CN113521278A - Female health care composition, preparation method and use method - Google Patents

Abstract

The invention relates to the field of antibodies and microbial preparations, and particularly discloses a female health-care composition, which comprises the following components: yolk antibody lyophilized powder, lactobacillus lyophilized powder, and acid buffer. The probiotic freeze-dried powder contains prebiotics, the yolk antibody can specifically and efficiently kill pathogenic microorganisms in the vagina, the lactobacillus can be planted and rapidly propagated in the vagina under the cooperation of the prebiotics, the balance of healthy microbial flora in the vagina is recovered, and the acidic buffer assists in maintaining the acidic environment in the vagina. The composition of the invention can kill main vaginal pathogenic bacteria, simultaneously synchronously utilize probiotics to quickly establish vaginal microbial flora ecology, does not generate drug resistance, has quick curative effect and good curative effect, and completely eliminates the defect that the disease condition is easy to relapse in the traditional treatment method.

Description

Female health care composition, preparation method and use method

Technical Field

The invention relates to the field of antibodies and microbial preparations, in particular to a female health-care composition, a preparation method and a use method.

Background

Vaginal infectious diseases commonly occur in women of all ages, which bring serious inconvenience to working and life and cause serious damage to physical and psychological health. Common vaginal infectious diseases include: bacterial vaginitis, trichomonas vaginitis, mycotic vaginitis and the like, and the root cause of the bacterial vaginitis, the trichomonas vaginitis, the mycotic vaginitis and the like is the result of the imbalance of vaginal microbial flora, the reduction of lactobacillus and the increase of other pathogenic bacteria (pathogenic protozoa). The abundance of lactobacillus in the vagina of healthy women reaches more than 95 percent, and comprises the following steps: lactobacillus crispatus, lactobacillus gasseri, lactobacillus jensenii, lactobacillus ferments glycogen into glucose and finally forms lactic acid, so that the pH in the vagina is reduced to below 4.0, and the propagation of a plurality of pathogenic bacteria is avoided; hydrogen peroxide, bacteriocin and other substances secreted by lactobacillus have the effect of killing pathogenic bacteria; meanwhile, the competitive adhesion of the lactobacillus in vaginal mucosa epithelial cells plays a role of biological barrier, and the propagation of pathogenic bacteria in the vagina is prevented. Due to the influence of internal and external factors such as antibiotics, age, living dietary habits, personal physique and the like, bacterial vaginitis is caused by the dominance of pathogenic bacteria such as gardnerella, proteus mirabilis, campylobacter mobilis and the like along with the decrease of lactobacillus in the vagina, the increase of PH in the vagina and the mass propagation of anaerobic bacteria and other pathogenic bacteria; the pH value reaches above 5.0, and the gel is suitable for the mass propagation of candida albicans and has dominance, so as to cause mycotic vaginitis (candida albicans vaginitis); the pH value is more than 5.0, and the trichomonas vaginalis preparation is suitable for mass propagation of trichomonas vaginalis in the vagina to cause trichomonas vaginitis. Gardner bacteria generally exist in the vagina, even if a healthy woman usually detects a small amount of gardner bacteria in the vagina, the gardner bacteria can decompose protein to generate amino acid, when the environment in the vagina is proper, the amino acid is decomposed into ammonia (or amine) by other anaerobic bacteria, the pH value in the vagina is increased, pathogenic bacteria are further propagated to generate fishy substances, and then various vaginal infectious diseases are caused.

At present, the treatment methods for common vaginal infectious diseases can be divided into 3 types: antibiotic therapy, probiotic replacement, and antibiotic + probiotic combination therapy. The antibiotic therapy mainly uses metronidazole, clindamycin and other medicines to kill anaerobic bacteria in the vagina in an oral or suppository mode, has quick curative effect, but can not quickly establish the ecological environment of healthy microbial flora in the vagina, and generally causes relapse within 3-12 months; the probiotic replacement method is to use active microbial inoculum of lactobacillus crispatus, lactobacillus gasseri, lactobacillus hophilus, sometimes lactobacillus rhamnosus, lactobacillus reuteri and lactobacillus delbrueckii to replace microorganisms in the vagina, but the curative effect is slow; the combined treatment of the antibiotics and the probiotics is to kill the microorganisms in the vagina by using the antibiotics at first and then culture healthy microbial flora in the vagina by using the probiotics, so that the treatment method is complicated, and meanwhile, the use of the antibiotics is easy to cause pathogenic bacteria to generate drug resistance and the problem of relapse of disease exists. Unlike the intestinal tract, the lower the vaginal microbial diversity is, the more the vagina can maintain its healthy state under normal conditions.

