High-activity lactobacillus rhamnosus probiotic microcapsule and application thereof

Abstract

The invention relates to the technical field of biological medicines, and discloses a high-activity lactobacillus rhamnosus probiotic microcapsule, which comprises the following components in parts by weight: lactobacillus rhamnosus: each gram of formula contains 1010 CFU; bifidobacterium: each gram of the formula contains 109 CFU; 10% to 20% of sodium alginate; 5%-15% of chitosan; 3%-8% of hydroxypropyl methyl cellulose; 3%-7% of pectin; 5% to 10% of fructo-oligosaccharide; 5%-10% of inulin; 0.1%-0.5% of vitamin C; and 0.1%-0.3% of vitamin E. Through double-layer and three-layer embedding technologies (such as compound use of sodium alginate and chitosan and compound use of pectin and gelatin), the tolerance of the probiotics in gastric acid and bile environments is remarkably improved, the probiotics are effectively protected through a multi-layer embedding structure, the survival rate of the probiotics in the gastric environment reaches up to 90%, and the release rate in the small intestine exceeds 75%.

Description

Technical Field

[0001] The invention relates to the technical field of biomedicine, and in particular to a high-activity Lactobacillus rhamnosus probiotic microcapsule and application thereof. Background Art

[0002] As people's health awareness increases, probiotics are gaining more and more attention as an effective way to regulate intestinal flora, enhance immunity and improve digestive health. At present, probiotic preparations on the market mainly exist in the form of powders, tablets, liquids and capsules, and commonly used probiotic species include Lactobacillus, Bifidobacterium, etc. However, existing probiotic preparations have some obvious deficiencies during use, especially in terms of survival rate, stability and release control in the intestine, which limit the actual health effects of probiotics.

[0003] First of all, most existing probiotic preparations lack effective protection measures, resulting in a low survival rate of probiotics in the harsh environment of the digestive tract, such as gastric acid and bile. The corrosive effects of gastric acid and bile salts kill a large number of probiotics before they reach the small intestine, greatly reducing the number of active bacteria that actually reach the intestine and play a role. Therefore, the bioavailability of probiotics is low and they cannot fully exert their due health functions. In order to solve this problem, some preparations use simple encapsulation techniques, such as single-layer coating, but these methods cannot provide sufficient protection in the complex gastrointestinal environment, and probiotics are still prone to inactivation.

[0004] Secondly, many probiotic preparations are not stable enough during storage and are easily inactivated by temperature, humidity and oxidation factors. Especially under normal storage conditions, the activity of probiotics is greatly reduced, resulting in the number of viable bacteria in the product failing to meet the product label requirements during the validity period, which seriously affects the quality and efficacy of probiotic products. Although attempts have been made to use antioxidants and prebiotics to improve the stability of probiotics in the prior art, they often only work for a short period of time and are difficult to maintain a high level of activity during long-term storage.

[0005] In addition, existing probiotic preparations generally lack effective control over the release of probiotics in the digestive tract, and cannot achieve targeted release and sustained effects of probiotics. Probiotics are usually released disorderly throughout the digestive tract, which not only reduces their effectiveness in specific intestinal segments, but may also lose some of their activity due to premature release. Traditional probiotic preparations fail to achieve precise controlled release, resulting in limited effects in specific intestinal areas, affecting their application in regulating intestinal health.

[0006] To this end, those skilled in the art provide a highly active Lactobacillus rhamnosus probiotic microcapsule and application thereof to solve the problems raised in the above background technology. Summary of the invention

[0007] In view of the shortcomings of the prior art, the present invention provides a highly active Lactobacillus rhamnosus probiotic microcapsule and application thereof, which solves the problems of the prior art in terms of survival rate, stability and release control in the intestine.

