Method for drying multi-layer fecal microbiota dropping pill

Abstract

The present application belongs to the technical field of pharmaceutical preparation. Provided is a method for drying a multi-layer fecal microbiota dropping pill. The drying method of the present application comprises: subjecting a multi-layer fecal microbiota dropping pill to centrifugal deoiling with a deoiling agent, followed by a separation treatment to obtain a deoiled dropping pill, wherein the deoiling agent is a mixture of fumed silica, soybean fiber and oat cellulose; subjecting the deoiled dropping pill to blast drying to obtain a pre-dried dropping pill; and subjecting the pre-dried dropping pill to negative-pressure drying to obtain a Huanglong dropping pill. According to the drying method provided by the present application, centrifugal deoiling, blast drying and negative-pressure drying are sequentially performed to further reduce the water activity of the Huanglong dropping pill. Data from the examples demonstrate that the obtained Huanglong dropping pill has a water activity (aw) of 0.08-0.14, exhibiting an excellent tolerance, particularly an in vitro simulated gastrointestinal tolerance, and an excellent stability.

Description

A drying method of fecal bacteria multi-layer dripping pills

[0001] The present application claims priority to the Chinese patent application No. 202411369203.8, filed on September 29, 2024, and entitled "A drying method of fecal bacteria multi-layer dripping pills", the content of which is incorporated herein by reference in its entirety. TECHNICAL FIELD

[0002] The present application relates to the technical field of pharmaceutical preparation, in particular to a drying method of fecal bacteria multi-layer dripping pills. BACKGROUND

[0003] Fecal microbiota transplantation (FMT) refers to transplanting functional flora in healthy human feces into the intestinal tract of patients to reconstruct new intestinal flora and achieve the treatment of intestinal and extra-intestinal diseases. It is used to treat intestinal diseases such as Clostridium difficile infection, intractable diarrhea, constipation, gastrointestinal inflammation, Crohn's disease, ulcerative gastrointestinal inflammation, and extra-intestinal diseases such as obesity, asthma, obesity, autism, depression, diabetes, and cancer.

[0004] The fecal microbiota transplantation access route is divided into three routes: upper digestive tract, middle digestive tract, and lower digestive tract. The upper digestive tract route mainly refers to oral administration of fecal bacteria capsules, which has the advantages of non-invasiveness and high patient acceptance. The middle digestive tract route mainly includes through a nasogastric tube, a nasojejunal tube, and a nasoduodenal tube, which is particularly suitable for patients with colon inflammation, but intubation causes discomfort to patients and the route cannot assess the intestinal mucosa of patients and collect tissue samples. The lower digestive tract route mainly includes colonoscopy and retention enema, which reduces patient discomfort compared with intubation, and due to the need for intestinal cleaning before the operation, the colon condition can be located and beneficial flora can be sent to the target site, but the disadvantage is that only the distal colon can be perfused. The three methods have similar treatment effects, and the oral administration of fecal bacteria capsules is the most acceptable to patients, so it also has the most extensive application prospect.

[0005] Huanglong dripping pills are seamless dripping pills prepared by using a new type of dripping pill preparation technology with fecal bacteria freeze-dried bacteria powder as raw material, and then drying the seamless dripping pills. The in-vitro disintegration test results of Huanglong dripping pills show that they are intact in an acidic environment with pH 1.2 for 2 hours and still have good integrity in a pH 6.8 environment for 4 hours, and the storage conditions are simple. Water activity is crucial for the efficacy of Huanglong dripping pills and storage. Huanglong dripping pills have high water activity, are inconvenient to store, and affect the efficacy. Drying the seamless dripping pills is the key step that affects the water activity of Huanglong dripping pills, and how to ensure the activity of freeze-dried bacteria powder in Huanglong dripping pills and low water activity is the research focus of existing Huanglong dripping pills.

[0006] CN115671067A discloses the use of Huanglong dripping pills for treating and preventing irritable bowel syndrome. The patent discloses dropping a plurality of layers of materials into a cooling liquid to obtain seamless dripping pills with a concentric structure, and then air-drying the obtained seamless dripping pills with a concentric structure at 15-20°C for 10-12 hours to remove the outer layer moisture, thereby obtaining the dripping pills. The water activity (aw) of the Huanglong dripping pills obtained by the above drying method is high, being 0.15-0.20.

