Bacteriostatic composition capable of balancing micro-ecology and preparation method thereof

Abstract

The invention relates to an antibacterial composition capable of balancing micro-ecology and a preparation method thereof, and the antibacterial composition is prepared by taking platycodon grandiflorum and iris tectorum roots as raw materials through fermentation and extraction. The content of polysaccharide and flavonoid components in the traditional Chinese medicine composition is increased through a fermentation technology, and enzyme generated by microorganisms in the fermentation process can hydrolyze macromolecular polysaccharide and generate micromolecular oligosaccharide, so that the prebiotic effect of the traditional Chinese medicine composition is improved; in plants, flavone can be combined with glucoside to form flavonoid glycoside, and the fermentation process can promote hydrolysis of glucosidic bonds, release flavonoid aglycones and improve the antibacterial effect.

Description

Technical Field

[0001] This invention belongs to the field of inflammation treatment technology, and specifically relates to an antibacterial composition that can balance the microecology and its preparation method. Background Technology

[0002] The female vagina contains a variety of microorganisms, forming a complex micro-ecosystem. Lactobacilli are the dominant bacteria, accompanied by a small number of other bacteria. These microorganisms interact, regulate, and depend on the host and the environment to maintain the dynamic balance of the vaginal microecology. However, under the influence of various internal and external factors, this balance can be disrupted, leading to changes in the flora and potentially causing various vaginal inflammations, resulting in abnormal vaginal discharge, vaginal itching, burning, and odor.

[0003] Common lactobacilli found in the vagina include *Lactobacillus curvatureii*, *Lactobacillus gasseri*, *Lactobacillus indolentus*, and *Lactobacillus janniae*. They maintain an acidic vaginal environment to prevent the colonization of pathogens. During their growth, lactobacilli release lactic acid, making the vagina a slightly acidic local environment. This low-pH acidic vaginal environment makes it difficult for many microorganisms to survive. For example, aerobic bacteria prefer a slightly alkaline environment, with pH values ​​often greater than 5 or even 6. Anaerobic bacteria, which cause bacterial vaginosis, prefer a slightly alkaline environment, but the pH value is often greater than 4.5. Trichomonas vaginalis also prefers a slightly alkaline environment.

[0004] In the recovery process from vaginal inflammation, restoring the number of lactobacilli and the acidic environment in the vagina, and quickly establishing the balance of the reproductive tract microecology, are crucial factors. Summary of the Invention

[0005] The technical problem to be solved by the present invention is to provide an antibacterial composition that can balance the microecology and its preparation method. The composition contains polysaccharides and flavonoids, which can effectively restore the microecological balance in the female vagina and prevent the occurrence of gynecological diseases.

[0006] This invention provides an antibacterial composition that can balance the microecology. The antibacterial composition is obtained by fermentation and extraction of Platycodon grandiflorus and Iris tectorum root.

[0007] Preferably, the mass ratio of the bellflower root to the iris root is 2-5:1-3.

[0008] This invention also provides a method for preparing an antibacterial composition that can balance the microecology, comprising the following steps:

[0009] After pulverizing the Platycodon grandiflorus and Iris tectorum root, sieve (40 mesh). Weigh 2-5 parts of Platycodon grandiflorus powder and 1-3 parts of Iris tectorum root powder according to the mass ratio, mix them, add 5-10 times the amount of water, stir evenly, and then sterilize. Inoculate with microbial spore suspension at 1-3% of the total weight of the medicinal materials (including Platycodon grandiflorus powder and Iris tectorum root powder), and culture at 28-37℃ for 2-4 days. After the culture is completed, add 10-30 times the amount of water for extraction. Filter, centrifuge, collect the supernatant, concentrate under reduced pressure, and freeze dry to obtain an antibacterial composition that can balance the microecology.

[0010] Preferably, the microorganism includes one or more of Aspergillus oryzae, Monascus purpureus, and Aspergillus niger.

[0011] Preferably, the extraction temperature is 80-100℃ and the extraction time is 1-2 hours.

[0012] Preferably, the centrifugation speed is 5000-10000 r / min.

