A kind of probiotic micro-pill and preparation method thereof

Abstract

The invention discloses a probiotic micro-pill and a preparation method thereof. The probiotic micro-pill comprises a pellet core and a coating layer wrapped outside the pellet core; the pellet core is composed of 30-80% of probiotic powder and 20-70% of auxiliary materials by weight, the probiotic powder is selected from two or more of plant lactobacillus, rhamnosus lactobacillus, paracasei lactobacillus and reuteri mucosa lactobacillus, and the total lactic acid bacteria content of the probiotic powder is ≥2.5×10 11 CFU / g. The present invention adopts a mild pellet preparation process to prepare the probiotic pellet product, avoiding the high-temperature heating process, effectively ensuring the bacterial load of the overall pellet product after preparation, and at the same time, the probiotic pellet can effectively isolate moisture and oxygen, solving the problem that the probiotics are afraid of water, so that the survival rate of the probiotics is high during storage and the shelf life of the product is extended. Acid-resistant ingredients are added to the coating layer of the pellet to solve the problem that the probiotics are not resistant to gastric acid after taking them, and ensure that the probiotics reach the intestines and colonize to exert corresponding effects.

Description

Technical Field

[0001] The invention belongs to the technical field of probiotic preparation, and particularly relates to a probiotic micropill and a preparation method thereof. Background Art

[0002] Probiotics are a class of active microorganisms that can colonize in the human body and are beneficial to the host. The intake of probiotics has many health benefits, such as: (1) adhering to the host's intestinal epithelium, regulating the balance of intestinal flora, reducing the adhesion of pathogenic bacteria, and inhibiting the proliferation or activity of pathogenic bacteria; (2) promoting host mucosal repair and enhancing systemic immune function; (3) degrading biomacromolecules and promoting gastrointestinal digestion and absorption. Therefore, probiotics have been used in ordinary foods, health products, medical foods and pharmaceutical products. Usually, only the number of live cells of probiotics in the food matrix needs to reach 10 6 ~10 7 CFU / mL, in order to fully exert its beneficial effects on the human body. At the same time, probiotics are very sensitive to adverse external environments. Oxygen, temperature, humidity, etc. have a great impact on the survival of probiotics, which can lead to a significant reduction in the number of live bacteria during production, transportation and sales. In addition, most probiotics cannot withstand the destruction of gastric acid, bile salts and various digestive enzymes after entering the digestive tract, and it is difficult to maintain a sufficient number of live bacteria to reach the intestines for colonization and thus play a role. Obviously, probiotic products are extremely susceptible to environmental influences during processing, storage and entry into the human gastrointestinal tract, and their survival and vitality decrease rapidly. Therefore, it is crucial to maintain the ability of probiotics to survive in food and the upper gastrointestinal tract and release them in the colon.

[0003] Prebiotics are substances that can be selectively utilized and transformed by intestinal flora under the premise of ensuring the health of the host. Currently, the widely used prebiotics mainly include inulin, oligofructose and oligogalactose. Prebiotics can be used as nutrients for probiotics to promote the proliferation of probiotics, and produce synergistic effects, thereby exerting their physiological functions, such as regulating the body's immunity, resisting pathogens, improving the intestinal environment, regulating lipid metabolism, and increasing mineral absorption. For example, patent document CN1545936A reports an edible composition containing active probiotics and a preparation method thereof, which, by weight percentage, includes 0.5-50% of live bacterial powder protected by microcapsules, 4.5-99.5% of prebiotics and 0-95% of food additives.

[0004] In recent years, different types of prebiotic preparations have received more and more attention, especially the construction of micro-pellet type embedding system, which can not only ensure the structural stability of prebiotics during food processing and storage, but also ensure its targeted release and effective absorption in the gastrointestinal digestive environment. After the probiotic micro-pellet preparation, it can be isolated from the external environment to a certain extent, and its tolerance to adverse environments can be improved. In addition, by selecting a wall material with special properties, the ability of probiotics to locate and slowly release in the intestine after microencapsulation can be given. In the prior art, such as patent document CN109498661A, a compound probiotic enteric-coated micro-pellet and a preparation method thereof are provided, which is composed of lactic acid bacteria, prebiotics, and enteric coating materials. The process is relatively extensive during the preparation process, and the number of viable bacteria of the product is not detected.

[0005] In fact, some products on the market claim to add a large amount of probiotics, but the results of live bacteria testing of probiotics at the factory usually show that the number is only 1.0 to 5.0 × 10 9 Billions of cfu / g, which makes it difficult to ensure that probiotics enter the gastrointestinal tract and survive the gastric acid and bile salt environment, so its activity is difficult to guarantee, and its efficacy naturally decreases or even becomes negligible.

[0006] In view of this, the present invention is proposed. Summary of the invention

[0007] In view of the shortcomings of the prior art, the purpose of the present invention is to provide a probiotic micropill and a preparation method thereof, which adopts a mild micropill preparation process, avoids a high-temperature heating process, and effectively ensures the bacterial load of the overall micropill product after preparation. At the same time, the probiotic micropill can effectively isolate moisture and oxygen, solve the problem that probiotics are afraid of water, so that the survival rate of probiotics is high during storage and the shelf life of the product is extended; acid-resistant ingredients are added to the coating layer of the micropill to solve the problem that probiotics are not resistant to gastric acid after taking them, and ensure that the probiotics reach the intestines and colonize before exerting their corresponding effects.

