Methods for preparing Choling-to, Choling extract, and Choling solid preparations

Abstract

The present invention relates to the pharmaceutical field, specifically to a method for preparing Zhuling Tang, Zhuling extract, and Zhuling solid formulation. The preparation method provided by the present invention includes the steps of: soaking four kinds of traditional Chinese medicines, including Zhuling, Poria, Zexiang, and Talc, in water to obtain a prepreg liquid; performing a first boiling extraction on the prepreg liquid to obtain a first aqueous extract and a medicinal residue; mixing the medicinal residue with water to perform a second boiling extraction to obtain a second aqueous extract; first concentrating the first aqueous extract to obtain a first concentrate; mixing the first concentrate and the second aqueous extract to perform a second concentration to obtain a second concentrate; and mixing the second concentrate with an aqueous solution of Erjiao to obtain Zhuling Tang. The preparation method of Zhuling Tang provided by the present invention comprehensively takes into account water extraction and concentration, ensuring that the extraction effect of the obtained Zhuling Tang is consistent with that of the reference sample of conventional Zhuling Tang preparations, and the content of active ingredients in the obtained Zhuling Tang is high, resulting in good medicinal efficacy.

Description

[Technical Field]

[0001] This application claims priority to a Chinese patent application filed with the China Patent Office on September 26, 2022, bearing application number CN202211174012.7 and entitled "Method for preparing Zhuling Tang, Zhuling Extract, and Zhuling Solid Preparations," the entire contents of which are incorporated herein by reference.

[0002] The present invention relates to the technical field of medicine, and more particularly to a method for preparing Choling-to, Choling extract, and Choling solid preparation. [Background technology]

[0003] Zhuling Tang is from prescription number 9 in the Catalogue of Ancient Classical Prescriptions (first batch) published by the State Administration of Traditional Chinese Medicine of China. The original text is the Treatise on Febrile Diseases (Eastern Han Dynasty, Zhang Zhongjing), which Zhuling Tang prescribed for the pathology of yin deficiency, water heat interaction. It is a fundamental prescription with clinical effectiveness among physicians of all times and the present, and will serve as a pioneer in the treatment of yin nourishing and water-activating for future generations, with great application value. According to the Chinese Herbal Medicine Registration Classification and Application Data Requirements, it is classified as new Chinese herbal medicine 3.1, a Chinese herbal compound preparation managed based on the Catalogue of Ancient Classical Prescriptions.

[0004] Currently, there is no granular preparation of the ancient classical prescription Zhuling Tang available on the Chinese market. According to the requirements of relevant regulations such as the "Simplified Registration, Examination and Approval Administration Regulations for Ancient Classical Prescription Chinese Herbal Compound Preparations" and the "Drug Registration Administration Method," the development of a modern preparation of Zhuling Tang requires using a reference sample of Zhuling Tang as the standard, and using modern pharmaceutical equipment to prepare the finished Zhuling Tang granules through processes such as extraction, purification, concentration, drying, granulation, and packaging.

[0005] Currently, the commercial production of traditional Chinese herbal medicines generally employs a process flow that includes aqueous or alcohol extraction, concentration, alcohol precipitation, high-temperature drying, grinding, and molding. This common production process has advantages such as simple equipment, low cost, and mature technology, making it very popular in actual industrialization. Unfortunately, when applied to the production of classical formulas, this production process currently has two obvious flaws. First, ancient classical formulas require extraction methods and solvents consistent with traditional decoctions, making alcohol precipitation clearly inappropriate. Second, the high-temperature drying process destroys some heat-labile components. Due to these two issues, the material basis of classical formulas changes during industrialized production, making them unable to meet the requirements of reference samples. Therefore, if the current traditional Chinese herbal medicine extraction process is used to commercialize ancient classical formulas, there is a risk of defects in the finished drug, i.e., the essential quality consistency between mass production and products obtained by traditional processes cannot be guaranteed. Summary of the Invention [Problem to be solved by the invention]

[0006] In view of this, the object of the present invention is to provide a method for preparing Zhuling Tang, Zhuling extract, and Zhuling solid preparations, and the Zhuling solid preparations provided by the present invention are basically consistent with the quality standards of the reference sample of Zhuling Tang, have good solubility, high bioavailability, are more convenient to take and carry than traditional decoctions, and have better stability. [Means for solving the problem]

[0007] In order to achieve the above object of the invention, the present invention provides the following technical solutions:

[0008] The present invention provides a method for preparing Choling-to, Soaking the four kinds of herbal medicines in water to obtain a prepreg liquid; performing a first boiling extraction on the prepreg liquid to obtain a first water extract and a residue; Mixing the residue with water to perform a second boiling extraction to obtain a second water extract; performing a first concentration on the first aqueous extract to obtain a first concentrate; Mixing the first concentrate and the second aqueous extract to perform a second concentration to obtain a second concentrate; mixing the second concentrate with an aqueous solution of Erjiao to obtain the Zhuling Tang; the four herbal medicines include Poria cocos, Poria cocos, Zelkova and talc, and during the soaking, the ratio of the mass of water to the total mass of the four herbal medicines is 14:1 or less; During the second boiling extraction, the ratio of the mass of water to the total mass of the four herbal medicines is 12:1 or less; The temperatures of the first condensation and the second condensation are independently 90° C. or less.

[0009] Preferably, during the soaking, the ratio of the mass of water to the total mass of the four herbal medicines is (8-14):1, and the soaking time is 40-60 minutes; The first boiling extraction includes a temperature-raising step by steam heating and a boiling and keeping warm step by steam heating, the keeping warm time of the boiling and keeping warm step is 40 to 90 minutes, the steam pressure in the boiling and keeping warm step is 0.1 MPa or less, and the steam pressure in the temperature-raising step is 0.2 MPa or less.

[0010] Preferably, during the second boiling extraction, the ratio of the mass of water to the total mass of the four herbal medicines is (6-12):1, and the second boiling extraction includes a steam heating temperature increase stage and a steam heating boiling and keeping warm stage, the keeping warm time of the boiling and keeping warm stage is 25-60 minutes, the steam pressure during the boiling and keeping warm stage is 0.1 MPa or less, and the steam pressure during the heating stage is 0.2 MPa or less.

[0011] Preferably, the mass ratio of the Poria cocos, Poria cocos, Zea mays and the talc is (0.7-1.3):(0.7-1.3):(0.7-1.3):(0.7-1.3).

[0012] Preferably, the first concentration and the second concentration are independently carried out under steam heating conditions; The temperatures of the first condensation and the second condensation are independently 70 to 90°C, the vacuum pressures of the first condensation and the second condensation are independently 0.03 to 0.06 MPa, the incubation times of the first condensation and the second condensation are independently 6 hours or less, and the vapor pressures of the first condensation and the second condensation are independently 0.01 to 0.05 MPa.

[0013] The present invention provides a method for preparing a polygala extract, The method includes concentrating the Choling Tang prepared by the preparation method described in the above technical solution to obtain the Choling extract, wherein the concentration temperature is 90°C or less, and the relative density of the Choling extract at 20°C is 1.12-1.16.

[0014] Preferably, the concentration is carried out under steam heating conditions; The temperature for the concentration is 70 to 90° C., the vacuum pressure for the concentration is 0.03 to 0.06 MPa, the heat retention time for the concentration is 6 hours or less, and the vapor pressure for the concentration is 0.01 to 0.05 MPa.

[0015] The present invention provides a method for preparing a solid preparation of Zhuling, the solid preparation including dry powder or granules, the method for preparing the dry powder includes the steps of: mixing Zhuling Tang prepared by the preparation method described in the above technical solution with an aqueous solution of auxiliary materials, and then concentrating the mixture to obtain a thick paste; and vacuum-drying the thick paste to obtain Zhuling dry powder, the mass ratio of the auxiliary materials to the Zhuling Tang is (1~7):50, and the mass of the Zhuling Tang is calculated as the total mass of Zhuling, Poria, Zeaxanthus, Talc and Erjiao; The method for preparing the granules includes dry granulating the dried powder of Choling to obtain Choling granules.

