Composition containing dendrobium officinale fermentation product extract and soybean fermentation product extract as well as preparation method and application thereof

Abstract

The invention discloses a composition containing a dendrobium officinale fermentation product extract and a soybean fermentation product extract as well as a preparation method and application thereof, and belongs to the field of biological medicine, the composition is prepared from the dendrobium officinale fermentation product extract, the soybean fermentation product extract and soybean peptide in a mass ratio of (1-2): (1-2): (0.2-0.6); the dendrobium officinale fermentation product extract and the soybean fermentation product extract are respectively obtained through extraction, fermentation, separation and purification; the fermented composition disclosed by the invention can be used for gynecological inflammation, oral inflammation and the like, can be combined with other components such as plants and natural compounds such as lactic acid, chlorhexidine gluconate and a pseudo-ginseng extract in a compatible manner according to different symptoms, and has the advantages of being capable of realizing industrial large-scale production, wide in human body application range, safe, effective, free of toxic and side effects and the like. The treatment cost can be reduced; the application scene is wide; the use is convenient; the environment-friendly effect is realized;

Description

Technical Field

[0001] The present invention relates to the field of biomedicine, and in particular to a composition containing a Dendrobium officinale fermentation product extract and a soybean fermentation product extract, a preparation method thereof and an application thereof. Background Art

[0002] Gynecological inflammation is a common gynecological disease. The current treatment method is mainly based on Western medicine using antibiotics, but the problems of drug resistance and normal flora imbalance caused by the use of antibiotics are becoming more and more serious. There are also reports on the use of traditional Chinese medicine preparations, but the current traditional Chinese medicine preparations have problems such as drug residues and uncertain effects, which need to be improved urgently. Traditional Chinese medicine lotions have shortcomings such as uncertain effects.

[0003] Dendrobium officinale Kimura & Migo is a plant of the genus Dendrobium in the Orchidaceae family. It is a traditional Chinese medicine that is both a medicine and food. "Shennong's Herbal Classic" commented that "Fenshan Dendrobium is sweet but not dry, chewed without residue, rich in flavor and rich in fat, beneficial to the stomach and fluid, but without the worry of cooling and hindering the spleen. It is indeed a supreme product." Dendrobium officinale has the effects of increasing immunity, improving gastrointestinal function, protecting the liver, and inhibiting tumors. It is a holy medicine for nourishing yin, has the effects of benefiting the stomach and promoting fluid, nourishing yin and clearing heat, contains antibacterial and anti-inflammatory ingredients, and promotes uterine detoxification. It plays a certain auxiliary therapeutic role in gynecological diseases such as female vaginitis, pelvic inflammatory disease, and endometritis. However, the current application method is mainly edible, and there are no reports on its use in the treatment of gynecological inflammation.

[0004] Soybean is also a common food and is currently widely used in cosmetics. Although there are reports of using soybean powder as one of the raw materials for gynecological inflammation, its role in the composition is unclear. Moreover, experiments have shown that using only soybean powder has little effect on gynecological inflammation. Summary of the invention

[0005] The purpose of the present invention is to provide a composition containing a fermentation product extract of Dendrobium officinale and a fermentation product extract of soybean to solve the above problems.

[0006] In order to achieve the above-mentioned purpose, the technical solution adopted by the present invention is as follows: a composition containing a Dendrobium officinale fermentation product extract and a soybean fermentation product extract, wherein the composition consists of a Dendrobium officinale fermentation product extract, a soybean fermentation product extract and a soybean peptide, wherein the mass ratio of the Dendrobium officinale fermentation product extract, the soybean fermentation product extract and the soybean peptide is (1-2): (1-2): (0.2-0.6).

[0007] After several years of research and testing, the inventors selected Dendrobium officinale and soybeans based on the traditional Chinese medicine fermentation process and combined modern biotechnology, and obtained Dendrobium officinale fermentation product extracts and soybean fermentation product extracts through crushing, solvent soaking, isothermal and isobaric extraction, coagulation and flocculation, ultrafiltration separation technology, etc. The new biological preparation obtained by mixing the obtained fermentation product extract with lactic acid, glucose acetate chlorhexidine, etc. has significant effects in inhibiting inflammation, repairing mucous membranes, increasing immunity, and improving skin.