[0007] SUMMARY

[0008] Therefore, the present application aims to provide a drying method for fecal bacteria multi-layer dripping pills. The Huanglong dripping pills obtained by the drying method provided by the present application have low water activity.

[0009] To achieve the above-mentioned application purposes, the present application provides the following technical solutions:

[0010] The present application provides a drying method for fecal bacteria multi-layer dripping pills, comprising the following steps:

[0011] providing fecal bacteria multi-layer dripping pills;

[0012] After centrifugal deoiling the fecal bacteria multi-layer dripping pills with a deoiling agent, separating the deoiled dripping pills; the deoiling agent is a mixture of fumed silica, soybean fiber and oat cellulose;

[0013] air-drying the deoiled dripping pills to obtain pre-dried Huanglong dripping pills;

[0014] negative pressure drying the pre-dried Huanglong dripping pills to obtain Huanglong dripping pills.

[0015] Preferably, the mass ratio of the fumed silica, soybean fiber and oat cellulose is 1:1-2:1-2.

[0016] Preferably, the mass ratio of the fecal bacteria multi-layer dripping pills and the deoiling agent is 1:1-3.

[0017] Preferably, the rotation speed of the centrifugal deoiling is 500-1200 rpm, and the time is 10-15 min.

[0018] Preferably, the separation time is 3-8 min.

[0019] The separation is performed in a drying drum, and the rotation speed of the drying drum is 5-10 rpm.

[0020] Preferably, the rotation speed of the air-drying is 4-10 rpm, and the time is 5.5-7.5 h; the air-drying air is sterile air.

[0021] The forced-air drying is carried out in a drying drum. The drum wall has holes with a diameter of 0.5 to 1.5 mm. The drying drum is filled with pure cotton oil-absorbing cloth.

[0022] The pure cotton oil-absorbing cloth is changed every 30 minutes during the first 60 minutes of the blower drying process.

[0023] Preferably, the water activity of the pre-dried Huanglong dripping pills is 0.15 to 0.2.

[0024] Preferably, the negative pressure drying process involves a pressure of 0–0.02 MPa, a temperature of 25–28°C, a humidity of 3–15%, and a time of 60–180 min.

[0025] Preferably, the negative pressure drying is carried out in a drying drum, and the rotation speed of the drying drum is 4 to 10 rpm;

[0026] The drying drum contains a desiccant, which is a mixture of color-changing silica gel and anhydrous calcium chloride, with a mass ratio of 1:1 to 3.

[0027] Preferably, the water activity of the Huanglong Dripping Pill is 0.08 to 0.14.

[0028] This application provides a drying method for multilayer fecal microorganism droplets, comprising the following steps: providing multilayer fecal microorganism droplets; centrifuging the multilayer fecal microorganism droplets with a degreasing agent to remove oil, followed by separation treatment to obtain de-oiled droplets; the degreasing agent is a mixture of fumed silica, soybean fiber, and oat cellulose; drying the de-oiled droplets with forced air to obtain pre-dried Huanglong droplets; and drying the pre-dried Huanglong droplets under negative pressure to obtain Huanglong droplets.

[0029] The drying method provided in this application involves centrifugal degreasing, forced-air drying, and negative pressure drying, which further reduces the water activity of Huanglong Dripping Pills. Data from the examples show that the resulting Huanglong Dripping Pills have a water activity (aw) of 0.08–0.14, exhibiting excellent tolerability, especially in in vitro simulated gastrointestinal tolerance, and also demonstrate excellent stability.

[0030] Furthermore, the drying method provided in this application is time-efficient and highly effective. Detailed Implementation

[0031] This application provides a method for drying multilayer droppings of fecal microorganisms, comprising the following steps:

[0032] Provides multi-layered fecal microbial droplets;

[0033] After centrifuging and degreasing the fecal microbial multilayer pellets with a degreasing agent, the pellets are separated to obtain degreased pellets; the degreasing agent is a mixture of fumed silica, soybean fiber and oat cellulose;

[0034] The deoiled dropping pills are subjected to air blast drying to obtain pre-dried Huanglong dropping pills;

[0035] The pre-dried Huanglong dropping pills are subjected to negative pressure drying to obtain Huanglong dropping pills.

