A probiotic agent for slowing down blood coagulation and its application in preparing sanitary napkin for preventing side leakage

Abstract

The present invention relates to a probiotic agent for slowing down blood coagulation and its application in preparing a sanitary napkin for preventing side leakage. The strain in the probiotic agent for slowing down blood coagulation is composed of Lactobacillus acidophilus. Lactobacillus acidophilus LA88 strain and Lactobacillus rhamnosus Lactobacillus rhamnosus LRa05 strain composition. The two strains can cooperate with each other, promote each other, and synergize in the effect of slowing down blood coagulation. They are further used to prepare blood absorption products such as sanitary napkin products, which can prolong the penetration and absorption time of the sanitary napkin absorption layer, promote the absorption effect, and prevent blood leakage. At the same time, the above two strains also have excellent inhibitory effects on common reproductive tract pathogens and can maintain a healthy colony environment of the reproductive tract. In addition, both bacteria are probiotics, the product is highly safe, and it is not easy to produce resistance.

Description

Technical Field

[0001] The invention belongs to the technical field of microorganisms and relates to a probiotic agent for slowing down blood coagulation and application of the probiotic agent in preparing a side leakage-proof sanitary napkin. Background Art

[0002] Sanitary napkins are a type of daily hygiene product for women. Their main function is to absorb blood and secretions discharged by women during menstruation or other periods to keep the vulva clean and comfortable. Sanitary napkins usually consist of three parts: the surface layer, the absorbent layer, and the bottom layer. The surface layer is in direct contact with the skin, so the breathability and skin-friendliness of the material need to be considered to reduce skin irritation and discomfort for women. It is usually made of soft non-woven fabrics or cotton materials. As the core part of the sanitary napkin, the absorbent layer needs to have strong absorption and water-locking capabilities, and is usually made of materials such as polymer absorbent resins. The bottom layer is made of leak-proof material and has good waterproof properties to prevent liquid leakage.

[0003] Probiotic sanitary napkins are traditional sanitary napkins with probiotic ingredients added. Probiotics refer to live bacterial preparations and their metabolites that can improve the balance of human microecology, play a beneficial role, and improve the health level and health status of the host. Probiotics are mainly colonized in the human intestine and reproductive system, and exert their effects in various ways: probiotics can stimulate intestinal immune cells, enhance the function of the intestinal mucosal immune barrier, and improve the body's resistance; probiotics can regulate the balance of intestinal flora, effectively inhibit the growth of harmful bacteria, increase the number of beneficial bacteria, and maintain the health of intestinal flora; probiotics can decompose indigestible substances in food and promote the absorption of nutrients; some acid-producing probiotics such as lactobacillus and lactobacillus acidophilus can ferment to produce organic acids, lower the pH of the intestine, and inhibit the growth of pathogenic bacteria; probiotics can produce antibacterial substances such as bacteriocins and hydrogen peroxide to antagonize the growth of pathogenic bacteria and protect intestinal health. In addition, probiotics can regulate vaginal microecology, and some sanitary napkin products with added probiotics have also been launched.

[0004] "Side leakage" refers to the phenomenon that during women's menstrual period, too much menstrual blood is discharged in a short period of time and the sanitary napkin cannot absorb it quickly, resulting in the leakage of menstrual blood. This phenomenon brings a lot of inconvenience and embarrassment to women's work and life. Therefore, it is very meaningful to develop more strategies to reduce the occurrence of "side leakage". Some appearance designs are applied to sanitary napkins to prevent side leakage, such as three-dimensional guardrails, anti-leakage grooves, and lengthened and widened tail wings. However, although the appearance design can play a certain role in preventing side leakage, the effect is limited. Summary of the invention

[0005] In view of the deficiencies of the prior art, the object of the present invention is to provide a probiotic agent for slowing down blood coagulation and use thereof in the preparation of a side leakage-proof sanitary napkin.

