A female health conditioning composition based on a complex of lactobacillus and medicinal mushroom extract

Abstract

The present application belongs to the technical field of composite microbial composition, and particularly relates to a female health conditioning composition based on Lactobacillus composite flora and medicinal fungus extract, which comprises composite lactic acid bacteria composed of Lactobacillus crispatus, Lactobacillus jensenii and Lactobacillus reuteri in a specific ratio, a prebiotic composite formed by galactooligosaccharide, fructooligosaccharide and inulin, extracts of Ganoderma lucidum, Hericium erinaceus, Poria cocos, Cordyceps sinensis and white birch fungus, and cranberry extract powder and pineapple powder. The lactic acid bacteria is formed into double-layer microcapsules by electrostatic spraying, and three types of functional components are constructed into a three-layer composite structure by layering and pressing, so as to finally form microcapsule powder. The present application has the characteristics of stable structure, synergistic components and remarkable conditioning effect by targeting regulating vaginal microecology, inhibiting inflammatory response and repairing mucosal barrier, and is suitable for female reproductive tract health management.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of composite microbial compositions, and particularly relates to a female health conditioning composition based on Lactobacillus composite flora and medicinal fungus extract. BACKGROUND

[0002] In recent years, with the popularization of health concepts and the enhancement of women's self-health awareness, the physiological balance and immune protection of women's private parts have gradually become the focus of nutritional intervention and functional food research. Especially in the aspects of probiotic conditioning, plant functional factor intervention and natural active substance integration, good prospects are shown. The microecological environment of the female reproductive tract is highly sensitive, and the maintenance of a healthy state depends on the synergistic action of multiple factor regulation mechanisms, therefore, the development of targeted nutritional compositions has attracted widespread attention.

[0003] In existing research, Lactobacillus probiotics are widely used for vaginal microecological reconstruction and conditioning due to their natural advantages in maintaining vaginal acidity and inhibiting pathogenic bacteria colonization. Some products use oral or local delivery methods to supplement Lactobacillus strains, and are supplemented with prebiotics such as GOS to improve colonization efficiency. In addition, medicinal fungi such as Ganoderma lucidum, Coriolus versicolor and Hericium erinaceus extracts are rich in polysaccharides, triterpenes and other active ingredients, and show multi-target effects in regulating the immune system and inhibiting inflammatory response. However, current market products are mostly single-function oriented, lacking comprehensive strategies that consider the balance of flora, mucosal defense and synergistic improvement of systemic immunity, resulting in limited use effect, poor sustainability, and difficulty in meeting women's long-term conditioning needs for private health in different physiological stages.

[0004] In the context of the continuous expansion of women's overall health maintenance needs, constructing a new private health conditioning scheme with multi-factor synergistic action has become an important direction for the development of functional dietary compositions. SUMMARY

[0005] To solve the above problems, the present application aims to provide a female health conditioning composition based on specific Lactobacillus flora and medicinal fungus extract, which comprises the following ingredients:

[0006] Composite Lactobacillus: composed of freeze-dried Lactobacillus crispatus, Lactobacillus jensenii and Lactobacillus reuteri in a mass ratio of 1.5-3:1-2:1-2;

[0007] Lactobacillus crispatus is purchased from China General Microbiological Culture Collection Center (CGMCC) with the preservation number of CGMCC No.25113; Lactobacillus jensenii is purchased from China General Microbiological Culture Collection Center (CGMCC) with the preservation number of CGMCC No.9439; and Lactobacillus reuteri is purchased from China General Microbiological Culture Collection Center (CGMCC) with the preservation number of CGMCC No.19746.

[0008] Prebiotics: a complex composed of three components of oligogalactose, oligofructose and inulin, wherein the mass fraction ratio of the three components is 60-70:15-25:15-25; the prebiotics is dissolved at 60 ℃ and cooled to 25 ℃, and then mixed with the complex lactic acid bacteria at a mass ratio of 1:2-3 to form gel particles, which are dried after solidification at 10 ℃ for 15 min;

[0009] Medicinal fungus extract: composed of Ganoderma lucidum, Hericium erinaceus, Poria cocos, Cordyceps sinensis and white birch fungus in a dry powder mass ratio of 20-30:15-25:10-20:5-15:5-10;

[0010] Plant extract: including cranberry extract powder and pineapple powder at a mass ratio of 1-2:1;

[0011] Auxiliary ingredients: including natural vitamin E and sodium ascorbate;

[0012] The composition is freeze-dried to form a three-layer structure solid powder, the inner layer is a mixed layer of complex lactic acid bacteria, prebiotics and auxiliary ingredients, the middle layer is a medicinal fungus extract layer, and the outer layer is a plant extract layer.

[0013] As a preferred technical solution, Lactobacillus crispatus, Lactobacillus jensenii and Lactobacillus reuteri are pre-cultured before mixing.

[0014] The culture medium is composed of glucose, yeast extract and casein hydrolysate, the culture temperature is controlled at 37 ℃, the time is 12 h, and the bacterial powder is obtained after centrifugal separation and freeze-drying treatment, the particle size of the bacterial powder is controlled in the range of 40 μm to 60 μm; the viable count of Lactobacillus crispatus after freeze-drying is controlled at ; the viable count of Lactobacillus jensenii after freeze-drying is controlled at ; and the viable count of Lactobacillus reuteri after freeze-drying is controlled at .

[0015] As a preferred technical solution, the complex lactic acid bacteria is treated by chitosan-sodium alginate double-layer embedding, and a double-layer microcapsule structure is formed by using electrostatic spraying method, and the particle size of the microcapsule is controlled to be 100-300 μm; the concentration of chitosan solution is 1.8%, the concentration of sodium alginate solution is 1.2%, the mixing and stirring rate is 200 rpm, the stirring time is 20 min, the spraying system uses a 0.3 mm nozzle and applies a voltage of 6-8 kV to generate particles, and the solidification solution is , and the solidification time is 10 min, and the particle size distribution of the obtained microcapsule is concentrated between 150 μm and 250 μm.

[0016] As a preferred technical solution, the three components of prebiotics are dissolved at 55 ℃ respectively, and the oligogalactose, oligofructose and inulin are mixed in the order of addition, and the stirring rate is 200 rpm, and after mixing, the mixture is naturally cooled to 25 ℃, and then mixed with the complex lactic acid bacteria powder to form gel particles with a particle size of 0.5-1.0 mm, and the gel particles are collected after being solidified at 10 ℃ for 20 min and vacuum dried.

[0017] As a preferred technical solution, the prebiotic complex is pretreated by spray drying before mixing, the inlet air temperature is 120 ℃, the outlet air temperature is 85 ℃, and the moisture content of the obtained powder after drying is not higher than 3%, and low-speed stirring is used when mixing with the bacteria powder, and the stirring time is 15 min to ensure uniform mixing of the powder.