[0016] Preferably, the vacuum drying is vacuum belt drying, and the operating parameters of the vacuum belt drying include: a material supply temperature of 75 to 85°C; a material supply rate of 2.0 to 3.5 L / h; a crawler travel speed of 150±50 mm / min; a material distribution motor rotation speed of 100 r / min; a swing arm angle of 12 to 25°C; heating temperature zones of a first zone of 95±5°C, a second zone of 96±5°C, and a third zone of 96±5°C, respectively; a vacuum degree of 0.097 to 0.1 MPa; a cutter speed of 10 to 20 seconds / cycle; and a cooling temperature of 30±5°C. The dry granulation is carried out in a granulator, and the operating parameters of the dry granulation include a pressure of the press roll of the granulator of 12 to 20 MPa, a rotation speed of the press roll of the granulator of 4 to 10 r / min, a vertical rotation speed of the screw feeder of the granulator of 25 to 40 r / min, a horizontal rotation speed of the screw feeder of the granulator of 70 to 120 r / min, and a mesh number of the sieve of 14 mesh.

[0017] The present invention provides a Zolin solid formulation prepared by the preparation method described in the above technical solution, wherein the dry paste ratio of the Zolin solid formulation is 18-23%, the total content of 23-acetylalisol B and 23-acetylalisol C in the Zolin solid formulation is 0.011-0.020 wt%, and the total content of donkey-derived polypeptide A1 and donkey-derived polypeptide A2 in the Zolin solid formulation is 0.091-0.18 wt%.

[0018] The present invention provides a method for preparing Choling Tang, which includes the steps of: soaking four kinds of Chinese herbal medicines in water to obtain a prepreg liquid; performing a first boiling extraction on the prepreg liquid to obtain a first aqueous extract and a medicinal residue; mixing the medicinal residue with water to perform a second boiling extraction to obtain a second aqueous extract; first concentrating the first aqueous extract to obtain a first concentrated liquid; and mixing the first concentrated liquid and the second aqueous extract to obtain a second aqueous extract. and mixing the second concentrated solution with an aqueous solution of Erjiao to obtain the Zhuling Tang, wherein the four herbal medicines include Zhuling, Poria, Zelkova and Talc, the ratio of the mass of the water to the total mass of the four herbal medicines is 14:1 or less, the ratio of the mass of the water to the total mass of the four herbal medicines is 12:1 or less, and the temperatures of the first concentration and the second concentration are independently 90°C or less. The preparation method for Zhuling Tang provided by the present invention comprehensively considers aqueous extraction and concentration, rationally designing the aqueous extraction step as a two-stage aqueous extraction, while rationally setting the amount of water used in each aqueous extraction step, aiming to achieve an active pharmaceutical ingredient extraction rate 20-30% higher than that of the reference sample, and setting the amount of water added to be no more than 14 times. The present invention also concentrates the extract at a low temperature (below 90°C), which is beneficial to protecting and preserving the active pharmaceutical ingredients and avoiding their destruction by high temperatures. In summary, the Zhuling Tang obtained by the preparation method provided by the present invention is guaranteed to be consistent with the extraction effect of the reference sample of conventional Zhuling Tang preparations, and the resulting Zhuling Tang has a high content of active ingredients and good medicinal efficacy.

[0019] Furthermore, in the present invention, the ratio of the mass of water to the total mass of the four herbs during the steeping process is (8-14):1, the steeping time is 40-60 minutes, and the first boiling extraction includes a steam heating temperature-raising step and a steam heating boiling and warming step, the warming time of the boiling and warming step is 40-90 minutes, the steam pressure during the boiling and warming step is 0.1 MPa or less, and the steam pressure during the temperature-raising step is 0.2 MPa or less. In order to pursue extraction rates during traditional modern herbal medicine extraction, each extraction often takes 1-2 hours, often extracting impurities such as fiber and starch, without increasing the extraction of active ingredients. Prolonged heating can destroy the active ingredients. Compared to traditional modern herbal medicine extraction methods, which require excessive extraction and consume high energy, the preparation method provided by the present invention, based on the addition of an appropriate amount of water during the first extraction process, shortens extraction time and increases extraction efficiency, further improving the quality and medicinal efficacy of Zhuling Tang products and reducing production costs.

[0020] Furthermore, in the present invention, the ratio of the mass of water to the total mass of the four herbs during the second boiling extraction is (6-12):1, and the second boiling extraction includes a steam heating temperature raising stage and a steam heating boiling warm keeping stage, the boiling warm keeping stage's keeping time is 25-60 minutes, the steam pressure during the boiling warm keeping stage is 0.1 MPa or less, and the steam pressure during the temperature raising stage is 0.2 MPa or less.Furthermore, based on the appropriate amount of water added during the second extraction process, the present invention shortens the extraction time and increases the extraction efficiency, further improving the quality and medicinal efficacy of Zhongling Tang products and reducing production costs.

[0021] Furthermore, in the present invention, the first and second condensations are independently performed under steam heating conditions, with the temperatures for the first and second condensations independently being 70-90°C, the vacuum pressures for the first and second condensations independently being 0.03-0.06 MPa, the incubation times for the first and second condensations independently being 6 hours or less, and the vapor pressures for the first and second condensations independently being 0.01-0.05 MPa. By employing vacuum concentration and controlling the temperature to 70-90°C, the present invention not only improves evaporation efficiency compared to conventional atmospheric pressure concentration methods, but also reduces destruction of the active drug ingredient.

[0022] The present invention provides a method for preparing Choling extract, which comprises concentrating Choling Tang prepared by the method described in the above technical solution to obtain the Choling extract, wherein the concentration temperature is 90°C or less, and the relative density of the Choling extract at 20°C is 1.12-1.16. In the present invention, since the concentration is carried out at 90°C or less, the destruction of active drug components can be reduced.

[0023] The present invention provides a method for preparing a Zhuling solid preparation, the solid preparation including dry powder or granules, the method for preparing the dry powder includes: mixing Zhuling Tang prepared by the preparation method described in the above technical solution with an aqueous solution of auxiliary materials, and then concentrating the mixture to obtain a thick paste; and vacuum-drying the thick paste to obtain Zhuling dry powder, the mass ratio of the auxiliary materials to the Zhuling Tang is (1~7):50, and the mass of the Zhuling Tang is calculated as the total mass of Zhuling, Poria, Zeaxanthus, talc and Euglena. The method for preparing the granules includes dry granulating the Zhuling dry powder to obtain Zhuling granules. The present invention improves on the traditional method of adding auxiliary materials to granules (adding them before granulation and mixing evenly), by shifting the addition time of the auxiliary materials to before concentration, and adding the aqueous solution of the auxiliary materials before concentration. This not only solves the problems of the viscous mixed thick paste (including gelatin, polysaccharides, etc.), clogging, and material loss due to tape adhesion during the drying process, but also reduces the loss of dried paste during the drying process. Compared with the traditional method of adding auxiliary materials before granulation, the preparation method provided by the present invention can more uniformly mix the auxiliary materials and Zhuling Tang. At the same time, the traditional high-temperature drying and spray drying methods are prone to agglomeration and destroy the active pharmaceutical ingredients. The preparation method of Zhuling solid preparations provided by the present invention uses vacuum drying, which has the advantages of low drying temperature, short drying time, low loss, and high efficiency. The obtained Zhuling solid preparations do not aggregate and destroy the active pharmaceutical ingredients during the drying process, and the various ingredients in the Zhuling solid preparations are uniformly distributed, resulting in good pharmaceutical efficacy.

[0024] The present invention provides a Zhongling solid preparation prepared by the preparation method described in the above technical solution, wherein the Zhongling solid preparation has a dry paste ratio of 18-23%, the total content of 23-acetylalisol B and 23-acetylalisol C in the Zhongling solid preparation is 0.011-0.020 wt%, and the total content of donkey-derived polypeptide A1 and donkey-derived polypeptide A2 in the Zhongling solid preparation is 0.091-0.18 wt%. The Zhongling solid preparation provided by the present invention is consistent with the quality standard of the reference sample of Zhongling Tang, has good solubility, high bioavailability, is more convenient to take and carry than traditional decoctions, and has better stability. [Brief explanation of the drawings]

[0025] [Figure 1] 1 is a flowchart of the preparation of Choling granules provided by an embodiment of the present invention. [Figure 2] FIG. 1 is an HPLC control characteristic spectrum of Choling granules. DETAILED DESCRIPTION OF THE INVENTION

[0026] The present invention provides a method for preparing Choling-to, Soaking the four kinds of herbal medicines in water to obtain a prepreg liquid; performing a first boiling extraction on the prepreg liquid to obtain a first water extract and a residue; Mixing the residue with water to perform a second boiling extraction to obtain a second water extract; performing a first concentration on the first aqueous extract to obtain a first concentrate; Mixing the first concentrate and the second aqueous extract to perform a second concentration to obtain a second concentrate; mixing the second concentrate with an aqueous solution of Erjiao to obtain the Zhuling Tang; the four herbal medicines include Poria cocos, Poria cocos, Zelkova and Talc, and the ratio of the mass of the water to the total mass of the four herbal medicines is 14:1 or less; The ratio of the mass of the water to the total mass of the four herbal medicines is 12:1 or less; The temperatures of the first condensation and the second condensation are independently 90° C. or less.

