A probiotic enteric-coated dry suspension and its preparation method and application

Abstract

The present invention provides a probiotic enteric dry suspension and its preparation method and application, relating to the technical field of pharmaceutical preparations. The probiotic enteric dry suspension comprises the following components in parts by mass: 1-8 parts of probiotic powder; 25-60 parts of microcapsule cross-linking system; 1-15 parts of suspending agent; 20-70 parts of filler; 0-0.1 parts of colorant; 0-1 parts of fragrance. The present invention utilizes the gelling property of sodium alginate to disperse the probiotic powder in water, and then encapsulates it to form a probiotic-sodium alginate complex. Citric acid and calcium carbonate react with each other to produce carbon dioxide bubbles that can prevent the probiotic-sodium alginate complex from agglomerating. At the same time, the released calcium ions and chitosan interact with the probiotic-sodium alginate complex, forming a dense probiotic microcapsule structure quickly and stably in one step, effectively protecting the probiotics in the microcapsule from damage by gastric acid and bile, improving the survival rate of the probiotics and achieving the enteric effect.

Description

Technical Field

[0001] The present invention relates to the technical field of pharmaceutical preparations, and in particular to a probiotic enteric-coated dry suspension, a preparation method thereof, and an application thereof. Background Art

[0002] Probiotics alter the composition of the host's microbial flora through colonization, resulting in single or mixed microorganisms that benefit host health. Probiotics play a vital role in maintaining the balance of the human intestinal microbiome, promoting nutrient absorption, and enhancing immunity. They are widely used in the prevention and treatment of various diseases, including digestive, immune, and metabolic disorders, and even neurological and psychiatric disorders. As people's understanding of probiotics deepens, market demand for probiotic preparations is also increasing.

[0003] As the organ with the highest abundance of human microbial flora, the intestines are well-suited for probiotic supplementation, improving intestinal function and contributing to the maintenance of human health. Despite their numerous benefits, the vast majority of probiotic strains, such as those from the genera Bifidobacterium, Lactobacillus, and Lactobacillus, have poor tolerance to the external environment. After oral administration, they are strongly affected by gastric acid and bile, leading to the death of numerous live bacteria and a significant decrease in the number of bacteria reaching the intestines, which in turn seriously affects their clinical efficacy.

[0004] To address the problem of probiotic survival during oral administration, researchers have developed a variety of probiotic microencapsulation technologies, including emulsification, spray drying, and extrusion. These technologies encapsulate probiotics in capsules made of different materials, effectively protecting them from damage by gastric acid and bile and enabling targeted release into the intestine. However, these microcapsule preparation technologies are generally complex, and the bacteria must withstand damage from physical factors such as heating, extrusion, and collision during processing, resulting in the death of a large number of live bacteria during the preparation process. Although the finished probiotic microcapsules have a high tolerance, the problem of large-scale loss of live bacteria during processing has not been effectively solved.

[0005] Chinese patent CN109453207A discloses a double-layer microcapsule of probiotics coated with selenized sodium alginate and selenized chitosan, a preparation method and its application, the preparation method comprising the following steps: treating sodium alginate and chitosan with sodium selenite to prepare selenized sodium alginate and selenized chitosan; embedding probiotics with selenized sodium alginate and selenized chitosan; using Ca 2+ Cross-linking was performed to obtain probiotic double-layer microcapsules. However, the viable bacterial survival rate after 2 hours was only 8.1%, and the survival rate of probiotics during the preparation process was not involved.

[0006] Chinese patent CN109232920A discloses a fish gelatin / sodium alginate double-network composite hydrogel, its preparation method, and the resulting probiotic microcapsules. These hydrogels address the shortcomings of fish gelatin and sodium alginate gels used as single-network hydrogels, which suffer from poor mechanical properties and stability, hindering probiotic encapsulation, resulting in low encapsulation efficiency and weak probiotic protection. However, they do not address the survival rate of probiotics during the preparation process.

[0007] Based on this, developing new probiotic preparations to solve the survival problem of probiotics during oral administration and improving the survival rate of probiotics during the preparation process are the research focuses of researchers in this field. Summary of the Invention

[0008] In response to the above problems, the present invention provides an enteric-coated probiotic dry suspension and a preparation method thereof. Suitable enteric-coated microcapsule materials are selected and the principle of microcapsule formation by emulsification is used for reference. Probiotic-sodium alginate gel is spontaneously formed during the dispersion of the preparation in water. The gel mass is dispersed into microparticles by the force of stirring and acid-base reaction. Calcium ions and chitosan released by the acid-base reaction further accelerate the formation of probiotic microcapsule particles. The probiotic microcapsule system is stabilized by a suspending agent, thereby achieving a one-step preparation of a probiotic enteric-coated dry suspension that is convenient for patients to directly take.

[0009] To achieve the above object, the technical solution adopted by the present invention is as follows:

[0010] In one aspect, the present invention provides a probiotic enteric-coated dry suspension comprising the following components in parts by mass:

[0011] Probiotic powder: 1-8 servings;

[0012] Microcapsule cross-linking system: 25-60 parts;

[0013] Suspending agent: 1-15 parts;

[0014] Filler: 20-70 parts;

[0015] Colorant: 0-0.1 parts;

[0016] And fragrance: 0-1 part.

[0017] Preferably, the following components are included in parts by mass:

[0018] Probiotic powder: 1-6 servings;

[0019] Microcapsule cross-linking system: 40-60 parts;

[0020] Suspending agent: 1-10 parts;

[0021] Filler: 40-60 parts;

[0022] Colorant: 0.01-0.05 parts;

[0023] And fragrance: 0.1-1 part.