[0018] As a preferred technical solution, the preparation process of the medicinal fungus extract includes four steps of crushing, extraction, concentration and drying; the crushing step crushes the raw material to a particle size of 80-100 μm; the extraction step is carried out at a solid-liquid ratio of 1:12, an ultrasonic frequency of 40 kHz, an extraction temperature of 60 ℃, an extraction time of 1 h; the centrifugal speed is 8000 rpm, and the time is 15 min, the solid content after concentration is controlled to be 25%-30%, the spray drying temperature is 150 ℃, and the atomization pressure is 0.5 MPa.

[0019] As a preferred technical solution, the cranberry extraction powder is prepared by using an ethanol immersion process with a volume fraction of 40%, the immersion temperature is 50 ℃, the immersion time is 45 min, and the powder is prepared after reverse osmosis concentration and-35 ℃ freeze drying, the pineapple powder is prepared by mechanical enzymatic hydrolysis, the enzymatic hydrolysis temperature is 45 ℃, the time is 30 min, and the powder particle size is less than 100 μm after low-temperature spray drying, and the two are mixed in a mass ratio of 1:1 to 2:1 and then sieved uniformly.

[0020] As a preferred technical solution, the three-layer composite solid powder is prepared by a segmented compression process, the inner layer of probiotics and prebiotics is compressed at a pressure of 2 MPa, the middle layer of fungi is compressed at a pressure of 1.5 MPa, and the outer layer of plants is compressed at a pressure of 1.2 MPa, and the thickness of each layer is controlled to be between 0.5 mm and 1.0 mm; the obtained solid block is crushed into a powder with a particle size of not greater than 300 mu m after cold air drying, and then the powder is uniformly coated with an edible hydroxypropyl methyl cellulose or shellac coating by a spray drying or freeze drying method to form a microcapsule structure.

[0021] As a preferred technical solution, natural vitamin E and sodium ascorbate are dissolved in an ethanol solution and then dispersed in a medicinal fungus extraction layer, the volume fraction of ethanol is 50%, the mixing and stirring rate is 120 rpm, and the stirring time is 20 min; the obtained product is packaged with a three-layer composite aluminum foil film, and before packaging, the product is replaced with nitrogen gas with a purity of 99.9%, the replacement pressure is 0.05 MPa, the sealing temperature is 120 DEG C, the sealing width is 10 mm, and the storage temperature after packaging is controlled to be 25 DEG C.

[0022] Beneficial effects:

[0023] By introducing the specific ratio co-formulation strategy of Lactobacillus crispatus, Lactobacillus jensenii and Lactobacillus reuteri, a composite probiotic system with multi-layer colonization mechanism and metabolic complementation ability is constructed, which breaks through the limitations of traditional single strain or two-strain formula in bacterial population stability and targeting. Among them, Lactobacillus reuteri can synthesize vitamin B12 and regulate the local pH of the mucosa, Lactobacillus jensenii has strong hydrogen peroxide production ability, and Lactobacillus crispatus is the dominant colonizing bacteria in the female vaginal flora. The synergistic effect of the three significantly enhances the comprehensive functions of maintaining the lactic acid environment, inhibiting pathogenic bacteria and repairing the vaginal epithelial barrier, and realizes the microecological targeted reconstruction of "bacteria regulating bacteria".

[0024] The composite bacterial flora is electrostatically sprayed and embedded by adopting the chitosan-sodium alginate double-layer microencapsulation technology, forming a double-layer structure microparticle with controllable particle size, which shows good protection and controlled release in normal temperature and gastric acid environment. Compared with single-layer embedding or non-embedding state, the double-layer structure not only significantly improves the bioavailability and survival ability of live bacteria in the stomach and intestines, but more importantly, it endows the probiotics with good structure programmability, so that the probiotics can be released on demand in the intestinal tract or urogenital axis, thereby improving the targeted efficiency of systemic conditioning, and is a key improvement in the bacterial flora delivery system.

[0025] In terms of dosage form construction, the present application innovatively constructs a "inner-middle-outer" three-layer compression composite structure, embeds probiotics / prebiotics, medicinal fungus extracts and plant extracts in different functional layers respectively, and realizes spatial separation and synergistic release of functional components through segmented compression. This structure not only optimizes the stability and synergistic relationship between components, avoids mutual degradation or antagonism, but also realizes the coupled release of the three mechanisms of probiotic protection, fungal regulation and plant anti-adhesion, significantly improving the biological effect intensity and sustainability of the product in the process of female vaginal microecological conditioning. BRIEF DESCRIPTION OF DRAWINGS

[0026] Figure 1 are photos of vaginal epithelial tissue staining of the embodiments of the present application, wherein a and b are from samples of experimental group one, c and d are from samples of experimental group two, and e and f are from samples of experimental group three;

[0027] Figure 2 are experimental results of vaginal flora quantitative analysis of the comparative experiments of the present application;

[0028] Figure 3 are experimental results of histological evaluation of vaginal epithelium of the comparative experiments of the present application;

[0029] Figure 4 are experimental results of determination of the level of local inflammation factors in vagina of the present application. DETAILED DESCRIPTION

[0030] In order to deepen the understanding of the present application, the present application will be further described in combination with embodiments, and the embodiments are only used to explain the present application and do not constitute a limitation on the protection scope of the present application.

[0031] Embodiment: Comparative experiments of the present application.

[0032] Experimental group one (T1): preferred parameter group:

[0033] The embodiment of the present experiment is aimed at verifying a preparation method of a female health conditioning composition based on specific Lactobacillus flora and medicinal fungus extracts and the stability of the composite structure and the synergistic effect of the components, and the implementation process is as follows:

[0034] S1, pre-culture and freeze-drying treatment of composite lactic acid bacteria:

[0035] Three strains were selected, Lactobacillus crispatus, Lactobacillus jensenii and Lactobacillus reuteri. After the quality verification, the strains were inoculated into the pre-culture medium, which was composed of 10 g / L glucose, 10 g / L yeast extract and 10 g / L casein hydrolysate, pH 6.5, sterilized at 121 ℃ for 20 min and cooled. The inoculation amount was 5%, and the logarithmic phase bacterial solution was collected after 12 h of shaking culture at 37 ℃. The bacterial solution was centrifuged at 8000 rpm for 15 min, and the supernatant was discarded. The bacterial cells were freeze-dried at -40 ℃ for 24 h, and the particle size of the obtained bacterial powder was controlled at about 50 μm. The viable bacterial count after freeze-drying was controlled at: L. crispatus 1 For For The three were mixed in a mass ratio of 2:1.5:1.5 and reserved.

