glyceryl monostearate, stir until completely dissolved, and obtain glycerol matrix for later use.

[0096] S12. Slowly add the pre-dispersed microcapsules to the glycerol matrix, shear at 5000 rpm for 10 min, then place in a vacuum drying oven and degas at 0.08 MPa for 60 min to obtain the feminine hygiene and antibacterial essential oil.

[0097] Comparative Example 2: A method for preparing a feminine cleansing and antibacterial essential oil, differing from Example 1 in that chitosan-β-cyclodextrin is used as the microcapsule wall material, specifically including the following steps:

[0098] S1, p-nitrobenzenesulfonation of β-cyclodextrin

[0099] Take 80 parts by weight of β-cyclodextrin and 100 parts by weight of p-nitrobenzenesulfonyl chloride, dissolve them in 150 parts of anhydrous DMSO, add 60 parts of triethylamine, and react at 55℃ for 1 h to obtain 6-Ns-β-CD activated solution.

[0100] S2. By weight, 100 parts of chitosan were dissolved in 2000 parts of 1% glacial acetic acid solution. After stirring at room temperature until completely dissolved, 6-Ns-β-CD activating solution was added, and the mixture was stirred at 55℃ for 20 hours. After the reaction was completed, the pH was adjusted to 9 with 0.1 mol / L NaOH, and a precipitate was formed. After filtration, the precipitate was redissolved in 6 wt% dilute acetic acid and transferred to a dialysis bag with a molecular weight cutoff of 8000 Da. After repeated dialyzing with deionized water 5 times to remove unreacted β-CD derivatives and impurities, the mixture was freeze-dried at -55℃ to obtain chitosan-β-cyclodextrin graft.

[0101] S3. By weight, take 100 parts of chitosan-β-cyclodextrin graft and dissolve them in 2000 parts of 1% glacial acetic acid solution. Stir at 300 rpm for 30 min at room temperature. Adjust the pH to 4.0 with 0.1 mol / L NaOH solution to obtain modified chitosan solution for later use.

[0102] S4. By weight, take 20 parts of oregano essential oil, 20 parts of thyme essential oil and 20 parts of sage essential oil, stir at 200 rpm for 10 minutes at room temperature to mix evenly, add 10 parts of emulsifier Tween-80 to obtain a compound essential oil; under high-speed shearing at 8000 rpm, slowly drip the compound essential oil into the modified chitosan solution, emulsify for 5 minutes to obtain an O / W type emulsion;

[0103] S5. By weight, take 12 parts of TPP and dissolve them in 2400 parts of deionized water to prepare a 0.5wt% TPP solution; under stirring at 400 rpm, slowly add the TPP solution dropwise to the O / W type emulsion at a rate of 1 mL / s. After the addition is complete, continue stirring for 30 min to obtain the microcapsule suspension.

[0104] S6. Centrifuge the obtained microcapsule suspension at 3000 rpm for 10 min, discard the supernatant, wash the obtained precipitate three times with deionized water, pre-freeze at -40℃ for 4 h, then heat it at -20℃ to 0℃ under vacuum of 10 Pa for 20 h, and finally dry it again at 25℃ under vacuum of 5 Pa for 6 h to obtain modified chitosan-essential oil microcapsules.

[0105] S7. By weight, take 60 parts of pharmaceutical grade glycerin, 15 parts of modified chitosan-essential oil microcapsules, 5 parts of glyceryl monostearate and 3 parts of Span-80.

[0106] S8. Mix the modified chitosan-essential oil microcapsules with Span-80, grind them in a mortar for 10 minutes to uniformly coat the surface of the microcapsules with wetting agent, and obtain pre-dispersed microcapsules for later use; heat pharmaceutical grade glycerol to 50°C, add glyceryl monostearate, stir until completely dissolved, and obtain glycerol matrix for later use.

[0107] S9. Slowly add the pre-dispersed microcapsules to the glycerol matrix, shear at 5000 rpm for 10 min, then place in a vacuum drying oven and degas at 0.08 MPa for 60 min to obtain the feminine hygiene and antibacterial essential oil.