[0027] In the present invention, unless otherwise specified, all raw materials are commercially available products familiar to those skilled in the art.

[0028] In the present invention, a prepreg solution is obtained by soaking four kinds of Chinese herbs, including Poria cocos, Zelkova, and Talc, in water, and the ratio of the mass of the water to the total mass of the four herbs is 14:1 or less.

[0029] In the present invention, the Zhuling root was purchased from Shandong Baimitang Traditional Chinese Medicine Pieces Co., Ltd.

[0030] In the present invention, the Poria cocos was purchased from Anhui Shentai Pharmaceutical Co., Ltd.

[0031] In the present invention, the alimentary candida was purchased from Shandong Baimitang Traditional Chinese Medicine Pieces Co., Ltd.

[0032] In the present invention, the talc was purchased from Quzhou Nankong Traditional Chinese Medicine Pieces Co., Ltd.

[0033] In the present invention, the above-mentioned glue was purchased from Dong Aer Glue Co., Ltd.

[0034] In the present invention, the mass ratio of the Poria cocos, Poria cocos, Zea mays and talc is preferably (0.7-1.3):(0.7-1.3):(0.7-1.3):(0.7-1.3), and more preferably 1:1:1:1.

[0035] In the present invention, the formulation of Choling-to is, in parts by mass, 750 parts of Choling, 750 parts of Poria, 750 parts of Zelkova, 750 parts of Talc, and 750 parts of Erjiao.

[0036] In the present invention, the method for processing the zhuling is preferably as follows: (1) sorting to remove impurities; (2) soaking at 5-35°C for 0.5-3 hours; (3) moistening at 5-35°C for 3-6 hours; (4) cutting into uniform, thick slices of 2-4 mm; and (5) drying at 70-80°C for 5-6 hours.

[0037] In the present invention, the method for processing Poria cocos is preferably as follows: (1) Selection to remove impurities.

[0038] In the present invention, the method for processing the zeaxanthin is preferably as follows: (1) sorting to remove impurities; (2) spraying and washing for 10-15 minutes; (3) soaking at 15-35°C for 5-8 hours; (4) moistening at 15-35°C for 18-36 hours; (5) cutting into uniform, thick slices of 2-4 mm; and (6) drying at 50-60°C for 5-6 hours.

[0039] In the present invention, the method for producing the talc is preferably as follows: (1) screening to remove impurities; (2) washing thoroughly; (3) drying at 70-80°C for 3-4 hours; and (4) crushing by crushing in a grinder and passing through an 80-mesh sieve for preparation.

[0040] In the present invention, the talc is preferably encapsulated.

[0041] In the present invention, the immersion is preferably carried out in an extraction tank.

[0042] In the present invention, the ratio of the mass of the water to the total mass of the four herbal medicines is 14:1 or less, preferably (8-12):1, more preferably (10-12):1.

[0043] In the present invention, the immersion temperature is preferably room temperature.

[0044] In the present invention, the immersion time is preferably 40 to 60 minutes, and more preferably 50 minutes.

[0045] After obtaining the prepreg liquid, the present invention performs a first boiling extraction on the prepreg liquid to obtain a first water extract and a dregs.

[0046] In the present invention, the specific implementation process of the first boiling extraction preferably includes opening the steam valve of the extraction tank, heating with heated steam, opening the external circulation of the extraction tank when the temperature reaches 90°C, starting timing when the temperature reaches boiling, maintaining the heated steam pressure stable, and performing the first boiling extraction. After the first boiling extraction is completed, closing the external circulation and releasing the medicinal liquid to obtain the first aqueous extract and medicinal residue.

[0047] In the present invention, the first boiling extraction preferably includes a temperature increasing step by steam heating and a boiling and keeping warm step by steam heating.

[0048] In the present invention, when the temperature of the poryophyllum prepreg liquid is increased by steam heating, the steam pressure during the temperature increase step is preferably 0.2 MPa or less.

[0049] In the present invention, during the first boiling extraction, the heating steam pressure in the boiling and keeping warm step is preferably 0.1 MPa or less, and more preferably 0.05 to 0.08 MPa.

[0050] In the present invention, the warming time in the boiling and warming step is preferably 40 to 90 minutes, more preferably 40 to 50 minutes.

[0051] In the present invention, after the first boiling extraction is completed, the first chemical solution obtained by the first boiling extraction is preferably subjected to post-treatment to obtain the first aqueous extract. In the present invention, the post-treatment preferably includes filtering the first chemical solution, centrifuging it, and microfiltrating the resulting liquid component to obtain the first aqueous extract. In the present invention, the filtration is preferably performed using a 120-mesh sieve. The centrifugation is preferably performed using a centrifuge, the separation speed of the centrifugation is preferably 30 to 40 L / min, and the sediment is preferably discharged every 20 minutes during the centrifugation. In the present invention, the microfiltration is preferably performed using a plate and frame filter, the pore size of the filter membrane of the plate and frame filter is preferably 5 to 15 μm.

[0052] After obtaining the residue, the present invention mixes the residue with water to perform a second boiling extraction to obtain a second water extract, and the ratio of the mass of the water to the total mass of the four herbal medicines is 12:1 or less.

[0053] In the present invention, during the second boiling extraction, the ratio of the mass of water to the total mass of the four herbal medicines is preferably (8-10):1.

[0054] In the present invention, the specific implementation process of the second boiling extraction preferably includes opening the steam valve of the extraction tank, heating with heated steam, opening the external circulation of the extraction tank when the temperature reaches 90°C, starting timing when the temperature reaches boiling, maintaining the heated steam pressure stable, and performing the second boiling extraction; after the second boiling extraction is completed, closing the external circulation and releasing the medicinal solution to obtain the second Chorizome water extract.

[0055] In the present invention, the second boiling extraction preferably includes a temperature increasing step by steam heating and a boiling and keeping warm step by steam heating.

[0056] In the present invention, when the temperature of the dregs mixture obtained by mixing the dregs and water is increased by steam heating, the heating steam pressure during the temperature increase step is preferably 0.2 MPa or less.

[0057] In the present invention, during the second boiling extraction, the heating steam pressure in the boiling and keeping warm step is preferably 0.1 MPa or less, and more preferably 0.05 to 0.08 MPa.

[0058] In the present invention, the incubation time in the boiling and incubation step during the second boiling extraction is preferably 25 to 60 minutes, more preferably 25 to 40 minutes.

[0059] In the present invention, after the second boiling extraction is completed, the second chemical solution obtained by the second boiling extraction is preferably post-treated to obtain the second aqueous extract. In the present invention, the post-treatment preferably includes filtering the second chemical solution, centrifuging it, and microfiltrating the resulting liquid component to obtain the second aqueous extract. In the present invention, the filtration is preferably performed using a 120-mesh sieve. The centrifugation is preferably performed using a centrifuge, the separation speed of the centrifugation is preferably 30 to 40 L / min, and the sediment is preferably discharged every 20 minutes during the centrifugation. In the present invention, the microfiltration is preferably performed using a plate and frame filter, the pore size of the filter membrane of the plate and frame filter is preferably 5 to 15 μm.

[0060] After obtaining the first aqueous extract and the second aqueous extract, the present invention performs a first concentration on the first aqueous extract to obtain a first concentrated liquid, and then mixes the first concentrated liquid and the second aqueous extract to perform a second concentration to obtain a second concentrated liquid.

[0061] In the present invention, the first concentration is preferably carried out under steam heating conditions, the temperature of the first concentration is preferably 70 to 90°C, more preferably 75 to 90°C, and most preferably 85°C, the vacuum pressure of the first concentration is preferably 0.03 to 0.06 MPa, and more preferably 0.04 to 0.05 MPa, the incubation time of the first concentration is preferably 6 hours or less, and the heated steam pressure of the first concentration is preferably 0.01 to 0.05 MPa, and more preferably 0.03 to 0.045 MPa.