[0036] In the present application, the raw materials used in the present application are preferably commercially available products unless otherwise specified.

[0037] The present application provides a fecal bacteria multilayer dropping pill.

[0038] In the present application, the fecal bacteria multilayer dropping pill refers to a non-dried Huanglong dropping pill, and the multilayer structure of the non-dried Huanglong dropping pill has already been formed, but it is not dried. The preparation method of the fecal bacteria multilayer dropping pill is not specifically limited in the present application, and the operation well known to those skilled in the art can be used.

[0039] After providing the fecal bacteria multilayer dropping pill, the present application uses a deoiling agent to centrifugally deoiling the fecal bacteria multilayer dropping pill, and then separates the deoiling agent to obtain deoiled dropping pills; the deoiling agent is a mixture of fumed silica, soybean fiber and oat cellulose.

[0040] In the present application, the mass ratio of the fumed silica, soybean fiber and oat cellulose is preferably 1:1-2:1-2, and further preferably 1:1:1. In the present application, the preparation method of the deoiling agent preferably comprises the following steps: mixing fumed silica, soybean fiber and oat cellulose, vacuum drying and cooling to obtain the deoiling agent, the temperature of the vacuum drying is preferably 40-60°C, and further preferably 50°C; the time is preferably 4-6h, and further preferably 5h, and the vacuum drying is preferably carried out in a vacuum drying box. In the present application, the mass ratio of the fecal bacteria multilayer dropping pill and the deoiling agent is preferably 1:1-3, and specifically preferably 1:1.

[0041] In the present application, the rotation speed of the centrifugal deoiling is preferably 500-1200rpm, and further preferably 800-1000rpm; the time is preferably 10-15min, and further preferably 11-14min, and more preferably 12-13min. In the present application, the centrifugal deoiling preferably comprises: loading the deoiling agent and the fecal bacteria multilayer dropping pill into a sterile bag, and then placing the sterile bag in a centrifuge for centrifugal deoiling.

[0042] In the present application, the time of the separation treatment is preferably 3-8 min. In the present application, the separation treatment is preferably performed in a drying drum, the drum wall of which is preferably provided with holes, the diameter of which is preferably 0.5-1.5 mm; the rotation speed of the drying drum is preferably 5-10 rpm. In the present application, the drum in the drying drum is preferably pretreated before use, the pretreatment preferably comprises boiling in boiling water and blow-drying in sequence, the time of the boiling in boiling water is preferably 30-45 min, the gas of the blow-drying is preferably sterile air, the pressure of the sterile air is preferably 0.2-0.3 MPa. In the present application, the separation treatment can separate the deoiling agent and the deoiled dropping pills.

[0043] After obtaining the deoiled dropping pills, the present application performs air blast drying on the deoiled dropping pills to obtain pre-dried Huanglong dropping pills.

[0044] In the present application, the rotation speed of the air blast drying is preferably 4-10 rpm, and the time is preferably 5.5-7.5 h. In the present application, the air blast gas of the air blast drying is preferably sterile air.

[0045] In the present application, the air blast drying is preferably performed in a drying drum, the drum wall of which is preferably provided with holes, the diameter of which is preferably 0.5-1.5 mm, and the drying drum is preferably provided with pure cotton oil-absorbing cloth. In the present application, the pure cotton oil-absorbing cloth is preferably pretreated before use, the pretreatment comprising steam sterilization and drying in sequence, the temperature of the steam sterilization is preferably 121℃, and the time is preferably 20 min; the temperature of the drying is preferably 45-50℃, and the time is preferably 2-3 h. In the present application, the drum in the drying drum is preferably pretreated before use, the pretreatment preferably comprising boiling in boiling water and blow-drying in sequence, the time of the boiling in boiling water is preferably 30-45 min, the gas of the blow-drying is preferably sterile air, and the pressure of the sterile air is preferably 0.2-0.3 MPa.

[0046] In the present application, the pure cotton oil-absorbing cloth is replaced every 30 min for the first 60 min of the air blast drying.

[0047] In the present application, the water activity (aw) of the pre-dried Huanglong dropping pills is preferably 0.15-0.2.

[0048] After obtaining the pre-dried Huanglong dropping pills, the present application performs negative pressure drying on the pre-dried Huanglong dropping pills to obtain Huanglong dropping pills.