[0013] This invention selects two medicinal materials: Platycodon grandiflorum and Iris root. Platycodon grandiflorum is the dried root of Platycodon grandiflorum (Jacq.) A. DC., a plant in the Campanulaceae family. The pharmacopoeia records that it has the effects of clearing the lungs, relieving sore throat, resolving phlegm, and draining pus. Platycodon grandiflorum is rich in saponins, phenolic acids, and polysaccharides, which have the effects of enhancing immunity, anti-inflammation, and anti-oxidation. Iris is a commonly used traditional Chinese medicine. Its main functions are: promoting blood circulation and removing blood stasis, dispelling wind and dampness, detoxifying, and eliminating stagnation. It is used for traumatic injuries, rheumatic pain, sore throat, abdominal distension due to food stagnation, and malaria; externally, it is used to treat boils, carbuncles, and external bleeding. Iris root contains flavonoids, lactones, coumarins, polysaccharides, phenolic tannins, organic acids, and other components.

[0014] Beneficial effects

[0015] (1) This invention increases the content of polysaccharides and flavonoids in the traditional Chinese medicine composition through fermentation process. During the fermentation process, the enzymes produced by microorganisms hydrolyze the macromolecular polysaccharides to produce small molecule oligosaccharides, thereby enhancing their prebiotic effect. In plants, flavonoids combine with glycosides to form flavonoid glycosides. The fermentation process can promote the hydrolysis of glycoside bonds, release flavonoid aglycones, and enhance the antibacterial effect.

[0016] (2) The raw materials of the composition of the present invention are derived from plants, which are mild and non-irritating. They can effectively inhibit the proliferation of harmful bacteria, promote the proliferation of lactobacilli, reduce the pH of the vaginal environment, restore the vaginal microecological balance, and will not produce drug resistance. Detailed Implementation

[0017] The present invention will be further illustrated below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Furthermore, it should be understood that after reading the teachings of this invention, those skilled in the art can make various alterations or modifications to the invention, and these equivalent forms also fall within the scope defined by the appended claims.

[0018] Example 1

[0019] After pulverizing the Platycodon grandiflorus and Iris tectorum root, pass them through a 40-mesh sieve. Weigh out 3 parts Platycodon grandiflorus powder and 2 parts Iris tectorum root powder by weight, mix them, add 5 times the amount of water, stir evenly, and then sterilize. Inoculate 2% of the total weight of the medicinal materials with Aspergillus oryzae (BNCC338380) spore suspension, and culture at 30℃ for 3 days. After the culture is completed, add 20 times the amount of water and extract at 100℃ for 2 hours. Filter, centrifuge (8000r / min) to collect the supernatant, concentrate under reduced pressure, and freeze dry to obtain an antibacterial composition that can balance the microecology.

[0020] Example 2

[0021] After pulverizing the Platycodon grandiflorus and Iris tectorum root, pass them through a 40-mesh sieve. Weigh out 2 parts Platycodon grandiflorus powder and 1 part Iris tectorum root powder by weight, mix them, add 5 times the amount of water, stir evenly, and then sterilize. Inoculate 2% of the total weight of the medicinal materials with Aspergillus oryzae (BNCC338380) spore suspension, and culture at 30℃ for 3 days. After the culture is completed, add 20 times the amount of water and extract at 100℃ for 2 hours. Filter, centrifuge (8000r / min) to collect the supernatant, concentrate under reduced pressure, and freeze dry to obtain an antibacterial composition that can balance the microecology.

[0022] Example 3

[0023] After pulverizing the Platycodon grandiflorus and Iris tectorum root, pass them through a 40-mesh sieve. Weigh out 3 parts Platycodon grandiflorus powder and 2 parts Iris tectorum root powder by weight, mix them, add 8 times the amount of water, stir evenly, and then sterilize. Inoculate 1% of the total weight of the medicinal materials with Aspergillus oryzae (BNCC338380) spore suspension and culture at 28℃ for 4 days. After the culture is completed, add 30 times the amount of water and extract at 100℃ for 2 hours. Filter, centrifuge (8000r / min) to collect the supernatant, concentrate under reduced pressure, and freeze dry to obtain an antibacterial composition that can balance the microecology.