[0036] S2, preparation of prebiotic compound and gel particle formation:

[0037] The three components, galactooligosaccharides, fructooligosaccharides and inulin, were weighed in a mass ratio of 65:20:15 and dissolved in hot water at 55 ℃, with a dissolution concentration of 10%. They were added in sequence and mixed by magnetic stirring at 200 rpm for 20 min. The solution was naturally cooled to 25 ℃, and then mixed with the lactic acid bacteria compound powder obtained in S1 in a mass ratio of 1:2.5. After stirring, a preliminary mixed gel was formed. The obtained gel was dropped through a 0.8 mm nozzle onto a 10 ℃ cooling platform, and the gelation time was controlled at 20 min. The obtained gel particle had a particle size of about 0.8 mm, which was collected after vacuum drying.

[0038] S3, compound lactic acid bacteria microencapsulation treatment:

[0039] In the chitosan (1.8% w / v) and sodium alginate (1.2% w / v) double solution, the gel particles obtained in S2 were added, and a double-layer microcapsule structure was formed by electrostatic spraying. The nozzle aperture was 0.3 mm, the spraying voltage was set to 7 kV, the mixing and stirring rate was 200 rpm, and the time was 20 min. The sprayed particles were solidified in 0.2 mol / L CaCl2 solution for 10 min, forming double-layer microcapsule lactic acid bacteria particles with an average particle size of about 200 μm.

[0040] S4, preparation of medicinal fungus extract:

[0041] Ganoderma lucidum, Hericium erinaceus, Poria cocos, Cordyceps sinensis and Inonotus obliquus in dry powder form were mixed in a mass ratio of 25:20:15:10:5, and then ground to a particle size of about 90 μm. The mixed powder was added to pure water at a solid-liquid ratio of 1:12, and the ultrasonic frequency was set to 40 kHz, the extraction temperature was 60 ℃, and the extraction time was 1 h. After centrifugation at 8000 rpm for 15 min, the supernatant was concentrated to a solid content of about 28%, and then spray dried (inlet temperature 150 ℃, atomization pressure 0.5 MPa) to obtain a brownish yellow powder.

[0042] S5, preparation of plant extract composite powder:

[0043] Cranberry pomace was extracted with 40% ethanol by volume, and the extraction temperature was set to 50 ℃ for 45 min. The obtained extract was concentrated by reverse osmosis and then freeze-dried at -35 ℃ to obtain cranberry extract powder. Pineapple was treated by enzymatic hydrolysis at a temperature of 45 ℃ for 30 min, and then low-temperature spray drying was performed. The obtained powder had a particle size of less than 100 μm. The two were mixed in a mass ratio of 2:1 and sieved through a 100 mesh screen.

[0044] S6, three-layer structure composite powder compression and drying:

[0045] The layers were divided as follows:

[0046] The inner layer was a composite lactic acid bacteria microcapsule, prebiotic compound, and sodium ascorbate, natural vitamin E blended layer, and the compression pressure was set to 2 MPa;

[0047] The middle layer was the medicinal fungus extract powder obtained in S4, and the compression pressure was 1.5 MPa;

[0048] The outer layer was the plant extract composite powder obtained in S5, and the compression pressure was 1.2 MPa.

[0049] The thickness of each layer was controlled to be 0.8 mm, and the overall thickness was about 2.4 mm. The three-layer composite solid tablets were obtained by multi-mode tabletting equipment for directional compression, and then dried at 10 ℃ for 12 h to obtain a solid powder with a particle size of not more than 300 μm.

[0050] S7, microcapsule coating and product processing:

[0051] The above composite powder was mixed with a 10% edible gelatin solution, and then spray dried to form a primary coating structure. Then, a hydroxypropyl methylcellulose (HPMC) solution was uniformly sprayed on the outer layer, and dried to form a stable coating on the surface, obtaining a microencapsulated powder.

[0052] Group two (T2):

[0053] The present experiment group adopts the minimum recommended ratio of each component in the composition and the lower limit parameter of the process to construct a composite structure and verify the preparation stability and basic function effect under a low dose.

[0054] S1, complex lactic acid bacteria pre-culture and freeze-drying treatment:

[0055] Lactobacillus crispatus, Lactobacillus jensenii and Lactobacillus reuteri were selected, and the culture medium was composed of 10 g / L glucose, 8 g / L yeast extract and 8 g / L casein hydrolysate. After sterilization, the culture was incubated at 37°C for 12 h. The bacterial cells were harvested by centrifugation at 8000 rpm for 15 min, and the freezing temperature was -40°C, and the freeze-drying time was 24 h. The particle size of the prepared bacterial powder was about 60 μm, and the viable bacterial count after freeze-drying was controlled at:

[0056] ; ; .

[0057] The three were mixed in a mass ratio of 1.5:1:1 and used.

[0058] S2, preparation of prebiotic compound and formation of gel particles:

[0059] Oligogalactose, oligofructose and inulin were taken as prebiotics, and were weighed in a mass ratio of 60:15:15, dissolved in hot water at 55°C to a concentration of 10%, and then mixed by stirring (speed 200 rpm). After cooling to 25°C, the compound bacterial powder in S1 was mixed in a mass ratio of 1:2 to form a colloid. The colloid was used to prepare gel particles with a diameter of about 0.5 mm by dropwise addition method, and was solidified at 10°C for 15 min, and then vacuum dried for standby.

[0060] S3, complex lactic acid bacteria microencapsulation treatment:

[0061] 1.8% chitosan solution and 1.2% sodium alginate solution were uniformly mixed under stirring at 200 rpm for 20 min; the gel particles obtained in S2 were added and electrostatically sprayed through a 0.3 mm nozzle, and a voltage of 6 kV was applied. The obtained droplets were dropped into 0.2 mol / L CaCl2 solution for solidification for 10 min, and double-layer microcapsule particles with a particle size of 150 μm were obtained.

[0062] S4, preparation of medicinal fungus extract:

[0063] Ganoderma lucidum, Hericium erinaceus, Poria cocos, Cordyceps sinensis, Inonotus obliquus were weighed according to the mass ratio of 20:15:10:5:5, mixed and crushed to a particle size of about 100 μm. The extraction solvent was water, the solid-liquid ratio was 1:12, the extraction temperature was 60 ℃, the ultrasonic frequency was 40 kHz, and the extraction time was 1 h. The extraction liquid was centrifuged at 8000 rpm for 15 min to remove impurities, and the supernatant was concentrated to a solid content of 25%. The extraction powder was obtained by spray drying (temperature 150 ℃, pressure 0.5 MPa).

[0064] S5, preparation of plant extract composite powder:

[0065] The cranberry extract powder was extracted with 40% ethanol solution at 50 ℃ for 45 min, and dry powder was obtained by reverse osmosis and −35 ℃ freeze-drying method. The pineapple pulp was mechanically enzymatically digested at 45 ℃ for 30 min and then low-temperature spray dried. The particle size of the obtained powder was less than 100 μm. The two were mixed in a mass ratio of 1:1 and sieved for use.

[0066] S6, three-layer composite powder compression and drying:

[0067] The composite bacteria obtained in S3 were blended with prebiotic powder, and appropriate amounts of sodium ascorbate and natural vitamin E to form the inner layer, and the compression pressure was 2 MPa.