[0008] To achieve the above objectives, the present invention is implemented by the following technical scheme: A highly active Lactobacillus rhamnosus probiotic microcapsule, comprising the following components in parts by weight:

[0009] Lactobacillus rhamnosus: 10^10 CFU per gram of formula;

[0010] Bifidobacterium: Each gram of formula contains 10^9 CFU;

[0011] Sodium alginate: 10%-20%;

[0012] Chitosan: 5%-15%;

[0013] Hydroxypropyl methylcellulose: 3% to 8%;

[0014] Pectin: 3% to 7%;

[0015] Fructooligosaccharides: 5% to 10%;

[0016] Inulin: 5% to 10%;

[0017] Vitamin C: 0.1% to 0.5%;

[0018] Vitamin E: 0.1%~0.3%.

[0019] Preferably, the preparation is prepared by double-layer or multi-layer embedding technology, the first layer uses sodium alginate and calcium ions to cross-link to form a gel layer, the second layer uses chitosan to form an acid-resistant coating layer, and the third layer uses hydroxypropyl methylcellulose to form an anti-oxidation protective film.

[0020] A method for preparing a highly active Lactobacillus rhamnosus probiotic microcapsule, which is used for a highly active Lactobacillus rhamnosus probiotic microcapsule, comprises the following steps:

[0021] S1. Probiotic culture and pretreatment: culture Lactobacillus rhamnosus and Bifidobacterium to the required bacterial concentration, collect the bacteria and dry them to prepare freeze-dried powder for standby use;

[0022] S2. Preparation of the inner layer of the microcapsule: mixing the dried probiotic powder with the sodium alginate solution to form a bacterial-sodium alginate suspension; spraying the suspension into a cross-linking bath to form sodium alginate gel microspheres;

[0023] S3. Preparation of chitosan coating layer: After washing the gel microspheres, they were immersed in a chitosan solution to form a coating layer;

[0024] S4. Formation of an outer protective film: Immersing the coated microcapsules in a hydroxypropyl methylcellulose solution to form an outer protective film;

[0025] S5. Drying and forming: The microcapsules are dried by a combination of spray drying and freeze drying to form stable microcapsule powders;

[0026] S6. Adding prebiotics and antioxidants: adding prebiotics and antioxidants to the dry microcapsule powder and mixing them evenly;

[0027] S7. Controlled-release coating and final formulation formation: coating the controlled-release agent on the outer surface of the microcapsules, performing final mixing, making a composite formulation in the form of granules or tablets, and sealing and storing.

[0028] Preferably, the probiotics in step S1 are dried by freeze drying at a temperature of -50°C to -20°C for a drying time of 12h to 48h.

[0029] Preferably, the concentration of the sodium alginate solution in step S2 is 1% to 3%, and a 0.2M calcium chloride solution is used in the cross-linking bath for 30 minutes.

[0030] Preferably, in step S3, the concentration of the chitosan solution is 0.5% to 2%, the coating time is 10 min to 30 min, and the chitosan is dissolved in 1% acetic acid solution.

[0031] Preferably, the concentration of the hydroxypropyl methylcellulose solution in step S4 is 1% to 2%, and the soaking time is 10 min to 15 min.

[0032] Preferably, in step S5, the air inlet temperature of the spray drying is 70°C to 90°C, the air outlet temperature is 50°C to 60°C, and the freeze drying temperature is -40°C to -20°C.

[0033] The invention discloses an application of a probiotic microcapsule composite preparation in a health product for regulating the balance of intestinal flora and enhancing immune function, which is used for a highly active Lactobacillus rhamnosus probiotic microcapsule. The health product is used for improving intestinal health, enhancing immune function, and preventing and alleviating intestinal discomfort symptoms such as diarrhea, constipation, and abdominal distension caused by intestinal flora imbalance.

[0034] The present invention provides a highly active Lactobacillus rhamnosus probiotic microcapsule and its application, which has the following beneficial effects:

[0035] 1. The present invention significantly improves the tolerance of probiotics in gastric acid and bile environments through double-layer and triple-layer encapsulation technology (such as the composite use of sodium alginate and chitosan, pectin and gelatin), ensuring its high survival rate in the digestive tract. The multi-layer encapsulation structure effectively protects the probiotics, making their survival rate in the gastric environment as high as 90%, and achieving a release rate of more than 75% in the small intestine. This design ensures the colonization and function of probiotics in the intestine, and significantly improves the bioavailability and health benefits of probiotics compared to traditional probiotic preparations.