[0024] Comparative Example 1

[0025] After pulverizing the Platycodon grandiflorus and Glycyrrhiza uralensis, pass them through a 40-mesh sieve. Weigh out 3 parts Platycodon grandiflorus powder and 2 parts Glycyrrhiza uralensis powder by weight, mix them, add 5 times the amount of water, stir evenly, and then sterilize. Inoculate 2% of the total weight of the medicinal materials with Aspergillus oryzae (BNCC338380) spore suspension and culture at 30℃ for 3 days. After the culture is completed, add 20 times the amount of water and extract at 100℃ for 2 hours. Filter, centrifuge (8000r / min) to collect the supernatant, concentrate under reduced pressure, and freeze dry to obtain the antibacterial composition.

[0026] Comparative Example 2

[0027] After pulverizing the Platycodon grandiflorus and Iris tectorum root, pass them through a 40-mesh sieve. Weigh out 1 part Platycodon grandiflorus powder and 5 parts Iris tectorum root powder according to the mass ratio, mix them, add 5 times the amount of water, stir evenly, and then sterilize. Inoculate 2% of the total weight of the medicinal materials with Aspergillus oryzae (BNCC338380) spore suspension, and culture at 30℃ for 3 days. After the culture, add 20 times the amount of water and extract at 100℃ for 2 hours. Filter, centrifuge (8000r / min) to collect the supernatant, concentrate under reduced pressure, and freeze dry to obtain the antibacterial composition.

[0028] Comparative Example 3

[0029] After pulverizing the Platycodon grandiflorus and Iris tectorum root, pass them through a 40-mesh sieve. Weigh out 3 parts Platycodon grandiflorus powder and 2 parts Iris tectorum root powder according to the mass ratio, mix them, add 20 times the amount of water, extract at 100℃ for 2 hours, filter and centrifuge (8000r / min) to collect the supernatant, concentrate under reduced pressure and freeze dry to obtain the antibacterial composition.

[0030] Test case

[0031] 1. The total flavonoid content was determined by ultraviolet spectrophotometry.

[0032] Accurately weigh 10.0 mg of rutin standard, dissolve it in 60% ethanol, and dilute to 50 mL to prepare a 0.2 mg / mL rutin standard solution. Take 0.0 mL, 0.5 mL, 1.0 mL, 2.0 mL, 2.5 mL, 3.0 mL, and 3.5 mL of the rutin standard solution respectively into 10 mL stoppered test tubes, add 60% ethanol solution to make up to 5 mL, add 0.3 mL of 5% sodium nitrite solution, shake well, and let stand for 6 min. Add 0.3 mL of 10% aluminum nitrate solution, shake well, and let stand for 6 min. Add 4 mL of 5% sodium hydroxide solution, shake well, and let stand for 15 min. Measure the absorbance at a wavelength of 510 nm. Plot a standard curve with rutin concentration on the x-axis and absorbance on the y-axis, obtaining the standard equation: y = 12.612x - 0.0354 (R² / 2π)² / 2π. 2=0.9977). Weigh 1g of each sample from the examples and comparative examples into a volumetric flask, dissolve in 60% ethanol and dilute to the mark, take 1ml of the above method for testing, and calculate the total flavonoid content in the sample according to the formula.

[0033] 2. The total polysaccharide content was determined by ultraviolet spectrophotometry.

[0034] Accurately weigh 10.0 mg of anhydrous glucose, dissolve it in distilled water, and dilute to a 100 mL volumetric flask to prepare a 100 mg / L glucose solution. Accurately pipette 0.2 mL, 0.4 mL, 0.6 mL, 0.8 mL, and 1.0 mL of the glucose standard solution into separate 20 mL stoppered test tubes. Add distilled water to a final volume of 1 mL, then add 1.0 mL of 5% phenol solution sequentially. Shake thoroughly, then quickly add 5.0 mL of concentrated sulfuric acid. Let stand for 10 min, shake thoroughly, and then incubate the test tubes in a 30°C water bath for 20 min. Measure the absorbance at 490 nm. Plot a standard curve with glucose concentration on the x-axis and absorbance on the y-axis. The standard equation is: y = 0.1503x + 0.0032 (R² / 2π)² / 2π. 2 =0.9998). Weigh 1g of the samples from the examples and comparative examples into a volumetric flask, dissolve in distilled water and bring to volume. Take 1ml of the sample and perform the test as described above, and calculate the total polysaccharide content in the sample according to the formula.