[0068] The medicinal fungus extract obtained in S4 was the middle layer, and the compression pressure was 1.5 MPa.

[0069] The plant extract mixed powder obtained in S5 was the outer layer, and the compression pressure was 1.2 MPa.

[0070] The thickness of each layer was controlled to be 0.5 mm, and the total thickness was about 1.5 mm. The layered compression process was used for molding. The obtained three-layer structure solid block was crushed into a powder not greater than 300 μm after cold air drying at 10 ℃ for 12 h.

[0071] S7, microcapsule coating and product processing:

[0072] The composite powder was blended with 10% edible gelatin solution, and the primary capsule structure was formed by freeze-drying. The outer layer was coated with hydroxypropyl methyl cellulose (HPMC) to form a complete coating. The obtained microcapsule powder had uniform particle size distribution, light yellow color, and good flowability.

[0073] Experimental group three (T3):

[0074] In this experimental group, the upper limit values of each key ingredient and process parameter were used to maximize the synergistic effect of probiotics, fungi and plant active ingredients in the composite formula. The processing stability and preparation performance under high dose ratio and high parameter setting were evaluated.

[0075] S1, pre-culture and freeze-drying of composite lactic acid bacteria:

[0076] Lactobacillus crispatus, Lactobacillus jensenii and Lactobacillus reuteri were inoculated into sterilized complex liquid medium (glucose 12 g / L, yeast extract 12 g / L, casein hydrolysate 12 g / L) respectively, and incubated at 37°C for 12 h. The bacterial cells were collected by centrifugation at 8000 rpm, pre-frozen at -40°C, and freeze-dried for 36 h. The particle size of the obtained bacterial powder was controlled at 40 μm. The viable bacterial counts were:

[0077] ; ; .

[0078] The three were mixed in a mass ratio of 3:2:2 to form a high proportion of complex bacterial powder.

[0079] S2, preparation of prebiotic complex and gel particle formation:

[0080] Take oligogalactose, oligofructose and inulin as prebiotics, and mix them in a mass ratio of 70:25:25. Dissolve them in hot water at 55°C to a concentration of 12%, and then add them to the mixture under magnetic stirring (stirring rate 200 rpm, time 30 min). After natural cooling to 25°C, mix the obtained complex lactic acid bacteria powder in a mass ratio of 1:3. Add the mixture dropwise to form particles with a particle size of about 1.0 mm, and then gel and solidify at 10°C for 20 min. Finally, use a vacuum dryer to dehydrate and shape the particles, and the obtained prebiotic complex gel particles are ready for use.

[0081] S3, lactic acid bacteria microencapsulation treatment (double-layer embedding):

[0082] Add the gel particles obtained in S2 to the 1.8% chitosan and 1.2% sodium alginate double solution, and mix them thoroughly by high-speed homogenization (200 rpm) for 20 min. Spray granulation is performed by an electrostatic spraying system (0.3 mm nozzle, voltage 8 kV). The liquid droplets enter the 0.2 mol / L CaCl2 solution and solidify for 10 min, forming double-layer embedded microcapsules with an average particle size of about 250 μm and clear complete membrane structure.

[0083] S4, preparation of medicinal fungus extract:

[0084] Ganoderma lucidum, Hericium erinaceus, Poria cocos, Cordyceps sinensis and white birch fungus were weighed according to the mass ratio of 30:25:20:15:10 of dry powder. The particle size was controlled at 80 μm. Ultrasonic-assisted extraction was used, with a solid-liquid ratio of 1:12, an extraction temperature of 60 ℃, a frequency of 40 kHz, and an extraction time of 1 h. The obtained extract was centrifuged at 8000 rpm for 15 min to obtain the supernatant, which was concentrated to a solid content of 30%, and then spray dried (temperature 150 ℃, atomization pressure 0.5 MPa) to obtain high-purity fungal extract powder.

[0085] S5, preparation of plant extract composite powder:

[0086] Cranberry extract was extracted with 40% ethanol as the extraction agent at 50 ℃ for 45 min, and the concentrated liquid was freeze-dried at −35 ℃ to retain the original anthocyanin components. The pineapple enzymatic treatment temperature was 45 ℃, and the time was 30 min. The particle size of the powder obtained by spray drying was less than 80 μm. The two were mixed in a mass ratio of 2:1, sieved through an 80-mesh sieve, and the powder uniformity was good.

[0087] S6, three-layer composite compression and drying treatment:

[0088] The lactic acid bacteria microcapsules in S3, the prebiotic gel powder in S2, sodium ascorbate, and natural vitamin E were mixed to form the inner layer, and the compression pressure was 2 MPa;

[0089] The medicinal fungal extract in S4 was the middle layer, and the compression pressure was 1.5 MPa;

[0090] The plant extract in S5 was the outer layer, and the compression pressure was 1.2 MPa.

[0091] The thickness of each layer was controlled at 1.0 mm, and the overall tablet thickness was 3.0 mm. The whole tablet was prepared by a three-stage layered compression molding process, and was crushed after drying for 16 h at 10 ℃. The particle size of the obtained composite powder was ≤300 μm.

[0092] S7, microcapsule coating process:

[0093] The obtained composite powder was mixed with 10% edible gelatin at a mass ratio of 1:1, and spray dried to form a preliminary capsule structure. The outer layer was coated with hydroxypropyl methyl cellulose (HPMC) by thin layer spraying, forming a complete film. The drying temperature was set to 45 ℃, and the obtained microcapsules had a concentrated particle size distribution and a smooth surface.

[0094] Comparative Example 1 (C1): Three strains of composite lactic acid bacteria were not used, and only a single strain was used.

[0095] This group is used to verify the impact on product structure and compounding effect under the condition of not using three-strain complex lactic acid bacteria, only using Lactobacillus crispatus.

[0096] S1, Preparation and treatment of lactic acid bacteria:

[0097] This group only uses Lactobacillus crispatus as the source of active bacteria, and does not use Lactobacillus jensenii and Lactobacillus reuteri. The strain used is a direct freeze-dried commercial powder, which has not been pre-cultured, and the number of live bacteria is The particle size of the bacterial powder is controlled at about 60 μm, and there is no additional purification treatment. The synergistic colonization and metabolic complementary effect that may be produced after three-strain compounding is omitted.

[0098] S2, Preparation of prebiotic compound and formation of gel particles:

[0099] According to the preferred ratio of 65:20:15, oligogalactose, oligofructose and inulin are weighed respectively and dissolved to 10% under the condition of 55 ℃ water bath. After cooling to 25 ℃, the Lactobacillus crispatus powder is mixed in a mass ratio of 1:2.5. The obtained mixed gel is treated by dropwise granulation to form gel particles with a particle size of about 0.8 mm. After cooling and solidification for 15 min and vacuum drying, the particles are collected.