[0024] More preferably, the following components are included in parts by mass:

[0025] Probiotic powder: 1-5 servings;

[0026] Microcapsule cross-linking system: 40-50 parts;

[0027] Suspending agent: 2-13 parts;

[0028] Filler: 40-50 parts;

[0029] Colorant: 0.01-0.02 parts;

[0030] And fragrance: 0.1-0.5 parts.

[0031] Preferably, the probiotics of the probiotic powder are selected from at least one of Bifidobacterium, Lactobacillus, Lactobacillus casei, Lactobacillus plantarum, Streptococcus and Lactococcus.

[0032] More preferably, the probiotics of the probiotic powder are selected from at least one of Bifidobacterium and Lactobacillus.

[0033] Preferably, the microcapsule cross-linking system comprises sodium alginate, oligochitosan, citric acid and calcium carbonate.

[0034] Preferably, the polymerization degree of the oligochitosan is 2-20.

[0035] Preferably, the microcapsule cross-linking system comprises the following components in parts by mass: 1-5 parts of sodium alginate, 1-5 parts of oligosaccharide chitosan, 15-35 parts of citric acid and 10-20 parts of calcium carbonate.

[0036] Further preferably, the microcapsule cross-linking system comprises the following components, calculated by weight: 1-4 parts of sodium alginate, 1-3 parts of oligochitosan, 20-30 parts of citric acid, and 10-18 parts of calcium carbonate.

[0037] More preferably, the microcapsule cross-linking system comprises the following components in parts by mass: 2-3 parts of sodium alginate, 1-1.5 parts of oligochitosan, 25-30 parts of citric acid and 14-16 parts of calcium carbonate.

[0038] Preferably, the filler is at least one selected from sucrose, lactose, glucose and maltodextrin.

[0039] Further preferably, the filler is at least one selected from sucrose, glucose and maltodextrin.

[0040] More preferably, the filler is a mixture of sucrose and maltodextrin, or a mixture of glucose and maltodextrin.

[0041] Preferably, the mass ratio of sucrose / glucose to maltodextrin is 10-20:25-30.

[0042] Preferably, the aroma is selected from at least one of strawberry flavor, orange flavor, apple flavor, grape flavor and banana flavor.

[0043] Preferably, the aromatic agent is selected from at least one of orange flavor and grape flavor.

[0044] Preferably, the colorant is selected from at least one of yellow iron oxide, red iron oxide and violet iron oxide.

[0045] Preferably, the suspending agent is at least one selected from xanthan gum, hydroxypropyl methylcellulose (HPMC), polyvinyl pyrrolidone (PVP) and gum arabic.

[0046] Further preferably, the suspending agent is selected from at least one of xanthan gum, hydroxypropyl methylcellulose (HPMC) and polyvinylpyrrolidone (PVP).

[0047] More preferably, the suspending agent is selected from any two of xanthan gum, hydroxypropyl methylcellulose (HPMC) and polyvinylpyrrolidone (PVP).

[0048] In another aspect, the present invention provides a method for preparing the above-mentioned enteric-coated probiotic dry suspension, comprising the following steps:

[0049] Step 1: Mix the probiotic powder and the filler in a mass ratio of 1:8-12 to obtain a mixture 1;

[0050] Step 2: mixing the fragrance, colorant, suspending agent and filler in a weight ratio of 1:8-12 to obtain mixture 2;

[0051] Step 3: mixing the microcapsule cross-linking system with the mixture 1 to obtain a mixture 3;

[0052] Step 4: Mix mixture 2, mixture 3 and the remaining filler to obtain the probiotic enteric-coated dry suspension.

[0053] Preferably, in step 1, the mass ratio is 1:10.

[0054] Preferably, in step 1, the mixing time is 1-5 minutes.

[0055] Preferably, in step 1, the step of sieving is further included after mixing, and the mesh size of the sieving is 50-80 mesh.

[0056] Preferably, in step 2, the mass ratio is 1:10.

[0057] Preferably, in step 2, the mixing time is 1-5 minutes.

[0058] Preferably, in step 2, the mixing step is further followed by sieving, and the sieving mesh size is 50-80 meshes.

[0059] Preferably, in step 3, the mixing is stirring for 1-10 minutes.

[0060] In another aspect, the present invention provides use of the above-mentioned enteric-coated probiotic dry suspension in the preparation of an oral probiotic preparation.

[0061] Compared with the prior art, the present invention has the following beneficial effects:

[0062] (1) The present invention selects suitable enteric-coated microcapsule materials and suspension systems through formulation design, so that the probiotic preparation can spontaneously form probiotic microcapsules in one step during the oral administration process, so that the probiotics have gastric acid resistance and enteric-coated properties. Compared with the traditional microcapsule preparation process, the complexity of the process is greatly reduced.

[0063] (2) Compared with traditional probiotic microcapsule preparation technology, this method can form stable probiotic microcapsule particles only by the force of stirring and acid-base reaction. The preparation process is gentle and causes less damage to the bacteria.