[0008] As a preferred technical solution, the mass ratio of the extract of Dendrobium officinale fermentation product, the extract of soybean fermentation product and the soybean peptide is 1:1:0.35. Such a ratio has a better effect.

[0009] The second object of the present invention is to provide a method for preparing the composition containing the extract of Dendrobium officinale fermentation product and soybean fermentation product extract, comprising the following steps:

[0010] (1) respectively taking Dendrobium officinale and soybean, respectively extracting and separating them with ethanol, thereby obtaining Dendrobium officinale fermentation matrix and soybean fermentation matrix;

[0011] (2) using the Dendrobium officinale fermentation substrate and the soybean fermentation substrate obtained in step (1) to ferment respectively using the same method to obtain Dendrobium officinale fermentation product base liquid and soybean fermentation product base liquid respectively;

[0012] (3) extracting and drying the base liquid of the Dendrobium officinale fermentation product and the base liquid of the soybean fermentation product obtained in step (2) respectively to obtain powdered Dendrobium officinale fermentation product extract and soybean fermentation product extract;

[0013] (4) Mixing the extract of Dendrobium officinale fermentation product and soybean fermentation product extract obtained in step (3) with soybean peptide to obtain.

[0014] As a preferred technical solution, in step (1), the extraction and separation method is: first, remove impurities from Dendrobium officinale and soybeans, crush them, soak them, and then extract them with 45vol% to 55vol% ethanol respectively to obtain extracts; then extract and separate, and pasteurize to obtain the product.

[0015] As a preferred technical solution, in step (2), the fermentation method is: adding 5% to 8% of the mass of the fermentation matrix of Lactobacillus lactis, carrying out sterile aerobic constant temperature fermentation at 28°C to 32°C for 24h to 30h, and after the fermentation is completed, carrying out constant temperature enzymatic hydrolysis for 2h to 4h to obtain the fermentation product base liquid, and pasteurizing.

[0016] As a preferred technical solution, in step (3), the extraction and drying method is: first, the base liquid of Dendrobium officinale fermentation product and the base liquid of soybean fermentation product are heated for pretreatment respectively, and then electrolytes or polymer flocculants are added respectively for coagulation or flocculation, and then they are filtered with hollow fiber ultrafiltration membranes respectively, and then the material on the membrane is dried to obtain.

[0017] The soybean peptide of the present invention can be a commercially available product or can be prepared by oneself; the preparation method is as follows:

[0018] Soybean meal was used as raw material and Bacillus subtilis was used as fermentation strain. Through solid-state two-step fermentation, 45.92% and 56.33% of protein were decomposed into small-molecule soybean peptides. Mixed bacteria were added, and the strains added were 6% Bacillus subtilis 1389, 3.5% Aspergillus oryzae A-9005, and 2.5% Aspergillus niger AS3.350 in the fermentation base. The preparation process of soybean peptides was optimized by liquid fermentation of soybean meal powder, achieving a high conversion rate of small-molecule polypeptides with a molecular weight distribution below 1000Da. Soybean peptides were prepared by microbial fermentation, which significantly improved the content and antioxidant capacity of short peptides.

[0019] A third object of the present invention is to provide a preparation or device made using the above-mentioned composition.

[0020] As a preferred technical solution, the preparation is in liquid dosage form, semisolid dosage form or foam dosage form.

[0021] A fourth object of the present invention is to provide the use of the above-mentioned preparation in preventing or treating gynecological inflammation.

[0022] A fifth object of the present invention is to provide the use of the above-mentioned preparation combination in preventing or treating oral inflammation.