[0100] S3, Without microencapsulation embedding treatment:

[0101] This group does not perform the double embedding step of chitosan and sodium alginate, and does not use an electrostatic spraying device for spray granulation. The probiotics are directly involved in the subsequent compression process in a naked state, exposed to the external environment.

[0102] S4, Preparation of medicinal fungus extract:

[0103] Ganoderma lucidum, Hericium erinaceus, Poria cocos, Cordyceps sinensis and white birch fungus are weighed according to the mass ratio of 25:20:15:10:5, and are crushed to a particle size of about 90 μm. Water extraction method is used, solid-liquid ratio is 1:12, ultrasonic extraction (frequency 40 kHz) is carried out at 60 ℃ for 1 h, and after centrifugation to remove the residue, it is concentrated to 28% solid, and then spray dried (temperature 150 ℃, pressure 0.5 MPa) to obtain brownish-brown extraction powder.

[0104] S5, Preparation of plant extract:

[0105] Cranberry was extracted by 40% ethanol at 50 ℃ for 45 min, followed by reverse osmosis concentration and freeze-drying at -35 ℃ to obtain the extraction powder. Pineapple was enzymatically hydrolyzed at 45 ℃ for 30 min, and then spray-dried to obtain a fine powder with a particle size of less than 100 μm. The two were mixed in a mass ratio of 2:1 and sieved for use.

[0106] S6, three-layer structure pressing process:

[0107] The group retains the three-layer structure form:

[0108] The inner layer is a blend of Lactobacillus crispatus + prebiotic gel particles and excipients, with a pressing pressure of 2 MPa;

[0109] The middle layer is a medicinal fungus extract, with a pressing pressure of 1.5 MPa;

[0110] The outer layer is a plant extract powder, with a pressing pressure of 1.2 MPa.

[0111] The thickness of each layer is controlled to be 0.8 mm, and the total thickness of the tablet is about 2.4 mm. After cold air drying at 10 ℃ for 12 h, the powder is crushed, and the particle size of the obtained powder is not more than 300 μm.

[0112] S7, microcapsule coating and drying treatment:

[0113] The composite powder is mixed with a 10% gelatin solution and spray-dried, and the outer layer is coated with hydroxypropyl methyl cellulose (HPMC) to form a primary encapsulated coating.

[0114] Comparative Example Two (C2): No microencapsulation protection treatment:

[0115] This group is used to verify the possible effects of not using microencapsulation technology on the survival rate and stability of probiotics.

[0116] S1, pre-culture and freeze-drying of complex lactic acid bacteria:

[0117] Three strains of bacteria, Lactobacillus crispatus, Lactobacillus jensenii, and Lactobacillus reuteri, were mixed in a mass ratio of 1.5:1:1. Each strain was inoculated into liquid medium (glucose 10 g / L, yeast extract 10 g / L, casein hydrolysate 10 g / L) and cultured at 37 ℃ for 12 h. The bacterial cells were collected by centrifugation, and freeze-dried for 24 h to obtain a complex bacterial powder with a particle size of 60 μm. The viable bacterial counts were: and .

[0118] S2, preparation of prebiotic complex and formation of gel particles:

[0119] Galacto-oligosaccharide, fructo-oligosaccharide, inulin were dissolved and mixed in the ratio of 60:15:15, cooled to 25℃, and mixed with the bacterial powder at a ratio of 1:2. The gel particle size was about 0.6 mm, cooled for 15 min and vacuum dried.

[0120] S3, without microencapsulation treatment:

[0121] Directly use the above naked probiotics and prebiotic particles, without chitosan-sodium alginate wrapping, and without electrostatic spraying treatment, omitting the double-layer embedding protection mechanism.

[0122] S4-S5: Fungal extract and plant extract powder preparation parameters consistent with experimental group T2.

[0123] S6, three-layer structure compression and drying:

[0124] The inner layer is a bacterial powder and prebiotic gel particle, with a compression pressure of 2 MPa;

[0125] The middle layer is a medicinal fungal extract, with a compression pressure of 1.5 MPa;

[0126] The outer layer is a plant extract powder, with a compression pressure of 1.2 MPa.

[0127] The thickness of each layer is controlled at 0.5 mm, and the total thickness is 1.5 mm. The obtained tablets are dried by cold air for 12 h and then crushed.

[0128] S7, microcapsule coating:

[0129] Still using gelatin spray drying to form an outer film, and applying a thin layer of HPMC coating, but the internal probiotics do not have a core protection structure.

[0130] Comparative Example Three (C3): No three-layer structure is constructed, only physical mixing and tabletting

[0131] This group is used to verify the impact on product structure integrity and functional stability if the three-layer structure is cancelled and only physical mixing and tabletting is performed.

[0132] S1, complex lactic acid bacteria pretreatment:

[0133] The same strains (L. crispatus, L. jensenii, L. reuteri) as T3 were used, with a mass ratio of 3:2:2. After cultivation, they were freeze-dried, and the viable bacterial count was: , and the powder particle size was 40 μm.

[0134] S2, prebiotic compound preparation:

[0135] GOS, FOS, inulin were mixed in the ratio of 70:25:25 and formed into gel particles, which were mixed with the fungi powder in the ratio of 1:3, with a particle size of 1.0 mm.

[0136] S3, using complete microcapsules:

[0137] After mixing the chitosan and sodium alginate solution, a double-layer embedding microcapsule was formed by electrostatic spraying (8 kV, 0.3 mm nozzle), with a particle size controlled at 250 μm.

[0138] S4-S5: The extraction process of medicinal fungi and plant extracts is consistent with T3, with higher purity.

[0139] S6, cancel the three-layer compression structure, and only mix and compress:

[0140] The lactic acid bacteria microcapsule powder, prebiotic gel particles, fungal extract, and plant extract were mixed in the mass ratio of 4:3:2:1, and directly compressed in one step, with a pressure of 1.5 MPa and a thickness of about 2.5 mm. There was no functional layer, and the ingredients were not evenly distributed.

[0141] S7, microcapsule coating and drying:

[0142] The whole powder was coated with 10% gelatin and spray-dried to form a microcapsule powder, and then coated with a layer of hydroxypropyl methylcellulose (HPMC) to form a complete coating.

[0143] Comparative Example Four (C4): Only two lactic acid bacterial strains (Lactobacillus crispatus and Lactobacillus jensenii) were used

[0144] This comparative example aims to verify the influence of the absence of Lactobacillus reuteri in the composition of the complex strain on the structural stability and functional synergy of the finished conditioning composition.

[0145] S1, pre-culture and freeze-drying of complex lactic acid bacteria:

[0146] Lactobacillus crispatus and Lactobacillus jensenii were selected as the two strains, and L. reuteri was removed from the original plan. The two strains were inoculated in the complex culture medium, and the culture medium formula was: glucose 10 g / L, yeast extract 10 g / L, casein hydrolysate 10 g / L, pH adjusted to 6.5, sterilized at 121 ℃ for 20 min. The strains were cultured at 37 ℃ for 12 h, then centrifuged at 8000 rpm for 15 min to harvest the bacteria. After pre-freezing at −40 ℃, vacuum freeze-drying was performed for 24 h to obtain dry bacterial powder, with a particle size controlled within 50 μm.