[0064] (2) The probiotic microcapsule particles formed during the oral administration process of the present invention are evenly distributed in the suspending agent, which can effectively reduce the possibility of particles adhering to the oral cavity or esophagus and reduce the difficulty of swallowing;

[0065] (3) In the present invention, the taste and color of the drug are adjusted by adding an appropriate amount of citric acid, as well as sweet fillers, colorants and fragrances, thereby masking the unpleasant odor caused by other excipients in the preparation, reducing irritation to the oral cavity and esophagus, and improving the patient's physical sensation when taking the drug. DETAILED DESCRIPTION

[0066] In order to make the technical means, creative features, purpose and effect of the present invention easy to understand, the present invention is further illustrated below in conjunction with specific embodiment, but the following embodiment is only a preferred embodiment of the present invention, not all. Based on the embodiment in the embodiment, other embodiments obtained by those skilled in the art without making creative work all fall within the protection scope of the present invention. It is worth noting that the raw materials used in the present invention are all common commercial products, and their source is not specifically limited. The technology and scientific terms used in the embodiment have the meaning commonly understood by those of ordinary skill in the art to which the present invention belongs.

[0067] Sources of raw materials used in the present invention:

[0068] Lactobacillus plantarum powder: The live bacteria fermentation liquid is obtained by step-by-step amplification of the strain through high-density fermentation. The bacteria are collected by centrifugation, suspended with a protective agent, and then freeze-dried and crushed.

[0069] Sodium alginate: purchased from Qingdao Mingyue Seaweed Group Co., Ltd.

[0070] Oligosaccharide: purchased from Shandong Hailongyuan Biotechnology Co., Ltd.

[0071] Citric acid: purchased from Hunan Jiudian Pharmaceutical Co., Ltd.

[0072] Tartaric acid: purchased from Changmao Biochemical Engineering Co., Ltd.

[0073] Calcium citrate: purchased from Hongyan Reagent Factory, Hedong District, Tianjin.

[0074] Calcium carbonate: purchased from Hunan Xinlvfang Pharmaceutical Co., Ltd.

[0075] Calcium chloride: purchased from Hongyan Reagent Factory, Hedong District, Tianjin.

[0076] Xanthan gum: purchased from Shanghai Aladdin Biochemical Technology Co., Ltd.

[0077] HPMC: SH-E30 was purchased from Anhui Shanhe Pharmaceutical Excipients Co., Ltd.

[0078] Sucrose: purchased from Hunan Jiudian Pharmaceutical Co., Ltd.

[0079] Maltodextrin: purchased from Weifang Shengtai Pharmaceutical Co., Ltd.

[0080] Mannitol: purchased from Qingdao Mingyue Seaweed Group Co., Ltd.

[0081] Sorbitol: purchased from Qingdao Mingyue Seaweed Group Co., Ltd.

[0082] Orange flavor: purchased from IFF International Flavors and Fragrances.

[0083] Yellow iron oxide: purchased from Ningbo Yipin Biotechnology Co., Ltd.

[0084] Example 1

[0085] A probiotic enteric-coated dry suspension, the specific formula of which is as shown in Table 1:

[0086] Table 1. Formulation of Example 1

[0087]

[0088] The preparation method is as follows:

[0089] Step 1: Accurately weigh each component material in 10kg batches;

[0090] Step 2: Disperse and mix Lactobacillus plantarum powder and maltodextrin at a weight ratio of 1:10 for 3-5 minutes, and then evenly pass through a 60-mesh sieve;

[0091] Step 3: Disperse and mix orange flavor, yellow iron oxide, HPMC, xanthan gum and maltodextrin in a weight ratio of 1:10 for 3-5 minutes, and then evenly pass through a 60-mesh sieve;

[0092] Step 4: Sodium alginate, oligochitosan, citric acid and calcium carbonate are sequentially added to the mixed powder obtained in step 2, and the mixture is evenly dispersed and then added to a mixer;

[0093] Step 5: Add the mixed powder obtained in step 3, sucrose and the remaining maltodextrin into a mixer and mix at a certain stirring speed for 5-10 minutes;

[0094] Step 6: Use a powder packaging machine to package and seal the mixed powder obtained in step 5 to obtain the orange-flavored probiotic enteric-coated dry suspension.

[0095] Example 2

[0096] A probiotic enteric-coated dry suspension, the specific formula of which is as shown in Table 2:

[0097] Table 2. Formulation of Example 2

[0098]

[0099] The preparation method is as follows:

[0100] Step 1: Accurately weigh each component material in 10kg batches;

[0101] Step 2: Disperse and mix Bifidobacterium longum powder and maltodextrin in a weight ratio of 1:10 for 3-5 minutes, and then evenly pass through a 60-mesh sieve;

[0102] Step 3: Disperse and mix orange flavor, purple iron oxide, HPMC, PVP and maltodextrin in a weight ratio of 1:10 for 3-5 minutes, and then evenly pass through a 60-mesh sieve;

[0103] Step 4: Sodium alginate, oligochitosan, citric acid and calcium carbonate are sequentially added to the mixed powder obtained in step 2, and the mixture is evenly dispersed and then added to a mixer;

[0104] Step 5: Add the mixed powder obtained in step 3, glucose and the remaining maltodextrin into a mixer and mix at a certain stirring speed for 5-10 minutes;

[0105] Step 6: Use a powder packaging machine to package and seal the mixed powder obtained in step 5 to obtain the grape-flavored probiotic enteric-coated dry suspension.

[0106] Comparative Example 1

[0107] A probiotic enteric-coated dry suspension, the specific formula of which is as shown in Table 3:

[0108] Compared with Example 1, only the formulation of Comparative Example 1 was changed.

[0109] Table 3. Formula of Comparative Example 1

[0110]

[0111] The preparation method is the same as that in Example 1.