[0008] In order to achieve the above object, the present invention adopts the following technical solution:

[0009] In a first aspect, the present invention provides a probiotic micro-pellet, comprising a pellet core and a coating layer wrapped around the pellet core;

[0010] In terms of weight percentage, the pellet core is composed of 30-80% probiotic powder and 20-70% auxiliary materials, wherein the probiotic powder is selected from two or more of Lactobacillus plantarum, Lactobacillus rhamnosus, Lactobacillus paracasei, and Lactobacillus reuteri, and the total lactic acid bacteria content of the probiotic powder is ≥2.5×10 11 CFU / g.

[0011] As a preferred embodiment of the technical solution of the present invention, the coating layer is composed of 10-65% methyl cellulose, 10-65% hydroxypropyl methyl cellulose, 5-20% sodium alginate, 0.5-5% magnesium stearate and 0.01-5% food coloring in weight percentage.

[0012] As a preferred embodiment of the technical solution of the present invention, the auxiliary materials are composed of microcrystalline cellulose, sodium carboxymethyl cellulose, maltodextrin and prebiotic components.

[0013] As a preferred embodiment of the technical solution of the present invention, the prebiotic component is selected from one or more of xylooligosaccharides, stachyose, galacto-oligosaccharides and inulin.

[0014] As a preferred embodiment of the technical solution of the present invention, the auxiliary materials are composed of the following by weight percentage: 5-20% microcrystalline cellulose, 0.5-5% sodium carboxymethyl cellulose, 30-50% maltodextrin, and 20-50% prebiotic components.

[0015] In a second aspect, the present invention also provides a method for preparing the above-mentioned probiotic micropills, comprising the following steps:

[0016] S1. Preparation of pellet core

[0017] S11, preparation of masterbatch: after prebiotic components and maltodextrin are uniformly mixed, purified water is added, and the mixture is granulated, dried, and sieved to obtain masterbatch; illustratively, granulation can be performed using a swing granulator, and specifically, a 36-50 mesh screen can be used for wet granulation;

[0018] S12, encapsulation of the probiotic layer: after adding the masterbatch obtained in step S11 to the granulator, start the fan and the turntable, mix the probiotic powder, microcrystalline cellulose and sodium carboxymethyl cellulose evenly, and then add them to the turntable, and spray purified water at the same time. After encapsulation, stop spraying, and continue to round the particles until the particle size increases; illustratively, the granulator can be a centrifugal granulator, the air intake frequency of the centrifugal granulator is 10-18 Hz, the rotation speed is 45-60 r / min, the pure water is sprayed by an atomizing spray gun, the flow rate is controlled at 20-60 mL / min, and the spray gun pressure is controlled at 0.10-0.35 MPa, and the final particle size of the particles is between 10 and 40 meshes;

[0019] S13, drying of the pellet core: turning on the heating device, drying and sieving the material in step S12 to obtain the pellet core; illustratively, before turning on the device, the speed of the turntable can be reduced (e.g., to 30-45 r / min) so that the material in step S12 maintains a flowing state;

[0020] S2. Coating

[0021] S21, preparation of coating solution: adding the coating layer raw materials into purified water according to the proportion, stirring evenly and filtering to obtain the coating solution for standby use;

[0022] S22, coating: adding the pellets obtained in step S13 into a fluidized bed, spraying the coating liquid into the fluidized bed, and performing coating treatment; illustratively, the fluidized bed may be preheated before adding the pellets; the inlet air temperature is set to 100-130° C., the outlet air temperature is set to 40-50° C., the fan frequency is 30-50 Hz, and the fluidized bed preheating time is 20-40 min;

[0023] S23, drying and sieving: After the coating weight gain reaches the predetermined range, stop spraying the coating liquid, dry and sieve to obtain the probiotic micro-pellets. It should be emphasized that before drying, the fan frequency can be reduced to 10-20Hz, and the drying process is stopped when the moisture content of the coated particles is ≤3.5%.

[0024] As a preferred embodiment of the technical solution of the present invention, in step S11, the drying temperature is 50-60° C., the drying time is 6-10 hours, and the mesh size of the sieve is 20-50 meshes;

[0025] In step S12, the air intake frequency of the fan is 10-18 Hz, and the speed is 40-60 r / min; purified water is sprayed by a spray gun, the flow rate is 20-60 mL / min, and the pressure is 0.10-0.35 MPa; the particles are rounded to 10-40 mesh;

[0026] As a preferred embodiment of the technical solution of the present invention, in step S13, the drying temperature is 45-55°C, the drying time is 30-60 minutes, and the sieving mesh number is 10-40 meshes.

[0027] As a preferred embodiment of the technical solution of the present invention, in step S21, the total concentration of the coating solution is 8 to 12 wt%.

[0028] As a preferred embodiment of the technical solution of the present invention, in step S22, the inlet air temperature is set to 100-130°C, the outlet air temperature is set to 40-50°C, the fan frequency is 30-50HZ, and the fluidized bed preheating time is 20-40min.