The egg yolk antibody has the characteristics of stable chemical property, high yield, low cost, strong specificity, no toxic or side effect and the like, is a research hotspot at present, and has been applied to the fields of cultivation and health-care food to a certain extent. The yolk antibody is an antibody which is extracted from an immunized egg and aims at a specific antigen, and is the most main immunoglobulin in yolk; the hen is immunized by the antigen prepared by vaginal pathogenic bacteria, the immune system of the hen generates antibody to eliminate the antigen, and the obtained antibody is selectively transferred to the yolk in the blood in the oviduct, namely the IGY antibody.

In the currently published patents, various methods are provided, such as treatment of certain vaginal microbial infections using egg yolk antibodies (CN 102329392A), vaginal microbial conditioning using prebiotics (CN 112336834A), and improvement of vaginal microbial balance using lactobacilli by oral or direct administration (CN 110917174A). However, the methods generally have the defects of single treatment mechanism and unsatisfactory effect, for example, the egg yolk antibody is not highly purified, and the mixed protein carried in the antibody can promote the propagation of harmful bacteria to cause the increase of the pH in the vagina, so that the method is not suitable for direct vaginal administration; the prebiotic or lactobacillus preparation is administered in order to promote the proliferation of the lactobacilli, but in the case of a preponderance of harmful bacteria in the vagina, the establishment of the dominance of the lactobacilli is very slow and difficult, and therefore the course of treatment is particularly long and the effect is greatly compromised.

Disclosure of Invention

The invention provides a female health-care composition, a preparation method and a use method thereof in order to make up for the defects of the prior art.

The invention is realized by the following technical scheme: a feminine care composition comprising: yolk antibody lyophilized powder, lactobacillus lyophilized powder, and acid buffer.

Furthermore, the mixture ratio of each part of the composition is as follows: 2-20 mg of yolk antibody, 50-150mg of lactobacillus lyophilized powder and 5-100 mg of acid buffer.

Furthermore, the mixture ratio of each part of the composition is as follows: 8-12 mg of yolk antibody, 80-120mg of lactobacillus lyophilized powder and 10-50 mg of acid buffer.

Further, the lactobacillus freeze-dried powder comprises lactobacillus and prebiotics, wherein the content of the lactobacillus is 0.1-100 hundred million/part, and optimally 1-10 hundred million/part; the lactobacillus includes Lactobacillus crispatus, Lactobacillus gasseri, Lactobacillus jensenii, preferably Lactobacillus crispatus.

Further, the acidic buffer comprises: d-sodium lactate buffer, L-sodium lactate buffer, sodium acetate buffer, sodium citrate buffer; the acidic buffer dilutes the solution 100-fold over a pH range of 3.5-4.0.

The invention also provides a preparation method of the female health-care composition, which comprises the following steps:

(1) culturing pathogenic bacteria from vagina, collecting thallus, and preparing inactivated antigen; the pathogenic bacteria include: any one or any combination of Gardner bacteria, Candida albicans, Proteus mirabilis and Campylobacter bacteria;

(2) immunizing laying hens with the antigen obtained in the step (1), wherein the dosage of each laying hen is 2-100 mg of antigen prepared by thalli, then carrying out boosting immunization once every 15-45 days, and eliminating the laying hens when the egg yolk antibody titer of laid eggs is reduced to 10 ten thousand;

(3) detecting the antibody titer of serum and egg yolk of the immunized laying hens, collecting eggs laid by the immunized laying hens, performing egg white separation, salt-free water dilution, acidification, centrifugal primary treatment, and separating and purifying by using a chromatographic column;

(4) carrying out ultrafiltration, desalination and concentration on the desorption solution, and freeze-drying to obtain yolk antibody freeze-dried powder;

(5) culturing Lactobacillus from healthy vagina, collecting thallus, adding prebiotics, and freeze drying to obtain Lactobacillus lyophilized powder

(6) Mixing the yolk antibody freeze-dried powder, the lactobacillus freeze-dried powder and the acid buffer to obtain the composition;

(7) the composition can be made into suppository, pill, capsule or powder.