[0023] Compared with the prior art, the advantages of the present invention are:

[0024] Compared with the existing suppositories, tablets, etc. of antibiotic drugs / chemical ingredients, and Chinese patent medicine lotion technology, the advantages of the present invention include:

[0025] (1) The preparation of the present invention, which is a combination of extracts of Dendrobium officinale fermentation products and soybean fermentation products, can effectively inhibit Escherichia coli, Staphylococcus aureus, etc. in the female vagina, and alleviate common gynecological inflammation problems;

[0026] (2) The combination of the combined preparation of the present invention and plants and natural compounds such as Panax notoginseng extract, tea polyphenols, chlorhexidine gluconate, etc. has a significant bactericidal effect on oral discomfort, gingival swelling and pain, oral ulcers, etc. caused by oral dental plaque, with a bactericidal rate of >99%;

[0027] (3) The combined preparation of the present invention will not produce drug resistance and side effects similar to long-term use of antibiotics, and can solve the problems of poor antibacterial effect of simple Chinese patent medicine preparations, drug residues, and drug toxicity. The preparation of the present invention can achieve industrial large-scale production, reduce treatment costs, have a wide range of application scenarios, be easy to use, be environmentally friendly, and have no toxic side effects. DETAILED DESCRIPTION

[0028] The present invention will be further described below in conjunction with embodiments.

[0029] Embodiment 1:

[0030] A composition containing a Dendrobium officinale fermentation product extract and a soybean fermentation product extract, the composition consisting of the Dendrobium officinale fermentation product extract, the soybean fermentation product extract and the soybean peptide, wherein the mass ratio of the Dendrobium officinale fermentation product extract, the soybean fermentation product extract and the soybean peptide is 1:1:0.35, and the preparation method thereof comprises the following steps:

[0031] (1) Dendrobium officinale and soybean are screened to remove impurities, and they are crushed to 200 mesh by ultrafine grinders, transported into extraction tanks by air flow, extracted and soaked with 50 vol% ethanol, the material-solvent volume ratio is 1:1.4, the extraction temperature is 36°C, the extraction working pressure is 1.5 MPa, each extraction is 60 minutes, and the extraction is performed 6 times. The extract of the initial extraction is pumped into a separation tank to separate the solvent and the liquid medicine, and the supernatant is extracted. The remaining extract is collected by countercurrent extraction and used as the solvent of the next batch of materials. The supernatant is adjusted to pH 5.5 by acid-base precipitation, solid-liquid separation is performed, and pasteurization is performed to obtain the base liquid of Dendrobium officinale fermentation product and the base liquid of soybean fermentation product respectively; the gasified solvent gas is compressed, condensed and liquefied by a compressor and can be recycled;

[0032] (2) fermentation: fermenting the base liquid of the Dendrobium officinale fermentation product and the base liquid of the soybean fermentation product obtained in step (1) respectively, wherein the fermentation method comprises: adding 5% to 8% of Lactobacillus lactis to the base liquid of the fermentation product according to the total weight of the fermentation raw materials, performing aseptic aerobic fermentation, and fermenting at a constant temperature of 30° C. for 25 hours. After the fermentation is completed, performing constant temperature enzymolysis for 3 hours to obtain the base liquid of the Dendrobium officinale fermentation product and the base liquid of the soybean fermentation product respectively;

[0033] (3) Extraction and purification of fermentation products:

[0034] Pretreatment of fermentation liquid: heat the base liquid of Dendrobium officinale fermentation product and soybean fermentation product to 32°C to improve the filtration performance of the liquid;

[0035] Coagulation and flocculation: Adding electrolytes or high molecular weight flocculants to the mature fermentation broth causes it to coagulate or flocculate, making it easier to be filtered out;

[0036] Extraction: The above-mentioned Dendrobium officinale fermentation product and soybean fermentation product were placed in sterilized containers respectively, and ultrafiltration separation and extraction technology was used to separate the target substance from the solution in the form of solid, and then freeze-dried to obtain powdered Dendrobium officinale fermentation product extract and soybean fermentation product extract. The component detection results are shown in Table 1;

[0037] Table 1 Composition of extracts of Dendrobium officinale fermentation products and soybean fermentation products

[0038]

[0039]

[0040] In the above table, "%" refers to mass percentage.