[0029] As a preferred embodiment of the technical solution of the present invention, in step S23, the coating weight gain range is 10-20%; and the sieving mesh number is 14-36 meshes.

[0030] In a third aspect, the present invention also hopes to use the prepared probiotic micropellets in the preparation of foods, nutritional dietary supplements, health foods or medicines; for example, in the preparation of pharmaceutical products for preventing and controlling obesity.

[0031] Compared with the prior art, the present invention has the following beneficial effects:

[0032] (1) In view of the problem that the existing preparation process is easily affected by processing factors such as high temperature and humidity, resulting in low survival rate and small number of live bacteria of probiotics, the present invention avoids the high-temperature heating process through a mild micro-pellet preparation process. The material used has strong acid resistance, which can enable the embedded lactic acid bacteria to pass through gastric acid smoothly to colonize in the intestine, thereby effectively improving the survival rate of probiotics.

[0033] (2) The moisture content of the probiotic micropellets prepared by the present invention is ≤3.5%, and the bacterial loading rate of the finished product can reach 1.0×10 11 CFU / g; the probiotic pellets obtained after preparation can effectively block the effects of temperature, humidity and oxygen on the activity of probiotics, which is beneficial to the maintenance of the number of live probiotics. Studies have shown that the probiotic pellets prepared by the present invention have a probiotic activity retention rate of more than 87% after being stored at 4°C for 12 months; the probiotic activity retention rate after being stored at 25°C for 12 months is more than 75%, which can effectively ensure the activity of probiotics, avoid the requirements of conventional cold chain transportation, low temperature storage and other harsh conditions for no loss of bacterial activity, reduce energy consumption, improve the production efficiency of enterprises and reduce production costs.

[0034] (3) The probiotic micropellet product prepared by the present invention can be widely used in food, nutritional dietary supplements, health foods and medicines.

[0035] (4) The method for preparing probiotic micro-pellets provided by the present invention has a simple process and is suitable for large-scale industrial production. During the production process, the production process is mild, the processing temperature of the probiotic raw materials does not exceed 55°C, and the heating time is short, which can reduce the bacterial loss rate of the probiotics during the entire micro-pellet processing process as much as possible. In addition, the probiotic micro-pellets have a beautiful appearance. By adding ingredients of different pigments, probiotic micro-pellets products with different colors can be prepared, which increases the diversity of probiotic preparation forms and can realize industrial production.

[0036] (5) The micropellet product prepared by the present invention can effectively regulate the intestinal function of the human body, affect the metabolism of the host by regulating the intestinal flora, and thus prevent and control obesity and reduce the occurrence of obesity complications.

[0037] In summary, the present invention optimizes the probiotic flora, auxiliary materials and coating materials, and combines them with a specific pill production process to obtain a micro-pill product with stable properties and high bacterial viability, which has great market sales prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0038] Figure 1 This is a product diagram of Example 1;

[0039] Figure 2 This is the product diagram of Example 2;

[0040] Figure 3 This is the product diagram of Example 3;

[0041] Figure 4 This is the product picture of Example 4. DETAILED DESCRIPTION

[0042] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0043] Unless otherwise specified, the chemical reagents used in the present invention are purchased from the market. Some examples of raw materials are as follows:

[0044] Table 1 Raw material purchase information

[0045]

[0046]

[0047] It should be particularly emphasized that the preparation methods of the culture medium, artificial gastrointestinal fluid and the sources of the strains involved in the test examples of the present invention are as follows:

[0048] (1) Preparation of MRS liquid culture medium and artificial gastrointestinal fluid

[0049] MRS liquid culture medium: 10g soy protein, 10g beef extract, 5g yeast extract, 20g glucose, 2g diammonium citrate, 5g sodium acetate, 2g dipotassium hydrogen phosphate, 0.58g magnesium sulfate, 0.25g manganese sulfate, 80mL Tween, 1000mL deionized water, pH 6.4, sterilization at 121℃ for 20min.

[0050] Preparation of artificial gastric juice: 8 mL of 9%-10% hydrochloric acid, dilute with water to a pH value of 3.0. Add 0.1 g of pepsin to every 10 mL of solution, mix well, and filter with a 0.2 μm sterile filter.

[0051] Preparation of artificial intestinal fluid: Weigh 4 g of potassium dihydrogen phosphate, add 250 mL of water to dissolve, adjust the pH to 6.8 with 0.4% sodium hydroxide solution, dilute to 500 mL with water, add 0.1 g of trypsin to every 10 mL of solution, shake the solution well, and filter with a 0.2 μm sterile filter.

[0052] (2) Standard strains and culture medium for stool culture

[0053] Enterococcus 44146, eosin-methylene blue agar; Enterobacter 9086, sodium azide-crystal violet-esculin agar; Lactobacillus 1794, LBS agar; Bifidobacterium 21065, BBL agar; Clostridium perfringens 64701, SPS agar.