[0062] In the present invention, the second concentration is preferably carried out under steam heating conditions, the temperature of the second concentration is preferably 70 to 90°C, more preferably 75 to 90°C, and most preferably 85°C, the vacuum pressure of the second concentration is preferably 0.03 to 0.06 MPa, and more preferably 0.04 to 0.05 MPa, the incubation time of the second concentration is preferably 6 hours or less, and the heated steam pressure of the second concentration is preferably 0.01 to 0.05 MPa, and more preferably 0.03 to 0.045 MPa.

[0063] In the present invention, the second concentrate obtained by the second concentration preferably has a relative density of 1.10 to 1.14 at 20° C. In the present invention, the relative density of the second concentrate is the density relative to water.

[0064] After obtaining the second concentrated solution, the present invention mixes the second concentrated solution with an aqueous solution of Erwinia japonica to obtain the Zhuling Tang.

[0065] In the present invention, the mass ratio of the glue to water in the aqueous solution of glue is preferably 1:(2 to 6), and more preferably 1:(2.5 to 5).

[0066] In the present invention, the method for preparing the aqueous solution of acrylic gelatin preferably includes dissolving the acrylic gel in water, sieving the solution, and the portion below the sieve being the aqueous solution of acrylic gelatin. In the present invention, the dissolution is preferably carried out under heating and stirring conditions, and the present invention does not particularly limit the stirring speed or time, as long as the stirring is continued until the acrylic gelatin is dissolved and a uniform acrylic gel solution is obtained. After the dissolution, the present invention preferably further includes removing bubbles and impurities from the upper layer of the dissolution system, and then sieving the solution. In the present invention, the size of the sieve used for sieving is preferably 100 mesh.

[0067] In the present invention, the specific process for mixing the second concentrated liquid and the aqueous gelatin solution is not particularly limited, as long as the materials are mixed uniformly.

[0068] The present invention provides a method for preparing a polygala extract, The method includes concentrating the Choling Tang prepared by the preparation method described in the above technical solution (hereinafter referred to as the third concentration) to obtain the Choling extract, wherein the temperature of the third concentration is 90°C or less, and the relative density of the Choling extract is 1.12 to 1.16 (20°C).

[0069] In the present invention, the third concentration is preferably carried out under steam heating conditions, the temperature of the third concentration is preferably 70 to 90°C, more preferably 75 to 90°C, and most preferably 85°C, the vacuum pressure of the third concentration is preferably 0.03 to 0.06 MPa, and more preferably 0.04 to 0.05 MPa, the incubation time of the third concentration is preferably 6 hours or less, and the heated steam pressure of the third concentration is preferably 0.01 to 0.05 MPa, and more preferably 0.03 to 0.045 MPa.

[0070] In the present invention, the relative density of the Choling extract is 1.12 to 1.16, and the relative density of the Choling extract is the relative density at 20° C. In the present invention, the relative density of the Choling extract is the density relative to water.

[0071] The present invention provides a method for preparing a solid preparation of Zhuling, the solid preparation including dry powder or granules, the method for preparing the dry powder includes the steps of: mixing Zhuling Tang prepared by the preparation method described in the above technical solution with an aqueous solution of auxiliary materials, and then concentrating the mixture to obtain a thick paste; and vacuum-drying the thick paste to obtain Zhuling dry powder, the mass ratio of the auxiliary materials to the Zhuling Tang is (1~7):50, and the mass of the Zhuling Tang is calculated as the total mass of Zhuling, Poria, Zeaxanthus, Talc and Erjiao; The method for preparing the granules includes dry granulating the dried powder of Choling to obtain Choling granules.

[0072] In the present invention, the method for preparing the dry powder includes the steps of mixing the aqueous solutions of Zhuling Tang and auxiliary materials prepared by the preparation method described in the above technical solution, and then concentrating (hereinafter referred to as the fourth concentration) to obtain a mixed thick paste, and vacuum drying the mixed thick paste to obtain Zhuling Tang dried powder, where the mass ratio of the auxiliary materials to the Zhuling Tang is (1~7):50, and the mass of the Zhuling Tang is calculated as the total mass of Zhuling, Poria, Zeaxanthus, talc and Erjiao.

[0073] In the present invention, the auxiliary material is preferably a pharmaceutically acceptable auxiliary material.

[0074] In the present invention, the auxiliary material is preferably at least one selected from the group consisting of dextrin, lactose, and soluble starch, and more preferably lactose.

[0075] In the present invention, the mass ratio of the auxiliary material to the Choling-to is (1-7):50, and more preferably (1.5-6.0):50.

[0076] In the present invention, the fourth concentration is preferably carried out under steam heating conditions, the temperature of the fourth concentration is preferably 70 to 90°C, more preferably 75 to 90°C, the vacuum pressure of the fourth concentration is preferably 0.03 to 0.06 MPa, more preferably 0.04 to 0.05 MPa, the incubation time of the fourth concentration is preferably 6 hours or less, and the heated steam pressure of the fourth concentration is preferably 0.01 to 0.05 MPa, more preferably 0.03 to 0.045 MPa.

[0077] In the present invention, the relative density of the mixed thick paste is preferably 1.12 to 1.16. The relative density of the mixed thick paste is the relative density at 20° C. In the present invention, the relative density of the mixed thick paste is the density relative to water.

[0078] In the present invention, after the fourth concentration, the present invention preferably pours the obtained mixed thick paste into a rotating barrel, weighs it, and prepares it for use.

[0079] In the present invention, the vacuum drying is preferably vacuum belt drying, and the operation parameters of the vacuum belt drying preferably include the following: The material supply temperature is preferably 75 to 85°C, more preferably 76 to 82°C, the material supply rate is preferably 2.0 to 3.5 L / h, more preferably 2.2 to 3.5 L / h, the crawler travel speed is preferably 150 ± 50 mm / min, more preferably 150 ± 20 mm / min, the rotation speed of the material distribution motor is preferably 100 r / min, the swing arm angle is preferably 12 to 25 °, more preferably 15 to 22 °, the heating temperature zones are preferably 95 ± 5 °C zone 1, 96 ± 5 °C zone 2, and 96 ± 5 °C zone 3, in that order, the vacuum degree is preferably 0.097 to 0.1 MPa, more preferably 0.098 to 0.09 MPa, the cutter speed is preferably 10 to 20 seconds / cycle, more preferably 11 to 18 seconds / cycle, and the cooling temperature is preferably 30 ± 5 °C.

[0080] In the present invention, after the vacuum drying, the powder is preferably collected using a rotary barrel and sealed and packaged to obtain the dried powder of Chorizome Rhizome for use.

[0081] In the present invention, the method for preparing the granules includes dry granulating the dried powder of Choling to obtain Choling granules.

[0082] The dry granulation is preferably carried out in a granulator, and the operating parameters of the dry granulation preferably include the following: the pressure of the press roll of the granulator is preferably 12 to 20 MPa, more preferably 13 to 16 MPa, the rotation speed of the press roll of the granulator is preferably 4 to 10 r / min, more preferably 5 to 8 r / min, the vertical rotation speed of the screw feeder of the granulator is preferably 25 to 40 r / min, more preferably 30 to 38 r / min, the horizontal rotation speed of the screw feeder of the granulator is preferably 70 to 120 r / min, more preferably 80 to 115 r / min, and the mesh number of the sieve is preferably 14 mesh.

[0083] In the present invention, it is preferred that the granules obtained by the dry granulation are sieved and then placed in a rotating barrel for use.

[0084] In the present invention, the granules are preferably packaged in a composite film at 6 g per bag, and the filling amount of 6 g per bag is controlled to 5.58 to 6.42 g.

[0085] The present invention provides a Zolin solid formulation prepared by the preparation method described in the above technical solution, wherein the dry paste ratio of the Zolin solid formulation is 18-23%, the total content of 23-acetylalisol B and 23-acetylalismol C in the Zolin solid formulation is 0.011-0.020 wt%, and the total content of donkey-derived polypeptide A1 and donkey-derived polypeptide A2 in the Zolin solid formulation is 0.091%-0.18 wt%.

[0086] In the present invention, the raw materials for preparing 1,000 parts of Choling granules, in terms of mass parts, preferably include 750 parts of Choling, 750 parts of Poria, 750 parts of Zelkova, 750 parts of Talc, 750 parts of Euglena, and 100 to 400 parts of auxiliary materials, and the mass of the auxiliary materials is more preferably 120 to 380 parts, and even more preferably 140 to 360 parts.

[0087] In a specific embodiment of the present invention, the raw materials for preparing 1000g of Zoling Granules preferably include 750g of Zoling, 750g of Poria Cocos, 750g of Zeaxanthin, 750g of Talc, 750g of Erwinia Japonica, and 100-400g of auxiliary materials.

