Probiotic enteric dry suspension as well as preparation method and application thereof

Abstract

The invention provides a probiotic enteric dry suspension as well as a preparation method and application thereof, and relates 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 to 60 parts of a microcapsule cross-linking system; 1-15 parts of a suspending aid; 20 to 70 parts of a filling agent; 0-0.1 part of a coloring agent; and 0-1 part of an aromatic. According to the invention, when the probiotic powder is dispersed in water by utilizing the gelling property of sodium alginate, the probiotic powder is wrapped to form a probiotic-sodium alginate compound, citric acid and calcium carbonate are subjected to an acid-base reaction, and generated carbon dioxide bubbles can prevent the probiotic-sodium alginate compound from agglomerating; meanwhile, the released calcium ions and chitosan interact with the probiotic-sodium alginate compound, a compact probiotic microcapsule structure is rapidly and stably formed in one step, the probiotics in the microcapsule are effectively protected from being damaged by gastric acid and bile, the survival rate of the probiotics is increased, and the enteric effect is achieved.

Description

Technical Field

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

[0002] Probiotics change the composition of the flora in a certain part of the host through colonization, thereby producing a single or mixed microorganism with a clear composition that is beneficial to the health of the host. Probiotics play an important role in maintaining the balance of the human intestinal microecology, promoting nutrient absorption, and enhancing immunity, and have been widely used in the prevention and treatment of various diseases, such as digestive system diseases, immune diseases, metabolic diseases, and even neurological and mental diseases. As people's understanding of probiotics deepens, the market demand for probiotic preparations is also increasing.

[0003] As the organ with the highest abundance of human microbial flora, the intestinal tract can be properly supplemented with probiotics to improve intestinal function and has a positive significance for maintaining human health. Although probiotics have many benefits, most probiotic strains such as Bifidobacterium, Lactobacillus, Lactobacillus and other strains have poor tolerance to the external environment. After oral administration, they will be strongly affected by gastric acid and bile, resulting in the death of a large number of live bacteria, and the number of bacteria reaching the intestine will drop significantly, which will seriously affect its clinical efficacy.

[0004] In order to solve 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 capsule shells of different materials, effectively protecting probiotics from damage by gastric acid and bile and achieving targeted release in the intestine. However, these microcapsule preparation technologies are generally complex, and the bacteria still need to withstand damage from physical factors such as heating, extrusion, and collision during the processing, resulting in a large number of live bacteria dying during the preparation process. Although the finished probiotic microcapsule products have a high tolerance, the problem of large-scale loss of live bacteria during the 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 application thereof, the preparation method comprising the following steps: treating sodium alginate and chitosan with sodium selenite to obtain selenized sodium alginate and selenized chitosan; embedding probiotics with selenized sodium alginate and selenized chitosan; using Ca 2+ The probiotic double-layer microcapsules were obtained by cross-linking. However, the survival rate of live bacteria 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 obtained probiotic microcapsules, which can solve the problems that fish gelatin gel and sodium alginate gel have poor mechanical properties and stability when used as single network gel, which is not conducive to the embedding of probiotics, resulting in low probiotic embedding rate and weak protection for probiotics. However, it does not involve 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 view of the above problems, the present invention provides a probiotic enteric dry suspension and a preparation method thereof, selects suitable enteric microcapsule materials and draws on the principle of microcapsule formation by emulsification method, so that probiotic-sodium alginate gel is spontaneously formed in the process of dispersion of the preparation in water, and the gel mass is dispersed into microparticles by the force of stirring and acid-base reaction, and calcium ions and chitosan released by the acid-base reaction further accelerate the formation of probiotic microcapsule particles, and the probiotic microcapsule system is stabilized by a suspending agent, so as to realize one-step preparation of a probiotic enteric dry suspension that is convenient for patients to take directly.

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

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

[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 degree of polymerization 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 oligochitosan, 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 selected from at least one of 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 fragrance 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 selected from at least one of 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 polyvinyl pyrrolidone (PVP).