Further, the dosage of the antigen in the step (2) is 6mg-16 mg.

Further, the prebiotics in the step (5) comprise: any one or any combination of D-galactose, D-glucose, D-fructose, lactose, trehalose, raffinose, mannose and D-mannitol.

The invention also provides a method of using a feminine care composition, comprising:

(1) the composition in form of suppository, pill or capsule is directly applied to vagina once a day;

(2) the composition pill or powder is dissolved in 2-5ml of cold boiled water, and injected into vagina with syringe once a day;

(3) the composition pill or powder is dissolved by cold boiled water in each dose, and externally genitalia is wiped every day;

(4) the composition pill or powder is dissolved in cold boiled water for use.

Compared with the prior art, the invention has the advantages that:

1. the IGY antibody generated by the invention has stronger heat resistance, acid resistance, anti-ion strength and certain enzyme degradation resistance; the IGY antibody has high affinity to mammals and good use safety;

2. in the traditional treatment method, even if healthy microecological flora is reestablished after antibiotics are stopped in the later stage, the intermediate has a blank period, and especially some drug-resistant pathogenic bacteria can be propagated on the machine, so that the treatment effect is influenced, and the disease relapse is caused. The highly purified egg yolk antibody is used for direct vaginal administration, main vaginal pathogenic bacteria are efficiently and specifically killed, probiotics are synchronously utilized to restore the ecological balance of microbial communities in the vagina, drug resistance is avoided, the disease condition is not relapsed, the curative effect is quick, the treatment effect is good, and the defect that the abundance of lactobacillus is greatly reduced while anaerobic bacteria and pathogenic bacteria (pathogenic protozoa) are removed by using antibacterial drugs in the traditional treatment method is completely eradicated;

3. the invention adopts a comprehensive treatment mechanism of targeted inactivation, multi-target synergy, environmental synergy, directional proliferation and multi-measure combination, and the designed composition formula has the characteristics of exact effect and quick curative effect. The acidic buffer agent rapidly changes the pH value in the vagina, so that the vagina environment is in a healthy pH environment with the pH value of 3.5-4.0, the activity of harmful bacteria is reduced, the minimum concentration level required by the coagulation inactivation of the antibody for the harmful bacteria is reduced, and the antibody effect is greatly improved; the vaginal diseases are embodied by certain characteristic disease symptoms (such as mycotic vaginitis), although characteristic harmful bacteria (such as candida albicans) of the disease are dominant in the vagina, other harmful bacteria also grow in a considerable amount, the invention adopts the antibody prepared by multiple antigens to carry out targeted efficient inactivation on various main harmful bacteria causing the vaginal diseases, thereby not only increasing the treatment range of the disease types, but also increasing the treatment effect of the specific symptom diseases; the antibody inactivates harmful bacteria, and the antagonistic action of the harmful bacteria on the growth of lactobacillus is eliminated; the acidic environment is suitable for the proliferation of lactobacillus, and under the coordination of prebiotics, the lactobacillus grows and breeds in large quantity and rapidly builds up a healthy vaginal micro-ecological environment; with the continuous improvement of the abundance of the lactobacillus, the continuous decline of the abundance of anaerobic bacteria and pathogenic bacteria, the killing effect of hydrogen peroxide, bacteriocin and lactic acid generated by the lactobacillus on the antibody-incompatible pathogenic bacteria (pathogenic protozoa) is generated, and the infectious vaginal diseases are thoroughly cured. The composition of the invention can obtain instant effect for the first time.

Detailed Description

The following embodiments of the present invention are provided, and it should be noted that the present invention is not limited to the following embodiments, and all equivalent changes based on the technical solutions of the present invention are within the protection scope of the present invention.