[0036] 2. The present invention improves the antioxidant capacity and intestinal colonization effect of probiotic microcapsules by introducing composite prebiotics and antioxidants. The added prebiotic ingredients such as oligofructose and inulin provide necessary nutritional support for probiotics and enhance their proliferation ability in the intestine, while the combination of vitamin C and vitamin E significantly improves the antioxidant protection ability. Experimental data show that the survival rate of probiotics in microcapsules with added antioxidants under oxidative stress conditions is 85%, which is significantly higher than that of traditional formulas. These improvements effectively extend the storage period and stability of probiotics, and are suitable for the market demand with high requirements for the quality of probiotics. .

[0037] 3. The present invention significantly improves the release behavior of probiotics by adding a controlled release agent to the outer protective film, reducing the release in the stomach and gradually releasing it in the small intestine and colon, thereby enhancing the targeting effect and sustained action ability. Animal experimental results show that the release rate of the controlled-release microcapsules of the present invention in the small intestine and colon is as high as 82%, which is much better than the unbalanced release pattern of traditional probiotic preparations. Through this optimized controlled-release design, probiotics can fully play their role in the target area, improving bioavailability and health effects. BRIEF DESCRIPTION OF THE DRAWINGS

[0038] Figure 1 It is a schematic diagram of the process of the present invention. DETAILED DESCRIPTION

[0039] The technical solution of the present invention will be described clearly and completely below in conjunction with the accompanying drawings of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0040] Please refer to the attached Figure 1 ;

[0041] Example 1: Preparation of double-layer embedded probiotic microcapsules

[0042] step:

[0043] Probiotic culture and pretreatment:

[0044] Lactobacillus rhamnosus was cultured in MRS medium to 10^10 CFU / mL. The bacteria were collected by centrifugation (4000 rpm, 10 minutes) and freeze-dried (-40°C, 24 hours) to prepare lyophilized powder.

[0045] Preparation of sodium alginate gel microspheres:

[0046] The probiotic freeze-dried powder was added to a 2% sodium alginate solution and stirred evenly. The suspension was sprayed into a 0.2M calcium chloride solution using a sprayer to form gel microspheres with a diameter of 100-200 microns, and the reaction lasted for 30 minutes.

[0047] Chitosan coating formation:

[0048] The gel microspheres were taken out, washed with distilled water, immersed in a 1% chitosan solution (dissolved in 1% acetic acid), and reacted for 20 minutes to form a coating layer.

[0049] Preparation of outer protective film:

[0050] The coated microspheres were immersed in a 1.5% hydroxypropyl methylcellulose (HPMC) solution for 15 minutes to form an outer protective film.

[0051] dry:

[0052] The dry microcapsule powder was obtained by spray drying (inlet air temperature 80°C, outlet air temperature 60°C) and freeze drying (-40°C, 12 hours).

[0053] Summary: Through the double-layer encapsulation design, the synergistic effect of sodium alginate and chitosan is utilized to form acid-resistant and enteric microcapsules, ensuring the high survival rate of Lactobacillus rhamnosus in the gastric acid environment. Through further coating with the outer layer of hydroxypropyl methylcellulose (HPMC) protective film, the intrusion of oxygen and moisture is prevented, and the mechanical strength and storage stability of the microcapsules are improved. The drying step combining spray drying and freeze drying effectively maintains the activity of probiotics. The experimental results show that the survival rate of the microcapsules in the simulated gastrointestinal environment is as high as 85%, and the release efficiency in the small intestine can reach 70%, which significantly improves the bioavailability of probiotics and is suitable for functional foods for intestinal health.

[0054] Example 2: Preparation of three-layer embedded probiotic microcapsules

[0055] step:

[0056] Probiotic culture and pretreatment:

[0057] Lactobacillus rhamnosus was cultured in MRS medium to 10^10 CFU / mL. The bacteria were collected by centrifugation (5000 rpm, 8 minutes) and freeze-dried (-50°C, 30 hours) to prepare lyophilized powder.