[0108] Comparative Example 3: A method for preparing a feminine cleansing and antibacterial essential oil, differing from Example 1 in that it uses dodecanoic acid-chitosan as the microcapsule wall material, specifically including the following steps:

[0109] S1. Take 100 parts by weight of chitosan (molecular weight of 50kDa, degree of deacetylation ≥90%), dissolve it in 2000 parts by volume of 1% glacial acetic acid solution, stir at 300 rpm for 2 hours at room temperature, and adjust the pH to 4 with 0.1 mol / L NaOH solution to obtain chitosan solution.

[0110] S2. Take 40 parts by weight of dodecyl aldehyde and 2 parts by weight of sodium dodecyl sulfonate; add dodecyl aldehyde to the chitosan solution obtained in S1, stir and emulsify for 20 min, add sodium dodecyl sulfonate, and stir and reflux at 95°C for 12 h.

[0111] S3. After the reflux reaction of S2 is completed, cool naturally to room temperature, add 20 parts of sodium borohydride under an ice-water bath, and continue to stir the reaction at room temperature for 2 hours. After the reaction is completed, adjust the pH to 9 with 0.1 mol / L NaOH solution until a precipitate is formed. Filter the precipitate and wash it repeatedly with deionized water until the washing solution is neutral. Dry it under vacuum at 60°C for 12 hours to obtain N-dodecyl chitosan.

[0112] S4. By weight, take 100 parts of N-dodecyl chitosan and dissolve it in 2000 parts of 1% glacial acetic acid solution. Stir at 300 rpm for 30 min at room temperature. Adjust the pH to 4.0 with 0.1 mol / L NaOH solution to obtain modified chitosan solution for later use.

[0113] S5. By weight, take 20 parts of oregano essential oil, 20 parts of thyme essential oil and 20 parts of sage essential oil, stir at 200 rpm for 10 minutes at room temperature to mix evenly, add 10 parts of emulsifier Tween-80 to obtain a compound essential oil; under high-speed shearing at 8000 rpm, slowly drip the compound essential oil into the modified chitosan solution, emulsify for 5 minutes to obtain an O / W type emulsion;

[0114] S6. By weight, take 12 parts of TPP and dissolve them in 2400 parts of deionized water to prepare a 0.5wt% TPP solution; under stirring at 400 rpm, slowly add the TPP solution dropwise to the O / W type emulsion at a rate of 1 mL / s. After the addition is complete, continue stirring for 30 min to obtain the microcapsule suspension.

[0115] S7. Centrifuge the obtained microcapsule suspension at 3000 rpm for 10 min, discard the supernatant, wash the obtained precipitate three times with deionized water, pre-freeze at -40℃ for 4 h, then heat it at -20℃ to 0℃ under vacuum of 10 Pa for 20 h, and finally dry it again at 25℃ under vacuum of 5 Pa for 6 h to obtain modified chitosan-essential oil microcapsules.

[0116] S8. By weight, take 60 parts of pharmaceutical grade glycerin, 15 parts of modified chitosan-essential oil microcapsules, 5 parts of glyceryl monostearate and 3 parts of Span-80.

[0117] S9. Mix the modified chitosan-essential oil microcapsules with Span-80, grind them in a mortar for 10 minutes to uniformly coat the surface of the microcapsules with wetting agent, and obtain pre-dispersed microcapsules for later use; heat pharmaceutical grade glycerol to 50°C, add glyceryl monostearate, stir until completely dissolved, and obtain glycerol matrix for later use.

[0118] S10. Slowly add the pre-dispersed microcapsules to the glycerol matrix, shear at 5000 rpm for 10 min, then place in a vacuum drying oven and degas at 0.08 MPa for 60 min to obtain the feminine hygiene and antibacterial essential oil.

[0119] I. Antibacterial test of feminine cleansing and antibacterial essential oil

[0120] (1) Take Candida albicans ATCC 10231 and prepare a solution with a concentration of 5×10⁻⁶ using sterile physiological saline. 6 Prepare a bacterial suspension at CFU / mL for later use;

[0121] (2) Take a sterile dry filter paper with a diameter of 5 mm, add 20 μL of feminine hygiene and antibacterial essential oil to each paper, and place it in a 37℃ oven for later use; set up a negative control group and a positive control group. Add an equal amount of sterile physiological saline to the negative control group and add an equal amount of clotrimazole solution with a concentration of 50 μg / mL to the positive control group. Also place them in a 37℃ oven for later use.