[0006] In order to achieve the purpose of the invention, the present invention adopts the following technical solutions:

[0007] In a first aspect, the present invention provides a probiotic for slowing blood coagulation, wherein the probiotic for slowing blood coagulation comprises a strain of Lactobacillus acidophilus with a deposit number of CGMCC No. 24109. Lactobacillus acidophilus LA88 strain and Lactobacillus rhamnosus with a deposit number of CGMCC No. 24377 Lactobacillus rhamnosus LRa05 strain composition.

[0008] The present invention has developed a new probiotic compounding method, namely, Lactobacillus acidophilus Lactobacillus acidophilus LA88 strain and Lactobacillus rhamnosus Lactobacillus rhamnosus The LRa05 strain was compounded and used, and it was found that the two strains can cooperate with each other, promote each other, and synergize in the effect of slowing blood coagulation. It is further used to prepare blood absorption products such as sanitary napkin products, which can slow blood coagulation, thereby prolonging the penetration time of menstrual blood, promoting the absorption effect of the sanitary napkin absorption layer on menstrual blood, and preventing blood leakage. At the same time, the above two strains also have excellent inhibitory effects on common reproductive tract pathogens, and can maintain a healthy colony environment of the reproductive tract. In addition, both bacteria are probiotics, the product is highly safe, and it is not easy to produce resistance.

[0009] Preferably, the ratio of the number of viable cells of the LA88 strain to that of the LRa05 strain is (2-10):1, for example 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, etc.; other specific point values ​​within this numerical range can be selected and will not be described one by one here.

[0010] Preferably, the content of live bacteria in the probiotic is not less than 1×10 11 CFU / g or 1×10 11 CFU / mL, for example 1×10 11 CFU / g (CFU / mL), 3×10 11 CFU / g (CFU / mL), 5×10 11 CFU / g (CFU / mL), 8×10 11 CFU / g (CFU / mL), 1×10 12 CFU / g (CFU / mL), 3×10 12 CFU / g (CFU / mL), 5×10 12 CFU / g (CFU / mL), 1×10 13 CFU / g (CFU / mL), etc. Other specific point values ​​within this numerical range can be selected and will not be described here one by one.

[0011] Preferably, the dosage form of the probiotics includes solutions, lyophilized powders, capsules, tablets or granules. The dosage form of the probiotics involved in the present invention is not limited, including the most commonly used solutions, lyophilized powders, or further prepared capsules, tablets or granules.

[0012] Preferably, the probiotic is in the form of a freeze-dried powder, which is prepared by the following method:

[0013] The LA88 strain and the LRa05 strain are inoculated into the culture medium respectively, activated and fermented in sequence to obtain fermentation broth; the fermentation broth is centrifuged to obtain bacterial mud; the bacterial mud is mixed with a protective agent and resuspended to obtain a resuspension; the resuspension is freeze-dried to obtain LA88 bacterial powder and LRa05 bacterial powder; the LA88 bacterial powder and the LRa05 bacterial powder are mixed according to the ratio of the number of live bacteria to obtain the probiotic agent.

[0014] Preferably, the protective agent is selected from any one or a combination of at least two of skim milk, gelatin, dextrin, gum arabic, dextran, sodium alginate, polyvinyl pyrrolidone, sucrose, lactose, trehalose, sorbitol or xylitol.

[0015] Preferably, the fermentation culture temperature is 35-40°C, for example, 35°C, 36°C, 37°C, 38°C, 39°C, 40°C, etc.; the fermentation time is 12-30 h, for example, 12 h, 15 h, 16 h, 18 h, 20 h, 24 h, 28 h, 30 h, etc.; other specific point values ​​within the above numerical range can be selected, and they will not be described one by one here.

[0016] Preferably, the inoculation amount is 1-5%, for example 1%, 2%, 3%, 4%, 5%, etc. Other specific point values ​​within the numerical range can be selected and will not be described here one by one.

[0017] Preferably, the culture medium is MRS medium.

[0018] In a second aspect, the present invention provides use of the probiotic agent described in the first aspect in enhancing the anti-side leakage effect of a sanitary napkin.