[0058] Preparation of sodium alginate and pectin composite gel microspheres:

[0059] The freeze-dried probiotic powder was added to a composite solution of 2% sodium alginate and 1% pectin, and then sprayed into a 0.1M calcium chloride solution to form gel microspheres.

[0060] Chitosan and gelatin composite coating layer formation:

[0061] After washing, the gel microspheres were immersed in a mixed solution of 1% chitosan and 0.5% gelatin and reacted for 20 minutes to form a second coating layer.

[0062] Preparation of outer protective film:

[0063] The coated microspheres were immersed in a mixed solution of 1.5% hydroxypropyl methylcellulose (HPMC) and 0.5% gelatin to form a three-layer protective film.

[0064] dry:

[0065] The three-layer embedded microcapsule powder was obtained by combining spray drying (inlet temperature 85°C, outlet temperature 65°C) and freeze drying (-45°C, 15 hours).

[0066] Summary: A three-layer encapsulation design is adopted. In addition to sodium alginate and chitosan, pectin and gelatin composite materials are added to form a multi-layer protection structure. This multi-layer design greatly enhances the protection ability and controlled release performance of the microcapsules in the complex digestive tract environment. Through the optimization of a series of coating layers, probiotics can maintain a 90% survival rate in gastric acid, while being gradually released in the small intestine, improving the colonization effect of active bacteria in specific parts. Compared with double-layer encapsulation, triple-layer encapsulation increases the survival rate and storage period of probiotics, showing a more excellent protection effect and a longer shelf life, which is suitable for the development of high-demand drugs and health products.

[0067] Example 3: Preparation of probiotic microcapsules assisted by composite prebiotics

[0068] step:

[0069] Probiotic culture and pretreatment:

[0070] Lactobacillus rhamnosus was cultured in MRS medium to 10^10 CFU / mL. The bacteria were collected by centrifugation (3000 rpm, 12 minutes) and freeze-dried (-30°C, 36 hours) to prepare lyophilized powder.

[0071] Preparation of sodium alginate gel microspheres:

[0072] The probiotic freeze-dried powder was added to a mixed solution of 1.5% sodium alginate and 2% oligofructose, and then sprayed into a 0.2M calcium chloride solution to form gel microspheres.

[0073] Chitosan coating formation:

[0074] After washing, the gel microspheres were immersed in a mixed solution of 0.75% chitosan and 1.5% inulin and reacted for 25 minutes to form a coating layer.

[0075] Preparation of outer protective film:

[0076] The coated microspheres were immersed in a 2% hydroxypropyl methylcellulose (HPMC) solution to form an outer protective film.

[0077] Drying and molding:

[0078] Stable microcapsules were formed by combining spray drying (inlet temperature 75°C, outlet temperature 55°C) and freeze drying (-35°C, 18 hours).

[0079] Summary: The composite prebiotic (oligofructose and inulin) auxiliary design is introduced to provide additional nutritional support for probiotics and enhance their proliferation ability and colonization effect in the intestine. Compared with the basic double-layer encapsulation, the composite design significantly improves the activity stability of probiotics in the simulated gastrointestinal environment, especially in the small intestine release stage, reaching a release rate of 78%. The addition of prebiotics promotes the reproduction and functional expression of probiotics, further optimizes the health benefits of microcapsules, and is particularly suitable for dietary supplements and functional food markets that hope to enhance immunity by improving the balance of intestinal flora.

[0080] Example 4: Preparation of antioxidant enhanced probiotic microcapsules

[0081] step:

[0082] Probiotic culture and pretreatment:

[0083] Lactobacillus rhamnosus was cultured in MRS medium to 10^10 CFU / mL. The bacteria were collected by centrifugation (3500 rpm, 10 minutes) and freeze-dried (-35°C, 20 hours) to prepare lyophilized powder.

[0084] Preparation of sodium alginate gel microspheres:

[0085] The probiotic freeze-dried powder was added to 1.8% sodium alginate solution and sprayed into 0.15M calcium chloride solution to form gel microspheres.

[0086] Chitosan coating formation:

[0087] After washing, the gel microspheres were immersed in a 1% chitosan solution and reacted for 15 minutes.