[0054] Example 1

[0055] In this embodiment, the raw materials for preparing a probiotic micro-pill provided are as follows:

[0056] Table 2 Example 1 Raw material amount

[0057]

[0058]

[0059] In this embodiment, a method for preparing the above-mentioned probiotic micropills is also provided, comprising the following steps:

[0060] S1. Preparation of pellet core

[0061] S11, preparation of masterbatch: prebiotic components and maltodextrin were mixed evenly and purified water was added to obtain a soft material, which was granulated by a swing granulator (36 mesh screen, wet granulation), dried (55° C., 8 h), and sieved through 50 mesh to obtain a masterbatch;

[0062] S12, coating of the probiotic layer: after adding the masterbatch prepared in step S11 into the centrifugal granulator, start the fan and the turntable (the air inlet frequency is 15 Hz, the speed is 50 r / min), mix the probiotic powder, microcrystalline cellulose and sodium carboxymethyl cellulose evenly and add them to the turntable, and at the same time spray purified water at 35 mL / min and 0.2 MPa pressure. When the mixed powder has been coated, stop spraying, and continue to round the particles until the particle size increases to about 35 mesh;

[0063] S13, drying of the pellet core: reducing the rotation speed of the turntable to about 30 r / min, so that the material in step S12 maintains a flowing state, turning on the heating device (the inlet air temperature is 50° C., the heating time is 45 min), drying the material in step S12, and sieving it through 35 meshes to obtain the pellet core;

[0064] S2. Coating

[0065] S21, preparation of coating solution: adding the coating layer raw materials into purified water according to the proportion, stirring evenly and filtering to obtain a 10wt% coating solution for standby use;

[0066] S22, coating: preheat the fluidized bed first, add the pellet cores obtained in step S13 into the fluidized bed, start the induced draft fan and the heater, spray the coating liquid, and perform coating treatment; wherein the wind temperature is set to 110°C, the outlet air temperature is set to 45°C, the fan frequency is 40Hz, and the preheating time is 30min;

[0067] S23, drying and sieving: when the coating weight gain reaches about 15%, stop spraying the coating liquid, reduce the fan frequency to 15 Hz, maintain the convection state, so that the dry air flow dries the coated particles, stop when the moisture content of the coated particles is ≤3.5%, and sieve through 32 mesh to obtain the probiotic microcapsules.

[0068] Example 2

[0069] In this embodiment, the raw materials for preparing a probiotic micro-pill provided are as follows:

[0070] Table 3 Example 2 Raw Material Amount

[0071]

[0072] In this embodiment, a method for preparing the above-mentioned probiotic micropills is also provided, comprising the following steps:

[0073] S1. Preparation of pellet core

[0074] S11, preparation of masterbatch: prebiotic components and maltodextrin were mixed evenly and purified water was added to obtain a soft material, which was granulated by a swing granulator (36 mesh screen, wet granulation), dried (55° C., 8 h), and sieved through 50 mesh to obtain a masterbatch;

[0075] S12, coating of the probiotic layer: after adding the masterbatch prepared in step S11 into the centrifugal granulator, start the fan and the turntable (the air inlet frequency is 15 Hz, the speed is 50 r / min), mix the probiotic powder, microcrystalline cellulose and sodium carboxymethyl cellulose evenly and add them to the turntable, and at the same time spray purified water at 35 mL / min and 0.2 MPa pressure. When the mixed powder has been coated, stop spraying, and continue to round the particles until the particle size increases to about 35 mesh;

[0076] S13, drying of the pellet core: reducing the rotation speed of the turntable to about 30 r / min, so that the material in step S12 maintains a flowing state, turning on the heating device (the inlet air temperature is 50° C., the heating time is 45 min), drying the material in step S12, and sieving it through 35 meshes to obtain the pellet core;

[0077] S2. Coating

[0078] S21, preparation of coating solution: adding the coating layer raw materials into purified water according to the proportion, stirring evenly and filtering to obtain a 10wt% coating solution for standby use;

[0079] S22, coating: preheat the fluidized bed first, add the pellet cores obtained in step S13 into the fluidized bed, start the induced draft fan and the heater, spray the coating liquid, and perform coating treatment; wherein the wind temperature is set to 110°C, the outlet air temperature is set to 45°C, the fan frequency is 40Hz, and the preheating time is 30min;

[0080] S23, drying and sieving: when the coating weight gain reaches about 15%, stop spraying the coating liquid, reduce the fan frequency to 15 Hz, maintain the convection state, so that the dry air flow dries the coated particles, stop when the moisture content of the coated particles is ≤3.5%, and sieve through 32 mesh to obtain the probiotic microcapsules.

[0081] Example 3

[0082] In this embodiment, the raw materials for preparing a probiotic micro-pill provided are as follows:

[0083] Table 4 Example 3 Raw material amount

[0084]

[0085]

[0086] In this embodiment, a method for preparing the above-mentioned probiotic micropills is also provided, comprising the following steps:

[0087] S1. Preparation of pellet core

[0088] S11, preparation of masterbatch: prebiotic components and maltodextrin were mixed evenly and purified water was added to obtain a soft material, which was granulated by a swing granulator (36 mesh screen, wet granulation), dried (55° C., 8 h), and sieved through 50 mesh to obtain a masterbatch;

[0089] S12, coating of the probiotic layer: after adding the masterbatch prepared in step S11 into the centrifugal granulator, start the fan and the turntable (the air inlet frequency is 15 Hz, the speed is 50 r / min), mix the probiotic powder, microcrystalline cellulose and sodium carboxymethyl cellulose evenly and add them to the turntable, and at the same time spray purified water at 35 mL / min and 0.2 MPa pressure. When the mixed powder has been coated, stop spraying, and continue to round the particles until the particle size increases to about 35 mesh;