[0019] In a third aspect, the present invention provides a sanitary napkin that promotes absorption and prevents side leakage, the sanitary napkin comprising a surface layer, an absorption layer and a bottom layer; the absorption layer is loaded with the probiotic agent described in the first aspect.

[0020] Preferably, a colored pattern is provided on the surface layer, and the projection of the distribution area of ​​the probiotics on the absorption layer on the surface layer falls into the colored pattern.

[0021] In order to feel the addition of probiotics more intuitively, certain areas on the surface of sanitary napkin products can be demarcated with colors to indicate the probiotic load area.

[0022] Preferably, the loading amount of the probiotic on the absorption layer is 1-5 mg / cm 2 .

[0023] Compared with the prior art, the present invention has the following beneficial effects:

[0024] The present invention has developed a new probiotic compounding method, namely, Lactobacillus acidophilus Lactobacillus acidophilus LA88 strain and Lactobacillus rhamnosus Lactobacillus rhamnosus The LRa05 strain was compounded and used, and it was found that the two strains can cooperate with each other, promote each other, and synergize in the effect of slowing blood coagulation. It is further used to prepare blood absorption products such as sanitary napkin products, which can slow blood coagulation, thereby prolonging the penetration time of menstrual blood, promoting the absorption effect of the sanitary napkin absorption layer on menstrual blood, and preventing blood leakage. At the same time, the above two strains also have excellent inhibitory effects on common reproductive tract pathogens, and can maintain a healthy colony environment of the reproductive tract. In addition, both bacteria are probiotics, the product is highly safe, and it is not easy to produce resistance.

[0025] Classification and naming of the LA88 strain involved in the present invention: Lactobacillus acidophilus Lactobacillus acidophilus The depository is the General Microbiology Center of China Microorganism Culture Collection Administration. The deposit date is December 15, 2021. The deposit number is CGMCC No. 24109. The address is: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

[0026] Classification and naming of the LRa05 strain involved in the present invention: Lactobacillus rhamnosus Lactobacillus rhamnosus The depository is the General Microbiology Center of China Microbiological Culture Collection Administration. The deposit date is January 24, 2022. The deposit number is CGMCC No. 24377. The address is: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 This is a result diagram of Example 3 verifying the effect of probiotics on promoting the absorption of menstrual blood by sanitary napkins. DETAILED DESCRIPTION

[0028] The technical solution of the present invention is further described below by specific implementation methods. It should be understood by those skilled in the art that the embodiments are only to help understand the present invention and should not be regarded as specific limitations of the present invention.

[0029] The classification and nomenclature of the LA88 strain involved in the following examples is Lactobacillus acidophilus Lactobacillus acidophilus The depository is the General Microbiology Center of China Microorganism Culture Collection Administration. The deposit date is December 15, 2021. The deposit number is CGMCC No. 24109. The address is: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

[0030] The classification and naming of the LRa05 strain involved in the following examples is Lactobacillus rhamnosus Lactobacillus rhamnosus The depository is the General Microbiology Center of China Microbiological Culture Collection Administration. The deposit date is January 24, 2022. The deposit number is CGMCC No. 24377. The address is: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

[0031] The culture medium formula involved in the following examples is as follows:

[0032] MRS medium: peptone 10g / L, beef extract 10g / L, glucose 20g / L, sodium acetate 2g / L, yeast powder 5g / L, diammonium hydrogen citrate 2g / L, K2PO4·3H2O 2.6g / L, MgSO4·7H2O 0.1g / L, MnSO4 0.05g / L, Tween 801mL / L, cysteine ​​hydrochloride 0.5g / L.

[0033] YPD liquid culture medium: 10 g / L glucose, 10 g / L peptone, 5 g / L yeast extract, 1000 mL water, pH 7.0; add 1.5% agar powder to the solid culture medium.

[0034] LB liquid culture medium: 10 g / L tryptone, 5 g / L yeast, 5 g / L NaCl, 1000 mL water, pH 7.2; add 1.5% agar powder to the solid culture medium.