[0088] Outer protective film and antioxidant added:

[0089] 0.2% vitamin C and 0.15% vitamin E were added to 1.5% hydroxypropyl methylcellulose (HPMC) solution to form a composite outer protective film.

[0090] Drying and molding:

[0091] Microcapsules with enhanced antioxidant function were obtained by combining spray drying (inlet air temperature 80°C, outlet air temperature 60°C) and freeze drying (-40°C, 24 hours).

[0092] Summary: By adding antioxidants (vitamin C and vitamin E) to the outer layer of the microcapsule, the antioxidant protection ability and storage stability of the probiotics are significantly enhanced. This design can not only effectively resist environmental oxidative stress, but also prolong the active storage period of the probiotics. Experimental data show that the survival rate of antioxidant-enhanced microcapsules is as high as 85% under oxidative stress conditions, which is much higher than the control group without added antioxidants. This improvement significantly reduces the oxidative inactivation of probiotics during storage and transportation, and is suitable for use in medical or high-end health care products with higher requirements, reflecting good innovation value.

[0093] Example 5: Preparation of multifunctional controlled-release probiotic microcapsules

[0094] step:

[0095] Probiotic culture and pretreatment:

[0096] Lactobacillus rhamnosus was cultured in MRS medium to 10^10 CFU / mL. The bacteria were collected by centrifugation (4000 rpm, 15 minutes) and freeze-dried (-45°C, 30 hours) to prepare lyophilized powder.

[0097] Preparation of sodium alginate gel microspheres:

[0098] The probiotic freeze-dried powder was added to a 2% sodium alginate solution and sprayed into a 0.1 M calcium chloride solution to form gel microspheres.

[0099] Chitosan coating formation:

[0100] After washing, the gel microspheres were immersed in a mixed solution of 0.8% chitosan and 1% gelatin for 30 minutes.

[0101] Outer protective film and controlled release agent addition:

[0102] 2% magnesium stearate was added to 1.2% hydroxypropyl methylcellulose (HPMC) solution to form a controlled-release protective film.

[0103] Drying and molding:

[0104] The multifunctional controlled-release microcapsules were obtained by combining spray drying (inlet temperature 90°C, outlet temperature 65°C) and freeze drying (-30°C, 20 hours).

[0105] Summary: The multifunctional controlled-release design is adopted, and the controlled-release agent magnesium stearate is added to the outer protective film to further optimize the release behavior of probiotics in the digestive tract. This design ensures that probiotics can reduce release in the gastric acid environment and gradually release in the small intestine and colon, achieving a small intestinal release rate of up to 82%. The multifunctional controlled-release microcapsules show excellent targeted release characteristics and lasting bioavailability, which helps the proliferation and action of probiotics in the target area. This design is very suitable for specific medical and functional food fields that require precise release and long-lasting effects, and meets the multi-level needs for intestinal health.

[0106] Summary: Five examples demonstrate the diverse applications of Lactobacillus rhamnosus probiotic microcapsules with different designs in improving activity, stability and functionality. The double-layer and triple-layer encapsulation designs provide basic acid resistance and enteric protection, while the composite prebiotics and antioxidant enhancement designs further enhance the intestinal proliferation effect and storage stability of probiotics. The multifunctional controlled release design improves the efficiency of probiotics in specific parts of the digestive tract by optimizing the release behavior. Comprehensive experimental results show that these different design schemes significantly improve the survival rate, bioavailability and functional performance of probiotics, showing excellent protection effects and wide application potential. Through targeted design for different application requirements, these highly active Lactobacillus rhamnosus probiotic microcapsules are not only suitable for use in the fields of food and health products, but also provide new technical directions and solutions for drug development, demonstrating the unique innovative value of the present invention in the field of probiotic preparations.

[0107] Test experiments and data

[0108] Experimental design

[0109] The following test experiments were designed to evaluate the performance of probiotic microcapsules under different embodiments:

[0110] Survival test: Testing the survival rate of probiotics in a simulated gastrointestinal environment.

[0111] Controlled release test: Testing the release behavior of probiotics under different pH conditions.

[0112] Antioxidant performance testing: Testing the antioxidant protection of probiotics under storage conditions.