[0090] S13, drying of the pellet core: reducing the rotation speed of the turntable to about 30 r / min, so that the material in step S12 maintains a flowing state, turning on the heating device (the inlet air temperature is 50° C., the heating time is 45 min), drying the material in step S12, and sieving it through 35 meshes to obtain the pellet core;

[0091] S2. Coating

[0092] S21, preparation of coating solution: adding the coating layer raw materials into purified water according to the proportion, stirring evenly and filtering to obtain a 10wt% coating solution for standby use;

[0093] S22, coating: preheat the fluidized bed first, add the pellet cores obtained in step S13 into the fluidized bed, start the induced draft fan and the heater, spray the coating liquid, and perform coating treatment; wherein the wind temperature is set to 110°C, the outlet air temperature is set to 45°C, the fan frequency is 40Hz, and the preheating time is 30min;

[0094] S23, drying and sieving: when the coating weight gain reaches about 15%, stop spraying the coating liquid, reduce the fan frequency to 15 Hz, maintain the convection state, so that the dry air flow dries the coated particles, stop when the moisture content of the coated particles is ≤3.5%, and sieve through 32 mesh to obtain the probiotic microcapsules.

[0095] Example 4

[0096] In this embodiment, the raw materials for preparing a probiotic micro-pill provided are as follows:

[0097] Table 5 Example 4 Raw material amount

[0098]

[0099] In this embodiment, a method for preparing the above-mentioned probiotic micropills is also provided, comprising the following steps:

[0100] S1. Preparation of pellet core

[0101] S11, preparation of masterbatch: prebiotic components and maltodextrin were mixed evenly and purified water was added to obtain a soft material, which was granulated by a swing granulator (36 mesh screen, wet granulation), dried (55° C., 8 h), and sieved through 50 mesh to obtain a masterbatch;

[0102] S12, coating of the probiotic layer: after adding the masterbatch prepared in step S11 into the centrifugal granulator, start the fan and the turntable (the air inlet frequency is 15 Hz, the speed is 50 r / min), mix the probiotic powder, microcrystalline cellulose and sodium carboxymethyl cellulose evenly and add them to the turntable, and at the same time spray purified water at 35 mL / min and 0.2 MPa pressure. When the mixed powder has been coated, stop spraying, and continue to round the particles until the particle size increases to about 35 mesh;

[0103] S13, drying of the pellet core: reducing the rotation speed of the turntable to about 30 r / min, so that the material in step S12 maintains a flowing state, turning on the heating device (the inlet air temperature is 50° C., the heating time is 45 min), drying the material in step S12, and sieving it through 35 meshes to obtain the pellet core;

[0104] S2. Coating

[0105] S21, preparation of coating solution: adding the coating layer raw materials into purified water according to the proportion, stirring evenly and filtering to obtain a 10wt% coating solution for standby use;

[0106] S22, coating: preheat the fluidized bed first, add the pellet cores obtained in step S13 into the fluidized bed, start the induced draft fan and the heater, spray the coating liquid, and perform coating treatment; wherein the wind temperature is set to 110°C, the outlet air temperature is set to 45°C, the fan frequency is 40Hz, and the preheating time is 30min;

[0107] S23, drying and sieving: when the coating weight gain reaches about 15%, stop spraying the coating liquid, reduce the fan frequency to 15 Hz, maintain the convection state, so that the dry air flow dries the coated particles, stop when the moisture content of the coated particles is ≤3.5%, and sieve through 32 mesh to obtain the probiotic microcapsules.

[0108] Comparative Example 1

[0109] In this comparative example, a probiotic product is provided, and the formula is as follows:

[0110] Table 6 Comparative Example 1 Formula

[0111]

[0112]

[0113] In this comparative example, a preparation method of the above-mentioned probiotic product is also provided, comprising: weighing according to the formula amount, and mixing in a three-dimensional mixer for 60 minutes.

[0114] Comparative Example 2

[0115] In this comparative example, a probiotic product is provided, and the formula is as follows:

[0116] Table 7 Comparative Example 2 Formulation

[0117] Probiotic Mix Powder Dosage / g Lactobacillus plantarum CCFM8661 20 Lactobacillus rhamnosus GloryLG12 25 Lactobacillus paracasei GloryLP16 15 Lactobacillus reuteri GloryLR15 15 Maltodextrin 45

[0118] In this comparative example, a preparation method of the above-mentioned probiotic product is also provided, comprising: weighing according to the formula amount, and mixing in a three-dimensional mixer for 60 minutes.

[0119] Test Example 1: Bacterial Activity Test

[0120] The products obtained in Examples 1 to 4 and Comparative Examples 1 to 2 were tested for bacterial activity, and the results are shown in Table 8.

[0121] Table 8 Preservation rate of bacterial activity after preparing probiotic products

[0122]

[0123] As can be seen from the above table, the preparation method of the probiotic micro-pill provided by the present invention has a live bacteria survival rate of more than 70.5% after being made into a micro-pill dosage form. The probiotic mixed powder product in the comparative example has a relatively large bacterial survival rate because it is only a simple raw material mixing, and there is no excessive overall loss. However, it can be seen that the probiotic micro-pill prepared by the present invention has a higher bacterial survival retention rate, which is higher than the general products on the market.