[0035] Table 1. Total flavonoids and total polysaccharides in the samples

[0036]

[0037] As can be seen from the results in the table above, the total flavonoid and total polysaccharide content in the extract obtained by this invention is significantly increased.

[0038] 3. Antibacterial test

[0039] The Oxford cup inhibition zone method was used to test the antibacterial effects of the compositions in the examples and comparative examples against Staphylococcus aureus, Escherichia coli, Candida albicans, and Lactobacillus. The concentration of the composition solution was 1%, and the test method was the beneficial bacteria ring method in WS / T 650. The results are shown in the table below.

[0040]

[0041] Note: An inhibition zone diameter > 7mm indicates antibacterial effect; otherwise, it indicates no antibacterial effect.

[0042] As shown in the table above, the extract of the present invention has good antibacterial ability against harmful bacteria such as Escherichia coli, Staphylococcus aureus and Candida albicans, and the antibacterial ability of the extract is improved after being processed by the process of the present invention.

[0043] 4. Test on the proliferation effect of lactobacilli

[0044] In conventional MRS medium, 1% of the compositions from the examples and comparative examples were used to replace glucose to prepare experimental media. A basal medium without a carbon source was used as a blank control group. All the above media were adjusted to pH 6.2 ± 0.2, dispensed, and sterilized in an autoclave at 121 °C for 20 min before use.

[0045] Dissolve an appropriate amount of lyophilized Lactobacillus curvature (BNCC135057) powder in sterile distilled water, spread it on a plate, and incubate at 37°C for 24 hours. Select a single colony and inoculate it into a conventional liquid MRS medium. Incubate at 37°C for 16 hours. Inoculate the activated strain into the experimental MRS medium at an inoculation rate of 1% and incubate at 37°C for 48 hours. Measure the OD value and pH value.

[0046]

[0047] The results in the table above show that, compared with the control group, the extract obtained by this invention can effectively increase the OD value of *Lactobacillus curvature*, promote its proliferation, and decrease the pH value of the solution, i.e., increase the lactic acid content. This has a promoting effect on the restoration of the vaginal microecology.

Claims

1. An antibacterial composition capable of balancing the microecology, characterized in that, The antibacterial composition is obtained by fermentation and extraction of Platycodon grandiflorus and Iris tectorum roots.

2. The antibacterial composition for balancing the microecology according to claim 1, characterized in that, The mass ratio of the bellflower root to the iris root is 2-5:1-3.

3. A method for preparing an antibacterial composition capable of balancing the microecology, characterized in that, Includes the following steps: After pulverizing the Platycodon grandiflorus and Iris tectorum root, sieve them, and weigh out 2-5 parts of Platycodon grandiflorus powder and 1-3 parts of Iris tectorum root powder according to the mass ratio. Mix them and add 5-10 times the amount of water. Stir well and then sterilize. Inoculate with microbial spore suspension at 1-3% of the total weight of the medicinal materials and culture at 28-37℃ for 2-4 days. After the culture is completed, add 10-30 times the amount of water for extraction. Filter, centrifuge and collect the supernatant. Concentrate under reduced pressure and freeze dry to obtain an antibacterial composition that can balance the microecology.

4. The preparation method according to claim 3, characterized in that, The microorganisms include one or more of Aspergillus oryzae, Monascus purpureus, and Aspergillus niger.

5. The preparation method according to claim 3, characterized in that, The extraction temperature is 80-100℃, and the extraction time is 1-2 hours.

6. The preparation method according to claim 3, characterized in that, The centrifugation speed is 5000-10000 r / min.

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