Example 1

The method for culturing the vaginal pathogenic bacteria comprises the following steps:

(1) culture of Gardnerella

The volume of the 30L full-automatic fermentation tank is 16L, and 320g of Brookfield broth culture medium is dissolved by deionized water and is constant to about 12L. Sterilizing at 121 deg.C for 20 min, cooling to 37 deg.C, automatically controlling temperature, and introducing binary gas (5% CO)₂ ，95% N₂) Introducing into a fermentation tank to replace air in the fermentation tank until the dissolved oxygen value is reduced to near zero and stable, and adjusting pH to 7.0 + -0.2 with liquid alkali; opening the inoculation port, adding 400ml of bovine serum, inoculating 200ml of the Gardner strain seed culture solution, screwing the inoculation port, and continuously introducing binary gas to reduce the dissolved oxygen value to be near zero and stable. The rotation speed of the stirrer was set at 100 rpm, and the culture was carried out for 24 hours. The culture solution was centrifuged at 10000rpm for 20 minutes at high speed, and 105.4g of the cells were collected.

(2) Culture of Campylobacter

30L of full-automatic fermentation tank, the metering volume is 16L, and 320g of Brookfield broth culture medium is added with deionized water to be about 12L. Sterilizing at 121 deg.C for 20 min, cooling to 37 deg.C, introducing nitrogen gas into the fermenter to replace air therein until dissolved oxygen value is reduced to near zero and stabilized, and adjusting pH to 6.8 + -0.1 with liquid alkali. Opening the inoculation port, adding 300ml of bovine serum, inoculating 200ml of the campylobacter mobilis seed culture solution, screwing the inoculation port, and continuously introducing nitrogen until the dissolved oxygen indicated value is stable near zero. The rotation speed of the stirrer was set at 100 rpm, and the culture was carried out for 72 hours. Centrifuging the culture solution at 10000rpm for 20 minutes at a high speed, and collecting 67.8g of thalli;

(3) culture of Proteus mirabilis

30L of full-automatic fermentation tank, the material volume is 16L, and the mixture ratio is carried out according to TB culture medium. Sterilizing at 121 deg.C for 20 min, cooling to 37 deg.C, and adjusting pH to 6.8 + -0.1 with ammonia water. 200ml of proteus mirabilis seed culture solution is added, the stirring speed is set as 100 r/min, the culture is carried out for 24h, and the PH is controlled to be 6.8 plus or minus 0.1 by ammonia water in the whole process. The culture solution was centrifuged at 10000rpm for 20 minutes at high speed, and 173.5g of the cells were collected.

(4) Candida albicans culture

30L of full-automatic fermentation tank, the material metering volume is 16L, and the following mixture ratio is adopted: 5g/L of peptone, 10g/L of yeast extract powder, 6g/L of ammonium dihydrogen phosphate, 0.43g/L of calcium sulfate, 8.4g/L of magnesium sulfate and 0.5ml/L of foam killer. Sterilizing at 121 deg.C for 20 min, cooling to 30 deg.C, pumping 640ml sterilized glucose solution with concentration of 50% into fermentation tank, and adjusting pH to 6.0 + -0.2 with ammonia water. Inoculating Candida albicans seed culture solution 600ml, setting stirring speed at 100 rpm, culturing for 20h, and controlling pH with ammonia water to 6.0 + -0.2. The culture solution was centrifuged at 10000rpm for 20 minutes at high speed, and 974.2g of the cells were collected.

Example 2

The preparation method of the lactobacillus crispatus freeze-dried powder comprises the following steps:

(1) according to the ratio of MRS culture medium: 10.0g/L of peptone, 20g/L of glucose, 10g/L of beef extract, 5g/L of yeast extract, 4.8g/L of anhydrous sodium acetate, 2.0g/L of diammonium citrate, 800.1 g/L of tween-800, 0.58g/L of magnesium sulfate, 0.28g/L of manganese sulfate, 2g/L of dipotassium hydrogen phosphate and single digestion of glucose solution. A 30L fermentation tank, the material metering volume is 16L, the sterilization is carried out for 20 minutes at the temperature of 121 ℃, the temperature is reduced to 37 ℃, the inoculation amount is 3 percent, the pH of ammonia water is automatically controlled to be 5.8, the stirring speed is 100 r/min, and the culture is carried out for 16 hours. Diluting the culture solution by 1500 times, dyeing the culture solution by a methylene blue reagent, directly counting the culture solution by a blood counting plate, centrifuging the culture solution for 20 minutes at 10000rpm of the culture solution with the number of lactobacillus crispatus being 33.2 hundred million/ml, and collecting 152.3g of thalli;