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

[0048] On the other hand, the present invention provides a method for preparing the above-mentioned probiotic enteric 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: Mix the aromatic agent, the colorant and the suspending agent with the filler in a weight ratio of 1:8-12 to obtain a mixture 2;

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

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

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

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

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

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

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

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

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

[0060] In another aspect, the present invention provides use of the above-mentioned probiotic enteric 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 the 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 the 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, and adding a sweet filler, a colorant and a fragrance, thereby masking the unpleasant odor caused by other auxiliary materials 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 explained below in conjunction with specific embodiments, but the following embodiments are only preferred embodiments of the present invention, not all. Based on the embodiments in the implementation mode, other embodiments obtained by those skilled in the art without making creative work all belong to 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 sources are not specifically limited. The technology and scientific terms used in the embodiments have the meanings 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, a protective agent is added to the suspension, 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, 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 & 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]

[0089] The preparation method is as follows:

[0090] Step 1: Accurately weigh each component material in 10kg batches;

[0091] Step 2: Disperse and mix the plant lactobacillus powder and maltodextrin at a weight ratio of 1:10 for 3-5 minutes, and then evenly pass through a 60-mesh sieve;

[0092] 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;

[0093] Step 4: adding sodium alginate, oligochitosan, citric acid and calcium carbonate to the mixed powder obtained in step 2 in sequence, and adding the mixture into a mixer after being evenly dispersed;

[0094] 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;

[0095] Step 6: Use a powder packaging machine to package and seal the mixed powder obtained in step 5 to obtain an orange-flavored probiotic enteric-coated dry suspension.

[0096] Example 2

[0097] A probiotic enteric-coated dry suspension, the specific formula of which is as shown in Table 2:

[0098] Table 2. Formulation of Example 2

[0099]

[0100] The preparation method is as follows:

[0101] Step 1: Accurately weigh each component material in 10kg batches;

[0102] Step 2: Disperse and mix the 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;

[0103] 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;

[0104] Step 4: adding sodium alginate, oligochitosan, citric acid and calcium carbonate to the mixed powder obtained in step 2 in sequence, and adding the mixture into a mixer after being evenly dispersed;

[0105] 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;

[0106] 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 dry suspension.

[0107] Comparative Example 1

[0108] A probiotic enteric dry suspension, the specific formula of which is shown in Table 3: Compared with Example 1, only the formula of Comparative Example 1 is 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 dry suspension, the specific formula of which is shown in Table 4 below: Compared with Example 1, only calcium carbonate is replaced by calcium chloride.

[0114] Table 4. Formula of Comparative Example 2

[0115]

[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 the 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;

[0120] 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;

[0121] Step 4: adding sodium alginate, oligochitosan, citric acid and calcium chloride to the mixed powder obtained in step 2 in sequence, and adding the mixture into a mixer after being evenly dispersed;

[0122] 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;

[0123] 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 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 the calcium carbonate was removed.

[0127] Table 5. Formula of Comparative Example 3

[0128]

[0129]

[0130] The preparation method is as follows:

[0131] Step 1: Accurately weigh each component material in 10kg batches;

[0132] Step 2: Disperse and mix the 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;

[0133] 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;

[0134] Step 4: Add sodium alginate, oligochitosan and citric acid to the mixed powder obtained in step 2 in sequence, and add the mixture into a mixer after being evenly dispersed;

[0135] 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;

[0136] 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 dry suspension.

[0137] Comparative Example 4

[0138] A probiotic enteric-coated dry suspension, the specific formula of which is as shown in Table 6:

[0139] Compared with Example 1, only citric acid was replaced by tartaric acid.

[0140] Table 6. Formula of Comparative Example 4

[0141]

[0142] The preparation method is as follows:

[0143] Step 1: Accurately weigh each component material in 10kg batches;

[0144] Step 2: Disperse and mix the plant lactobacillus powder and maltodextrin at a weight ratio of 1:10 for 3-5 minutes, and then evenly pass through a 60-mesh sieve;

[0145] 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;

[0146] Step 4: adding sodium alginate, oligochitosan, 1% (w / v) tartaric acid and calcium carbonate to the mixed powder obtained in step 2 in sequence, dispersing evenly and adding to a mixer;

[0147] 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;

[0148] Step 6: Use a powder packaging machine to package and seal the mixed powder obtained in step 5 to obtain an orange-flavored probiotic enteric-coated dry suspension.

[0149] Comparative Example 5

[0150] A probiotic enteric-coated dry suspension, the specific formula of which is as shown in Table 7:

[0151] Compared with Example 1, only citric acid was replaced by calcium citrate.