[0112] Comparative Example 2

[0113] A probiotic enteric-coated dry suspension, the specific formula of which is as shown in Table 4:

[0114] Compared with Example 1, only calcium carbonate was replaced by calcium chloride.

[0115] Table 4. Formula of Comparative Example 2

[0116]

[0117] The preparation method is as follows:

[0118] Step 1: Accurately weigh each component material in 10kg batches;

[0119] Step 2: Disperse and mix Lactobacillus plantarum powder and maltodextrin at a weight ratio of 1:10 for 3-5 minutes, and then evenly pass through a 60-mesh sieve;

[0120] Step 3: Disperse and mix orange flavor, yellow iron oxide, HPMC, xanthan gum and maltodextrin in a weight ratio of 1:10 for 3-5 minutes, and then evenly pass through a 60-mesh sieve;

[0121] Step 4: Sodium alginate, oligochitosan, citric acid and calcium chloride are sequentially added to the mixed powder obtained in step 2, and the mixture is evenly dispersed and then added to a mixer;

[0122] Step 5: Add the mixed powder obtained in step 3, sucrose and the remaining maltodextrin into a mixer and mix at a certain stirring speed for 5-10 minutes;

[0123] Step 6: Use a powder packaging machine to package and seal the mixed powder obtained in step 5 to obtain the orange-flavored probiotic enteric-coated dry suspension.

[0124] Comparative Example 3

[0125] A probiotic enteric-coated dry suspension, the specific formula of which is as shown in Table 5:

[0126] Compared with Example 1, only calcium carbonate was removed.

[0127] Table 5. Formula of Comparative Example 3

[0128]

[0129] The preparation method is as follows:

[0130] Step 1: Accurately weigh each component material in 10kg batches;

[0131] Step 2: Disperse and mix Lactobacillus plantarum powder and maltodextrin at a weight ratio of 1:10 for 3-5 minutes, and then evenly pass through a 60-mesh sieve;

[0132] Step 3: Disperse and mix orange flavor, yellow iron oxide, HPMC, xanthan gum and maltodextrin in a weight ratio of 1:10 for 3-5 minutes, and then evenly pass through a 60-mesh sieve;

[0133] Step 4: Sodium alginate, oligochitosan and citric acid are sequentially added to the mixed powder obtained in step 2, and after being evenly dispersed, the mixture is added to a mixer;

[0134] Step 5: Add the mixed powder obtained in step 3, sucrose and the remaining maltodextrin into a mixer and mix at a certain stirring speed for 5-10 minutes;

[0135] Step 6: Use a powder packaging machine to package and seal the mixed powder obtained in step 5 to obtain the orange-flavored probiotic enteric-coated dry suspension.

[0136] Comparative Example 4

[0137] A probiotic enteric-coated dry suspension, the specific formula of which is as shown in Table 6:

[0138] Compared with Example 1, only citric acid was replaced by tartaric acid.

[0139] Table 6. Formula of Comparative Example 4

[0140]

[0141] The preparation method is as follows:

[0142] Step 1: Accurately weigh each component material in 10kg batches;

[0143] Step 2: Disperse and mix Lactobacillus plantarum powder and maltodextrin at a weight ratio of 1:10 for 3-5 minutes, and then evenly pass through a 60-mesh sieve;

[0144] Step 3: Disperse and mix orange flavor, yellow iron oxide, HPMC, xanthan gum and maltodextrin in a weight ratio of 1:10 for 3-5 minutes, and then evenly pass through a 60-mesh sieve;

[0145] Step 4: Sodium alginate, oligochitosan, 1% (w / v) tartaric acid and calcium carbonate are sequentially added to the mixed powder obtained in step 2, dispersed evenly and then added to a mixer;

[0146] Step 5: Add the mixed powder obtained in step 3, sucrose and the remaining maltodextrin into a mixer and mix at a certain stirring speed for 5-10 minutes;

[0147] Step 6: Use a powder packaging machine to package and seal the mixed powder obtained in step 5 to obtain the orange-flavored probiotic enteric-coated dry suspension.

[0148] Comparative Example 5

[0149] A probiotic enteric-coated dry suspension, the specific formula of which is as shown in Table 7:

[0150] Compared with Example 1, only citric acid was replaced by calcium citrate.

[0151] Table 7. Formula of Comparative Example 5

[0152]

[0153] The preparation method is as follows:

[0154] Step 1: Accurately weigh each component material in 10kg batches;

[0155] Step 2: Disperse and mix Lactobacillus plantarum powder and maltodextrin at a weight ratio of 1:10 for 3-5 minutes, and then evenly pass through a 60-mesh sieve;

[0156] Step 3: Disperse and mix orange flavor, yellow iron oxide, HPMC, xanthan gum and maltodextrin in a weight ratio of 1:10 for 3-5 minutes, and then evenly pass through a 60-mesh sieve;

[0157] Step 4: Sodium alginate, oligochitosan, calcium citrate and calcium carbonate are sequentially added to the mixed powder obtained in step 2, and the mixture is evenly dispersed and then added to a mixer;

[0158] Step 5: Add the mixed powder obtained in step 3, sucrose and the remaining maltodextrin into a mixer and mix at a certain stirring speed for 5-10 minutes;

[0159] Step 6: Use a powder packaging machine to package and seal the mixed powder obtained in step 5 to obtain the orange-flavored probiotic enteric-coated dry suspension.

[0160] Comparative Example 6

[0161] A probiotic enteric-coated dry suspension, the specific formula of which is as shown in Table 8:

[0162] Compared with Example 1, only the ratio of the components of the microcapsule cross-linking system was changed.