[0122] (3) Use a sterile cotton swab to dip the bacterial suspension and spread it evenly on the surface of the Sabouraud dextrose agar plate three times. After each application, rotate the plate 60°. Finally, spread the sterile cotton swab around the edge of the plate, cover the plate, and let it dry at room temperature for 5 minutes.

[0123] (4) Place 4 filter paper pieces treated with antibacterial essential oil, 1 negative control group filter paper piece, and 1 positive control group filter paper piece on the surface of a Sabouraud agar plate. The center of each sample piece should be at least 25 mm apart and at least 15 mm away from the periphery of the plate. After placing the filter paper pieces, use sterile tweezers to gently press them so that they are firmly attached to the surface of the plate.

[0124] (5) Place the plates obtained in step 4 in a constant temperature incubator at 37℃ for 16-18 hours. Measure the diameter of the inhibition zone using calipers, with the outer edge of the inhibition zone as the boundary. Set up 3 parallel plates for each sample, and repeat the test 3 times. Record the diameter (mm) of each inhibition zone. An inhibition zone diameter > 7 mm is considered to have antibacterial effect, and ≤ 7 mm is considered to have no antibacterial effect. The test results are shown in Table 1.

[0125] Table 1. Antibacterial test results of feminine hygiene and antibacterial essential oils

[0126]

[0127] As shown in Table 1, the diameter of the inhibition zone in Examples 1-3 was significantly larger than that in Comparative Example 1 (P < 0.01). The antibacterial activity of the feminine hygiene and antibacterial essential oils obtained in Examples 1-3 was significantly higher than that in Comparative Example 1. The combination of oregano, thyme, and lemon balm essential oils produced a significant synergistic effect. Comparative Example 1 did not use lemon balm essential oil, and its antibacterial activity decreased significantly.

[0128] II. Biomembrane Inhibition Rate Test

[0129] (1) Take Candida albicans ATCC 10231 and prepare a bacterial suspension using yeast extract peptone glucose liquid medium (YPD). Adjust the concentration of the bacterial suspension to 1×10⁻⁶. 6 CFU / mL;

[0130] (2) Take a 96-well cell culture plate, add 180 μL of bacterial suspension to each well, and then add 20 μL of the fecal antibacterial oil obtained in Examples 1-3 and Comparative Example 1 respectively. Set the final concentration of fecal antibacterial oil in each group to be 500, 250, 125, 62.5, 31.25 and 15.625 μg / mL. Add an equal volume of clotrimazole solution with a final concentration of 50 μg / mL to the positive control group. Add an equal volume of sterile physiological saline to the negative control group. Set 3 replicates for each group. Place the 96-well plate in a constant temperature incubator at 37℃ and incubate for 72 h.

[0131] (3) After the culture is completed, discard the culture medium in the wells and gently wash each well three times with sterile PBS buffer to remove unadhered airborne bacteria; add 200 μL of 0.1 wt% crystal violet staining solution to each well, stain at room temperature for 15 min, discard the staining solution, and wash three times with sterile PBS buffer to remove unbound crystal violet; after the 96-well plate is inverted and air-dried, add 95 wt% anhydrous ethanol to each well and shake to dissolve at room temperature for 10 min to fully dissolve the crystal violet bound to the biofilm;

[0132] (4) Place the 96-well plate in a microplate reader and measure the absorbance (OD value) of each well at 570 nm. Calculate the biofilm inhibition rate (%) = (1-OD value) （实验组） / OD （阴性对照组） ) × 100%.

[0133] Table 2 Biofilm inhibition rate of feminine cleansing and antibacterial essential oils

[0134]

[0135] By comparing 96-well plates, the minimum inhibitory concentration (MIC) of the feminine hygiene antibacterial oil obtained in each example was determined to be 125 μg / mL, and the MIC of the feminine hygiene antibacterial oil obtained in Comparative Example 1 was 250 μg / mL. In order to more intuitively compare the effects of the feminine hygiene antibacterial oils obtained in each example and the comparative example, Table 2 compares the inhibition rate of the feminine hygiene antibacterial oils obtained in Examples 1-3 and Comparative Example 1 on Candida albicans biofilm at the same sample concentration (250 μg / mL).