[0088] In the present invention, the mass of the auxiliary material is more preferably 120 to 380 g, and even more preferably 140 to 360 g. The auxiliary material is preferably at least one selected from the group consisting of dextrin, lactose, and soluble starch, and more preferably lactose.

[0089] In the present invention, the standard of the Choling granules is preferably 6 g per bag.

[0090] In the present invention, the Choling granules are preferably packaged in a pharmaceutical composite film.

[0091] The present invention also provides a method for researching and evaluating the preparation method of the Zhongling solid formulation. The quality standard of the Zhongling Tang reference sample is used as the benchmark, and the dry paste rate is used as the main evaluation index, supplemented by content measurement indexes in stages. In the extraction, extract post-treatment (purification), and concentration stages of the preparation method of the Zhongling solid formulation, the contents of 23-acetylalisol B and 23-acetylalisol C are preferably used as auxiliary indexes. In the vacuum drying and dry granulation stages, the contents of 23-acetylalisol B and 23-acetylalisol C and the content of characteristic polypeptides are preferably used as auxiliary indexes. The present invention preferably starts tracking the changes in these indexes from the addition of the raw medicinal materials for the Zhongling solid formulation, and systematically evaluates the preparation method of the Zhongling solid formulation.

[0092] In the present invention, the method for detecting the dry paste ratio preferably includes the following steps:

[0093] (1) Constant weight of weighing bottle Open the lid of a clean weighing bottle, dry it in an electric fan oven at 105°C for 3 hours, close the lid, transfer it to a dryer, cool it for 30 minutes, weigh it accurately, dry it at the same temperature for 1 hour, cool it for 30 minutes, and then weigh it again. The difference between two consecutive weighings must not exceed 0.3 mg.

[0094] (2) Measurement The intermediate product is measured by taking an appropriate amount of the uniformly mixed test substance, placing it in a constant-weight weighing bottle, weighing it accurately, evaporating it in a water bath until it is almost dry, transferring it to a dryer and drying it at 105°C for 5 hours, removing it from the dryer, cooling it for 30 minutes, and weighing it accurately. It is then dried for another hour, removed from the dryer, cooled it for 30 minutes, and weighed again. The difference between the two weighings must not exceed 5 mg.

[0095] The detection method for the finished product shall be according to the drying method, Method 2 of General Principle 0832 of the 2020 edition of the Chinese Pharmacopoeia.

[0096] (3) Calculation Intermediate products:

number

number

[0097] In the present invention, the content of 23-acetylalisol B and 23-acetylalisol C is preferably determined according to high performance liquid chromatography (General Principle 0512 of the 2020 edition of the Chinese Pharmacopoeia).

[0098] Chromatography conditions and system suitability test: Octadecylsilane-bonded silica gel was used as the packing material, acetonitrile was used as the mobile phase A, and 0.2% phosphoric acid solution was used as the mobile phase B. Gradient elution was performed (0-15 min, 55-70% A; 15-30 min, 70-80% A; 30-35 min, 80-95% A; 35-45 min, 95% A). The detection wavelengths were 208 nm and 246 nm, and the column temperature was 30°C. The theoretical plate number, calculated using 23-acetylalisol B, should be greater than 3,000.

[0099] Preparation of standard solution: Weigh out appropriate amounts of 23-acetylalisol B standard substance and 23-acetylalisol C standard substance, and add methanol to prepare a mixed standard solution containing 3 μg of 23-acetylalisol B and 2.5 μg of 23-acetylalisol C per mL.

[0100] Preparation of test substance solution: Accurately weigh out an appropriate amount of sample and place it in a 50 mL volumetric flask. Add an equal amount of acetonitrile, weigh, and ultrasonicate for 30 minutes. Allow to cool, then add 50% acetonitrile up to the line, shake well, and filter. Take 25 mL of the filtrate and evaporate to dryness. Dissolve the residue in 50% acetonitrile, make the volume to 10 mL, filter, and take the next filtrate to obtain the solution.

[0101] Measurement method: 10 μL of the standard solution and test substance solution are precisely drawn up, injected into a liquid chromatograph, and measured.

[0102] In the present invention, the method for measuring the content of characteristic polypeptides is preferably performed according to high performance liquid chromatography-mass spectrometry (General Rules 0512 and 0431 of the 2020 edition of the Chinese Pharmacopoeia).

[0103] Chromatography, mass spectrometry conditions and system suitability test: Octadecylsilane-bonded silica gel was used as the packing material (chromatography column inner diameter 2.1 mm), acetonitrile was used as the mobile phase A, and 0.1% formic acid solution was used as the mobile phase B. Gradient elution (0–15 min, 5% → 14% A) was performed at a flow rate of 0.3 mL / min.

[0104] Multiple reaction monitoring (MRM) was performed using a triple quadrupole mass spectrometer in electrospray ionization (ESI) positive ion mode, and the monitored ion pairs are shown in Table 1.

[0105] JPEG2025531477000004.jpg45170

[0106] The theoretical plate number should be 4000 or more, calculated for the donkey-derived polypeptide A1 peak.

[0107] Preparation of standard substance solution: Take appropriate amounts of donkey-derived polypeptide A1 standard substance and donkey-derived polypeptide A2 standard substance, weigh them accurately, and add 1% ammonium bicarbonate solution to prepare a mixed solution containing 2.5 μg of donkey-derived polypeptide A1 standard substance and 2.5 μg of donkey-derived polypeptide A2 standard substance per mL.

[0108] Preparation of test substance solution: Take an appropriate amount of sample, weigh it accurately, place it in a 25mL volumetric flask, add 20mL of 1% ammonium bicarbonate solution, and sonicate for 30 minutes (power 250W, frequency 40kHz). Dilute to the mark with 1% ammonium bicarbonate solution and shake well. Take 200µL of the above solution, place it in a microcentrifuge tube, add 200µL of 5mg / mL trypsin solution, add 600µL of 1% ammonium bicarbonate solution, shake well, and let it digest enzymatically at a constant temperature of 37°C for 12 hours. Filter and collect the filtrate.

[0109] Measurement method: 1mL, 2mL, 5mL, 10mL, 20mL, and 25mL of the standard solution are accurately measured and placed in a 50mL volumetric flask, and 1% ammonium bicarbonate solution is added to dilute to the graduated mark to prepare the calibration curve solution. 5μL of each of the calibration curve solutions and the test substance solution at different concentrations are accurately aspirated and injected into a high-performance liquid chromatography-mass spectrometer. A calibration curve is created with the peak area of ​​the standard substance as the ordinate and the concentration of the standard substance as the abscissa. The amounts corresponding to donkey-derived polypeptide A1 and donkey-derived polypeptide A2 in the test substance solution are read from the calibration curve and calculated.

[0110] The present invention further provides quality control standards for the Zhuling solid preparations described in the above technical solutions, including HPLC characteristic spectrum of Zhuling solid preparations, identification of characteristic polypeptides, content determination, and dry paste rate.

[0111] The quality control standards for the solid formulation of Choling preferably include the properties of granules, particle size, solubility, filling amount difference, and microbial limit requirements.

[0112] In the present invention, the formulation of the Choling granules is preferably Choling 750 parts, Poria 750 parts, Zeaxanthin 750 parts, Talc 750 parts, and Erwinia 750 parts by mass.

[0113] In the present invention, the preparation method of the Choling granules is carried out with reference to the preparation method of Choling granules provided by the above technical solution.

[0114] In the present invention, the properties of the Choling granules are light brown to dark brown granules, and are slightly bitter and slightly sweet.

[0115] In the present invention, the method for identifying the Zhuling granules is as follows: (1) 4 g of Zhuling granule powder is taken, 50 mL of diethyl ether is added, ultrasonicated for 10 minutes, filtered, the filtrate is evaporated to dryness, and 1 mL of methanol is added to dissolve the residue to prepare the test substance solution. Another 1 g of Zhuling medicinal material and a control Poria corymbine are taken and prepared in the same manner. The test is performed according to thin-layer chromatography (General Principle 0502 of the 2020 edition of the Chinese Pharmacopoeia). 2-5 μL of the control Poria corymbine solution and Zhuling medicinal material solution, and 20-50 μL of test substance solution A are absorbed and spotted on the same silica gel G thin-layer plate. The sample is developed with cyclohexane-ethyl acetate-formic acid (8:1:0.1) as a developing agent, dried, and then directly subjected to fluorescence at 365 nm, or sprayed with 10% sulfuric acid ethanol, dried, and then subjected to fluorescence at 365 nm. In the chromatogram of the test substance, a spot of the same color appears at a position corresponding to that of the control drug.

