Plant composition with effect of regulating micro-ecological balance and application of plant composition in female private products

Abstract

The invention relates to a plant composition with a micro-ecological balance adjusting effect and application of the plant composition in female private products, and belongs to the technical field of biology. The preparation method of the plant polysaccharide fermentation product comprises the following steps: S1, extracting a plant composition with water to obtain a plant water extract; the plant composition is prepared from echinacea purpurea and circium japonicum; s2, filtering and concentrating the plant water extract in the step S1 to obtain a plant water extract concentrated solution; s3, carrying out alcohol precipitation on the plant water extract concentrated solution obtained in the step S2, and collecting the precipitate to obtain crude plant polysaccharide; and S4, fermenting the plant crude polysaccharide by using lactobacillus plantarum and lactobacillus delbrueckii to obtain a plant polysaccharide fermentation product. The echinacea and circium japonicum composition, the lactobacillus plantarum and the lactobacillus delbrueckii have a synergistic effect, polysaccharide substances are decomposed into oligosaccharide, disaccharide or monosaccharide, and by means of the prebiotics effect, female vaginal flora can be adjusted, and the micro-ecology of private parts can be balanced.

Description

Technical Field

[0001] This invention relates to the field of biotechnology, and in particular to a plant composition that regulates the balance of the microecology and its application in feminine hygiene products. Background Technology

[0002] The normal vaginal microecological system in women is composed of various microorganisms, with more than 50 types of vaginal microorganisms currently identified. Lactobacilli are the dominant bacteria in the normal vaginal microecological flora, accounting for more than 95% of all vaginal resident bacteria. Lactobacilli produce substances such as hydrogen peroxide and lactic acid through metabolism, maintaining a slightly acidic vaginal microenvironment. In this slightly acidic environment, the growth of most pathogenic and parasitic bacteria is inhibited. Therefore, lactobacilli are important probiotics in the vagina, possessing strong affinity for host cell receptors, coordinating and restricting the excessive growth of other flora, and locally regulating vaginal immune function. Simultaneously, lactobacilli can prevent other pathogenic bacteria from adhering to vaginal epithelial cells and inhibit the invasion and growth of pathogenic bacteria by activating the body's immune system, thereby maintaining the vaginal microecological balance. Various types of lactobacilli can be detected in the vagina, including *Lactobacillus curvatureii*, *Lactobacillus jenny*, *Lactobacillus gasseri*, *Lactobacillus indolentus*, and *Lactobacillus johnsonii*, among which *Lactobacillus curvatureii* is the dominant strain, accounting for 54%.

[0003] When the vaginal microecological balance is disrupted, the risk of invasion by pathogens such as bacteria and fungi increases significantly, potentially leading to various vaginal infections and even malignant tumors, such as vaginitis, cervicitis, endometritis, pelvic inflammatory disease, high-risk HPV infection, and cervical lesions. However, common feminine hygiene products on the market, such as silver ions and chlorhexidine acetate, have the problem of non-selective sterilization. While these sterilizers kill harmful bacteria, they also kill beneficial bacteria such as lactobacilli. Prolonged use may lead to an imbalance in the vaginal microecology, thereby causing various gynecological problems. Summary of the Invention

[0004] The purpose of this invention is to overcome the shortcomings of the prior art and provide a plant composition that has the function of regulating the microecological balance and its application in feminine intimate products.

[0005] To achieve the above objectives, the technical solution adopted by the present invention is as follows: In a first aspect, the present invention provides a method for preparing a plant polysaccharide fermentation product, the method comprising the following steps: S1: Extract the plant composition with water to obtain a plant water extract; the plant composition consists of Echinacea purpurea and Cirsium japonicum; S2: Filter and concentrate the plant water extract from step S1 to obtain a concentrated plant water extract solution; S3: The concentrated plant water extract from step S2 is subjected to alcohol precipitation, and the precipitate is collected to obtain crude plant polysaccharides. S4: Ferment plant crude polysaccharides with Lactobacillus plantarum and Lactobacillus delbrueckii to obtain plant polysaccharide fermentation products.