[0152] Table 7. Formula of Comparative Example 5

[0153]

[0154] The preparation method is as follows:

[0155] Step 1: Accurately weigh each component material in 10kg batches;

[0156] Step 2: Disperse and mix the plant lactobacillus powder and maltodextrin at a weight ratio of 1:10 for 3-5 minutes, and then evenly pass through a 60-mesh sieve;

[0157] 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;

[0158] Step 4: adding sodium alginate, oligochitosan, calcium citrate and calcium carbonate to the mixed powder obtained in step 2 in sequence, and adding the mixture into a mixer after being evenly dispersed;

[0159] 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;

[0160] Step 6: Use a powder packaging machine to package and seal the mixed powder obtained in step 5 to obtain an orange-flavored probiotic enteric-coated dry suspension.

[0161] Comparative Example 6

[0162] A probiotic enteric-coated dry suspension, the specific formula of which is as shown in Table 8:

[0163] Compared with Example 1, only the ratio of the components of the microcapsule cross-linking system is changed.

[0164] Table 8. Formula of Comparative Example 6

[0165]

[0166] The preparation method is the same as that in Example 1.

[0167] Comparative Example 7

[0168] A probiotic enteric-coated dry suspension, the specific formula of which is as shown in Table 9:

[0169] Compared with Example 1, only the suspending agent was changed.

[0170] Table 9. Formula of Comparative Example 7

[0171]

[0172]

[0173] The preparation method is as follows:

[0174] Step 1: Accurately weigh each component material in 10kg batches;

[0175] Step 2: Disperse and mix the plant lactobacillus powder and maltodextrin at a weight ratio of 1:10 for 3-5 minutes, and then evenly pass through a 60-mesh sieve;

[0176] 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;

[0177] Step 4: adding sodium alginate, oligochitosan, citric acid and calcium carbonate to the mixed powder obtained in step 2 in sequence, and adding the mixture into a mixer after being evenly dispersed;

[0178] 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;

[0179] Step 6: Use a powder packaging machine to package and seal the mixed powder obtained in step 5 to obtain an orange-flavored probiotic enteric-coated dry suspension.

[0180] Comparative Example 8

[0181] A probiotic enteric dry suspension, the specific formula is as shown in Table 10:

[0182] Compared to Example 1, only the filler was changed.

[0183] Table 10. Formula of Comparative Example 8

[0184]

[0185] The preparation method is as follows:

[0186] Step 1: Accurately weigh each component material in 10kg batches;

[0187] Step 2: Disperse and mix the Lactobacillus plantarum powder and sorbitol at a weight ratio of 1:10 for 3-5 minutes, and then evenly pass through a 60-mesh sieve;

[0188] 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;

[0189] Step 4: adding sodium alginate, oligochitosan, citric acid and calcium carbonate to the mixed powder obtained in step 2 in sequence, and adding the mixture into a mixer after being evenly dispersed;

[0190] 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;

[0191] Step 6: Use a powder packaging machine to package and seal the mixed powder obtained in step 5 to obtain an orange-flavored probiotic enteric-coated dry suspension.

[0192] Comparative Example 9

[0193] A probiotic microcapsule, wherein the probiotic addition amount and the microcapsule cross-linking system formula are consistent with those in Example 1.

[0194] Probiotic microcapsules were prepared according to the conventional probiotic-sodium alginate-chitosan microcapsule preparation method. The preparation method is:

[0195] 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 plant lactobacillus powder to liquid A (100 mL, 2 wt% sodium alginate solution) and mix thoroughly, then use a sterile syringe to squeeze dropwise into the mixed solution of liquid B (150 mL, 1 wt% oligochitosan, citric acid and calcium chloride), control the distance between the needle tip and the surface of solution B to about 50 cm during instillation and continue to stir the solution, after all liquid A is instilled, let the mixed solution stand and harden for 30 minutes to form gel balls, filter, rinse with sterile water and collect the probiotic microcapsules.

[0196] Test Example 1: Probiotic Survival Rate in the Preparation Process of Probiotic Enteric Coated Dry Suspension

[0197] 3.00 g of the probiotic enteric dry suspension 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 ratio of 10 times. 1 mL of the diluted sample was taken at an appropriate dilution ratio, and the total number of viable bacteria in the dry suspension was determined by the pouring method. After anaerobically culturing at 37 ° C. for 48 h on MRS agar medium, the sample was counted.

[0198] Survival rate (%) = number of viable bacteria in dry suspension / theoretical number of viable bacteria added × 100%.