[0041] Main testing methods for ingredients:

[0042] a. Polysaccharides: using the anthrone-sulfuric acid method;

[0043] b. Dendrobium alkaloids, flavonoids, and phenolic acids: using high performance liquid chromatography (HPLC);

[0044] c. Amino acid detection method: photometric analysis;

[0045] (4) Soybean peptide, purchased from Xi'an Lavia Biotechnology Co., Ltd.;

[0046] (5) The extract of the Dendrobium officinale fermentation product obtained in step (3) and the soybean fermentation product extract and the soybean peptide obtained in step (4) are mixed uniformly in a mass ratio of 1:1:0.35, placed in a sterilized sealed can and dissolved with purified water to a concentration of 15.7wt% after dissolution, and allowed to stand for 8h, lactic acid is added at a mass ratio of 0.8% of the total material after dissolution, and all are uniformly mixed to obtain a semi-finished product; and filling and packaging are performed under sterile conditions to obtain a finished product.

[0047] Embodiment 2:

[0048] Compared with Example 1, in step (5), the mass ratio of the extract of Dendrobium officinale fermentation product, the extract of soybean fermentation product and the soybean peptide is 2:1:0.6, and the rest is the same as Example 1.

[0049] Embodiment 3:

[0050] Compared with Example 1, in step (5), the mass ratio of the extract of Dendrobium officinale fermentation product, the extract of soybean fermentation product, and the soybean peptide is 1:2:0.2, and the rest is the same as Example 1.

[0051] Comparative Example 1

[0052] Compared with Example 1, this comparative example does not add the extract of Dendrobium officinale fermentation product, and the rest is the same as Example 1.

[0053] Comparative Example 2

[0054] Compared with Example 1, this comparative example does not add soybean fermentation product extract, and the rest is the same as Example 1.

[0055] Comparative Example 3

[0056] Compared with Example 1, this comparative example does not add soybean peptide, and the rest is the same as Example 1.

[0057] General performance test:

[0058] Evaluation basis: GB15979-2002 "Standard for Disposable Hygiene Products", "Technical Specification for Disinfection" 2002 Edition, "Technical Specification for Cosmetic Safety" 2015 Edition, WS628-2018 "Technical Requirements for Hygiene and Safety Evaluation of Disinfection Products", Enterprise Standard Q / CDSF02-2022. After testing:

[0059] 1. pH value: 5.76

[0060] 2. Lactic acid content and stability: Inspection basis: According to GB5009.157-2016. The lactic acid content of the sample Jieyin antibacterial liquid was measured to be 0.982%; after being stored under stable conditions (temperature 37C-40C, relative humidity>75%, and stored for 90 days), the lactic acid content was 0.966%, and the decline rate was 1.63%. It complies with the relevant requirements of the 2002 edition of the "Disinfection Technical Specifications" and the enterprise standard Q / CDSF02-2022, and the validity period of the sample storage can be set at 2 years.

[0061] 3. Lead, arsenic and mercury: Conducted in accordance with the method specified in the 2015 edition of the "Technical Specifications for Safety of Cosmetics". After determination, the lead content of the sample Jieyin antibacterial liquid was <1.5mg / Kg, the arsenic content was <0.010mg / Kg, and the mercury content was <0.002mg / Kg, which met the relevant requirements of the 2015 edition of the "Technical Specifications for Safety of Cosmetics".

[0062] 4. Microbiological index test: It is carried out in accordance with Appendix B of GB15979-2002 "Hygiene Standard for Disposable Sanitary Products". After testing, the microbiological index test results of the sample Jieyin Antibacterial Liquid meet the relevant requirements of GB15979-2002 "Hygiene Standard for Disposable Sanitary Products".

[0063] 5. Laboratory antibacterial test:

[0064] 1. Equipment:

[0065] 1.1 Test strains: Escherichia coli (8099), cultured for 5 generations, Candida albicans (ATCC10231), cultured for 5 generations, Staphylococcus aureus (ATCC6538), cultured for 4 generations, provided by China Culture Collection Center;

[0066] 1.2 Ordinary nutrient agar medium and Sabouraud agar medium were provided by Beijing Luqiao Technology Co., Ltd.;

[0067] 1.3 0.03 mol / L phosphate buffer solution (PBS);

[0068] 1.4 Carrier: filter paper (2.0cm*3.0cm), sterile graduated pipette, sterile plate, sterile test tube, constant temperature incubator, etc.