[0035] The following method for preparing the probiotic bacterial liquid is as follows: the seed liquid of the LA88 strain or the LRa05 strain is inoculated into the MRS culture medium at an inoculation amount of 3% of the total mass of the culture medium, and cultured at 37° C. to obtain a culture liquid; the culture liquid is centrifuged to obtain bacterial cells; and the bacterial cells are resuspended with PBS solution.

[0036] The following is a method for preparing the probiotic powder: the seed liquid of the LA88 strain or the LRa05 strain is inoculated into the MRS culture medium at an inoculation amount of 3% of the total mass of the culture medium, and cultured at 37°C to obtain a culture solution; the culture solution is centrifuged to obtain bacterial cells; the bacterial cells are resuspended with a lyophilization protective agent (a trehalose aqueous solution with a concentration of 100 g / L), the mass ratio of the lyophilization protective agent to the bacterial cells is 2:1, and a resuspension is obtained; the resuspension is freeze-dried by a vacuum freezing method to obtain freeze-dried powders of the two bacteria. Example 1

[0037] Preparation of menstrual blood simulant:

[0038] GB / T 8939-2018 records a standard synthetic test solution that simulates the physical properties of human menstrual blood (distilled water or deionized water: 860 mL; sodium chloride: 10.00 g, chemically pure; sodium carbonate: 40.00 g, chemically pure; glycerol: 140 mL, chemically pure; sodium benzoate: 1.00 g, chemically pure; color (edible coloring): appropriate amount, chemically pure; sodium carboxymethyl cellulose: about 5 g, chemically pure; standard vehicle: 1% (volume ratio)), which has fluidity and absorption characteristics similar to menstrual blood.

[0039] In this example, we use sterile glass beads to defibrinate pig blood. Add 10 glass beads to a conical flask and sterilize it under high temperature and high pressure. Add 100 mL of fresh pig blood to the conical flask and shake it clockwise for 10 minutes. Fibrin is adsorbed onto the surface of the glass beads to achieve defibrination of the blood. The defibrinated pig blood is filtered and sterilized using a 0.22 μm filter membrane to produce sterile defibrinated pig blood. Here, we compare sterile defibrinated pig blood with a standard synthetic test solution in terms of density, viscosity, and surface tension.

[0040] Density: Take 1 mL of pig blood and weigh it on a 1 / 10,000 balance. The weight divided by the volume is the density of pig blood. After multiple measurements, the density of pig blood is (1.05±0.35) g / cm 3 , which is in line with the density range of the standard synthetic test solution in GB / T 8939-2018.

[0041] Viscosity: Clean and dry the No. 4 viscosity cup to ensure it is clean and dry. Place the pig blood sample in a 23°C environment in advance and wait for its temperature to be consistent with room temperature. Adjust the viscometer to a horizontal state, plug the nozzle with your hand and pour the pig blood sample into the viscosity cup to remove bubbles. Quickly remove your finger and start timing, and stop timing when the sample has completely flowed out. The viscosity of pig blood was determined to be (12.1±0.45) s by experiment, which is in line with the viscosity range of the standard synthetic test solution in GB / T 8939-2018.

[0042] Surface tension: Use a liquid surface tension meter to measure the surface tension of the pig blood sample. Clean the instrument, add the sample, adjust the parameters to meet the measurement requirements and perform the measurement. The surface tension of the pig blood was measured to be (34±2) mN / m, which meets the surface tension requirements of the standard synthetic test solution in GB / T 8939-2018.

[0043] In summary, the pig blood samples treated as above can be used as menstrual blood simulation fluid for subsequent performance studies. Example 2

[0044] This example verifies the effect of probiotics in slowing down blood coagulation:

[0045] A control group (no bacterial powder was added) and bacterial powder intervention groups were set up: LA88 group (LA88 bacterial powder was added), LRa05 group (LRa05 bacterial powder was added), LA88+LRa05 group 1 (LA88 bacterial powder and LRa05 bacterial powder were added with a live bacteria count ratio of 2:1), and LA88+LRa05 group 2 (LA88 bacterial powder and LRa05 bacterial powder were added with a live bacteria count ratio of 10:1).