[0199] The survival rates of probiotics in the dry suspension preparation processes of Examples 1-2 and Comparative Examples 1-8 are shown in Table 11 below. The survival rates of probiotics in the preparation processes of Examples 1-2 are all higher than 95%, which are better than the survival rates of live probiotics in Comparative Examples 1-8, indicating that the formulation has less damage to probiotics.

[0200] Table 11. Probiotic survival rate

[0201]

[0202]

[0203] Test Example 2 Probiotics Encapsulation Rate after Adding Water to Probiotics Enteric Coated Dry Suspension

[0204] 5 g of the dry suspension 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, and stirring was stopped after a stable suspension was formed. All the suspensions were transferred to a centrifuge tube, centrifuged at 3000 rpm for 5 min to discard the upper solution, and then PBS was added to resuspend and wash 3 times. The cleaned probiotic microcapsules were all transferred to a triangular flask equipped with glass beads and fixed to 100 mL of PBS solution. After 200 rpm shaking and homogenizing for 15 min, gradient dilution was performed according to a 10-fold ratio, and a suitable dilution was selected to draw 1 mL of diluted sample. The pouring method was used, and the probiotic embedding rate after anaerobic culture at 37 ° C for 48 h on MRS agar medium was counted to determine the probiotic embedding rate after the dry suspension was mixed with water.

[0205] Embedding rate (%) = number of live bacteria in probiotic microcapsules / number of live bacteria in dry suspension × 100%.

[0206] The probiotic encapsulation rates of the probiotic enteric dry suspensions of Examples 1-2 and Comparative Examples 1-8 after adding water and mixing are shown in Table 12 below. The probiotic encapsulation rates of Examples 1-2 are 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 excessively high proportion of probiotic powder added, which exceeded the encapsulation limit of the cross-linking system of the enteric dry suspension microcapsule, part of the bacteria could not effectively form microcapsules, resulting in the embedding rate dropping to 61.1%, 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 could not fully contact the microcapsule cross-linking system and form microcapsule particles with uniform particle size, resulting in the embedding rate dropping 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, and only a small amount of probiotics were 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 from the suspension, and the probiotics were not fully exposed to the cross-linking system of the microcapsule and formed microcapsule particles with uniform particle size. Citric acid was replaced by tartaric acid. Although the effervescent cross-linking system was not destroyed, the acidity of tartaric acid was too strong, and the live bacteria were highly damaged, so the encapsulation rate was reduced to 63.6%; 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, 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 rate 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 calcium carbonate failed to react completely, resulting in the encapsulation rate being reduced to 68.5%; in comparative example 7, the suspending agent was adjusted to affect the state of the suspension system, thereby affecting the formation process of the microcapsules, and the encapsulation rate was reduced to 63.1%; in comparative example 8, the filler was adjusted to also have a certain impact on the suspension system, resulting in the encapsulation rate being reduced to 70.3%.

[0207] Table 12. Probiotic encapsulation rate

[0208]

[0209] Test Example 3 Determination of sedimentation volume ratio of probiotic enteric-coated dry suspension and observation of suspension state

[0210] According to the inspection method of the sedimentation volume ratio of suspension in the 2020 edition of the Chinese Pharmacopoeia, 2.5 g of the probiotic enteric dry suspension of Examples 1-2 and Comparative Examples 1-8 were weighed respectively, added to a stoppered measuring cylinder, added water to a constant volume of 50 mL, plugged, shaken vigorously for 1 minute, and 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:

[0211] Sedimentation volume ratio = H / H0.

[0212] The sedimentation volume ratio is required to be no less than 0.90.

[0213] The suspension state and sedimentation volume ratio of the probiotic enteric 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 requirements for the suspension sedimentation volume ratio specified in the Chinese Pharmacopoeia. Comparative Examples 1-8 adjusted the formulation prescription, which 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 suspension, poor stability of the suspension system, and failed to meet the requirement of the Chinese Pharmacopoeia that the sedimentation volume ratio should be not less than 0.90.

[0214] Table 13. Suspension state and sedimentation volume ratio of probiotic dry suspension

[0215]

[0216]

[0217] Test Example 4 Tolerance of Probiotic Enteric Dry Suspension Microcapsules in Simulated Gastric Juice

[0218] 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 into 45 mL of simulated gastric fluid respectively, mixed thoroughly for 15 s, and shaken on a constant temperature shaker at 37°C and 200 r / min. The number of viable bacteria in the initial and 120 min simulated gastric acid samples was determined and the survival rate was calculated.