[0049] In the present application, the pressure of the negative pressure drying is preferably 0-0.02 MPa, the temperature is preferably 25-28℃, the humidity is preferably 3-15%, and the time is preferably 60-180 min.

[0050] In the present application, the negative pressure drying is preferably carried out in a drying drum, and the rotation speed of the drying drum is preferably 4-10 rpm.

[0051] In the present application, the drying drum is preferably loaded with a drying agent, which is preferably a mixture of color-changing silica gel and anhydrous calcium chloride, and the mass ratio of the color-changing silica gel and the anhydrous calcium chloride is preferably 1:1-3. In the present application, the particle size of the color-changing silica gel and the anhydrous calcium chloride is independently preferably 5-8 mm. In the present application, the preparation method of the drying agent preferably comprises the following steps: mixing color-changing silica gel and anhydrous calcium chloride, drying to obtain the drying agent; and the drying temperature is preferably 150-170°C, and the time is preferably 6-8 h.

[0052] After the negative pressure drying, the present application preferably further comprises screening the obtained material. In the present application, the screening is preferably carried out in an anaerobic glove box.

[0053] In the present application, the water activity of the Huanglong dripping pills is preferably 0.08-0.14; and the particle size of the Huanglong dripping pills is preferably 1.8-2.5 mm.

[0054] The drying method of the fecal bacteria multi-layer dripping pills provided by the present application will be described in detail below in combination with examples, but they should not be understood as limiting the scope of protection of the present application.

[0055] Example 1

[0056] First, 200 g of fumed silica, 200 g of soybean fiber and 200 g of oat cellulose are mixed, and then vacuum dried at 50°C for 5 h, cooled to room temperature and loaded into a sterile bag.

[0057] Then, 600 g of the fecal bacteria multi-layer dripping pills prepared by the dripping machine are loaded into a sterile bag, and centrifugal deoiling is carried out at 1000 rpm for 12 min in a centrifuge.

[0058] The drying drum is pretreated: the drying drum is boiled in boiling water for 45 min, and then dried with 0.3 MPa sterile filtered air.

[0059] After centrifugal deoiling, the material in the sterile bag is poured into the pretreated drying drum (the hole is 0.5-1.5 mm), and the deoiling agent is removed by screening after 5 min of separation at a rotation speed of 6 rpm, to obtain deoiled dripping pills.

[0060] The deoiled drop pills were placed in a drying drum (the drying drum was pretreated before use, the pretreatment was that the drying drum was boiled in boiling water for 45 min, then dried with 0.3 MPa sterile filtered air; and pure cotton oil-absorbing cloth was loaded in the drying drum, the pure cotton oil-absorbing cloth was pure cotton oil-absorbing cloth that was sterilized by steam at 121 ℃ for 20 min and dried at 45 ℃ for 2.5 h), and dried at 6 rpm for 6.5 h under the condition of blowing sterile air, wherein the pure cotton oil-absorbing cloth was replaced every 30 min for the first 60 min, to obtain pre-dried Huanglong drop pills, and the water activity of the pre-dried Huanglong drop pills was detected, and the result was that the water activity aw was 0.158.

[0061] 200 g of discolored silica gel (particle size 6 mm) and 200 g of anhydrous calcium chloride (particle size 6 mm) were mixed, dried at 115 ℃ for 2.5 h and cooled to room temperature to obtain a drying agent; the drying agent was loaded into a drying drum (the drying drum was pretreated before use, the pretreatment was that the drying drum was boiled in boiling water for 45 min, then dried with 0.3 MPa sterile filtered air), the temperature of the drying drum was controlled at 28 ℃, the humidity was controlled at 5%, the rotation speed was 5 rpm, vacuum was drawn to ensure that the pressure of the drying drum was negative pressure 0.02 MPa, and negative pressure drying was performed for 120 min to obtain Huanglong drop pills, and the water activity of the obtained Huanglong drop pills was detected, and the result was that the water activity aw of the obtained Huanglong drop pills was 0.092.

[0062] The final Huanglong drop pills were sieved in an anaerobic glove box to obtain Huanglong drop pills 1 with a particle size of 1.8-2.5 mm.