[0116] (2) Take 2 g of this product powder, add 50 mL of methanol, sonicate for 10 minutes, filter, evaporate the filtrate, and dissolve the residue in 1 mL of methanol to prepare the test substance solution. Another 1 g of Zea mays (Zeniella zea) control solution was prepared using the same method. Test according to thin-layer chromatography (General Rule 0502, 2020 Edition of the Chinese Pharmacopoeia), absorbing 2 μL of the control solution and 5 μL of the test substance solution, spotting them onto the same silica gel G thin-layer plate, developing with cyclohexane-ethyl acetate (1:1) as the eluent, drying, spraying with 2% vanillin sulfuric acid solution, and heating at 105°C until the spots become clear. The chromatogram of the test substance should show spots of the same color at the positions corresponding to those of the control substance.

[0117] (3) Accurately weigh 0.1 g of sample powder into a 25 mL volumetric flask, add 20 mL of 1% ammonium bicarbonate solution, and sonicate for 30 minutes (power 250 W, frequency 40 kHz). Dilute to the mark with 1% ammonium bicarbonate solution and shake well. Transfer 200 μL of the above solution to a microcentrifuge tube, add 200 μL of 1 mg / mL trypsin solution, and add 600 μL of 1% ammonium bicarbonate solution. Shake well and allow to enzymatically hydrolyze at 37°C for 12 hours to obtain the test substance solution. Take 0.1 g of another gelatinized control drug and prepare a control drug solution using the same method. [Content Measurement] Test according to the chromatography and mass spectrometry conditions for the characteristic polypeptide item. The mass-to-charge ratio (m / z) of 539.8 (double charge) → 612.4 and m / z of 539.8 (double charge) → 923.8 were selected as detection ion pairs. Take the reference gelatinous drug solution and inject 5 μL. The signal-to-noise ratio of the MRM chromatographic peaks measured according to the above detection ion pairs should be greater than 3:1.

[0118] Five microliters of the test substance solution is drawn up and injected into a high-performance liquid chromatography-mass spectrometer for measurement. The ion flow chromatogram of the test substance, extracted with the mass-to-charge ratio (m / z) 539.8 (double charge) → 612.4 and m / z 539.8 (double charge) → 923.8 as the detection ion pairs, should simultaneously show chromatographic peaks that match the retention times of the control drug.

[0119] In the present invention, the inspection of the Zhuling Granules should comply with the provisions of each item related to the granule category (General Principle 0104 of the 2020 edition of the Chinese Pharmacopoeia).

[0120] In the present invention, the dry paste ratio of the Zhuling granules is preferably measured according to the moisture determination method (General Rule 0832 of the 2020 edition of the Chinese Pharmacopoeia), and should be 18.0% to 23.0%.

[0121] In the present invention, the method for measuring the exudates of the Zhuling granules is preferably as follows: the Zhuling granules are finely ground, and about 2 g of them are taken and precisely weighed, and 100 mL of ethanol is precisely added. The content of the alcohol-soluble exudates is measured according to the hot immersion method in the section on the measurement of alcohol-soluble exudates (General Principles 2201 of the 2020 edition of the Chinese Pharmacopoeia), which should be 12.0% to 23.5%.

[0122] In the present invention, the characteristic spectrum of the Zhuling Granules is measured according to high performance liquid chromatography (General Principles 0512 of the 2020 edition of the Chinese Pharmacopoeia).

[0123] Chromatography conditions and system suitability test: Octadecylsilane-bonded silica gel was used as the packing material, acetonitrile was used as mobile phase A, and 0.2% phosphoric acid solution was used as mobile phase B. Gradient elution was performed according to the specifications in Table 2, the detection wavelength was 208 nm, and the column temperature was 30°C. The theoretical plate number calculated based on the peak of Alisol B should be greater than 2000.

[0124] JPEG2025531477000005.jpg41170

[0125] Preparation of reference substance solution: Accurately weigh Alisol A, Alisol B, and 23-acetylalisol B, and dissolve them in acetonitrile to prepare a mixed standard substance solution with mass concentrations of 40 μg / mL Alisol A, 25 μg / mL Alisol B, and 23 μg / mL 23-acetylalisol B.

[0126] Preparation of test substance solution: Take 1.0 g of sample powder, weigh it accurately, and place it in a stoppered Erlenmeyer flask. Add 50 mL of acetonitrile accurately, seal it, sonicate for 30 minutes (power 800 W, frequency 40 kHz), allow to cool, filter, evaporate the filtrate to dryness, add 80% acetonitrile to the residue, dissolve, adjust the volume to 5 mL, and filter to obtain the test substance solution.

[0127] Measurement method: 20 μL each of the reference substance solution and the test substance solution is precisely aspirated, injected into a liquid chromatograph, and measured.

[0128] The characteristic spectrum of the test substance should show eight characteristic peaks, three of which should correspond to the retention times of the corresponding reference substance peaks. The peak corresponding to the Alisol B reference substance peak is the S peak. The relative retention times of each characteristic peak and the S peak are calculated, and the relative retention times should be within ±10% of the specified values. The specified values ​​are 0.61 (Peak 1), 1.00 (Peak 2), 1.28 (Peak 3), 1.31 (Peak 4), 1.34 (Peak 5), 1.52 (Peak 6), 1.57 (Peak 7), and 1.81 (Peak 8). The control characteristic spectrum is shown in Figure 2. In Figure 2, Peak 1 corresponds to Alisol A, Peak S corresponds to Alisol B, and Peak 3 corresponds to 23-acetylalisol B.

[0129] In the present invention, the amino acid content of the Zhuling Granules is determined according to high performance liquid chromatography (General Principle 0512 of the 2020 edition of the Chinese Pharmacopoeia).

[0130] Chromatography conditions and system suitability test: Octadecylsilane-bonded silica gel was used as the packing material, and acetonitrile-0.1 mol / L sodium acetate solution (adjusted to pH 6.5 with acetic acid) (7:93) was used as mobile phase A, and acetonitrile-water (4:1) was used as mobile phase B. Gradient elution was performed according to the specifications in Table 3, with a flow rate of 0.8 mL / min, a detection wavelength of 254 nm, and a column temperature of 43°C. The theoretical plate number, calculated at the L-hydroxyproline peak, should be greater than 5,000.

[0131] JPEG2025531477000006.jpg45170

[0132] Preparation of standard solution: Take appropriate amounts of L-hydroxyproline, glycine, alanine, and proline, weigh them accurately, and add 0.1 mol / L hydrochloric acid solution to prepare a mixed solution containing 80 μg of L-hydroxyproline, 0.16 mg of glycine, 70 μg of alanine, and 0.12 mg of proline per mL.

[0133] Preparation of test substance solution: 0.2 g of powder was accurately weighed and placed in a 25 mL volumetric flask. 20 mL of 0.1 mol / L hydrochloric acid solution was added, and ultrasonicated for 30 minutes (power 500 W, frequency 40 kHz). Allow to cool, and then add 0.1 mol / L hydrochloric acid solution up to the line and shake well. 2 mL was accurately weighed and placed in a 10 mL ampoule. 2 mL of hydrochloric acid was added, and hydrolyzed at 150°C for 1 hour. Allow to cool, then transferred to an evaporating dish and washed with 10 mL of water in several portions. The washed solution was then transferred to an evaporating dish and evaporated to dryness. The residue was dissolved in 0.1 mol / L hydrochloric acid solution, transferred to a 25 mL volumetric flask, and added 0.1 mol / L hydrochloric acid solution up to the line. Shake well to obtain the desired solution.

[0134] Accurately measure 5 mL of each of the standard solution and test substance solution into 25 mL volumetric flasks, add 2.5 mL of 0.1 mol / L phenylisothiocyanate (PITC) in acetonitrile and 2.5 mL of 1 mol / L triethylamine in acetonitrile, shake well, leave at room temperature for 1 hour, then add 50% acetonitrile up to the graduation mark and shake well. Take 10 mL, add 10 mL of n-hexane, shake, leave for 10 minutes, remove the lower layer, filter, and collect the filtrate.

[0135] Measurement method: After derivatization, 5 μL of each of the standard solution and test substance solution is precisely aspirated, injected into a liquid chromatograph, and measured.