[0147] The viable count after lyophilization is controlled as follows:

[0148] .

[0149] Mix the two strains in a mass ratio of 2:1 to form a double-strain composite lactic acid bacteria powder for standby.

[0150] S2, preparation of prebiotic complex and formation of gel particles:

[0151] Dissolve oligogalactose, oligofructose, and inulin in hot water at 55°C (solution concentration 10%) at a mass ratio of 65:20:15, respectively, add them in turn and stir for 20 min (rate 200 rpm), and then mix thoroughly and cool naturally to 25°C. Mix the cooled solution with the double-strain powder obtained in S1 at a mass ratio of 1:2.5, and granulate the obtained gel by dropwise addition, controlling the particle size at about 0.8 mm. After 20 min of cooling and solidification at 10°C, collect the vacuum-dried product.

[0152] S3, microencapsulation treatment (complete execution):

[0153] Embed the prebiotic gel particles obtained in S2 in a double-layer microcapsule using a 1.8% chitosan and 1.2% sodium alginate double-liquid system. Mix the system by stirring at 200 rpm for 20 min, and then granulate by electrostatic spraying system (nozzle aperture 0.3 mm, voltage 8 kV). The sprayed droplets fall into the solution, solidify for 10 min, and form double-layer embedded microcapsules with a particle size of about 250 μm.

[0154] S4, preparation of medicinal fungus extract:

[0155] Mix ganoderma lucidum, hericium erinaceus, poria cocos, cordyceps sinensis, and white birch fungus at a mass ratio of 25:20:15:10:5, and pulverize to 90 μm. Perform water extraction at a solid-liquid ratio of 1:12, a temperature of 60°C, an ultrasonic frequency of 40 kHz, and an extraction time of 1 h. Centrifuge the extraction liquid at 8000 rpm for 15 min, concentrate to 28% solids, and then spray dry (150°C, 0.5 MPa) to form a powder.

[0156] S5, preparation of plant extract:

[0157] Extract cranberry using 40% ethanol solvent at 50°C for 45 min, and then perform reverse osmosis and -35°C freeze-drying to obtain a powder. Enzymatically hydrolyze pineapple at 45°C for 30 min, and then spray dry the obtained material to obtain a powder with a particle size of less than 100 μm. Mix the two powders at a mass ratio of 2:1 and sieve to the same particle size.

[0158] S6, three-layer composite structure compression and drying:

[0159] The inner layer is a double-strain complex lactic acid bacteria microcapsule + prebiotic colloidal particles + vitamin E and sodium ascorbate mixture, and the compression pressure is 2 MPa;

[0160] The middle layer is a medicinal fungus extract powder, and the compression pressure is 1.5 MPa;

[0161] The outer layer is a plant extract composite powder, and the compression pressure is 1.2 MPa;

[0162] The thickness of each layer is controlled at 0.8 mm, and the total thickness is 2.4 mm. The segmented compression molding equipment is used to sequentially compress into a tablet-shaped structure. The obtained three-layer solid block is dried by 10 ℃ cold air for 12 h, and then crushed into a fine powder below 300 μm.

[0163] S7, microcapsule coating treatment:

[0164] The obtained powder is blended with a 10% edible gelatin solution, and spray dried to form a preliminary coating structure. Further, hydroxypropyl methyl cellulose (HPMC) is sprayed to form a dense surface coating, and the drying temperature is 45 ℃, to form a microcapsule structure.

[0165] Animal comparison experiment design scheme (for verifying the vaginal health conditioning effect):

[0166] Experimental purpose: The experiment aims to establish a female mouse vaginal flora imbalance model, evaluate the biological effects of the "female health conditioning composition based on specific Lactobacillus flora and medicinal fungus extract" in improving vaginal microecology, reducing local inflammation level, repairing vaginal mucosal barrier, etc., and verify its application value in the field of female reproductive tract health.

[0167] Experimental materials and methods:

[0168] 1. Animal model:

[0169] Experimental animals: SPF grade female ICR mice, 8 weeks old, weighing 22 ± 2 g;

[0170] Number of animals: a total of 90;

[0171] Raising conditions: clean environment, temperature 22-25 ℃, relative humidity 55%-65%, light / dark cycle 12 h / 12 h, free feeding and drinking;

[0172] Modeling method: The antibiotic combined with glucocorticoid intervention method is used to construct a vaginal flora imbalance model, simulating the human vaginal microecological disorder state.

[0173] The animals are randomly divided into the following 9 groups (10 in each group). The specific grouping is shown in Table 1:

[0174] Table 1 Animal grouping

[0175] Group Treatment Name Treatment Regimen Overview N Blank Group No modeling, no drug M Model Blank Group Modeling, no intervention after T1 Example 1 Group Drug composition T1 T2 Example 2 Group Drug composition T2 T3 Example 3 Group Drug composition T3 C1 Comparative Example 1 Group Missing multi-strain co-formulation C2 Comparative Example 2 Group Un-embedded treatment C3 Comparative Example 3 Group Non-three-layer compressed structure C4 Comparative Example 4 Group Remove Lactobacillus reuteri

[0176] Dosing method and dosage:

[0177] Dosing cycle: continuous intervention for 14 days;

[0178] Dosing method: gavage, once a day;

[0179] Dosing dose: human equivalent dose equivalent dose according to body weight, 0.15 mL / 10 g of body weight;

[0180] The control group N and the model group M were given the same amount of distilled water.

[0181] Detection items and evaluation methods

[0182] 1. Quantitative analysis of vaginal flora:

[0183] Smear staining observation: collect vaginal secretion samples, perform gram staining and microscopic observation;

[0184] Colony count: count the number of Lactobacillus and conditional pathogenic bacteria (including Escherichia coli and Candida albicans) unit colony forming units (CFU) using selective medium;

[0185] Flora ratio analysis: calculate the ratio of Lactobacillus to pathogenic bacteria to evaluate the microecological balance.

[0186] 2. Histological evaluation of vaginal epithelium:

[0187] HE staining: observe the epithelial thickness, cell arrangement, and inflammatory cell infiltration by taking paraffin sections of vaginal tissue;

[0188] PAS staining: evaluate mucus secretion and goblet cell distribution.

[0189] 3. Determination of local inflammatory factor levels in vagina:

[0190] ELISA detection: collect vaginal lavage fluid or local tissue homogenate, and detect the concentrations of TNF-α, IL-1β, and IL-6 inflammatory factors.