[0069] 2. Methods:

[0070] 2.1 Sample treatment: Take 5 mL of the sample solution prepared in Example 1 and Comparative Examples 1-3, and directly conduct the test according to Appendix C4 of GB15979-2002 "Sanitary Standards for Disposable Sanitary Products" and "Technical Specifications for Disinfection 2002 Edition";

[0071] 2.2 Antibacterial test No suspension test. The test temperature is 20℃+1℃, and the test is repeated three times;

[0072] 3. Results:

[0073] Under the test temperature of 20℃, the average antibacterial rates of the samples tested against Escherichia coli (8009), Candida albicans (ATCC10231), and Staphylococcus aureus (ATCC6538) are as follows:

[0074] Table 2 Average inhibition rate of Escherichia coli by different samples at different time (%)

[0075]

[0076]

[0077] Table 3 Average inhibition rate of Staphylococcus aureus by different samples at different time (%)

[0078] 2min 5min 10min 20min Example 1 52.52 57.77 61.44 66.53 Comparative Example 1 6.31 6.79 7.35 7.56 Comparative Example 2 15.03 16.11 16.76 17.35 Comparative Example 3 35.15 36.19 38.98 43.37

[0079] Table 4 Average inhibition rate of Candida albicans by different samples at different time (%)

[0080] 2min 5min 10min 20min Example 1 52.86 55.78 60.56 64.54 Comparative Example 1 5.11 5.73 6.13 6.99 Comparative Example 2 17.53 17.86 18.79 20.31 Comparative Example 3 38.32 40.19 45.56 50.36

[0081] Conclusion: The results show that after 20 minutes of exposure, the antibacterial rate of the sample against Escherichia coli (8009), Candida albicans (ATCC10231) and Staphylococcus aureus (ATCC6538) can reach more than 50%, which shows that it has antibacterial effect and meets the relevant requirements of the 2002 edition of the Technical Specifications for Disinfection.

[0082] Vaginal mucosal irritation test:

[0083] 1) Equipment

[0084] a. Sample solution prepared in Example 1

[0085] b. Experimental animals: 6 female New Zealand rabbits weighing 2.0-2.5 kg were provided by Shaanxi Junxing Biotechnology Co., Ltd., license number: SYXK (Shaanxi) 2018-007, and the source of feed and bedding was the same as above;

[0086] c. Animal room environment: temperature: 22-25C, relative humidity: 45%-55%.

[0087] 2) Methods:

[0088] a. Conduct the experiment according to the method in 2.3.5 of the 2002 edition of Technical Specifications for Disinfection.

[0089] b. Test substance dosage: 2 mL / mouse.

[0090] c. Test method: 6 New Zealand rabbits in non-estrus were randomly divided into 2 groups, namely the test group and the control group, with 3 rabbits in each group. The animals were fixed in supine position, and the test group was injected with 2 mL of the test substance through the vagina, and the control group was given the same volume of normal saline. The animals were fixed in supine position for 4 hours to allow the test substance to fully contact the vaginal mucosa. After the test, the animals were returned to the cage for breeding. After 24 hours, the animals were killed and the vaginal tissue was removed. The vascular congestion, edema and other phenomena were carefully observed with the naked eye. If there were any changes, pathological histological examination was performed;

[0091] 3) Test results: The vaginal mucosa of rabbits in the test group was observed to be normal in morphology, without congestion, swelling, or leukocyte infiltration;

[0092] 4) The average score of the sample solution prepared in Example 1 on the irritation reaction of the vaginal mucosa of rabbits was 0. According to the grading standard of vaginal mucosal irritation reaction in the 2002 edition of the "Technical Specifications for Disinfection" of the Ministry of Health, the original vaginal antibacterial solution was non-irritating to the vaginal mucosa of rabbits, which met the relevant requirements of the 2002 edition of the "Technical Specifications for Disinfection".