[0219] The tolerance of the probiotic microcapsules spontaneously formed by the dry suspension of Examples 1-2 and Comparative Examples 1-8 and water in simulated gastric fluid is shown in Table 14. The survival rate of the unencapsulated Lactobacillus plantarum powder in simulated gastric fluid is only 12.7%. The survival rates of the probiotic microcapsules of Examples 1-2 are all higher than 90%, which meets the requirements. In Comparative Example 1, due to the excessively high proportion of probiotic powder added, the encapsulation limit of the cross-linking system of the enteric dry suspension microcapsule was exceeded, which affected the embedding rate and also had an adverse effect on 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 by 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 had poor integrity and the survival rate was 64.6%, which did not meet the requirements; 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, 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 by tartaric acid. Although the effervescent system was not destroyed, the acidity of tartaric acid was too strong, which caused great damage to the bacterial powder. Even with the protection of the microcapsules, the survival rate was still low. In comparative example 5, citric acid was replaced by calcium citrate, which destroyed the effervescent reaction in the cross-linking system of the microcapsule on the one hand, and reduced the solubility of calcium citrate in the non-acidic environment on the other hand, and reduced the number of free calcium ions, affecting the integrity of the probiotic microcapsule structure, and the survival rate was 30.8%, which did not meet the requirements; in comparative example 6, the cross-linking system of the microcapsule was adjusted, resulting in an imbalance in the ratio of citric acid and calcium carbonate, 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, and the spontaneous formation of probiotic microcapsules could not maintain a good suspension state, which had a certain impact on its formation and the integrity of the microcapsules, resulting in a survival rate reduced to 78.7% and 83.0%, which did not meet the requirements.

[0220] Table 14. Tolerance of probiotic microcapsules in gastric juice

[0221]

[0222]

[0223] Test Example 5 Release of probiotic enteric-coated dry suspension in simulated intestinal fluid

[0224] 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 into 45 mL of simulated intestinal fluid respectively, mixed thoroughly for 15 s, shaken on a constant temperature shaker at 37°C and 200 r / min, and the number of viable bacteria in the simulated intestinal fluid samples was measured at 30 min, 60 min, 90 min, and 120 min.

[0225] The release of the probiotic microcapsules spontaneously formed by the dry suspensions of Examples 1-2 and Comparative Examples 1-8 and water in intestinal fluid is shown in Table 15 below. The probiotic microcapsules of Examples 1-2 have good intestinal release characteristics, and the probiotics in the microcapsules can be continuously released within 1 hour in the simulated artificial intestinal fluid, and have the characteristics of intestinal targeted sustained release. Comparative Example 1-8 has poor microcapsule structural integrity, resulting in rapid release within 30 minutes in the simulated artificial intestinal fluid, and the sustained release effect cannot be achieved.

[0226] Table 15. Release in intestinal fluid

[0227]

[0228]

[0229] Test Example 6 Taste and Swallowing Experience Evaluation

[0230] 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.

[0231] The taste and swallowing experience of the probiotic enteric dry suspensions of Examples 1-2 and Comparative Examples 1-8 are shown in Table 16 below. The taste evaluation and swallowing experience of Examples 1-2 are good. Due to the high proportion of probiotic powder added in Ratio 1, the unencapsulated powder particles are dispersed in the suspension system, and the granular feel is obvious when taken, which does not meet the requirements; Comparative Example 2 replaces calcium carbonate with calcium chloride, and citric acid is not neutralized, resulting in a higher acidity of the suspension and a sour taste. In addition, there is a lack of dispersion 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; Comparative Example 3 removes calcium carbonate, and citric acid is not neutralized, resulting in a higher acidity of the suspension and a sour taste. In addition, due to the lack of calcium ion participation, the probiotic microcapsule structure is loose, the suspension is viscous, the bacterial powder particles are exposed, and the granular feel is obvious, which does not meet the requirements; Comparative Example 4 replaces citric acid with tartaric acid. Although part of the tartaric acid is neutralized, the tartaric acid itself has a strong sour taste and a strong irritation after entering the mouth, which does not meet the requirements; Comparative Example 5 replaces citric acid with calcium citrate, the number of dissolved calcium ions is reduced, most of the probiotics are not wrapped, and the granular feel is obvious. In addition, the lack of acidifiers makes the taste sweet, which does not meet the requirements; Comparative Example 6 adjusts the microcapsule cross-linking system, resulting in an imbalance in the ratio of citric acid to calcium carbonate, and the citric acid is completely consumed, and the taste is sweet. Comparative Examples 7-8 adjust the suspension system and filling excipients, resulting in a low viscosity of the suspension, which cannot well cover the granular feel of the probiotic microcapsules, and does not meet the requirements.