[0191] The experimental results are shown in Tables 2 to 4:

[0192] Table 2 Experimental results of quantitative analysis of vaginal flora

[0193] Group Lactobacillus quantity (Log CFU / mL) Pathogenic bacteria quantity (Log CFU / mL) Lactobacillus / pathogenic bacteria ratio Blank group N 7.65±0.23 2.31±0.19 3.31±0.28 Model group M 4.13±0.32 5.76±0.27 0.72±0.14 Example T1 7.12±0.26 2.68±0.22 2.66±0.25 Example T2 6.75±0.29 3.01±0.24 2.24±0.20 Example T3 7.38±0.21 2.45±0.20 3.01±0.30 Comparative example C1 5.52±0.30 4.10±0.28 1.15±0.18 Comparative example C2 5.31±0.26 4.25±0.31 1.00±0.16 Comparative example C3 5.78±0.22 3.89±0.24 1.49±0.14 Comparative example C4 5.40±0.27 4.01±0.26 1.21±0.15

[0194] Table 3 Experimental results of histological evaluation of vaginal epithelium

[0195] Group Epithelial integrity score (0-5 points) Inflammatory cell infiltration score (0-5 points) PAS staining positive area (%) Blank group N 4.8±0.2 0.4±0.1 87.3±3.1 Model group M 2.1±0.3 3.9±0.2 38.5±2.6 Example T1 4.6±0.2 0.9±0.2 82.7±2.8 Example T2 4.3±0.3 1.2±0.2 78.6±3.4 Example T3 4.7±0.1 0.7±0.1 85.9±2.3 Comparative example C1 3.1±0.3 2.6±0.3 61.2±2.7 Comparative example C2 3.0±0.2 2.8±0.2 58.5±3.1 Comparative example C3 3.4±0.2 2.3±0.3 64.9±2.5 Comparative example C4 3.2±0.2 2.5±0.3 60.7±2.9

[0196] Table 4 Results of vaginal local inflammation factor level determination experiment

[0197] Group TNF-α IL-1β IL-6 Blank group N 21.3±1.5 18.6±1.2 35.9±2.1 Model group M 68.5±3.2 55.7±2.8 92.1±3.5 Example T1 26.1±1.6 21.7±1.4 41.2±2.2 Example T2 29.5±1.8 24.3±1.7 47.3±2.5 Example T3 24.7±1.3 20.8±1.5 39.6±1.9 Comparative example C1 42.3±2.5 37.9±2.0 65.4±3.0 Comparative example C2 44.8±2.7 39.5±2.3 68.1±3.1 Comparative example C3 38.7±2.2 34.1±2.1 59.6±2.8 Comparative example C4 41.5±2.4 36.7±2.2 62.3±2.9

[0198] Data analysis

[0199] To verify the comprehensive conditioning effect of the "female health conditioning composition based on specific Lactobacillus flora and medicinal fungus extract" of the present application in regulating vaginal microecology, repairing epithelial mucosa, inhibiting inflammatory response and improving secretion indicators, a comparative experiment was carried out using a female mouse vaginal flora imbalance model, mainly detecting flora distribution, histological changes, inflammation factor level and secretion characteristics, and the experimental results are shown as follows:

[0200] 1. Lactobacillus recovery and flora balance improvement effect:

[0201] As shown in Table 2 and Figure 2 , the number of Lactobacillus in the model group M mice was significantly reduced (4.13 log CFU / mL), while the number of pathogenic bacteria was significantly increased (5.76 log CFU / mL), and the ratio of Lactobacillus / pathogenic bacteria was reduced to 0.72, showing a state of flora imbalance. In contrast, Examples T1-T3 can significantly increase the number of Lactobacillus and inhibit the colonization of pathogenic bacteria, among which T3 is the most optimal, with the number of Lactobacillus reaching 7.38 log CFU / mL and the number of pathogenic bacteria being only 2.45 log CFU / mL, with a ratio of 3.01, close to the blank group (3.31). Although Comparative Examples C1-C4 have some improvement, they are far lower than the implementation group, especially C1 (lack of complex strains) and C2 (without embedding), which have poor flora recovery, indicating that the complex multi-strain configuration and microcapsule protection are key technical elements for maintaining flora balance.

[0202] 2. Epithelial tissue integrity and mucosal structure repair effect:

[0203] As shown in Table 3, Figure 1 , Figure 3 Under HE and PAS staining, the model group showed obvious thinning of the epithelium, disorderly arrangement of cells and a large number of inflammatory cell infiltration, and the PAS positive staining area was also significantly reduced, indicating impaired barrier function. Examples T1-T3 significantly improved the epithelial integrity score and inhibited inflammatory infiltration, with a score of 4.7 in T3 group and only 0.7 in inflammatory infiltration, and a PAS positive area of 85.9%, indicating that it has a good repair and promotion effect on mucosal structure. In the comparative examples, C1 and C2 have lower scores and still have obvious inflammatory cell infiltration, indicating that the use of single bacteria or lack of embedding treatment significantly weakens the mucosal repair effect.

[0204] 3. Inflammatory factor level regulation ability:

[0205] As shown in Table 4 and Figure 4 Table 5, the local TNF-a, IL-1β and IL-6 of the model group were significantly increased, indicating that the inflammatory state was activated. Examples T1-T3 can significantly reduce the level of the above inflammatory factors, and the decrease of T3 group is the largest, and TNF-a is reduced to 24.7 pg / mL, close to the normal group (21.3 pg / mL). The inflammation suppression ability of Comparative Examples C1-C4 is weaker than that of the implementation group as a whole, especially C2 and C4, and the decrease is not significant, indicating that the double-layer embedding and complete flora configuration have a synergistic enhancement effect on inflammation regulation.

[0206] Comprehensive conclusion:

[0207] The above experimental results show that the female health conditioning composition of the present application is superior to the comparative example group in many physiological indicators, and has the following comprehensive advantages:

[0208] The configuration of the three strains of complex lactic acid bacteria has the characteristics of synergistic colonization and mutual metabolic promotion, which can significantly improve the colonization ability of probiotics;

[0209] The double-layer microcapsule embedding structure effectively improves the stability of the bacterial body and the survival rate through the gastrointestinal tract;

[0210] The three-layer structure compression ensures the functional distribution and slow-release release of the ingredients, which is helpful for the synergistic effect of medicinal fungi and plant extracts;

[0211] The synergistic effect of the ingredients can inhibit the growth of pathogenic bacteria, reduce inflammatory factors, repair the barrier function of the vagina, and improve the secretion index.

[0212] Therefore, the composition provided by the present application has significant technical effects in promoting vaginal flora balance, improving mucosal health, regulating local immunity, etc., and has good clinical application prospect.

[0213] The above shows and describes the basic principles, main features and advantages of the present application. Those skilled in the art should understand that the present application is not limited to the above examples, and the above examples and descriptions in the specification are only to illustrate the principles of the present application. Without departing from the spirit and scope of the present application, various changes and improvements can be made to the present application, and these changes and improvements all fall within the scope of the claimed present application. The scope of protection of the present application is defined by the appended claims and their equivalents.