(2) dissolving 300g of lactose and 30g of D-mannitol in 1500ml of deionized water, adding the collected thalli, stirring to uniformly suspend the thalli, pre-freezing at-20 to-40 ℃, and freeze-drying by using a freeze dryer to obtain 341.6g of freeze-dried powder; the methylene blue reagent is stained, a blood counting chamber is used for counting, the viable count of the lactobacillus crispatus is 1419.7 hundred million/g of freeze-dried powder, and the yield of the freeze-dried viable count is 91.3 percent.

3000g of lactose which is crushed to more than 200 meshes is uniformly mixed with the freeze-dried powder to obtain the lactobacillus freeze-dried powder.

Example 3

The preparation method of the yolk antibody comprises the following steps:

(1) mixing 8g of Gardner bacteria thallus, 2g of Candida albicans wet thallus and 2g of Proteus mirabilis, re-suspending with 10 times of PBS buffer solution with pH7.4, adding formaldehyde with final concentration of 0.5%, and inactivating at 37 ℃ for 36 h; centrifuging to take the thallus, washing 3 times by using PBS buffer solution, and finally resuspending the thallus by using 240ml of PBS buffer solution;

(2) adding about 250ml of Freund's complete adjuvant into the inactivated bacterial suspension in the step (1), and emulsifying by using an emulsifier to obtain about 500ml of immune antigen emulsion; injecting the antibiotic-free laying hens of about 150 days old on the back by a syringe in four-point subcutaneous injection, wherein the dosage of each laying hen is 0.5 ml; performing secondary immunization after 15 days, performing tertiary immunization after 15 days, and performing the same immunization method and adjuvant dosage as the first immunization; then, the immunity is strengthened once every 1 month, and the dosage of the immune antigen emulsifier for each laying hen is 0.25 ml; when the egg yolk antibody titer of the produced eggs is reduced to 10 ten thousand, the batch of laying hens is eliminated;

(3) after the third immunization, the titer of antibodies of serum and egg yolk of the immune layer chicken is detected by an ELISA detection method, and when the titer of the antibodies of the serum and the egg yolk of the immune layer chicken exceeds 10 ten thousand, the eggs are collected for antibody extraction;

(4) taking 40 eggs laid by immunized laying hens, washing with clear water, sterilizing with 75% ethanol solution for 30min, washing with clear water, separating with yolk separator to remove egg white, collecting yolk with volume of 412ml and IGY content of 12.86 mg/ml;

(5) diluting yolk to 4000ml with purified water, stirring at room temperature for 30min, slowly adjusting pH to 5.0 with 0.2M hydrochloric acid, and standing at 4 deg.C for 12 h; centrifuging at 10000rpm/min for 30min with high speed centrifuge, collecting supernatant 389.6ml, total protein content of 2.62mg/ml, and IGY content of 1.24 mg/ml.

(6) Adjusting the concentration of the centrifugal supernatant to 0.8M by using ammonium sulfate, adjusting the pH to 8.0 by using sodium hydroxide, adsorbing by using a theta 30X 400 resin column, wherein the filler is 150ml of thiophilic affinity filler, the functional group is 2-mercaptopyridine, and the sampling flow rate is 450 ml/h; after loading, 450ml of buffer solution of ammonium sulfate, pH 8.0 and 0.8M was used to wash the suspension at a flow rate of 450 ml/h. Discarding the column passing liquid;

(7) the linear elution was carried out using a pH 8.0 ammonium sulfate solution, with a concentration range of 0.8M to 0M and an eluent volume of 1200 ml. The detection points are 0.6M, 0.4M, 0.08M, 0.02M and 0M, 586ml of eluent of the 0.4M-0.02M elution section is collected, the protein content of the eluent is 5.12mg/ml, the IGY content is 4.95mg/ml, and the purity is 96.7%;

(8) ultrafiltering the eluate with 8000Da organic membrane for desalting, adding deionized water 3-5 times (400 ml each time) until the conductivity is less than 30ms/cm, and concentrating to obtain 285ml concentrate;

(9) and (5) freeze-drying the concentrated solution. The freeze-drying conditions were 0.1mbar, -50 ℃, 26-40 hours. Obtain 2856.3mg of freeze-dried powder with the titer of 1166.4 ten thousand titer/g.