[0046] Weigh 0.2 g of the single bacterial powder or mixed bacterial solution of each group and dissolve it in 1 mL of water to prepare a 20% bacterial solution, then add 1 mL of milk to prepare different milk bacterial suspensions as different bacterial powder intervention groups; the control group prepared milk and tap water in a 1:1 volume ratio to prepare milk dilutions. Each group took 500 μL of pig blood samples into a 2 mL sterile centrifuge tube and added 500 μL of the above different milk bacterial suspensions or milk dilutions. The difference in the time required for blood coagulation between different groups was observed and recorded. The results are shown in Table 1.

[0047] Table 1

[0048]

[0049] It can be seen from the data results in Table 1 that, compared with the control group, the probiotic agent involved in the present invention has an excellent effect of slowing down blood coagulation, and LA88 bacterial powder and LRa05 bacterial powder have a synergistic effect in the above-mentioned efficacy. Example 3

[0050] This example verifies the effect of probiotics on promoting the absorption of menstrual blood by sanitary napkins:

[0051] A control group (no bacterial powder intervention) and bacterial powder intervention groups were set up: LA88 group (intervention with LA88 bacterial powder), LRa05 group (intervention with LRa05 bacterial powder), and LA88+LRa05 group (intervention with LA88 bacterial powder and LRa05 bacterial powder with a live bacteria count ratio of 5:1).

[0052] 6 mL of normal menstrual blood simulation solution was prepared according to the volume ratio of pig blood sample: tap water: milk = 2:1:1 as the control group; different 20% bacterial powder aqueous solutions were prepared with bacterial powder from each group, and then 6 mL of menstrual blood simulation solution containing bacterial powder was prepared according to the volume ratio of pig blood sample: 20% bacterial powder aqueous solution: milk = 2:1:1 as each bacterial powder intervention group. The normal menstrual blood simulation solution and the menstrual blood simulation solution containing bacterial powder in each group were uniformly added to the surface of the same commercially available sanitary napkin product without probiotics using a syringe at the same speed (6 mL / min) to simulate the process of diffusion and absorption of menstrual blood.

[0053] Observe 5 minutes after the addition is completed. The results are as follows Figure 1 As shown in the figure, it can be seen that compared with the control group, the probiotic agent involved in the present invention can make menstrual blood penetrate into the absorption layer of the sanitary napkin more quickly and diffuse in the absorption layer, and the diffusion area of ​​the surface of the sanitary napkin is small, that is, it has a better effect of promoting absorption and preventing side leakage, and LA88 bacterial powder and LRa05 bacterial powder have a synergistic effect in the above-mentioned effects. Example 4

[0054] This example verifies the inhibition evaluation test of probiotics on common vaginal bacteria:

[0055] (1) Preparation of indicator bacterial suspension: Gardnerella ATCC 14018, Escherichia coli ATCC 25922, Candida albicans ATCC 10231, and Staphylococcus aureus ATCC 25923 were activated on plate culture medium, and bacterial mosses were selected to prepare bacterial suspension in physiological saline. The concentration of the bacterial suspension was adjusted to 10 8 CFU / mL;

[0056] (2) Antibacterial experiment: BHI medium, eosin-methylene blue agar medium, YPD agar medium, and LB agar medium with 5% defibrinated sheep blood were cooled to about 55°C and mixed with suspensions of Gardnerella, Escherichia coli, Salmonella, and Staphylococcus aureus in sequence until the number of viable indicator bacteria reached 10. 6 CFU / mL, and then quickly poured into the plate with the Oxford cup placed in advance. After the culture medium cooled and solidified, the Oxford cup was taken out and 200 μL of LA88 bacterial solution or LRa05 bacterial solution (the number of viable bacteria was 10 8 CFU / mL), and then the diameter of the inhibition zone was measured after culturing at 37℃ for 24 h. The results are shown in Table 2.