[0124] Test Example 2: Probiotic Stability Test

[0125] The probiotic products prepared in Examples 1 to 4 and Comparative Examples 1 to 2 were stored under refrigeration (4° C.) and room temperature (25° C.) for 12 months, respectively, and the survival rate of the probiotics in the samples was measured at regular intervals. The results are shown in Tables 9 and 10 below.

[0126] It can be seen from the table that the probiotics in the embodiments after storage at 4°C for 12 months have a live retention rate of 87.7% to 88.8%, and the probiotics in the comparative example have a live retention rate of 49.0% to 51.6%; the probiotics in the embodiments after storage at 25°C for 12 months have a live retention rate of 75.6% to 77.4%, and the probiotics in the comparative example have a live retention rate of 10.4% to 15.0%.

[0127] Table 9 Stability changes of probiotic products under refrigerated (4°C) storage conditions

[0128]

[0129] Table 10 Stability changes of probiotic products under room temperature (25°C) storage conditions

[0130]

[0131]

[0132] Test Example 3: Probiotic micropellets gastric juice and intestinal juice release performance test

[0133] 4 g of each of the probiotic preparations obtained in Comparative Examples 1-2 and Examples 1-2 were placed in 200 mL of artificial gastric juice at 37° C. and a speed of 100 r / min. After 120 min, the mixture was taken out and transferred to 200 mL of artificial intestinal juice, also at 37° C. and a speed of 100 r / min, and stirred for 120 min. The observation results were recorded and shown in Table 11.

[0134] It can be seen from the table that the probiotic pellets prepared in the present invention can withstand the effects of artificial gastric juice and artificial intestinal juice for a long time, indicating that the product prepared in the embodiment has stable performance. In comparison, the comparative example has a shorter tolerance time and poorer performance.

[0135] Table 11 Release performance test of probiotic micropellets in gastric juice and intestinal juice

[0136]

[0137]

[0138] Test Example 4: Bile salt resistance and artificial gastric juice tolerance test of probiotic micropellets

[0139] Add ox bile salt to MRS liquid culture medium to make its concentration 0% (control), 0.1%, 0.3%, 0.5%, respectively, as bile salt liquid culture medium. Take 1.5g of the probiotic preparation obtained in the above Examples 1 to 4 and Comparative Examples 1 to 2, respectively, place them in 250mL stoppered conical flasks, add 100mL of bile salt liquid culture medium of different concentrations, stir magnetically, place in a 37°C constant temperature incubator for 24h, and then determine the number of viable bacteria by bacterial counting method. The results of the bile salt resistance characteristics of the probiotic micropellet preparation are shown in Table 12.

[0140] As can be seen from the table, the probiotic products of the embodiment have good growth conditions at bile salt concentrations of 0.1% and 0.3%, and there is no significant difference with the control group (0%); when the probiotics grow in a bile salt concentration of 0.5%, the number of viable bacteria is significantly reduced. It can be seen that the probiotic microcapsules can tolerate a bile salt concentration of 0.3%. The probiotic product of the comparative example has a significantly reduced number of viable bacteria at a bile salt concentration of 0.3%, and its bile salt resistance is poor.

[0141] Table 12 The number of viable bacteria of probiotic micropellets cultured in different concentrations of bile salt for 24 hours

[0142]

[0143]

[0144] Note: The calculation of survival rate is based on the number of viable bacteria at 0% bile salt concentration.

[0145] 1.5 g of the probiotic pellet preparations obtained in Examples 1 to 4 and Comparative Examples 1 to 2 were placed in 200 mL of artificial gastric juice, respectively, and cultured at 120 r / min in a shaker at 37° C. Bacteria were counted at 0 h, 1 h, and 3 h. The results of the tolerance of the probiotic pellet preparations to artificial gastric juice are shown in Table 13.

[0146] As can be seen from the table, the number of live bacteria in the probiotic product of the embodiment is basically unaffected when grown in artificial gastric juice for 1 hour, and its number of live bacteria decreases significantly after growing for 3 hours. It can be seen that the probiotic micro-pellet preparation can resist the digestion of artificial gastric juice. However, the number of live bacteria in the probiotic product of the comparative example decreases significantly when grown in artificial gastric juice for 1 hour, and the resistance to artificial gastric juice is poor.

[0147] Table 13 Number of viable bacteria of probiotic micropellets in artificial gastric juice

[0148]

[0149] Note: The calculation of survival rate is based on the number of live bacteria at 0h.

[0150] Experimental Example 5: Effect of probiotic pellets on the body weight of mice

[0151] 75 clean-grade female BALB / c mice (Suzhou Slake Experimental Animal Technology Service Company), weighing about 25g, were randomly divided into 5 groups, 15 in each group. After 1 week of adaptive feeding, a 28-day (4-week) feeding experiment of mouse probiotic micropellet preparation feed was carried out. The feeding environment temperature was 22±1°C, the humidity was 50±10%, and the mice were free to eat and drink water during the whole feeding process according to 12h light / 12h dark. The specific grouping and treatment conditions are shown in Table 14.