Example 4:

a method for preparing a female health composition comprises the following steps:

(1) taking 100mg of the antibody freeze-dried powder in the embodiment 3, 56mg of the lactobacillus crispatus freeze-dried powder in the embodiment 2, 400mg of a D-sodium lactate buffer and 1000mg of D-galactose, and mixing to obtain the composition, wherein each part of the composition is about 156 mg;

(2) the composition can be made into suppository, pill, capsule or powder, or made into various dosage forms by adding other adjuvants.

Example 5:

a method of using a feminine care composition comprising:

(1) the composition in form of suppository, pill or capsule is directly applied to vagina once a day;

(2) the pill or powder is dissolved in about 2-5ml of cold boiled water for each part, and is injected into the vagina by a syringe once a day;

(3) pills or powder, each part is dissolved by proper cool boiled water, and the external genitalia is wiped every day;

(4) the pills or the powder are dissolved by proper cold boiled water for each part, and are used on sanitary articles.

Example 6:

according to the previous antibody agglutination reaction experiment, 1ml of mixed vaginal harmful bacteria with the OD600 value of 1 is found to have the minimum antibody mass of about 11.5mg required for complete agglutination and inactivation. This example is based on this experimental data and a functional examination of the compositions of the invention and their components was conducted.

MRS culture medium was used to treat 3 mixed harmful bacteria: the gardner bacteria, the curvularia mobilis and the candida albicans are mixed and cultured, the culture conditions are treated according to anaerobic bacteria, and the growth of various bacteria is not influenced at the moment. Mixed harmful bacteria are inoculated into the MRS culture medium, so that the final OD600 reaches about 0.01, and the OD600 is taken as a reference; the required minimum antibody flocculation concentration under the control condition is 0.115mg antibody/ml, the antibody concentration is the standard antibody concentration, and the investigation level concentration is set above and below the standard antibody concentration; inoculating lactobacillus gasseri in the culture medium to ensure that the final concentration of lactobacillus OD600 reaches 0.01; the other components are considered to be the increase and decrease of different components or raw materials on the basis of the control. The culture period is 24h, and the detection results are shown in table 1:

TABLE 1 results of incubation of additive compositions and their components

Processing combinations/results	PH	Gardnerella/cfu/ml	Curvulus mobilis cfu/ml	Candida albicans cfu/ml	Lactobacillus gasseri cfu/ml
						Control	7.0	2.4*10↑8	1.6*10↑8	1.1*10↑8	0
Lactobacillus gasseri	7.0	3.4*10↑7	2.6*10↑7	4.3*10↑7	2.8*10↑8
						Lactobacillus gasseri + acidic buffer	3.68	5.6*10↑6	3.9*10↑6	1.5*10↑7	8.9*10↑8
1/4*IGY	7.0	2.3*10↑6	5.6*10↑5	7.4*10↑6	0
						2*IGY	7.0	0	0	0	0
1/4 IGY + Lactobacillus gasseri	7.0	1.2*10↑6	2.1*10↑5	2.4*10↑6	1.4*10↑9
						1/4 IGY + acidic buffer	3.64	0	0	0	0
1/4 IGY + acid buffer + Lactobacillus gasseri	3.72	0	0	0	2.1*10↑9
						1/50 composition prebiotics (MRS) glucose	3.67	0	0	0	1.9*10↑9
1/50 composition	3.67	0	0	0	2.9*10↑9

The culture result shows that when 3 kinds of harmful bacteria and the lactobacillus gasseri are co-cultured, the mutual antagonism phenomenon exists, when the PH is down-regulated and cultured, the growth of the 3 kinds of harmful bacteria is obviously inhibited, and the growth of the lactobacillus gasseri is enhanced; the antibody IGY has obvious inactivation effect on harmful bacteria, has stronger inhibition effect on the harmful bacteria when the IGY is in an extremely low concentration, strengthens the inactivation effect of the IGY on the harmful bacteria when the culture PH is in an acidic condition, and reduces the lowest flocculation experiment concentration level, and the analysis reason is that the activity of the harmful bacteria is reduced in a low-PH environment, and the harmful bacteria can be inactivated by the IGY with lower concentration; under the condition of co-culture with the composition, the antibody ¸ acidic buffer ¸ probiotics ¸ and the prebiotics have a synergistic and obvious synergistic effect.