[0093] Clinical trial example 1:

[0094] (1) The products prepared in the above-mentioned Examples 1-3 and Comparative Examples 1-3 were tested for a cumulative test time of more than 300 days:

[0095] (2) A total of 6912 subjects were randomly divided into 7 groups, including 1000 subjects in each of Example 1, Example 2, Example 3, Comparative Example 1, Comparative Example 2, and Comparative Example 3, and the remaining 912 subjects served as a blank control group;

[0096] The scope of subjects was selected with the cooperation of public welfare organizations: five schools, hospitals / community health centers, postpartum recovery, residential communities, medical beauty institutions, etc., as follows:

[0097] 1) Adopting an open, targeted test design method: covering a wide range of people

[0098] 2) Test subjects and selection criteria: (a) age range 25-65 years old, female; (b) voluntary participation in the test;

[0099] 3) Exclusion criteria: (a) patients with severe gynecological diseases: adnexitis, pelvic inflammatory disease, erosive cervicitis, etc.; (b) patients with syphilis, HPV, etc.; (c) patients who have used antibiotics within three weeks; (d) patients who are not suitable for testing;

[0100] (3) Test method: Users with mild inflammation, middle-aged and elderly vaginitis, dryness, etc. should clean their genitals and push the test drug deep into the vagina before going to bed every night. They should lie on their backs and keep the drug for 20-30 minutes until it is completely absorbed. One course of treatment is 15 days. Two courses of treatment are tested.

[0101] (4) Efficacy criteria: (1) Significantly effective, effective: medication symptoms and signs improved, with a score of ≥92.5%; (3) Ineffective, no response: medication symptoms and signs did not improve significantly, with a score reduction of <7.5%.

[0102] (5) Results: (1) Total effective rate of comprehensive efficacy: The total effective rate of comprehensive efficacy of 3,000 subjects using Examples 1-3 over 90 days was 92.5% (N=13). The difference between the groups was compared using the chi-square test with continuous correction, and the P value was >5.76.

[0103] The significant / effective ratios of each group are shown in Table 5:

[0104] Table 5 Proportion of markedly effective / effective in each test group

[0105]

[0106] Example 4

[0107] An oral antibacterial liquid, comprising: adding 0.144% of chlorhexidine gluconate, 0.3% of tea polyphenols, and 0.4% of Panax notoginseng extract (purchased from Yunnan Yunke Pharmaceutical Co., Ltd.) to the sample liquid prepared in Example 1;

[0108] 1. Stability determination of the above oral antibacterial solution (chemical method)

[0109] 1.1. Sample: oral antibacterial solution;

[0110] 1.2. Instruments and equipment: 10ml brown polytetrafluoroethylene burette; electronic balance; Shanghai Boxun SPX-250B-Z biochemical incubator; perchloric acid standard titration solution, concentration 0.1018mol / L;

[0111] 1.3. Method: Test basis: "Technical Specifications for Disinfection" (2002 edition) 2.2.3.2, 2.2.1.12 Method 1; Storage conditions: The sample is well sealed and placed in a 37°C constant temperature box for 3 months (90 days); the test environment temperature is 20°C and the relative humidity is 60%.

[0112] 1.4. Results: The content of the active ingredient chlorhexidine gluconate before storage was 0.144% (w / w), and the content of the active ingredient chlorhexidine gluconate after storage was 0.135% (w / w).

[0113] 2. Heavy metal (lead, arsenic, mercury) detection:

[0114] 2.1 Sample: Oral antibacterial solution

[0115] 2.2. Instruments and equipment: Atomic absorption spectrophotometer, Agilent / 240Z; Atomic fluorescence spectrophotometer, Haiguang / AFS-8510; Electronic balance.

[0116] 2.3. Standard solutions of lead, arsenic and mercury, all with a concentration of 100 mg / L.

[0117] 2.4. Inspection basis: Technical Specifications for Safety of Cosmetics (2015 edition).

[0118] 2.5. The test environment temperature is 29℃ and the relative humidity is 54%.

[0119] 2.6. Results: Lead, arsenic and mercury were not detected in the oral antibacterial solution of batch number 231114.

[0120] 3. Microbiological indicators:

[0121] 3.1 Equipment: Sample: oral antibacterial liquid, Instruments and equipment: intelligent constant temperature and humidity chamber BSC-250, biochemical incubator SPX-250B-Z

[0122] 3.2 Inspection basis: GB 15979-2002 Appendix B

[0123] The test environment temperature is 18-20℃ and the relative humidity is 45-60%.

[0124] 3.3 Results: The total bacterial colony count and fungal colony count of the sample were both <20 CFU / g. No coliform bacteria, Staphylococcus aureus, Pseudomonas aeruginosa and hemolytic Streptococcus were detected, which met the requirements of GB15979-2002 "Standards for Disposable Sanitary Products".