[0113] Test experiment data table:

[0114]

[0115]

[0116] Comparative Experimental Design and Data

[0117] Comparative experiment 1: Effects of different numbers of encapsulation layers on the activity of probiotics

[0118] Embedding layer number Survival rate (stomach) Survival rate (small intestine) Storage stability (days) Single layer 60％ 50％ 100 Double Layer 85％ 70％ 180 Three-layer 90％ 75％ 200

[0119] Comparative experiment 2: Effects of different antioxidant additions on the stability of probiotics

[0120]

[0121] Comparative experiment 3: Effects of different controlled-release agents on the release behavior of probiotics

[0122] Embedding layer number Survival rate (stomach) Survival rate (small intestine) Storage stability (days) Single layer 60％ 50％ 100 Double Layer 85％ 70％ 180 Three-layer 90％ 75％ 200

[0123] Summarize

[0124] Through these test experiments and comparative experiments, the results show that the high-activity Lactobacillus rhamnosus probiotic microcapsules of the present invention have superior survival rate, controlled release performance and antioxidant protection effect, which are significantly better than other types of probiotic preparations. Experimental data show that by optimizing the number of embedding layers, adding antioxidants and designing a controlled release system, the stability and bioavailability of probiotics in different environments can be significantly improved, which reflects the innovation and practicality of the present invention.

[0125] Animal experiment design: To evaluate the effect of highly active Lactobacillus rhamnosus probiotic microcapsules in regulating intestinal health;

[0126] Experimental animals

[0127] Experimental subjects: Male C57BL / 6 mice

[0128] Quantity: 60 (average weight 20±2g)

[0129] Grouping: Randomly divided into three groups, 20 in each group

[0130] Control group (GroupA): given normal saline;

[0131] Traditional formula group (Group B): given traditional Lactobacillus rhamnosus probiotic preparation;

[0132] The microcapsule group of the present invention (Group C): administered with highly active Lactobacillus rhamnosus probiotic microcapsules; Experimental results

[0133] 1. Weight change (mean ± standard deviation, g):

[0134]

[0135] 2. Changes in intestinal flora (increase rate of Lactobacillus and decrease rate of Escherichia coli):

[0136]

[0137] 3. Level of inflammatory factors (pg / mL):

[0138] Group TNF-α IL-6 IL-10 Control group (A) 45.2±5.3 38.1±4.8 20.5±2.7 Traditional formula group (B) 32.8±4.1 27.5±3.6 28.4±3.1 Microcapsule group (C) of the present invention 25.1±3.7 18.9±2.9 35.7±3.4

[0139] 4. Immune organ index (organ weight / body weight, mg / g):

[0140] Group Spleen index Thymus index Control group (A) 3.5±0.3 1.2±0.2 Traditional formula group (B) 4.1±0.4 1.5±0.3 Microcapsule group (C) of the present invention 4.8±0.5 1.8±0.3

[0141] Experimental Summary

[0142] The experimental results show that the high-activity Lactobacillus rhamnosus probiotic microcapsules (GroupC) of the present invention are superior to traditional probiotic formulas (GroupB) and control groups (GroupA) in multiple indicators. Specifically, the microcapsules of the present invention show significant advantages in mouse weight gain, intestinal beneficial bacteria (Lactobacillus) proliferation and harmful bacteria (Escherichia coli) inhibition, proving its excellent efficacy in regulating the balance of intestinal flora; in terms of inflammatory factor levels, the microcapsule group of the present invention shows lower TNF-α and IL-6 concentrations and higher anti-inflammatory factor IL-10 concentrations, indicating its superiority in reducing inflammatory response and enhancing immune function; in addition, the immune organ index (spleen index and thymus index) is also significantly higher than other groups, further proving the excellent effect of the microcapsules of the present invention in enhancing immune system function. In general, the high-activity Lactobacillus rhamnosus probiotic microcapsules of the present invention are significantly superior to traditional probiotic preparations in regulating intestinal health and enhancing immune function, showing its innovation and application potential in the field of probiotics.