[0232] Table 16. Taste and swallowing experience evaluation results

[0233]

[0234]

[0235] Test Example 7 Stability Test

[0236] 100 g of each dry suspension of Examples 1-2 and Comparative Examples 1-8 were taken and packed into aluminum-plastic bags at 10 g per bag and sealed, and an accelerated stability test was carried out for 3 months at a temperature of 40° C.±2° C. and a relative humidity of 75%±5%.

[0237] The stability of the probiotic enteric dry suspension of Examples 1-2 and Comparative Examples 1-8 is shown in Table 17 below. The accelerated stability survival rate of the probiotic enteric dry suspension samples of Examples 1-2 exceeds 85% for 3 months, which meets the requirements. The change in the formulation of Comparative Examples 1-8 reduces the protective effect of the probiotics therein, and the accelerated stability survival rate of live bacteria does not meet the evaluation standard requirement of 75%.

[0238] Table 17. Accelerated stability test results

[0239]

[0240]

[0241] Comparison of Experimental Example 7 with Conventional Probiotic Microcapsule Preparation Process

[0242] Probiotic microcapsules were prepared according to the method of Comparative Example 9, and their embedding rate, 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 by the enteric-coated dry suspension in Example 1.

[0243] The embedding rate, gastric acid resistance and intestinal release characteristics of the probiotic microcapsules of Example 1 and Comparative Example 9 are compared, 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 rate, gastric acid resistance and intestinal sustained-release targeted release characteristics, but the preparation process is significantly simplified, which is convenient for implementation and reduces the cost of probiotic microcapsule preparation.

[0244] Table 18. Comparison of Example 1 and Comparative Example 9

[0245] Sample name Embedding rate Gastric acid resistance survival rate Intestinal release time Example 1 85.3％ 91.6％ Gradually releases completely within 1 hour Comparative Example 8 89.2％ 92.0％ Gradually releases completely within 1 hour

[0246] 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 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.

2. The probiotic enteric dry suspension according to claim 1, characterized in that: The probiotics of the probiotic powder are selected from at least one of bifidobacterium, lactobacillus, lactobacillus casei, lactobacillus plantarum, streptococcus and lactococcus.

3. The probiotic enteric dry suspension according to claim 1, characterized in that: The microcapsule cross-linking system comprises sodium alginate, oligochitosan, citric acid and calcium carbonate.

4. The probiotic enteric dry suspension according to claim 1, characterized in that: The microcapsule cross-linking system comprises the following components, calculated by weight: 1-5 parts of sodium alginate, 1-5 parts of chitosan, 15-35 parts of citric acid and 10-20 parts of calcium carbonate.

5. The probiotic enteric dry suspension according to claim 1, characterized in that: The filler is selected from at least one of sucrose, lactose, glucose and maltodextrin.

6. The probiotic enteric dry suspension according to claim 1, characterized in that: The aromatic agent is selected from at least one of strawberry flavor, orange flavor, apple flavor, grape flavor and banana flavor.

7. The probiotic enteric dry suspension according to claim 1, characterized in that: The colorant is selected from at least one of yellow iron oxide, red iron oxide and purple iron oxide.

8. The probiotic enteric dry suspension according to claim 1, characterized in that: The suspending agent is selected from at least one of xanthan gum, hydroxypropyl methylcellulose, polyvinyl pyrrolidone and gum arabic.

9. The method for preparing the probiotic enteric dry suspension according to any one of claims 1 to 8, 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: Mix the aromatic agent, the colorant and the suspending agent with the filler in a weight ratio of 1:8-12 to obtain a mixture 2; Step 3: mixing the microcapsule cross-linking system with the mixture 1 to obtain a mixture 3; Step 4: Mix the mixture 2, the mixture 3 and the remaining filler to obtain the probiotic enteric dry suspension.

10. Use of the probiotic enteric dry suspension according to any one of claims 1 to 8 in the preparation of oral probiotic preparations.

Images