[0063] The preparation method of the fecal bacteria multi-layer drop pills comprises the following steps:

[0064] 1. Outer layer matrix

[0065] The outer layer matrix formula is: gelatin (freezing power 160) 25 wt%, glycerol 1.5 wt%, sucrose 4 wt%, water to 100 wt%. The outer layer matrix is prepared by soaking the gelatin with an appropriate amount of water (overnight), adding glycerol and sucrose, stirring to dissolve, and adding water to the full amount.

[0066] 2. Middle layer matrix

[0067] The middle layer matrix formula is: hardened fat (melting point 39-43 ℃, actually 43 ℃) 93 wt%, natural vitamin E oil 3 wt%, calcium chloride 4 wt%. The middle layer matrix is prepared by mixing the materials uniformly and melting at 60 ℃, and homogenizing with a 5000 r / min homogenizer.

[0068] 3. Inner layer capsule material

[0069] Inner layer capsule material formula: bioactive substances (such as probiotics, intestinal flora, enzymes, polypeptides, the addition amount is 10-30%; the intestinal flora powder obtained by removing impurities and freeze-drying the feces of healthy volunteer B in this embodiment is added in an amount of 20 wt%), natural vitamin E oil 1.8 wt%, dipotassium hydrogen phosphate-dipotassium phosphate (1:5) 1.2 wt%, vegetable fat (melting point 37-42℃, actually 38℃) to 100% (addition amount 60-80%, adjusted according to the addition amount of bioactive substances). Preparation of inner layer capsule material: melt the vegetable fat, add natural vitamin E oil and micronized dipotassium hydrogen phosphate-dipotassium phosphate, mix well, then add bioactive substances, homogenize with a 5000 r / min homogenizer, keep warm and stir to avoid sedimentation.

[0070] 4. Pelletizing

[0071] The weight ratio of the outer layer / middle layer / inner layer is 1:3:1.5. The inner layer material, the middle layer material and the outer layer material are dripped from the inner, middle and outer layers of the three-layer dripping head (the dripping head temperature is 65℃) into the cooling liquid (liquid paraffin) at 18-20℃ to form seamless dripping pills with a three-layer concentric structure (the obtained dripping pills have a diameter of 2.5mm), that is, the fecal bacteria multilayer dripping pills are obtained.

[0072] Comparative Example 1

[0073] 600g of the fecal bacteria multilayer dripping pills prepared by the dripping machine are dried at 18℃ for 11h to obtain Huanglong dripping pills 2.

[0074] Among them, the fecal bacteria multilayer dripping pills are taken from the fecal bacteria multilayer dripping pills prepared in Example 1, and the batch is consistent.

[0075] Test Example 1

[0076] Stability test of Huanglong dripping pills at 25℃ (0-28d)

[0077] Packaging machine: back-sealed granule packaging machine, model DXDK50IIE1, manufacturer Zhuhai Ruichuang Science and Technology Development Co., Ltd.

[0078] Aluminum foil bag: material PET12 / AL7 / PET12 / PE80, Zhejiang Jinshi Packaging Co., Ltd.

[0079] Deoxidizer: effective substance content 99%, batch number 20210201, Nanjing Weina Freshness Agent Factory.

[0080] Drying agent: effective substance content 99%, batch number 20210201, Nanjing Weina Freshness Agent Factory.

[0081] Dripping and drying date of the dripping pills: 20210727.

[0082] Storage condition of drop pills: (25℃±2℃ / 60%RH±5%RH).

[0083] Results are shown in Table 1 and Table 2.

[0084] Table 1 Water activity test results of Huanglong drop pills 0-28d at 25℃

[0085] Table 2 Moisture test results of Huanglong drop pills 0-28d at 25℃

[0086] Comparing Table 1 and Table 2, it can be seen that the water activity of Huanglong drop pills 1 obtained in Example 1 is lower than that of Huanglong drop pills 2 obtained in Comparative Example 1, indicating that the Huanglong drop pills 1 obtained in Example 1 has good resistance.

[0087] Test Example 2

[0088] Microbial test

[0089] Buffer: 0.9% NaCl solution containing 0.05% tween-80 (121℃-15min sterilization).

[0090] Preparation of Huanglong drop pill solution: 0.5g Huanglong drop pills were added to 49.5g sterilized buffer and homogenized at 18000rpm for 80s.