[0136] As shown in Table 2, the feminine hygiene and antibacterial essential oils obtained in Examples 1-3 showed an inhibition rate of 83.7% against the Candida albicans biofilm at 250 μg / mL, which was significantly higher than the 35.6% of Comparative Example 1 (P < 0.01). This indicates that, under the synergistic effect of thyme essential oil, Examples 1-3 can effectively attack and destroy the biofilm structure of Candida albicans, exposing planktonic cells, which is beneficial for the further attack of oregano essential oil and thyme essential oil on the Candida albicans cell membrane.

[0137] III. Simulated vaginal flushing retention rate test

[0138] (1) Take fresh pig vaginal tissue, remove the external connective tissue, cut it longitudinally to expose the mucosal surface, wash it with physiological saline, cut it into 3cm×3cm mucosal pieces, fix it in the diffusion pool, place it with the mucosal surface facing up in a 37℃ constant temperature water bath for equilibration for 30min.

[0139] (2) Take 100mg of each of the vaginal cleansing and antibacterial essential oils obtained in Examples 1-3 and Comparative Examples 1-3, apply them evenly to the surface of the mucosa, and let it stand at room temperature for 10min; use a peristaltic pump to control the simulated vaginal secretions to continuously rinse the mucosa surface at a flow rate of 1.0mL / min; take out the mucosa after rinsing for 0.5, 1, 2, 4, 6, 8 and 12h respectively;

[0140] (3) Gently absorb the free liquid on the surface of the mucosa with filter paper, cut it into small pieces and place it in a 10 mL centrifuge tube. Add 5 mL of ethyl acetate, extract by ultrasonication at 200 W and 40 kHz for 30 min, centrifuge at 4000 rpm for 10 min, take the supernatant, repeat the extraction twice, combine the supernatants, filter with a 0.22 μm organic filter membrane to obtain the test solution, analyze by gas chromatography, calculate the initial drug loading and the residual drug on the surface of the mucosa after rinsing, and calculate the retention rate (%) of the antibacterial essential oil; the test results are shown in Table 3.

[0141] Table 3. Retention rate test results of feminine cleansing and antibacterial essential oils

[0142]

[0143] The feminine cleansing and antibacterial essential oil obtained in Example 1 still maintained a drug retention rate of 58.3% after rinsing for 12 hours; while in Comparative Example 2, which lacked dodecyl, the retention rate dropped to 52.1% after rinsing for 2 hours and only 12.6% remained after 12 hours. The lack of the anchoring effect of the dodecyl hydrophobic group in dodecyl significantly reduced the retention rate of the feminine cleansing and antibacterial essential oil; Comparative Example 3, which lacked β-cyclodextrin, had only 31.2% remaining after rinsing for 2 hours. The lack of the sustained-release effect of β-cyclodextrin caused the essential oil to be released in large quantities after the microcapsule wall material swelled in the acidic environment and was quickly lost with the flushing of secretions.

[0144] The specific embodiments of the present invention described above do not constitute a limitation on the scope of protection of the present invention. Any other corresponding changes and modifications made in accordance with the technical concept of the present invention should be included within the scope of protection of the claims of the present invention.

Claims

1. A method for preparing a feminine cleansing and antibacterial essential oil, characterized in that, Includes the following steps: S11. By weight, take 60-80 parts of pharmaceutical grade glycerin, 15-25 parts of modified chitosan-essential oil microcapsules, 5-8 parts of glyceryl monostearate and 3-6 parts of Span80. S12. Mix the modified chitosan-essential oil microcapsules with Span80 and grind them to obtain pre-dispersed microcapsules; heat pharmaceutical grade glycerin to 50~60℃, add glyceryl monostearate, and stir until completely dissolved to obtain a glycerin matrix; S13. Slowly add the obtained pre-dispersed microcapsules into the glycerol matrix, shear at high speed for 10-15 min, and degas at 0.08-0.095 MPa to obtain the feminine hygiene and antibacterial essential oil. The modified chitosan is a ternary graft compound of dodecanal-chitosan-β-cyclodextrin.