[0006] This invention selects a plant composition (echinacea and thistle) with a specific ratio of components and ferments the plant composition with specific strains (Lactobacillus plantarum and Lactobacillus delbrueckii). The resulting plant polysaccharide fermentation product has a regulatory effect on the vaginal flora of women and has a synergistic effect, further balancing the microecology of the private parts.

[0007] Further, in step S1, the weight ratio of Echinacea purpurea and Cirsium japonicum in the plant composition is: Echinacea purpurea: Cirsium japonicum = 1: (1~3).

[0008] Further, in step S1, the weight ratio of water to plant composition is: water: plant composition = (19~21):1.

[0009] Furthermore, the ratio of water to plant composition is 20:1.

[0010] Further, in step S1, the plant composition is extracted with water at 94-96°C for 1.9-2.1 hours.

[0011] Furthermore, in step S1, the plant composition is extracted with water at 95°C for 2 hours.

[0012] Furthermore, in step S2, filtration is performed using a 300-500 mesh filter.

[0013] Furthermore, in step S2, filtration is performed using a 400-mesh filter.

[0014] Furthermore, in step S2, vacuum concentration is carried out at 28~32℃.

[0015] Furthermore, in step S2, the concentration is carried out under reduced pressure at 30°C.

[0016] Furthermore, in step S3, the alcohol used for alcohol precipitation is ethanol, and the concentration of the ethanol is 70%~80% v / v.

[0017] In a specific embodiment of the present invention, the concentration of ethanol is 80% v / v.

[0018] Furthermore, in step S4, the bacterial concentration of *Lactobacillus plantarum* and *Lactobacillus delbrueckii* is 0.8 × 10⁻⁶. 8 ~1.2×10 8 CFU / mL.

[0019] Furthermore, in step S4, the bacterial concentration of *Lactobacillus plantarum* and *Lactobacillus delbrueckii* is 1 × 10⁻⁶.8 CFU / mL.

[0020] In this invention, *Lactobacillus plantarum* HCS03-001 and *Lactobacillus delbrueckii* BNCC362385 can be selected.

[0021] Further, in step S4, the ratio of *Lactobacillus plantarum* or *Lactobacillus delbrueckii* bacterial solution to plant crude polysaccharide is: *Lactobacillus plantarum* or *Lactobacillus delbrueckii* bacterial solution : plant crude polysaccharide = (1~2) mL : (5~20) g.

[0022] Furthermore, in step S4, the ratio of *Lactobacillus plantarum* or *Lactobacillus delbrueckii* bacterial solution to crude plant polysaccharide is (1~2) mL: 20 g.

[0023] As a preferred embodiment of the present invention, the ratio of *Lactobacillus plantarum* bacterial solution to crude plant polysaccharide is 2 mL: 20 g, and the ratio of *Lactobacillus delbrueckii* bacterial solution to crude plant polysaccharide is 1 mL: 20 g.

[0024] Further, in step S4, the crude plant polysaccharide is prepared into a crude plant polysaccharide solution and then fermented. In the crude plant polysaccharide solution, the mass ratio of crude plant polysaccharide to solvent is 1:(4.8~5.2).

[0025] Furthermore, in step S4, the mass ratio of the plant crude polysaccharide to the solvent is 1:5.

[0026] Furthermore, in step S4, the solvent for preparing the plant crude polysaccharide solution can be water.

[0027] Furthermore, in step S4, the crude plant polysaccharide solution is sterilized before fermentation; any conventional sterilization method in the art is acceptable. In a specific embodiment of the present invention, sterilization is performed at 121°C for 15-20 minutes, preferably 15 minutes.

[0028] Furthermore, in step S4, the fermentation temperature is 30~38℃ and the fermentation time is 48~96h.