[0057] Table 2

[0058]

[0059] It can be seen from the data results in Table 2 that the probiotics involved in the present invention have excellent inhibitory effects on common vaginal pathogens.

[0060] The applicant declares that the present invention illustrates the technical solution of the present invention through the above-mentioned embodiments, but the present invention is not limited to the above-mentioned embodiments, that is, it does not mean that the present invention must rely on the above-mentioned embodiments to be implemented. Those skilled in the art should understand that any improvement of the present invention, equivalent replacement of various raw materials of the product of the present invention, addition of auxiliary components, selection of specific methods, etc., all fall within the protection scope and disclosure scope of the present invention.

[0061] The preferred embodiments of the present invention are described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the technical concept of the present invention, a variety of simple modifications can be made to the technical solution of the present invention, and these simple modifications all belong to the protection scope of the present invention.

[0062] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any suitable manner without contradiction. In order to avoid unnecessary repetition, the present invention will not further describe various possible combinations.

Claims

1. An application of a probiotic agent in enhancing the anti-side leakage effect of a sanitary napkin, characterized in that: The strain in the probiotic is Lactobacillus acidophilus with a deposit number of CGMCC No. 24109. Lactobacillus acidophilus LA88 strain and Lactobacillus rhamnosus with a deposit number of CGMCC No. 24377 Lactobacillus rhamnosus LRa05 strain composition; the ratio of the number of live bacteria of the LA88 strain to the LRa05 strain is (2-10):1; the content of live bacteria in the probiotic agent is not less than 1×10 11 CFU / g or 1×10 11 CFU / mL.

2. The use according to claim 1, characterized in that: The dosage form of the probiotics includes solution, lyophilized powder, capsule, tablet or granule.

3. The use according to claim 2, characterized in that: The dosage form of the probiotic is a freeze-dried powder, which is prepared by the following method: The LA88 strain and the LRa05 strain are inoculated into the culture medium respectively, activated and fermented in sequence to obtain fermentation broth; the fermentation broth is centrifuged to obtain bacterial mud; the bacterial mud is mixed with a protective agent and resuspended to obtain a resuspension; the resuspension is freeze-dried to obtain LA88 bacterial powder and LRa05 bacterial powder; the LA88 bacterial powder and the LRa05 bacterial powder are mixed according to the ratio of the number of live bacteria to obtain the probiotic agent.

4. The use according to claim 3, characterized in that: The protective agent is selected from any one of skim milk, gelatin, dextrin, gum arabic, dextran, sodium alginate, polyvinyl pyrrolidone, sucrose, lactose, trehalose, sorbitol or xylitol, or a combination of at least two thereof.

5. A sanitary napkin that promotes absorption and prevents side leakage, characterized in that: The absorption-promoting and side leakage-proof sanitary napkin comprises a surface layer, an absorption layer and a bottom layer; the absorption layer is loaded with a probiotic; the strain in the probiotic is Lactobacillus acidophilus with a preservation number of CGMCC No. 24109 Lactobacillus acidophilus LA88 strain and Lactobacillus rhamnosus with a deposit number of CGMCC No. 24377 Lactobacillus rhamnosus LRa05 strain composition; the ratio of the number of live bacteria of the LA88 strain to the LRa05 strain is (2-10):1; the content of live bacteria in the probiotic agent is not less than 1×10 11 CFU / g or 1×10 11 CFU / mL.

6. The absorption-promoting and side leakage-preventing sanitary napkin according to claim 5, characterized in that: The surface layer is provided with a colored pattern, and the projection of the distribution area of ​​the probiotics on the absorption layer on the surface layer falls into the colored pattern.

7. The absorption-promoting and side leakage-preventing sanitary napkin according to claim 5, characterized in that: The loading amount of the probiotic on the absorption layer is 1-5 mg / cm 2 .

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