[0125] 4. Laboratory antibacterial test:

[0126] 1. Test sample: oral antibacterial solution of Example 4

[0127] 2. Test strains: Escherichia coli (8099) and Staphylococcus aureus (ATCC6538), provided by the Food Safety Strain Collection Center of Guangdong Food Microbiological Safety Engineering Technology Research and Development Center, and the fifth generation of fresh slant culture was used for the test;

[0128] 3. Culture medium: nutrient agar medium;

[0129] 4. Method: Test basis: GB15979-2002 Appendix C1;

[0130] The test environment temperature was 19-21°C and the relative humidity was 45-60%. The test was repeated 3 times.

[0131] Preparation of test solution: stock solution;

[0132] 5. Results: The antibacterial rate data of the samples against Escherichia coli and Staphylococcus aureus are listed in Tables 6 and 7 respectively.

[0133] Table 6 Antibacterial effect of samples on Escherichia coli

[0134] The number of colonies in the control group 2min 5min 10min 20min 1 <![CDATA[2.53×10 4 ]]> >99.6％ >99.6％ >99.6％ >99.6％ 2 <![CDATA[3.02×10 4 ]]> >99.67％ >99.67％ >99.67％ >99.67％ 3 <![CDATA[2.38×10 4 ]]> >99.58％ >99.58％ >99.58％ >99.58％ average value <![CDATA[2.64×10 4 ]]> >99.62％ >99.62％ >99.62％ >99.62％

[0135] Table 7 Antibacterial effect of samples on Staphylococcus aureus

[0136] The number of colonies in the control group 2min 5min 10min 20min 1 <![CDATA[5.21×10 4 ]]> >99.81％ >99.81％ >99.81％ >99.81％ 2 <![CDATA[4.63×10 4 ]]> >99.78％ >99.78％ >99.78％ >99.78％ 3 <![CDATA[5.45×10 4 ]]> >99.82％ >99.82％ >99.82％ >99.82％ average value <![CDATA[5.10×10 4 ]]> >99.62％ >99.80％ >99.80％ >99.80％

[0137] 5. Safety test: Acute eye irritation test:

[0138] 1. Sample: oral antibacterial solution of Example 4

[0139] 2. Experimental animal information: Breed: 3 New Zealand rabbits, Pizhou Oriental Breeding Co., Ltd., production license: SCXK (Su) 2022-0004. Grade: ordinary grade, animal certificate number: NO.320728240100011665. Female, weight range: 2.0-2.5Kg, rabbit feed, feed source: Henan Huanyu Hekang Biotechnology Co., Ltd., feed production license number: SCXK (Yu) 2020-0004, feed certificate number: NO.4109782400032755.

[0140] Animal room environment: 16 - 26°C, relative humidity: 40% - 70%, Experimental Animal License Number: SYXK(E)2022 - 0132.

[0141] 3. Main reagents: 0.9% NaCl injection

[0142] 4. Inspection basis: "Disinfection Technical Specification" (2002 edition), 2.3.1 Acute eye irritation test.

[0143] 5. Preparation and administration method of the test substance: Preparation method of the test substance: Directly use 0.1 ml of the original test sample solution for the test.

[0144] 6. Administration method of the test sample: Conduct according to the acute eye irritation test method in "Disinfection Technical Specification" (2002 edition), 2.3.1; Check the eyes of the experimental rabbits 24 h before the test, and those with abnormalities are excluded. During the test, absorb 0.1 ml of the original test sample solution and drop it into the conjunctival sac of one eye of the rabbit, and use normal saline for the other eye as a normal control. After dropping the test substance, passively close the eye for 4 s, and rinse with normal saline after 30 s.

[0145] 7. Precautions: At 1 h, 24 h, 48 h, 72 h, 7 d, 14 d, and 21 d after eye drops, observe the damage and recovery of the conjunctiva, iris, and cornea of the rabbit with the naked eye and record. If no irritation reaction occurs within 72 h, or on the 7th day or the 14th day, the eye irritation reaction completely recovers, the test can be terminated in advance.