[0029] Furthermore, in step S4, the fermentation temperature is 37℃ and the fermentation time is 72h, resulting in a better microbial regulation effect of the obtained plant polysaccharide fermentation product.

[0030] Further, in step S4, the fermentation speed is 180~220 r / min. In a specific embodiment of the present invention, the fermentation speed is 200 r / min.

[0031] Secondly, the present invention provides a plant polysaccharide fermentation product, which is prepared by the aforementioned preparation method.

[0032] Thirdly, the present invention provides the application of the plant polysaccharide fermentation product in the preparation of a product for regulating the vaginal flora in women.

[0033] The plant polysaccharide fermentation product inhibits pathogenic bacteria and promotes the proliferation of beneficial bacteria.

[0034] In a specific embodiment of the present invention, the plant polysaccharide fermentation product inhibits at least one of Escherichia coli, Staphylococcus aureus and Candida albicans, and promotes the proliferation of Lactobacillus curvature.

[0035] Fourthly, the present invention provides a product for regulating the vaginal flora of women, the product containing the plant polysaccharide fermentation product.

[0036] Compared with the prior art, the beneficial effects of the present invention are as follows: This invention extracts polysaccharides from a combination of echinacea and thistle, and then ferments them with Lactobacillus plantarum and Lactobacillus delbrueckii. The echinacea and thistle combination, along with Lactobacillus plantarum and Lactobacillus delbrueckii, have a synergistic effect, breaking down polysaccharides into oligosaccharides, disaccharides, or monosaccharides. Through prebiotic action, this promotes the proliferation of Lactobacillus curvature and inhibits the growth of harmful bacteria (Escherichia coli, Staphylococcus aureus, and Candida albicans), which is beneficial for regulating the vaginal flora and balancing the microecology of the private parts. Detailed Implementation

[0037] To better illustrate the purpose, technical solution, and advantages of this invention, the invention will be further described below with reference to specific embodiments. Unless otherwise specified, other materials and reagents used in the embodiments are commercially available.

[0038] Lactobacillus plantarum HCS03-001 is sourced from Australian Koch (Beijing) Technology Co., Ltd. Lactobacillus delbrueckii BNCC362385 was sourced from Australian Koch (Beijing) Technology Co., Ltd. Bifidobacterium BNCC232112 was derived from Australian Koch (Beijing) Technology Co., Ltd. Escherichia coli 8099 was derived from the China Industrial Microbial Culture Collection Center; Staphylococcus aureus ATCC6538 was obtained from the Guangzhou Microbiology Analysis and Testing Center; Candida albicans ATCC10231 was obtained from Guangzhou Microbiology Analysis and Testing Center; Lactobacillus acidophilus GDMCC1.208 was obtained from the Guangdong Provincial Microbial Culture Collection Center; Lactobacillus curvature BNCC135057 was sourced from Beina Chuanglian Biotechnology Co., Ltd.

[0039] Example 1 (1) Take Echinacea and Cirsium japonicum respectively, crush them, and pass them through a 40-mesh sieve to obtain Echinacea powder and Cirsium japonicum powder.

[0040] (2) Weigh 200g of echinacea powder and 600g of thistle powder, mix them evenly to obtain a mixed powder. The weight ratio of echinacea powder to thistle powder is echinacea powder: thistle powder = 1:3.

[0041] (3) Add water to the mixed powder, the weight of water being 20 times the weight of the mixed powder, and heat and extract at 95°C for 2 hours to obtain the water extract.

[0042] (4) Filter the water extract through a 400-mesh filter, and concentrate the filtrate under reduced pressure at 30°C to obtain a concentrated solution.

[0043] (5) Add ethanol to the concentrate, adjust the ethanol concentration to 80% (v / v), filter, collect the precipitate, and dry it to obtain crude polysaccharide.