[0163] Table 8. Formula of Comparative Example 6

[0164]

[0165] The preparation method is the same as that in Example 1.

[0166] Comparative Example 7

[0167] A probiotic enteric-coated dry suspension, the specific formula of which is as shown in Table 9:

[0168] Compared with Example 1, only the suspending agent was changed.

[0169] Table 9. Formula of Comparative Example 7

[0170]

[0171] The preparation method is as follows:

[0172] Step 1: Accurately weigh each component material in 10kg batches;

[0173] Step 2: Disperse and mix Lactobacillus plantarum powder and maltodextrin at a weight ratio of 1:10 for 3-5 minutes, and then evenly pass through a 60-mesh sieve;

[0174] Step 3: Disperse and mix orange flavor, yellow iron oxide, microcrystalline cellulose, gelatin, and maltodextrin in a weight ratio of 1:10 for 3-5 minutes, and then evenly pass through a 60-mesh sieve;

[0175] Step 4: Sodium alginate, oligochitosan, citric acid and calcium carbonate are sequentially added to the mixed powder obtained in step 2, and the mixture is evenly dispersed and then added to a mixer;

[0176] Step 5: Add the mixed powder obtained in step 3, sucrose and the remaining maltodextrin into a mixer and mix at a certain stirring speed for 5-10 minutes;

[0177] Step 6: Use a powder packaging machine to package and seal the mixed powder obtained in step 5 to obtain the orange-flavored probiotic enteric-coated dry suspension.

[0178] Comparative Example 8

[0179] A probiotic enteric-coated dry suspension, the specific formula of which is as shown in Table 10:

[0180] Compared to Example 1, only the filler was changed.

[0181] Table 10. Formula of Comparative Example 8

[0182]

[0183] The preparation method is as follows:

[0184] Step 1: Accurately weigh each component material in 10kg batches;

[0185] Step 2: Disperse and mix Lactobacillus plantarum powder and sorbitol in a weight ratio of 1:10 for 3-5 minutes, and then evenly pass through a 60-mesh sieve;

[0186] Step 3: Disperse and mix orange flavor, yellow iron oxide, HPMC, xanthan gum and sorbitol in a weight ratio of 1:10 for 3-5 minutes, and then evenly pass through a 60-mesh sieve;

[0187] Step 4: Sodium alginate, oligochitosan, citric acid and calcium carbonate are sequentially added to the mixed powder obtained in step 2, and the mixture is evenly dispersed and then added to a mixer;

[0188] Step 5: Add the mixed powder obtained in step 3, mannitol and the remaining sorbitol into a mixer and mix at a certain stirring speed for 5-10 minutes;

[0189] Step 6: Use a powder packaging machine to package and seal the mixed powder obtained in step 5 to obtain the orange-flavored probiotic enteric-coated dry suspension.

[0190] Comparative Example 9

[0191] A probiotic microcapsule, wherein the probiotic addition amount and the microcapsule cross-linking system formula are consistent with those in Example 1.

[0192] Probiotic microcapsules were prepared according to the conventional probiotic-sodium alginate-chitosan microcapsule preparation method. The preparation method is as follows:

[0193] Prepare liquid A: 100 mL, 2% sodium alginate solution; liquid B: 150 mL, a mixed solution of 1 wt% oligochitosan, citric acid and calcium chloride. Add 1 g of Lactobacillus plantarum powder to liquid A (100 mL, 2 wt% sodium alginate solution) and mix thoroughly. Use a sterile syringe to squeeze it dropwise into the mixed solution of liquid B (150 mL, 1 wt% oligochitosan, citric acid and calcium chloride). During the instillation, control the distance between the needle tip and the surface of solution B to about 50 cm and continuously stir the solution. After all the liquid A is instilled, let the mixture stand and harden for 30 minutes to form gel balls. After filtering, rinse with sterile water and collect the probiotic microcapsules.

[0194] Test Example 1: Probiotic Survival Rate in the Preparation Process of Probiotic Enteric-coated Dry Suspension

[0195] 3.00 g of the enteric-coated probiotic dry suspensions of Examples 1-2 and Comparative Examples 1-8 were accurately weighed and added to a conical flask containing 27 mL of PBS buffer solution and glass beads. After homogenization on a shaker at 200 rpm for 15 min, a gradient dilution was performed at a 10-fold ratio. 1 mL of the diluted sample was drawn and anaerobically cultured on MRS agar medium at 37 ° C for 48 h using the pouring method. The total number of viable bacteria in the dry suspension was determined.

[0196] Survival rate (%) = number of viable bacteria in dry suspension / theoretical number of viable bacteria added × 100%.

[0197] The survival rates of probiotics during the preparation of the dry suspensions of Examples 1-2 and Comparative Examples 1-8 are shown in Table 11 below. The survival rates of probiotics during the preparation of Examples 1-2 were all higher than 95%, which were better than the survival rates of live probiotics in Comparative Examples 1-8, indicating that the formulation caused less damage to the probiotics.

[0198] Table 11. Probiotic survival rate

[0199]

[0200] Test Example 2: Encapsulation rate of probiotics after mixing probiotic enteric-coated dry suspension with water

[0201] 5g of the dry suspensions of embodiment 1-2 and comparative example 1-8 are accurately weighed respectively, poured into 100mL warm water and slowly stirred for 5-10min, and stopped stirring after forming a stable suspension. Whole suspensions are transferred to centrifuge tubes, and after being centrifuged at 3000rpm for 5min and discarding the upper solution, PBS is added and resuspended for cleaning 3 times. The probiotic microcapsules after cleaning are all transferred to a triangular flask equipped with glass beads and settled to 100mL PBS solution. After 200rpm shaking table mixing and homogenizing for 15 min, gradient dilution is carried out according to 10 times of ratios. Suitable dilution is selected to draw 1mL diluted sample. Using the pouring method, count after 37°C of anaerobic culture for 48h on MRS agar medium, and measure the probiotic embedding rate after the dry suspension is mixed with water.