[0091] Culture medium: ① Plate Count Agar (PCA) (HB0101, Haibo Biotechnology Co., Ltd.); ② Wilkins-Chalgren Agar (WCA) (HB0261, Haibo Biotechnology Co., Ltd.); ③ MRS medium (027315, Guangdong Huan Kai Microbial Science and Technology Co., Ltd.), which needs to add matching reagents SR0370 Mupirocin lithium salt and cysteine hydrochloride according to the instruction.

[0092] Culture method: After the prepared Huanglong drop pill solution was diluted with buffer to the appropriate concentration, it was coated: PCA was used to culture total aerobic bacteria, and after coating, it was cultured at 37℃; WCA was used to culture total anaerobic bacteria, and after coating, it was cultured anaerobically at 37℃; MRA was used to culture Bifidobacterium, and after coating, it was cultured anaerobically at 37℃. Plate counting was performed after 48h, and the results are shown in Table 3 and Table 4.

[0093] Among them, Bifidobacterium refers to the national standard GB4789.34-2016, which is cultured using MRS medium + Mupirocin lithium salt and cysteine hydrochloride.

[0094] Table 3 Microbial test results of Huanglong drop pills 2 at 25℃, 0-28d

[0095] Table 4 Microbial test results of Huanglong drop pills 1 at 25℃, 0-28d

[0096] As can be seen from Table 3 and Table 4, the stability of Huanglong Dropping Pills 1 obtained by the drying method provided in the application is better than that of Huanglong Dropping Pills 2 obtained by Comparative Example 1.

[0097] Test Example 3

[0098] In vitro simulated gastrointestinal tolerance test

[0099] The third method (small cup method) of "0931 Dissolution and Release Determination Method" in Chinese Pharmacopoeia 2020 Edition was used for dissolution test, 200 mL dissolution medium was used, the rotation speed was 50 rpm, the droppings were transferred to simulated intestinal fluid after 2 h of simulated gastric fluid, and sampling was performed after 4 h, and the survival rate of microorganisms after dissolution treatment of the test sample was determined according to the method of "Probiotic Viable Count Detection Method" (the relative survival rate calculated based on the 0 h microorganism survival rate without dissolution treatment was 100%, and the average value of 6 dissolution cups was represented).

[0100] The preparation method of the dissolution medium simulated gastric fluid is as follows: 3.84 mL of hydrochloric acid is taken, about 800 mL of water is added, 10 g of pepsin is shaken, and water is added to dilute to 1000 mL.

[0101] The preparation method of the dissolution medium simulated intestinal fluid is as follows: 6.8 g of potassium dihydrogen phosphate is taken, 500 mL is dissolved, and the pH value is adjusted to 6.8 with 0.1 mol / L sodium hydroxide solution; another 10 g of pancreatin and 3 g of bile salt are taken, and water is added to dissolve, then the two liquids are mixed, and water is added to dilute to 1000 mL.

[0102] Table 5 Huanglong Dropping Pills in vitro simulated gastrointestinal tolerance test

[0103] Huanglong Dropping Pills 2 was treated with pH 1.2 gastric acid solution and enzyme for 2 h, and the total anaerobic bacteria survival rate was 79.1%, and then treated with pH 6.8 phosphate and trypsin buffer for 4 h, and the survival rate was 39.2%.

[0104] Huanglong Dropping Pills 2 was treated with pH 1.2 gastric acid solution and enzyme for 2 h, and the total aerobic bacteria survival rate was 66.8%, and then treated with pH 6.8 phosphate and trypsin buffer for 4 h, and the survival rate was 37.4%.

[0105] Huanglong Dropping Pills 2 was treated with pH 1.2 gastric acid solution and enzyme for 2 h, and the survival rate of Bifidobacterium was 81.6%, and then treated with pH 6.8 phosphate and trypsin buffer for 4 h, and the survival rate was 50.1%.

[0106] The total anaerobic bacteria survival rate of Huanglong Dropping Pills 1 treated by pH 1.2 gastric acid solution and enzyme for 2 h was 82.6%, and the survival rate was 41.1% after treated by pH 6.8 phosphate and trypsin buffer for 4 h.

[0107] The total aerobic bacteria survival rate of Huanglong Dropping Pills 1 treated by pH 1.2 gastric acid solution and enzyme for 2 h was 71.2%, and the survival rate was 39.9% after treated by pH 6.8 phosphate and trypsin buffer for 4 h.