[0136] In the present invention, the amount of gelatin contained in each bag is 0.25 to 0.55 g of L-hydroxyproline, 0.50 to 1.05 g of glycine, 0.20 to 0.45 g of alanine, and 0.30 to 0.60 g of proline.

[0137] In the present invention, the characteristic polypeptide of the Zhuling Granules is measured according to high performance liquid chromatography-mass spectrometry (General Principles 0512 and 0431 of the 2020 edition of the Chinese Pharmacopoeia).

[0138] Chromatography, mass spectrometry conditions and system suitability test: Octadecylsilane-bonded silica gel was used as the packing material (chromatography column inner diameter 2.1 mm), acetonitrile was used as mobile phase A, and 0.1% formic acid solution was used as mobile phase B. Gradient elution was performed according to the specifications in Table 4, with a flow rate of 0.3 mL / min.

[0139] JPEG2025531477000007.jpg29170

[0140] Multiple reaction monitoring (MRM) was performed using a triple quadrupole mass spectrometer in electrospray ionization (ESI) positive ion mode, and the monitored ion pairs are shown in Table 5.

[0141] JPEG2025531477000008.jpg45170

[0142] The theoretical plate number should be 4000 or more, calculated for the donkey-derived polypeptide A1 peak.

[0143] Preparation of standard substance solution: Take appropriate amounts of donkey-derived polypeptide A1 standard substance and donkey-derived polypeptide A2 standard substance, weigh them accurately, and add 1% ammonium bicarbonate solution to prepare a mixed solution containing 2.5 μg of donkey-derived polypeptide A1 standard substance and 2.5 μg of donkey-derived polypeptide A2 standard substance per mL.

[0144] Preparation of test substance solution: 0.1 g of sample powder was weighed accurately and placed in a 25 mL volumetric flask. 20 mL of 1% ammonium bicarbonate solution was added. Sonicate for 30 minutes (power 250 W, frequency 40 kHz). Dilute to the mark with 1% ammonium bicarbonate solution and shake well. 200 μL of the above solution was placed in a microcentrifuge tube. 200 μL of 1 mg / mL trypsin solution was added, followed by 600 μL of 1% ammonium bicarbonate solution. Shake well and allow to enzymatically hydrolyze at 37°C for 12 hours. Filter and collect the filtrate.

[0145] Measurement method: 1mL, 2mL, 5mL, 10mL, 20mL, and 25mL of the standard solution are accurately measured and placed in a 50mL volumetric flask, and 1% ammonium bicarbonate solution is added to dilute to the graduated mark to prepare the calibration curve solution. 5μL of each of the calibration curve solutions and the test substance solution at different concentrations are accurately aspirated and injected into a high-performance liquid chromatography-mass spectrometer. A calibration curve is created with the peak area of ​​the standard substance as the ordinate and the concentration of the standard substance as the abscissa. The amounts corresponding to donkey-derived polypeptide A1 and donkey-derived polypeptide A2 in the test substance solution are read from the calibration curve and calculated.

[0146] In the present invention, the total amount of donkey-derived polypeptide A1 and donkey-derived polypeptide A2 contained in each bag should be 5.5 to 11 mg.

[0147] In the present invention, the functions and main indications of Zhuling Granules are as follows: induce diuresis, nourish yin, and clear heat. It is used for treating people with poor urination, fever, thirst and urge to drink, irritability and insomnia, cough, nausea, diarrhea, white or faintly yellow tongue coating, weak pulse, bloody stool, and painful, astringent urination, difficulty in urinating, and lower abdominal pain.

[0148] In the present invention, the dosage and administration of the Choling Granules is one bag taken with hot water three times a day.

[0149] In the present invention, the standard of the Choling granules is 6 g per bag.

[0150] In the present invention, the storage method of the Choling granules is sealed.

[0151] The following will clearly and completely describe the technical solutions of the present invention with reference to the embodiments of the present invention. Obviously, the described embodiments are only some of the embodiments of the present invention, and not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without any creative work fall within the protection scope of the present invention.

[0152] In the examples and comparative examples of the present invention, The basic ingredients of the decoction pieces: Poria cocos, Poria cocos, talc, and Ejiao, are consistent with the basic ingredients listed in the 2020 edition of the Chinese Pharmacopoeia, and Alisma orientale (Sam.) Juzep., a plant of the Alismaceae family and the genus Alisma.

[0153] Each herbal medicine is first prepared before use.

[0154] The process for processing zhuling is as follows: (1) Sorting to remove impurities; (2) Soaking at 5-35°C for 0.5-3 hours; (3) Moisturizing at 5-35°C for 3-6 hours; (4) Cutting into uniform, thick slices of 2-4 mm; (5) Drying at 70-80°C for 5-6 hours.

[0155] The process of processing Poria cocos is as follows: (1) Selection to remove impurities.

[0156] The processing method for zeaxanthin is as follows: (1) Sorting to remove impurities; (2) Spraying and washing for 10-15 minutes; (3) Soaking at 15-35°C for 5-8 hours; (4) Moisturizing at 15-35°C for 18-36 hours; (5) Cutting into uniform, thick slices of 2-4 mm; (6) Drying at 50-60°C for 5-6 hours.

[0157] The talc is produced by the following methods: (1) sorting to remove impurities; (2) washing thoroughly; (3) drying at 70-80°C for 3-4 hours; (4) crushing in a grinder and passing through an 80-mesh pharmaceutical sieve.

[0158] Example 1 The raw materials for the Zoling granules prepared in the examples of the present invention are 750g of Zoling, 750g of Poria cocos, 750g of Zeaxanthin, 750g of Talc, 750g of Erwinia gelatinosa, and 100-400g of auxiliary materials.

[0159] The origins of the raw materials are as follows: Boar: Purchased from Shandong Baimido Chinese Medicine Pill Co., Ltd. Poria cocos: Purchased from Anhui Shen-Tai Pharmaceutical Co., Ltd. Zebian: Purchased from Shandong Baimido Chinese Medicine Pill Co., Ltd. Talc: Purchased from Quzhou South Kong Medicinal Co., Ltd. Agluo: Purchased from Higashia Agluo Co., Ltd. Auxiliary materials: Purchased from Zhenjiang Kangfu Biological Engineering Co., Ltd.

[0160] 7.5 kg each of Poria cocos, Poria cocos, Zeaxanthus annuus, and Talc (bao decoction) were weighed and placed in an extraction tank. 8 to 14 times the amount of water as the pieces were added, and the mixture was soaked for 50 minutes to obtain a prepreg liquid. The steam valve of the extraction tank was opened, and the temperature was raised at a heating steam pressure of less than 0.20 MPa. When the temperature reached 90°C, the external circulation was opened, and timing began when the temperature reached a boil. The steam pressure was stabilized at 0.05 to 0.08 MPa and maintained for 40 to 50 minutes. The external circulation (first decoction) was then closed, and the first liquid was released to obtain the residue. The first liquid was passed through a 120-mesh filter and pumped into a separator for separation. The separation speed was controlled at 30 to 40 L / min, and the residue was discharged approximately every 20 minutes and centrifuged in a centrifuge. Each centrifuged liquid was passed through a plate-frame filter (filter membrane pore size 5 to 15 μm) and filtered to obtain the first aqueous extract.

[0161] The residue was added with water at a mass 6 to 12 times the amount of medicinal material, and heated at a heating steam pressure of less than 0.20 MPa. When the temperature reached 90°C, the external circulation was opened. When the temperature reached boiling, timing began. The steam pressure was stabilized at 0.05 to 0.08 MPa and maintained for 25 to 40 minutes. The external circulation (second decoction) was then closed, and the second medicinal liquid was released. The second medicinal liquid was passed through a 120-mesh filter and pumped into a separator for separation. The separation speed was controlled at 30 to 40 L / min, and the residue was discharged approximately every 20 minutes and centrifuged in a centrifuge. The centrifuged liquid from each run was passed through a plate-frame filter (filter membrane pore size 5 to 15 μm) and filtered to obtain the second aqueous extract.