[0202] Embedding rate (%) = number of viable bacteria in probiotic microcapsules / number of viable bacteria in dry suspension × 100%.

[0203] The probiotic encapsulation rates of the enteric-coated probiotic dry suspensions of Examples 1-2 and Comparative Examples 1-8 after mixing with water are shown in Table 12 below. The probiotic encapsulation rates of Examples 1-2 were 85.3% and 84.6%, respectively, indicating that the formulation can achieve one-step microencapsulation preparation of probiotics. In Comparative Example 1, due to the high proportion of probiotic powder added, the encapsulation limit of the cross-linking system of the enteric dry suspension microcapsule was exceeded, resulting in some bacteria being unable to effectively form microcapsules, causing the embedding rate to drop to 61.1%, which did not meet the requirements; in Comparative Example 2, calcium carbonate was replaced by calcium chloride, which destroyed the effervescent reaction in the microcapsule cross-linking system. Only manual slow stirring was relied upon, and the probiotics could not fully contact the microcapsule cross-linking system and form microcapsule particles with uniform particle size, resulting in the embedding rate being reduced to 57.9%; in Comparative Example 3, after calcium carbonate was removed, it was difficult to form a stable sodium alginate-chitosan-calcium ion microcapsule structure due to the lack of calcium ions. Only a small amount of probiotics was embedded in the unstable microcapsules. At the same time, a large amount of citric acid in the suspension also caused great damage to the activity of the bacteria, and its embedding rate was only 9.45%; in Comparative Example 4, citric acid was removed. Citric acid was replaced with tartaric acid. Although the effervescent cross-linking system was not destroyed, the acidity of tartaric acid was too strong, which caused high damage to live bacteria, and thus the encapsulation efficiency was reduced to 63.6%; in Comparative Example 5, citric acid was replaced with calcium citrate. On the one hand, the effervescent reaction in the microcapsule cross-linking system was destroyed, and on the other hand, the solubility of calcium citrate in a non-acidic environment was reduced, the number of free calcium ions was reduced, and the encapsulation efficiency was reduced to 22.9%; in Comparative Example 6, the microcapsule cross-linking system was adjusted, resulting in an imbalance in the ratio of citric acid and calcium carbonate, and the calcium carbonate failed to react completely, resulting in the encapsulation efficiency being reduced to 68.5%; in Comparative Example 7, the suspending agent was adjusted, which affected the state of the suspension system and thus the microcapsule formation process, and the encapsulation efficiency was reduced to 63.1%; in Comparative Example 8, the filler was adjusted, which also had a certain impact on the suspension system, resulting in the encapsulation efficiency being reduced to 70.3%.

[0204] Table 12. Probiotic encapsulation rate

[0205]

[0206] Test Example 3 Determination of Sedimentation Volume Ratio of Probiotic Enteric-Coated Dry Suspension and Observation of Suspension State

[0207] According to the inspection method of the sedimentation volume ratio of the suspension in the 2020 edition of the Chinese Pharmacopoeia, 2.5 g of the probiotic enteric-coated dry suspension of Examples 1-2 and Comparative Examples 1-8 were weighed respectively, added to a stoppered graduated cylinder, and water was added to a constant volume of 50 mL. The stopper was sealed and shaken vigorously for 1 minute. The starting height H0 of the suspension was recorded. After standing for 3 hours, the final height H of the suspension was recorded, and the state of the suspension was observed. The sedimentation volume ratio was calculated as follows:

[0208] Sedimentation volume ratio = H / H0.

[0209] The sedimentation volume ratio is required to be no less than 0.90.

[0210] The suspension states and sedimentation volume ratios of the probiotic enteric-coated dry suspensions of Examples 1-2 and Comparative Examples 1-8 after adding water and mixing are shown in Table 13 below. The microcapsule particle size and distribution in the suspensions of Examples 1 and 2 are uniform, and the sedimentation volume ratio meets the sedimentation volume ratio requirements for suspensions specified in the Chinese Pharmacopoeia. In Comparative Examples 1-8, adjustments to the formulations affected the spontaneous formation process of the probiotic microcapsules and the suspension system to varying degrees, resulting in uneven distribution of the probiotic microcapsule particles in the suspensions and poor suspension stability, failing to meet the Chinese Pharmacopoeia requirement that the sedimentation volume ratio be no less than 0.90.

[0211] Table 13. Suspension state and sedimentation volume ratio of probiotic dry suspension

[0212]

[0213] Test Example 4 Tolerance of Probiotic Enteric-Coated Dry Suspension Microcapsules in Simulated Gastric Fluid

[0214] The probiotic microcapsules of Examples 1-2 and Comparative Examples 1-8 were obtained according to the method of Experimental Example 2. 5 g of bacterial powder raw material was used as a control and added to 45 mL of simulated gastric fluid, respectively. The mixture was thoroughly mixed for 15 s and shaken on a constant temperature shaker at 37 ° C and 200 rpm. The number of viable bacteria in the initial and 120 min simulated gastric acid samples was measured and the survival rate was calculated.