The above-described embodiment is only one of the preferred embodiments of the present invention, and general changes and substitutions by those skilled in the art within the technical scope of the present invention are included in the protection scope of the present invention.

Claims (9)

1. A feminine care composition, characterized by: the composition comprises the following components: yolk antibody lyophilized powder, lactobacillus lyophilized powder, and acid buffer.

2. A feminine care composition according to claim 1, wherein: the mixture ratio of each part of the composition is as follows: 2-20 mg of yolk antibody, 50-150mg of lactobacillus lyophilized powder and 5-100 mg of acid buffer.

3. A feminine care composition according to claim 1, wherein: the mixture ratio of each part of the composition is as follows: 8-12 mg of yolk antibody, 80-120mg of lactobacillus lyophilized powder and 10-50 mg of acid buffer.

4. A feminine care composition according to claim 1, wherein: the lactobacillus freeze-dried powder comprises lactobacillus and prebiotics, wherein the content of the lactobacillus is 0.1-100 hundred million/part, and the optimal content is 1-10 hundred million/part; the lactobacillus includes Lactobacillus crispatus, Lactobacillus gasseri, Lactobacillus jensenii, preferably Lactobacillus crispatus.

5. A feminine care composition according to claim 1, wherein: the acidic buffer comprises: d-sodium lactate buffer, L-sodium lactate buffer, sodium acetate buffer, sodium citrate buffer; the acidic buffer dilutes the solution 100-fold over a pH range of 3.5-4.0.

6. A process for the preparation of a feminine care composition according to any one of claims 1 to 5, wherein: the method comprises the following steps:

(1) culturing pathogenic bacteria from vagina, collecting thallus, and preparing inactivated antigen; the pathogenic bacteria include: any one or any combination of Gardner bacteria, Candida albicans, Proteus mirabilis and Campylobacter bacteria;

(2) immunizing laying hens with the antigen obtained in the step (1), wherein the dosage of each laying hen is 2-100 mg of antigen prepared by thalli, then carrying out boosting immunization once every 15-45 days, and eliminating the laying hens when the egg yolk antibody titer of laid eggs is reduced to 10 ten thousand;

(3) detecting the antibody titer of serum and egg yolk of the immunized laying hens, collecting eggs laid by the immunized laying hens, performing egg white separation, salt-free water dilution, acidification, centrifugal primary treatment, and separating and purifying by using a chromatographic column;

(4) carrying out ultrafiltration, desalination and concentration on the desorption solution, and freeze-drying to obtain yolk antibody freeze-dried powder;

(5) culturing Lactobacillus from healthy vagina, collecting thallus, adding prebiotics, and freeze drying to obtain Lactobacillus lyophilized powder

(6) Mixing the yolk antibody freeze-dried powder, the lactobacillus freeze-dried powder and the acid buffer to obtain the composition;

(7) the composition can be made into suppository, pill, capsule or powder.

7. A method of preparing a feminine care composition according to claim 6, wherein: the dosage of the antigen in the step (2) is 6mg-16 mg.

8. A method of preparing a feminine care composition according to claim 6, wherein: the prebiotics comprise: any one or any combination of D-galactose, D-glucose, D-fructose, lactose, trehalose, raffinose, mannose and D-mannitol.

9. A method of using a feminine care composition according to any one of claims 1-5, wherein: the method comprises the following steps:

(1) the composition in form of suppository, pill or capsule is directly applied to vagina once a day;

(2) the composition pill or powder is dissolved in 2-5ml of cold boiled water, and injected into vagina with syringe once a day;

(3) the composition pill or powder is dissolved by cold boiled water in each dose, and externally genitalia is wiped every day;

(4) the composition pill or powder is dissolved in cold boiled water for use.