[0044] (6) Weigh 20g of crude polysaccharide, add 100g of water, mix and dissolve, sterilize at 121℃ for 15min, cool to room temperature, and inoculate 2mL of Lactobacillus plantarum (HCS03-001) seed solution (1×10⁻⁶) in a clean bench. 8 CFU / mL) and 1 mL of Lactobacillus delbrueckii (BNCC362385) seed culture (1×10⁻⁶ CFU / mL) 8 The product was obtained by fermenting at 37℃ and 200r / min for 72h with CFU / mL.

[0045] (7) Sterilize the fermentation product (121℃, 15min), centrifuge at 10000r / min for 10min, collect the supernatant, concentrate under reduced pressure at 30℃, freeze dry to obtain plant polysaccharide fermentation product.

[0046] Example 2 In step (2) of Example 1, the weights of both echinacea powder and thistle powder are 300g, that is, the weight ratio of echinacea powder to thistle powder is echinacea powder: thistle powder = 1:1. Other preparation methods remain unchanged, and plant polysaccharide fermentation products are obtained.

[0047] Example 3 In step (6) of Example 1, the content of Lactobacillus plantarum seed liquid is 1 mL and the content of Lactobacillus delbrueckii seed liquid is 2 mL. Other preparation methods remain unchanged to obtain plant polysaccharide fermentation products.

[0048] Example 4 In step (6) of Example 1, the fermentation time is 48 hours, and other preparation methods remain unchanged to obtain plant polysaccharide fermentation products.

[0049] Comparative Example 1 In step (2) of Example 1, the mixed powder contains only 200g of Echinacea powder and no thistle powder. Other preparation methods remain unchanged to obtain plant polysaccharide fermentation products.

[0050] Comparative Example 2 In step (2) of Example 1, the mixed powder contains only 600g of thistle powder and no echinacea powder. Other preparation methods remain unchanged to obtain plant polysaccharide fermentation products.

[0051] Comparative Example 3 In step (2) of Example 1, 200g of licorice powder was used to replace 200g of echinacea powder, while other preparation methods remained unchanged, to obtain plant polysaccharide fermentation products.

[0052] Comparative Example 4 In step (6) of Example 1, the content of Lactobacillus plantarum seed liquid is 3 mL, without Lactobacillus delbrueckii seed liquid, and other preparation methods remain unchanged to obtain plant polysaccharide fermentation products.

[0053] Comparative Example 5 In step (6) of Example 1, the content of Lactobacillus delbrueckii seed liquid is 3 mL, without Lactobacillus plantarum seed liquid, and other preparation methods remain unchanged to obtain plant polysaccharide fermentation products.

[0054] Comparative Example 6 In step (6) of Example 1, the seed liquid of Lactobacillus delbrueckii was replaced with the seed liquid of Bifidobacterium (BNCC232112), while the other preparation methods remained unchanged, and the plant polysaccharide fermentation product was obtained.

[0055] Comparative Example 7 In Example 1, the crude polysaccharide was not fermented; only the crude polysaccharide was used for subsequent experiments.

[0056] Test Example 1 I. Experimental Methods The plant polysaccharide fermentation products of Examples 1-4 and Comparative Examples 1-6, and the crude polysaccharide of Comparative Example 7 were prepared into test solutions with a concentration of 1% (w / v) using sterile water. The antibacterial effects of the test solutions on Staphylococcus aureus, Escherichia coli, Candida albicans, and Lactobacillus were tested. The test method was carried out in accordance with 5.1.1 of "WS / T 650-2019 Evaluation Method for Antibacterial and Antimicrobial Effects".

[0057] II. Experimental Results As shown in Table 1, the plant polysaccharide fermentation products prepared in Examples 1-4 of this invention exhibit antibacterial activity against *Escherichia coli* 8099, *Staphylococcus aureus* ATCC6538, and *Candida albicans* ATCC10231, but not against *Lactobacillus acidophilus* GDMCC1.208. The plant polysaccharide fermentation products of Comparative Examples 1-6 and the crude polysaccharide of Comparative Example 7 show weaker antibacterial effects against *Escherichia coli* 8099, *Staphylococcus aureus* ATCC6538, and *Candida albicans* ATCC10231 than those of Examples 1-4, and some even show no antibacterial activity.