[0215] The tolerance of the probiotic microcapsules spontaneously formed from the dry suspensions of Examples 1-2 and Comparative Examples 1-8 with water in simulated gastric fluid is shown in Table 14 below. The survival rate of the unencapsulated Lactobacillus plantarum powder in simulated gastric fluid was only 12.7%. The survival rates of the probiotic microcapsules of Examples 1-2 were all above 90%, meeting the requirements. In Comparative Example 1, due to the excessive addition ratio of probiotic powder, the encapsulation limit of the cross-linking system of the enteric dry suspension microcapsule was exceeded, which affected the embedding rate and the integrity of the microcapsules. The survival rate was 80.2%, which did not meet the requirements. In Comparative Example 2, calcium carbonate was replaced with calcium chloride, which destroyed the effervescent reaction in the microcapsule cross-linking system. Only manual slow stirring was relied on, and the probiotics and the microcapsule cross-linking system could not be fully dispersed and cross-linked. The microcapsule structure integrity was poor, and the survival rate was 64.6%, which did not meet the requirements. In Comparative Example 3, after removing calcium carbonate, it was difficult to form a stable sodium alginate-chitosan-calcium ion microcapsule structure due to the lack of calcium ions, and the bacteria could not be protected from gastric acid damage. The survival rate was only 15.7%, which did not meet the requirements. In Comparative Example 4, citric acid was replaced with tartaric acid. Although tartaric acid did not destroy the effervescent system, it was too acidic and caused great damage to the bacterial powder. Even with the protection of the microcapsules, the survival rate still dropped to 73.2%, which did not meet the requirements. In Comparative Example 5, citric acid was replaced by calcium citrate. On the one hand, the effervescent reaction in the microcapsule cross-linking system was destroyed. On the other hand, the solubility of calcium citrate in the non-acidic environment was reduced, the number of free calcium ions was reduced, and the integrity of the probiotic microcapsule structure was affected. The survival rate was 30.8%, which did not meet the requirements. In Comparative Example 6, the microcapsule cross-linking system was adjusted, resulting in an imbalance in the ratio of citric acid and calcium carbonate. The calcium carbonate failed to react completely, the calcium ion content was insufficient, and some microcapsules lacked a stable sodium alginate-chitosan-calcium ion structure, resulting in a survival rate reduced to 73.1%, which did not meet the requirements. In Comparative Examples 7-8, the suspension system and filling excipients were adjusted, resulting in a low viscosity of the suspension. The spontaneous formation of probiotic microcapsules could not maintain a good suspension state, which had a certain impact on their formation and microcapsule integrity, resulting in a survival rate reduced to 78.7% and 83.0%, which did not meet the requirements.

[0216] Table 14. Tolerance of probiotic microcapsules in gastric juice

[0217]

[0218] Test Example 5 Release of Probiotic Enteric-coated Dry Suspension in Simulated Intestinal Fluid

[0219] The probiotic microcapsules of Examples 1-2 and Comparative Examples 1-8 were obtained according to the method of Experimental Example 2, and 5 g of bacterial powder raw material was used as a control. They were added to 45 mL of simulated intestinal fluid, mixed thoroughly for 15 s, and shaken on a constant temperature shaker at 37 ° C and 200 rpm. The number of viable bacteria in the simulated intestinal fluid samples was measured after 30 min, 60 min, 90 min, and 120 min.

[0220] The release properties of the probiotic microcapsules spontaneously formed from the dry suspensions of Examples 1-2 and Comparative Examples 1-8 and water in intestinal fluid are shown in Table 15 below. The probiotic microcapsules of Examples 1-2 exhibited excellent intestinal release characteristics, with the probiotics continuously released within one hour in simulated intestinal fluid, demonstrating intestinal-targeted sustained release. However, the probiotic microcapsules of Comparative Example 1-8 exhibited poor structural integrity, resulting in rapid release within 30 minutes in simulated intestinal fluid, failing to achieve sustained release.

[0221] Table 15. Release in intestinal fluid

[0222]

[0223] Test Example 6 Taste and Swallowing Experience Evaluation

[0224] 5 g of the dry suspensions of Examples 1-2 and Comparative Examples 1-8 were accurately weighed, poured into 100 mL of warm water, and slowly stirred for 5-10 min. Stirring was stopped after a stable suspension was formed, and the taste and swallowing experience tests were performed.

[0225] The taste and swallowing experience of Example 1-2 and Comparative Example 1-8 probiotic enteric-coated dry suspension dissolved in water are shown in Table 16 below. The taste evaluation and swallowing experience of Example 1-2 are better. In Ratio 1, due to the high proportion of probiotic powder added, the unencapsulated powder particles are dispersed in the suspension system, and the granularity is obvious when taken, which does not meet the requirements; Comparative Example 2 replaces calcium carbonate with calcium chloride, and the citric acid is not neutralized, resulting in a higher acidity of the suspension and a sour taste. In addition, there is a lack of the dispersing effect of the effervescent reaction, and there are more large particles in the suspension, which affects swallowing, and the evaluation results do not meet the requirements; After the calcium carbonate is removed in Comparative Example 3, the citric acid is not neutralized, resulting in a higher acidity of the suspension and a sour taste. Furthermore, the lack of calcium ions resulted in a loose structure of the probiotic microcapsules, a viscous suspension, exposed bacterial powder particles, and a noticeable graininess, which did not meet the requirements. Comparative Example 4 replaced citric acid with tartaric acid. Although some of the tartaric acid was neutralized, the tartaric acid itself had a strong sour taste and a strong irritation upon ingestion, which did not meet the requirements. Comparative Example 5 replaced citric acid with calcium citrate, which reduced the amount of dissolved calcium ions, leaving most of the probiotics unencapsulated and resulting in a noticeable graininess. Furthermore, the lack of an acidulant resulted in a sweet taste, which did not meet the requirements. Adjustments to the microcapsule cross-linking system in Comparative Example 6 resulted in an imbalance in the ratio of citric acid to calcium carbonate, completely consuming the citric acid and resulting in a sweet taste. Adjustments to the suspension system and filler excipients in Comparative Examples 7-8 resulted in a low suspension viscosity, which failed to effectively mask the graininess of the probiotic microcapsules and did not meet the requirements.