[0108] The Bifidobacterium survival rate of Huanglong Dropping Pills 1 treated by pH 1.2 gastric acid solution and enzyme for 2 h was 86.7%, and the survival rate was 52.6% after treated by pH 6.8 phosphate and trypsin buffer for 4 h.

[0109] The Huanglong Dropping Pills 1 obtained by the drying method provided in the application has better tolerance effect than the Huanglong Dropping Pills 2 obtained by the comparison example 1.

[0110] The above only describes the preferred embodiments of the application, and it should be pointed out that, for those skilled in the art, without departing from the principles of the application, a number of improvements and refinements can be made, and these improvements and refinements should also be considered as the protection scope of the application.

Claims

1. A drying method of a multi-layered dropping pill of coprozoa, characterized by, The method comprises the following steps: Providing fecal bacteria multilayer drop pills; After centrifugal deoiling of the fecal bacteria multilayer drop pills by using a deoiling agent, the deoiled drop pills are obtained by separation treatment; the deoiling agent is a mixture of fumed silica, soybean fiber and oat cellulose; The deoiled drop pills are subjected to air blowing drying to obtain pre-dried Huanglong drop pills; The pre-dried Huanglong drop pills are subjected to negative pressure drying to obtain Huanglong drop pills.

2. The drying method according to claim 1, characterized in that, The mass ratio of the fumed silica, soybean fiber and oat cellulose is 1:1-2:1-2.

3. The drying method according to claim 1 or 2, characterized in that, The preparation method of the deoiling agent comprises the following steps: mixing fumed silica, soybean fiber and oat cellulose, and obtaining by vacuum drying and cooling; the temperature of the vacuum drying is 40-60°C, and the time is 4-6h.

4. The drying method according to claim 1 or 2, characterized in that, The mass ratio of the fecal bacteria multilayer drop pills and the deoiling agent is 1:1-3.

5. The drying method according to claim 1, characterized in that, The rotation speed of the centrifugal deoiling is 500-1200rpm, and the time is 10-15min.

6. The drying method according to claim 1 or 5, characterized in that, The centrifugal deoiling comprises: loading the deoiling agent and the fecal bacteria multilayer drop pills into a sterile bag, and then placing the sterile bag in a centrifuge for centrifugal deoiling.

7. The drying method according to claim 1, characterized by, The separation treatment is performed for 3-8min; The separation treatment is performed in a drying drum, and holes with a diameter of 0.5-1.5mm are arranged on the wall of the drying drum, and the rotation speed of the drying drum is 5-10rpm.

8. The drying method according to claim 1, characterized by, The rotation speed of the air blowing drying is 4-10rpm, the time of the air blowing drying is 5.5-7.5h, and the air blowing gas of the air blowing drying is sterile air; The air blowing drying is performed in a drying drum, and holes with a diameter of 0.5-1.5mm are arranged on the wall of the drying drum, and pure cotton oil absorption cloth is loaded in the drying drum; The pure cotton oil absorption cloth is replaced every 30min in the first 60min of the air blowing drying.

9. The drying method according to claim 1 or 8, characterized in that, The water activity of the pre-dried Huanglong drop pills is 0.15-0.

2.

10. The drying method according to claim 1, characterized in that, The pressure of the negative pressure drying is 0-0.02MPa, the temperature is 25-28°C, the humidity is 3-15%, and the time is 60-180min.

11. The drying method according to claim 1 or 10, characterized in that, The negative pressure drying is performed in a drying drum, and the rotation speed of the drying drum is 4-10rpm; The drying drum is loaded with a drying agent, and the drying agent is a mixture of color-changing silica gel and anhydrous calcium chloride, and the mass ratio of the color-changing silica gel and the anhydrous calcium chloride is 1:1-3.

12. The drying method according to claim 11, characterized in that, The particle size of the color-changing silica gel and the anhydrous calcium chloride is independently 5-8mm.

13. The drying method according to claim 11, characterized in that, The preparation method of the drying agent comprises the following steps: mixing color-changing silica gel and anhydrous calcium chloride, and drying to obtain the drying agent; the temperature of the drying is 150-170°C, and the time is 6-8h.

14. The drying method according to claim 1, characterized in that, The water activity of the Huanglong drop pills is 0.08-0.14.

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