Claims

1. A women's health conditioning composition based on a specific Lactobacillus flora and medicinal fungal extracts, characterized in that, Includes the following ingredients: Compound lactic acid bacteria: composed of freeze-dried Lactobacillus curvaturei, Lactobacillus janniae, and Lactobacillus reuteri mixed in a mass ratio of 1.5-3:1-2:1-2; Lactobacillus curvature, after freeze-drying, the viable count was controlled at 1×10⁻⁶. 9 ~5×10 10 CFU / g; Lactobacillus japonicus, after freeze-drying, the viable count was controlled at 5 × 10⁻⁶. 8 ~2×10 10 CFU / g; Lactobacillus reuteri, after freeze-drying, the viable count was controlled at 5×10⁻⁶. 8 ~2×10 10 CFU / g; Among them, *Lactobacillus curli* was purchased from the China General Microbiological Culture Collection Center (CGMCC), accession number CGMCC No. 25113; *Lactobacillus janniae* was purchased from the China General Microbiological Culture Collection Center (CGMCC), accession number CGMCC No. 9439; and *Lactobacillus reuteri* was purchased from the China General Microbiological Culture Collection Center (CGMCC), accession number CGMCC No. 19746. Prebiotics: A complex composed of three components: galactooligosaccharides, fructooligosaccharides, and inulin, wherein the mass fraction ratio of the three components is 60-70:15-25:15-25; the prebiotics are dissolved at 60 °C and cooled to 25 °C, then mixed with compound lactic acid bacteria at a mass ratio of 1:2-3 to form gel particles, which are then cured at 10 °C for 15 min and dried. Medicinal fungal extract: composed of Ganoderma lucidum, Hericium erinaceus, Poria cocos, Cordyceps sinensis and Chaga mushroom in a dry powder mass ratio of 20-30:15-25:10-20:5-15:5-10; Plant extracts: including cranberry extract powder and pineapple powder in a mass ratio of 1 to 2:1; Supporting ingredients: including natural vitamin E and sodium ascorbate; The composition is freeze-dried to form a three-layer solid powder, with the inner layer being a mixture of compound lactic acid bacteria, prebiotics and auxiliary ingredients, the middle layer being a medicinal fungal extract layer, and the outer layer being a plant extract layer.

2. The female health conditioning composition based on a specific Lactobacillus flora and medicinal fungal extracts according to claim 1, characterized in that, The three strains of Lactobacillus curvature, Lactobacillus janniae, and Lactobacillus reuteri were pre-cultured separately before being mixed. The culture medium consisted of glucose, yeast extract and casein hydrolysate. The culture temperature was controlled at 37 °C for 12 h. After culture, the bacterial powder was obtained by centrifugation and freeze-drying. The particle size of the bacterial powder was controlled in the range of 40 μm to 60 μm.

3. The female health conditioning composition based on a specific Lactobacillus flora and medicinal fungal extracts according to claim 1, characterized in that, The compound lactic acid bacteria were encapsulated in a chitosan-sodium alginate double layer and then formed into a double-layer microcapsule structure by electrostatic spraying. The microcapsule particle size was controlled between 100 and 300 μm. The chitosan solution concentration was 1.8%, the sodium alginate solution concentration was 1.2%, the mixing and stirring rate was 200 rpm, the stirring time was 20 min, the spray system used a 0.3 mm nozzle and applied a voltage of 6 kV to 8 kV to generate microparticles, the curing solution was 0.2 mol / L CaCl2, the curing time was 10 min, and the resulting microcapsule particle size distribution was concentrated between 150 μm and 250 μm.

4. The female health conditioning composition based on a specific Lactobacillus flora and medicinal fungal extracts according to claim 1, characterized in that, The three prebiotic components were dissolved at 55 °C, and the galactooligosaccharides, fructooligosaccharides and inulin were added in sequence and mixed together. The stirring rate is 200 rpm. After mixing, the mixture is naturally cooled to 25 ℃ and then mixed with compound lactic acid bacteria powder to form gel particles with a particle size of 0.5 mm to 1.0 mm. The gel particles are then collected after being cured at 10 ℃ for 20 min and vacuum dried.

5. The female health conditioning composition based on a specific Lactobacillus flora and medicinal fungal extracts according to claim 1, characterized in that, The prebiotic complex was pretreated by spray drying before mixing. The inlet air temperature was 120 ℃ and the outlet air temperature was 85 ℃. The moisture content of the powder obtained after drying was no more than 3%. When mixing with the bacterial powder, a low-speed stirring method was used for 15 min to ensure uniform mixing of the powder.

6. The female health conditioning composition based on a specific Lactobacillus flora and medicinal fungal extracts according to claim 1, characterized in that, The preparation process of medicinal fungal extracts includes four steps: pulverization, extraction, concentration, and drying. The pulverization step pulverizes the raw material to a particle size of 80 μm to 100 μm; the extraction step is carried out with a solid-liquid ratio of 1:12, an ultrasonic frequency of 40 kHz, an extraction temperature of 60 ℃, and an extraction time of 1 h. The centrifugation speed was 8000 rpm for 15 min, the solid content after concentration was controlled at 25% to 30%, the spray drying temperature was 150 ℃, and the atomization pressure was 0.5 MPa.

7. The female health conditioning composition based on a specific Lactobacillus flora and medicinal fungal extracts according to claim 1, characterized in that, Cranberry extract powder was prepared by ethanol extraction with a volume fraction of 40% at 50°C for 45 min. After reverse osmosis concentration and freeze-drying at −35°C, the powder was obtained. Pineapple powder was prepared by mechanical enzymatic hydrolysis at 45°C for 30 min. After low-temperature spray drying, the powder particle size was less than 100 μm. The two were mixed at a mass ratio of 1:1 to 2:1 and then sieved uniformly.

8. The female health conditioning composition based on a specific Lactobacillus flora and medicinal fungal extracts according to claim 1, characterized in that, The three-layer composite solid powder was prepared by a segmented pressing process. The inner layer of probiotics and prebiotics was pressed at a pressure of 2 MPa, the middle layer of fungi at a pressure of 1.5 MPa, and the outer layer of plants at a pressure of 1.2 MPa. The thickness of each layer was controlled between 0.5 mm and 1.0 mm. The resulting solid block was dried by cold air and then pulverized into powder with a particle size of no more than 300 μm.

9. The female health conditioning composition based on a specific Lactobacillus flora and medicinal fungal extracts according to claim 1, characterized in that, Natural vitamin E and sodium ascorbate were dissolved in an ethanol solution and then dispersed in the medicinal fungal extract layer. The ethanol volume fraction was 50%, the mixing speed was 120 rpm, and the stirring time was 20 min. The resulting product was packaged with a three-layer composite aluminum foil film. Before packaging, it was purged with nitrogen gas of 99.9% purity at a pressure of 0.05 MPa. The sealing temperature was 120 ℃ and the sealing width was 10 mm. After packaging, the storage temperature was controlled at 25 ℃.

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