[0143] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A highly active Lactobacillus rhamnosus probiotic microcapsule, characterized in that: The composition comprises the following components in parts by weight: Lactobacillus rhamnosus: 10^10 CFU per gram of formula; Bifidobacterium: Each gram of formula contains 10^9 CFU; Sodium alginate: 10%-20%; Chitosan: 5%-15%; Hydroxypropyl methylcellulose: 3% to 8%; Pectin: 3% to 7%; Fructooligosaccharides: 5% to 10%; Inulin: 5% to 10%; Vitamin C: 0.1% to 0.5%; Vitamin E: 0.1%~0.3%.

2. A highly active Lactobacillus rhamnosus probiotic microcapsule according to claim 1, characterized in that: The preparation is prepared by double-layer or multi-layer embedding technology, wherein the first layer uses sodium alginate and calcium ions to cross-link to form a gel layer, the second layer uses chitosan to form an acid-resistant coating layer, and the third layer uses hydroxypropyl methylcellulose to form an anti-oxidation protective film.

3. A method for preparing a highly active Lactobacillus rhamnosus probiotic microcapsule, used for the highly active Lactobacillus rhamnosus probiotic microcapsule according to any one of claims 1-2, characterized in that: The following steps are involved: S1. Probiotic culture and pretreatment: culture Lactobacillus rhamnosus and Bifidobacterium to the required bacterial concentration, collect the bacteria and dry them to prepare freeze-dried powder for standby use; S2. Preparation of the inner layer of the microcapsule: mixing the dried probiotic powder with the sodium alginate solution to form a bacterial-sodium alginate suspension; spraying the suspension into a cross-linking bath to form sodium alginate gel microspheres; S3. Preparation of chitosan coating layer: After washing the gel microspheres, they were immersed in a chitosan solution to form a coating layer; S4. Formation of an outer protective film: Immersing the coated microcapsules in a hydroxypropyl methylcellulose solution to form an outer protective film; S5. Drying and forming: The microcapsules are dried by a combination of spray drying and freeze drying to form stable microcapsule powders; S6. Adding prebiotics and antioxidants: adding prebiotics and antioxidants to the dry microcapsule powder and mixing them evenly; S7. Controlled-release coating and final formulation formation: coating the controlled-release agent on the outer surface of the microcapsules, performing final mixing, making a composite formulation in the form of granules or tablets, and sealing and storing.

4. The method for preparing a highly active Lactobacillus rhamnosus probiotic microcapsule according to claim 3, characterized in that: In step S1, the probiotics are dried by freeze drying at a temperature of -50°C to -20°C for a drying time of 12h to 48h.

5. The method for preparing a highly active Lactobacillus rhamnosus probiotic microcapsule according to claim 3, characterized in that: In step S2, the concentration of the sodium alginate solution is 1% to 3%, and a 0.2M calcium chloride solution is used in the cross-linking bath for 30 minutes.

6. The method for preparing a highly active Lactobacillus rhamnosus probiotic microcapsule according to claim 3, characterized in that: In the step S3, the concentration of the chitosan solution is 0.5% to 2%, the coating time is 10 min to 30 min, and the chitosan is dissolved in 1% acetic acid solution.

7. The method for preparing a highly active Lactobacillus rhamnosus probiotic microcapsule according to claim 3, characterized in that: In step S4, the concentration of the hydroxypropyl methylcellulose solution is 1% to 2%, and the soaking time is 10 min to 15 min.

8. The method for preparing a highly active Lactobacillus rhamnosus probiotic microcapsule according to claim 3, characterized in that: In the step S5, the air inlet temperature of the spray drying is 70°C to 90°C, the air outlet temperature is 50°C to 60°C, and the freeze drying temperature is -40°C to -20°C.

9. Application of a probiotic microcapsule composite preparation in a health product for regulating the balance of intestinal flora and enhancing immune function, for a highly active Lactobacillus rhamnosus probiotic microcapsule as claimed in any one of claims 1 to 2, characterized in that: The health product is used to improve intestinal health, enhance immune function, and prevent and alleviate intestinal discomfort symptoms such as diarrhea, constipation, and bloating caused by intestinal flora imbalance.

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