[0146] 8. Result evaluation method: Score the acute irritation reactions of the rabbit cornea, iris, and conjunctiva, and calculate the "average score" of each animal in four aspects of corneal damage, iris damage, conjunctival congestion, and conjunctival edema at three different observation times (24 h, 48 h, 72 h) (that is, the sum of the scores of each animal at 24 h, 48 h, and 72 h divided by the number of observations * 3). Judge the irritation intensity of the test substance to the eyes according to the average score and recovery time of corneal, iris, and conjunctival congestion and edema of the animals, in accordance with the eye irritation reaction grading standard.

[0147] 9. Results: The eye irritation reaction score is shown in Table 8. The average scores of corneal damage, iris damage, conjunctival congestion, and conjunctival edema of 3 animals are 0 on average.

[0148] Table 8 Eye irritation test score table

[0149]

[0150]

[0151] Conclusion: The oral antibacterial liquid stock solution of Example 4 is non-irritating to the acute eye irritation intensity of New Zealand rabbits, which meets the requirements of the "Technical Specification for Disinfection" (20022 edition).

[0152] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. A composition containing a Dendrobium officinale fermentation product extract and a soybean fermentation product extract, characterized in that: The composition consists of a Dendrobium officinale fermentation product extract, a soybean fermentation product extract and a soybean peptide, wherein the mass ratio of the Dendrobium officinale fermentation product extract, the soybean fermentation product extract and the soybean peptide is (1-2): (1-2): (0.2-0.6).

2. The composition containing the extract of Dendrobium officinale fermentation product and soybean fermentation product extract according to claim 1, characterized in that: The mass ratio of the Dendrobium officinale fermentation product extract, the soybean fermentation product extract and the soybean peptide is 1:1:0.

35.

3. The method for preparing the composition containing the extract of Dendrobium officinale fermentation product and soybean fermentation product extract according to claim 1 or 2, characterized in that: The steps include: (1) respectively taking Dendrobium officinale and soybean, respectively extracting and separating them with ethanol, thereby obtaining Dendrobium officinale fermentation matrix and soybean fermentation matrix; (2) using the Dendrobium officinale fermentation substrate and the soybean fermentation substrate obtained in step (1) to ferment respectively using the same method to obtain Dendrobium officinale fermentation product base liquid and soybean fermentation product base liquid respectively; (3) extracting and drying the base liquid of the Dendrobium officinale fermentation product and the base liquid of the soybean fermentation product obtained in step (2) respectively to obtain powdered Dendrobium officinale fermentation product extract and soybean fermentation product extract; (4) Mixing the extract of Dendrobium officinale fermentation product and soybean fermentation product extract obtained in step (3) with soybean peptide to obtain.

4. The method according to claim 3, characterized in that In step (1), the extraction and separation method is: firstly, remove impurities from Dendrobium officinale and soybeans, crush them, soak them, and then extract them with 45vol% to 55vol% ethanol respectively to obtain extracts; then extract and separate them, and pasteurize them to obtain the extracts.

5. The method according to claim 3, characterized in that: In step (2), the fermentation method is: adding 5% to 8% of the mass of the fermentation matrix of Lactobacillus lactis, carrying out sterile aerobic constant temperature fermentation at 28°C to 32°C for 24h to 30h, and after the fermentation is completed, carrying out constant temperature enzymolysis for 2h to 4h to obtain the fermentation product base liquid, and pasteurizing.

6. The method according to claim 3, characterized in that In step (3), the extraction and drying method is: firstly, the base liquid of Dendrobium officinale fermentation product and the base liquid of soybean fermentation product are respectively subjected to heating pretreatment, and then electrolytes or polymer flocculants are respectively added for coagulation or flocculation, and then they are respectively filtered with hollow fiber ultrafiltration membranes, and then the membrane material is dried to obtain.

7. A preparation or device made using the composition of claim 1 or 2.

8. The preparation according to claim 7, characterized in that The preparation is in the form of a liquid dosage form, a semisolid dosage form or a foam dosage form.

9. Use of the preparation according to claim 7 in preventing or treating gynecological inflammation.

10. Use of the preparation according to claim 7 in preventing or treating oral inflammation.

Images