[0152] All animal feeds are special feeds, specially customized and produced by Nantong Trophi Technology Co., Ltd. in Nantong, Jiangsu Province. The control group mouse feed is a special feed made by adding the probiotic micro-pellet preparation obtained in Comparative Example 1 (the proportion of probiotic micro-pellets is 3%), the 1st group of mouse feed is a special feed made by adding the probiotic micro-pellet preparation obtained in Example 1 (the proportion of probiotic micro-pellets is 3%), the 2nd group of mouse feed is a special feed made by adding the probiotic micro-pellet preparation obtained in Example 2 (the proportion of probiotic micro-pellets is 3%), the 3rd group of mouse feed is a special feed made by adding the probiotic micro-pellet preparation obtained in Example 3 (the proportion of probiotic micro-pellets is 3%), and the 4th group of mouse feed is a special feed made by adding the probiotic micro-pellet preparation obtained in Example 4 (the proportion of probiotic micro-pellets is 3%). All of the above feeds are stored in vacuum sealed bags and stored at -20°C until use.

[0153] Table 14 Grouping and treatment of mice

[0154]

[0155] The weight of mice (fasting) was measured every 7 days and the data were recorded. The results are shown in Table 15. The weight of each group of mice increased every week. Compared with the control group, the weight gain of mice in the probiotic pellet preparation group slowed down. It can be seen that the probiotic diet in the experimental group can delay the weight gain of animals to a certain extent.

[0156] Table 15 Mouse weight change results

[0157]

[0158]

[0159] Note: No mice died.

[0160] Experimental Example 6: Effects of probiotic pellets on intestinal flora of mice

[0161] Before administering the test substance, 0.1 g of mouse feces was collected aseptically, added with a certain amount of sterile saline, homogenized by electric current, and diluted 10-fold to 10 -8 , select appropriate dilutions from the stock solution and inoculate on eosin-methylene blue agar, sodium azide-crystal violet-esculin agar, LBS agar, BBL agar, and SPS agar medium. After cultivation, the colonies were counted by colony morphology, Gram staining, biochemical reaction, etc., and the number of bacteria per gram of wet feces was calculated, and the logarithm was taken for statistical calculation. 24 hours after the last administration of the test substance, feces were taken aseptically and the intestinal flora was detected using the same method as above. The statistical results are shown in Table 16.

[0162] The number of intestinal bacteria colonies per gram of wet feces in the control group did not change significantly between 0 and 28 days. The content of bifidobacteria in the probiotic pellet feed group was significantly higher than that on 0 day and also significantly higher than that in the control group, and the content of other indicator bacteria did not differ significantly.

[0163] Table 16 Results of mouse intestinal flora detection

[0164]

[0165] Experimental Example 7: Effect of probiotic pellets on cecal weight in mice

[0166] Although the cecum of animals is shorter than other intestines, it is rich in microorganisms and plays a great role in the digestion and absorption of food. Therefore, when the cecum is developed, the digestive performance of the animal body will be relatively improved. On the 7th, 14th, 21st and 28th days, 3 mice in each group were randomly killed and the cecum was collected and weighed. The data of cecal weight were summarized and calculated, and the results are shown in Table 17.

[0167] It can be found from the table that feeding probiotics had a tendency to increase the cecum weight of mice during the entire experimental period, and the probiotics-fed groups were significantly higher than the control group.

[0168] Table 17 Results of changes in mouse cecum weight

[0169]

[0170] Test Example 8: Effect of probiotic micropellets on human body fat

[0171] 100 volunteers aged 24-52 years old, with a body mass index (BMI) of 24-30, a body fat rate of 23-30% for men, and a body fat rate of 29-35% for women were selected as test subjects. Pregnant women, those with severe gastrointestinal diseases, and those with diabetic ketoacidosis in the past three months were excluded from the test subjects. The test subjects were randomly divided into 4 groups, with 25 in each group. During the test, the test subjects maintained good living and eating habits and did not take hypoglycemic and lipid-lowering drugs.

[0172] The probiotic microcapsules prepared in Examples 1 to 4 of the present invention were respectively administered to 4 groups of test subjects, with the administration method of 2 g / time, twice a day, directly orally. The efficacy was evaluated after taking it continuously for 1 month. The efficacy evaluation index was as follows: the changes in the body fat percentage of the test subjects before and after the test were recorded, and the body fat percentage change rate was calculated. The body fat percentage change rate = (body fat percentage before the test - body fat percentage after the test) / body fat percentage before the test × 100%. The efficacy evaluation standard: if the body fat percentage change rate is ≥5%, it is markedly effective; if the body fat percentage change rate is 1%≤<5%, it is effective; if the body fat percentage change rate is <1%, it is ineffective. The total effective rate was calculated based on the markedly effective and effective results. The test results are shown in Table 18.

[0173] Table 18 Population test results

[0174]

[0175]

[0176] Among them, the body fat percentage calculation formula for adult women is as follows:

[0177] Parameter a = waist circumference (cm) × 0.74;

[0178] Parameter b = body weight (kg) × 0.082 + 34.89;

[0179] Body fat weight (kg) = a - b;

[0180] Body fat percentage = (body fat weight ÷ body weight) × 100%.