2. The method for preparing a feminine cleansing and antibacterial essential oil according to claim 1, characterized in that, The modified chitosan-essential oil microcapsules were prepared according to the following method: S21. By weight, take 20-25 parts of oregano essential oil, 20-25 parts of thyme essential oil and 15-20 parts of sage essential oil, stir and mix evenly at room temperature, add 10-15 parts of Tween-80 to obtain a compound essential oil; slowly drip the obtained compound essential oil into the modified chitosan solution, and shear at high speed for 5-10 minutes to obtain an O / W type emulsion; S22. Add 0.5wt% TPP solution dropwise to the O / W type emulsion while stirring. After the addition is complete, continue stirring for 30-60 min to obtain a microcapsule suspension. After centrifugation, discard the supernatant. Wash the obtained precipitate 3-4 times with deionized water, pre-freeze at -40℃ for 4 h, then gradually increase the temperature from -20℃ to 0℃ and dry at 10 Pa for 20 h, and finally dry at 25℃ and 5 Pa for 6 h to obtain modified chitosan-essential oil microcapsules.

3. The method for preparing a feminine cleansing and antibacterial essential oil according to claim 2, characterized in that, The modified chitosan solution is prepared according to the following method: Take 100-120 parts by weight of dodecaldehyde-chitosan-β-cyclodextrin ternary graft compound, dissolve it in 2000-2500 parts of 1% glacial acetic acid solution, stir at room temperature until completely dissolved, and adjust the pH to 4.0-5.0 to obtain modified chitosan solution.

4. The method for preparing a feminine cleansing and antibacterial essential oil according to claim 3, characterized in that, The dodecanoic acid-chitosan-β-cyclodextrin ternary graft compound was prepared by the following method: S41. By weight, take 100 parts of N-dodecyl chitosan and dissolve it in 2000~3000 parts of 1% glacial acetic acid solution. Stir at room temperature until completely dissolved, then add 6-Ns-β-CD activating solution and stir at 55~65℃ for 20~28h. S42. After the reaction is complete, adjust the pH to 9-10 to precipitate the product. Dissolve the precipitate again in dilute acetic acid and then transfer it to a dialysis bag with a molecular weight cutoff of 8000 Da. Dialyze the product 5-6 times with deionized water and freeze-dry to obtain the dodecaldehyde-chitosan-β-cyclodextrin ternary graft compound.

5. The method for preparing a feminine cleansing and antibacterial essential oil according to claim 4, characterized in that, In step S41, the 6-Ns-β-CD activating solution is prepared according to the following method: S51. By weight, take 80-120 parts of β-cyclodextrin, 100-150 parts of p-nitrobenzenesulfonyl chloride, 150-250 parts of anhydrous DMSO and 60-100 parts of triethylamine; S52. Dissolve β-cyclodextrin and p-nitrobenzenesulfonyl chloride together in anhydrous DMSO, add triethylamine, and react at 55~65℃ for 1~2h to obtain 6-Ns-β-CD activated solution.

6. The method for preparing a feminine cleansing and antibacterial essential oil according to claim 4, characterized in that, In step S41, the N-dodecyl chitosan is prepared according to the following method: S61. By weight, take 40-60 parts of dodecyl aldehyde and 2-5 parts of sodium dodecyl sulfonate; add dodecyl aldehyde to chitosan solution, stir for 20-25 min, add sodium dodecyl sulfonate, and reflux at 95-100℃ for 8-12 h. S62. After the reflux reaction is completed, the mixture is naturally cooled to room temperature. Under an ice-water bath, 20-30 parts of sodium borohydride are added, and the mixture is stirred at room temperature for 2-4 hours. After the reaction is completed, the pH is adjusted to 9-10. After the precipitate is formed, it is filtered. The precipitate is washed with deionized water until the washing solution is neutral. After vacuum drying, N-dodecyl chitosan is obtained.

7. The method for preparing a feminine cleansing and antibacterial essential oil according to claim 6, characterized in that, In step S61, the chitosan solution is prepared according to the following method: Take 100 parts by weight of chitosan and dissolve it in 2000-2500 parts of 1-2% glacial acetic acid solution. Stir at room temperature for 2-4 hours and adjust the pH to 4-6 to obtain a chitosan solution.

8. The method for preparing a feminine cleansing and antibacterial essential oil according to claim 7, characterized in that, The chitosan has a molecular weight of 50~200kDa and a degree of deacetylation ≥90%.

9. The method for preparing a feminine cleansing and antibacterial essential oil according to claim 2, characterized in that, The mass ratio of modified chitosan in the modified chitosan solution to TPP in the 0.5wt% TPP solution is 6~10:

1.

10. The feminine cleansing and antibacterial essential oil prepared by the preparation method according to any one of claims 1 to 9.

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