[0226] Table 16. Taste and swallowing experience evaluation results

[0227]

[0228] Test Example 7 Stability Test

[0229] 100 g of each dry suspension of Examples 1-2 and Comparative Examples 1-8 were respectively packaged into aluminum-plastic bags at a rate of 10 g per bag and sealed. An accelerated stability test was performed for 3 months at a temperature of 40°C ± 2°C and a relative humidity of 75% ± 5%.

[0230] The stability of the enteric-coated probiotic dry suspensions of Examples 1-2 and Comparative Examples 1-8 is shown in Table 17 below. The accelerated stability three-month survival rate of the enteric-coated probiotic dry suspension samples of Examples 1-2 exceeded 85%, meeting the requirements. The formulation changes in Comparative Examples 1-8 reduced the protective effect of the probiotics therein, and the accelerated stability viable bacterial survival rates did not meet the evaluation standard requirement of 75%.

[0231] Table 17. Accelerated stability test results

[0232]

[0233] Experimental Example 7 Comparison with conventional probiotic microcapsule preparation process

[0234] Probiotic microcapsules were prepared according to the method of Comparative Example 9, and their embedding efficiency, gastric acid resistance and intestinal release capacity were measured according to the methods of Test Examples 1, 4 and 5, respectively, and compared with the microcapsules spontaneously formed from the enteric-coated dry suspension in Example 1.

[0235] Comparing the embedding efficiency, gastric acid resistance, and intestinal release characteristics of the probiotic microcapsules of Example 1 and Comparative Example 9, as shown in Table 18 below, the probiotic microcapsules spontaneously formed based on the formulation prescription design in Example 1 are basically consistent with the probiotic microcapsules produced by the conventional probiotic microcapsule preparation method in terms of embedding efficiency, gastric acid resistance, and intestinal sustained-release targeted release characteristics, but the preparation process is significantly simplified, which facilitates implementation and reduces the cost of probiotic microcapsule preparation.

[0236] Table 18. Comparison of Example 1 and Comparative Example 9

[0237]

[0238] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, rather than to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions of the technical solution of the present invention by ordinary technicians in this field do not deviate from the essence and scope of the technical solution of the present invention.

Claims

1. A probiotic enteric-coated dry suspension, characterized in that: According to the weight percentage, it contains the following components: Probiotic powder: 1-8 servings; Microcapsule cross-linking system: 25-60 parts; Suspending agent: 1-15 parts; Filler: 20-70 parts; Colorant: 0-0.1 parts; and fragrance: 0-1 part; The microcapsule cross-linking system comprises the following components, calculated by weight: 1-4 parts of sodium alginate, 1-3 parts of oligosaccharide chitosan, 20-30 parts of citric acid, and 10-18 parts of calcium carbonate; The suspending agent is selected from any two of xanthan gum, hydroxypropyl methylcellulose, and polyvinyl pyrrolidone; The filler is a mixture of sucrose and maltodextrin or a mixture of glucose and maltodextrin; The probiotics of the probiotic powder are selected from at least one of Bifidobacterium, Lactobacillus, Lactobacillus casei, Lactobacillus plantarum, Streptococcus and Lactococcus; The preparation method of the probiotic enteric-coated dry suspension comprises the following steps: Step 1: Mix the probiotic powder and the filler in a mass ratio of 1:8-12 to obtain a mixture 1; Step 2: mixing the fragrance, colorant, suspending agent and filler in a weight ratio of 1:8-12 to obtain mixture 2; Step 3: mixing the microcapsule cross-linking system with the mixture 1 to obtain a mixture 3; Step 4: Mix mixture 2, mixture 3 and the remaining filler to obtain the probiotic enteric-coated dry suspension.

2. The probiotic enteric-coated dry suspension according to claim 1, wherein The aromatic agent is selected from at least one of strawberry flavor, orange flavor, apple flavor, grape flavor and banana flavor.

3. The probiotic enteric-coated dry suspension according to claim 1, wherein The colorant is selected from at least one of yellow iron oxide, red iron oxide and violet iron oxide.

4. The method for preparing the enteric-coated probiotic dry suspension according to any one of claims 1 to 3, characterized in that: The following steps are involved: Step 1: Mix the probiotic powder and the filler in a mass ratio of 1:8-12 to obtain a mixture 1; Step 2: mixing the fragrance, colorant, suspending agent and filler in a weight ratio of 1:8-12 to obtain mixture 2; Step 3: mixing the microcapsule cross-linking system with the mixture 1 to obtain a mixture 3; Step 4: Mix mixture 2, mixture 3 and the remaining filler to obtain the probiotic enteric-coated dry suspension.

5. Use of the probiotic enteric-coated dry suspension according to any one of claims 1 to 3 in the preparation of an oral probiotic preparation.

Images