[0058] Table 1 Note: In Table 1, if the diameter of the inhibition zone is >7mm, it is considered to have antibacterial effect; otherwise, it has no antibacterial effect.

[0059] Test Example 2 I. Experimental Methods The test solution from Test Example 1 was used to replace the glucose in the MRS liquid medium to prepare the experimental medium. The MRS liquid medium without a carbon source was used as the blank control medium. The pH of all the above media was adjusted to 6.2±0.2, and after being dispensed, they were sterilized in an autoclave at 121℃ for 20 min and set aside for later use.

[0060] Lyophilized Lactobacillus curvature (BNCC135057) powder was dissolved in sterile distilled water, spread on MRS agar plates, and incubated at 37°C for 24 hours. Single colonies were selected and inoculated into MRS liquid medium, and incubated at 37°C for 16 hours to obtain activated Lactobacillus curvature.

[0061] Activated Lactobacillus curvature was inoculated into experimental and blank control media at a rate of 1% (w / v) and cultured at 37°C for 48 h. The OD value of Lactobacillus curvature and the pH value of the culture medium were measured.

[0062] II. Experimental Results As shown in Table 2, the OD values ​​of *Lactobacillus curvature* in Examples 1-4 were all greater than those in Comparative Examples 2-7, and the OD values ​​of *Lactobacillus curvature* in Examples 1-3 were all greater than those in Comparative Example 1.

[0063] Table 2 Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit the scope of protection of the present invention. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the essence and scope of the technical solutions of the present invention.

Claims

1. A method for the production of a plant polysaccharide fermentation product, characterized in that, The preparation method of the plant polysaccharide fermentation product comprises the following steps: S1: extracting a plant composition with water to obtain a plant water extract; the plant composition is composed of Echinacea purpurea and Cirsium japonicum; S2: filtering and concentrating the plant water extract of step S1 to obtain a plant water extract concentrate; S3: alcohol precipitating the plant water extract concentrate of step S2, collecting the precipitate, and obtaining a plant crude polysaccharide; S4: fermenting the plant crude polysaccharide with Lactobacillus plantarum and Lactobacillus delbrueckii to obtain a plant polysaccharide fermentation product.

2. The production method according to claim 1, characterized by, In step S1, the weight ratio of Echinacea purpurea to Cirsium japonicum in the plant composition is: Echinacea purpurea:Cirsium japonicum=1:(1~3).

3. The preparation method according to claim 1, characterized in that, In step S1, the weight ratio of water to the plant composition is: water:plant composition=(19~21):

1.

4. The method of claim 1, wherein, In step S1, the plant composition is extracted with water at 94~96℃ for 1.9~2.1h.

5. The preparation method according to claim 1, characterized in that, In step S4, the concentration of the Lactobacillus plantarum and Lactobacillus delbrueckii bacterial solution was 0.8 x 10 8 CFU / mL. 8 CFU / mL.

6. The production method according to claim 5, wherein In step S4, the ratio of Lactobacillus plantarum or Lactobacillus delbrueckii liquid to the plant crude polysaccharide is: Lactobacillus plantarum or Lactobacillus delbrueckii liquid:plant crude polysaccharide=(1~2)mL:(5~20)g.

7. The preparation method according to claim 1, characterized in that, In step S4, the fermentation temperature is 30~38℃, and the fermentation time is 48~96h.

8. A plant polysaccharide fermentation product, characterized in that, The plant polysaccharide fermentation product is prepared by the preparation method of any one of claims 1~8.

9. The use of the plant polysaccharide fermentation product of claim 8 in the preparation of a product for regulating female vaginal flora.

10. A product for modulating the bacterial flora of the female vagina, characterized in that, The product contains the plant polysaccharide fermentation product of claim 8.

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