[0181] The formula for calculating body fat percentage for adult males is as follows:

[0182] Parameter a = waist circumference (cm) × 0.74;

[0183] Parameter b = body weight (kg) × 0.082 + 44.74;

[0184] Body fat weight (kg) = a - b;

[0185] Body fat percentage = (body fat weight ÷ body weight) × 100%.

[0186] All the above test subjects completed the test process, and no gastrointestinal discomfort occurred during the use, and no adverse reactions such as diarrhea and flatulence occurred. In addition, the excretion was regular every day during the use. At the same time, it has a good fat-reducing effect. Probiotics help burn fat, and reducing body fat percentage also helps reduce the risk of diabetes.

[0187] Obviously, the probiotic pellets prepared in the examples of the present application have application prospects in food and / or health products, can effectively regulate the intestinal function of the human body, affect the metabolism of the host by regulating the intestinal flora, control body fat, and thus prevent and control obesity and reduce the occurrence of obesity complications.

[0188] The present invention illustrates the technical concept of the present invention through the above-mentioned embodiments, but the present invention is not limited to the above-mentioned embodiments, that is, it does not mean that the present invention must rely on the above-mentioned embodiments to be implemented. Those skilled in the art should understand that any improvement of the present invention, equivalent replacement of individual raw materials of the product of the present invention, addition of auxiliary components, selection of specific methods, etc., all fall within the protection scope and disclosure scope of the present invention.

Claims

1. A probiotic micro-pellet, characterized in that: It comprises a pellet core and a coating layer wrapped around the pellet core; In terms of weight percentage, the pellet core is composed of 30-80% probiotic powder and 20-70% auxiliary materials, wherein the probiotic powder is selected from two or more of Lactobacillus plantarum, Lactobacillus rhamnosus, Lactobacillus paracasei, and Lactobacillus reuteri, and the total lactic acid bacteria content of the probiotic powder is ≥2.5×10 11 CFU / g.

2. A probiotic micro-pellet according to claim 1, characterized in that: In terms of weight percentage, the coating layer is composed of 10-65% methylcellulose, 10-65% hydroxypropyl methylcellulose, 5-20% sodium alginate, 0.5-5% magnesium stearate and 0.01-5% food coloring.

3. A probiotic micro-pellet according to claim 1, characterized in that: The auxiliary materials consist of microcrystalline cellulose, sodium carboxymethyl cellulose, maltodextrin and prebiotic components.

4. A probiotic micro-pellet according to claim 3, characterized in that: The prebiotic component is selected from one or more of xylooligosaccharides, stachyose, galacto-oligosaccharides and inulin.

5. The probiotic micro-pellet according to claim 3, characterized in that: In terms of weight percentage, the composition of the auxiliary materials is as follows: 5-20% microcrystalline cellulose, 0.5-5% sodium carboxymethyl cellulose, 30-50% maltodextrin, and 20-50% prebiotic components.

6. A method for preparing the probiotic pellets according to any one of claims 1 to 5, characterized in that: The steps include: S1. Preparation of pellet core S11, preparation of masterbatch: the prebiotic components and maltodextrin are uniformly mixed, purified water is added, and the mixture is granulated, dried, and sieved to obtain the masterbatch; S12, coating of the probiotic layer: after adding the masterbatch prepared in step S11 into the granulator, start the fan and the turntable, mix the probiotic powder, microcrystalline cellulose and sodium carboxymethyl cellulose evenly, add them into the turntable, and spray purified water at the same time. After coating, stop spraying and continue to round the particles; S13, drying of the pellet core: turning on the heating device, drying and sieving the material in step S12 to obtain the pellet core; S2. Coating S21, preparation of coating solution: adding the coating layer raw materials into purified water according to the proportion, stirring evenly and filtering to obtain the coating solution for standby use; S22, coating: adding the pellet cores obtained in step S13 into a fluidized bed, spraying the coating liquid into the fluidized bed, and performing coating treatment; S23, drying and sieving: After the coating weight gain reaches a predetermined range, stop spraying the coating liquid, dry and sieve to obtain the probiotic micropellets.

7. The preparation method according to claim 6, characterized in that: In step S11, the drying temperature is 50-60°C, the drying time is 6-10 hours, and the mesh size of the sieve is 20-50 meshes; In step S12, the air intake frequency of the fan is 10-18 Hz, and the rotation speed is 40-60 r / min; purified water is sprayed by a spray gun, with a flow rate of 20-60 mL / min and a pressure of 0.10-0.35 MPa; and the particles are rounded to 10-40 mesh.

8. The preparation method according to claim 6, characterized in that: In step S13, the drying temperature is 45-55°C, the drying time is 30-60 minutes, and the sieving mesh size is 10-40 meshes.

9. The preparation method according to claim 6, characterized in that: In step S21, the total concentration of the coating solution is 8-12 wt %; In step S22, the air inlet temperature is set to 100-130°C, the air outlet temperature is set to 40-50°C, the fan frequency is 30-50HZ, and the fluidized bed preheating time is 20-40min; In step S23, the coating weight gain range is 10-20%; the sieving mesh number is 14-36 meshes.

10. Use of the probiotic pellets according to claims 1 to 5 or the probiotic pellets prepared by the method according to any one of claims 6 to 9 in the preparation of food, nutritional dietary supplements, health food or medicine.

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