[0162] The second aqueous extract is sucked into a vacuum concentrator, the steam valve is opened, heating is started, the temperature is controlled to 70-90°C, the vacuum pressure is 0.03-0.06 MPa, the steam pressure is 0.03-0.05 MPa, and the heat retention time for the concentration is set to 6 hours or less to obtain a first concentrated liquid. The first concentrated liquid and the first aqueous extract are mixed and sucked into a vacuum concentrator, the steam valve is opened, heating is started, the temperature is controlled to 70-90°C, the vacuum pressure is 0.03-0.06 MPa, the steam pressure is 0.01-0.05 MPa, and the heat retention time for the concentration is set to 6 hours or less to obtain a second concentrated liquid. The relative density of the second concentrate is 1.10~1.14 (20℃). For use, weigh out 7.5 kg of gelatin glue, add 2~6 times the mass of water of the gelatin glue, heat to boiling and stir constantly to dissolve the gelatin glue into a uniform gelatin glue solution, and sieve through a 100 mesh sieve to obtain an aqueous gelatin glue solution. For use, weigh out 1.0~4.0 kg of auxiliary materials, put the auxiliary materials into a stainless steel barrel, add 2~6 times the mass of water of the auxiliary materials, and stir to completely dissolve, to obtain an aqueous solution of auxiliary materials. For use,

[0163] The second concentrate, the aqueous gelatin solution, and the aqueous solution of auxiliary materials are sucked into a single-effect concentrator, the steam valve is opened, heating is started, the temperature is controlled at 70-90°C, the vacuum pressure is 0.03-0.06MPa, the steam pressure is 0.01-0.05MPa, and the concentration warming time is set to 6 hours or less until a mixed thick paste with a relative density of 1.12-1.16 (20°C) is obtained, which is then poured into a rotating barrel, weighed, and prepared for use.

[0164] The vacuum belt dryer was turned on and the mixed thick paste was vacuum dried to produce powder. The operating parameters were as follows: material supply temperature was 75-85°C, material supply rate was 2.0-3.5 L / h, crawler speed was 150±50 mm / min, swing arm angle was 12-25°, material distribution motor speed was 100 r / min, first, second, and third heating temperatures were 95±5°C, 96±5°C, and 96±5°C, respectively, cooling temperature was 30±5°C, cutter speed was 10-20 s / cycle, and vacuum level was 0.097-0.1 MPa. The dried porcini powder was obtained. The powder was collected in a rotating barrel, sealed, and packaged for use.

[0165] The dried Zoloft granules were weighed and transferred to a granulator. The pressure of the press roll was adjusted to 12-20 MPa, the screw feeder rotation speed was adjusted to 25-40 r / min vertically and 70-120 r / min horizontally, and the press roll rotation speed to 4-10 r / min horizontally. The sieve mesh was set to 14 mesh. Zoloft granules were obtained, sieved, and placed in a rotating barrel for use. The Zoloft granules were packaged in 6g bags using composite film, with the filling weight per bag controlled at 5.58g-6.42g.

[0166] Example 2 Study on the preparation process of Choling granules

[0167] 1. Purpose of the experiment The quality standards of the standard sample of Choreito were used as the benchmark, and the dry paste rate, 23-acetylalisol B and 23-acetylalisol C contents were used as evaluation indicators, which were determined by considering the number of extractions, extraction time, and amount of water added for the Choreito preparation.

[0168] 2. Experimental Design Zhuling Tang tea pieces (excluding Ejiao, containing 55.2g of Zhuling, Poria, Zeaxanthus, and Talc) were placed in a round-bottom flask, and an appropriate amount of water was added. After 50 minutes of steeping, the tea was placed in an intelligent thermostatic jacket and refluxed to obtain the extract. The process parameters were determined by detecting the dry paste ratio and the contents of 23-acetylalisol B and 23-acetylalisol C in the decoction. The extraction parameters are shown in Table 6.

[0169] The number of extractions, extraction time, and amount of water added were determined in a single factor experiment.

[0170] JPEG2025531477000009.jpg48170

[0171] 3. Experimental Method 3.1 Experimental method for considering the number of extractions: Zhuling Tang pieces (excluding Ejiao, Zhuling, Poria, Zeaxanthus, and Talc were each 55.2g) were placed in a round-bottom flask, and an appropriate amount of water was added. After soaking for 50 minutes, they were placed in an intelligent constant temperature electric heating jacket and reflux extracted three times. The first time, 14 times the amount of water was added and the extraction was continued for 45 minutes. The second and third times, 12 times the amount of water was added and the extraction was continued for 30 minutes. The extracts obtained each time were used to detect the dry paste rate and indicator components.

[0172] 3.2 Experimental method for investigating extraction time: Zhuling Tang pieces (excluding Ejiao, Zhuling, Poria, Zeaxanthus, and Talc were each 55.2g) were placed in a round-bottom flask, and an appropriate amount of water was added. After soaking for 50 minutes, they were placed in an intelligent constant temperature electric heating jacket and reflux extracted three times. The extraction times are shown in Table 6. The first time, 14 times the amount of water was added, and the second and third times, 12 times the amount of water was added.

[0173] 3.3 Experimental method for considering the amount of water added for extraction: Zhuling Tang pieces (excluding Ejiao, Zhuling, Poria, Zeaxanthus, and Talc were each 55.2g) were placed in a round-bottom flask, and an appropriate amount of water was added. After soaking for 50 minutes, the pieces were placed in an intelligent constant temperature electric heating jacket and reflux extracted twice. The extraction time was determined according to "3.2", and the amount of water added is shown in Table 6.

[0174] 4. Experimental Results 4.1 Consideration of sampling frequency is shown in Table 7.

[0175] JPEG2025531477000010.jpg90170

[0176] The results in Table 7 show the following: (1) Dry paste rate: The dry paste rate of the first extraction of Zhuling Granules (3.65%) was lower than the requirement for the reference sample (dry paste rate of 4.60%). The total dry paste rate of the first and second extractions (5.60%) was 21.7% higher than that of the reference sample, and the total dry paste rate of the first, second, and third extractions (6.80%) was 50% higher than that of the reference sample. From the perspective of compensating for losses in later processes (approximately 20% to 30%) and energy losses during production in modern factories, two extractions were selected.

[0177] (2) The contents of the indexes 23-acetylalisol B and 23-acetylalisol C in the first extraction were both higher than those in the reference sample. Under the current water addition conditions, the index components were completely extracted, and the number of extractions did not affect the index components.

[0178] 4.2 Extraction time considerations are shown in Table 8.

[0179] JPEG2025531477000011.jpg153170

[0180] The results in Table 8 show that extending the extraction time did not significantly increase the dry paste rate or content index, and the dry paste rate of the first two extractions at three different extraction times exceeded 80% of the total of the third extraction. The extraction was performed twice, with the extraction times being 45 and 30 minutes, respectively.

[0181] 4.3 Consideration of the amount of water added is shown in Table 9.

[0182] JPEG2025531477000012.jpg172170

[0183] The results in Table 9 show the following: (1) Dry paste ratio: With increasing water content, the dry paste ratio gradually increased. The total dry paste ratios of the first and second extractions of batches 006-010 were 11.1%, 19.3%, 23.0%, 28.7%, and 27.6% higher than those of the reference sample, respectively. The water content of the first extraction was 8-14 times, with no significant difference in dry paste ratio (RSD% less than 5). To compensate for losses in later processes (approximately 20%-30%), the water content was 10-14 times.

[0184] (2) Content index: The index content gradually increased with increasing extraction ratio. Under different water addition conditions, the total content index of the first and second extractions was higher than that of the reference sample. The content of 23-acetylalisol B remained essentially unchanged at 10 and 12 times water addition, exceeding 50% of the reference sample. The content of 23-acetylalisol C did not differ significantly under different water addition conditions, exceeding 50% of the reference sample in both cases, which could compensate for losses in subsequent processes. Taking into consideration both extraction efficiency and compensating for losses in subsequent processes, the water addition ratio was selected to be 10 to 14 times.

[0185] 5. Experimental Conclusion In summary, the extraction process parameters for Choling Granules were two extractions: the first time, 10-14 times the amount of water was added and the tea was boiled for 40-90 minutes; the second time, 8-12 times the amount of water was added and the tea was boiled for 25-60 minutes.

[0186] Example 3 Consideration of concentration methods

[0187] 1. Purpose of the experiment The influence of the concentration temperature and time of Zhuling granules on the dry paste rate, index component content and characteristic spectrum was considered, and the range of Zhuling granules concentration process parameters was determined.

[0188] 2. Experimental Method 2.1 Consideration of concentration temperature 1 L of the extract from the Choling granules was weighed and then concentrated under reduced pressure in a rotary evaporator. The changes in the dry paste rate and the contents of the index components after concentration for 2 hours at 70, 80, 90, and 95°C were examined.

[0189] 2.2 Consideration of concentration time 1L of the extract from the Choling granules was weighed and then concentrated under reduced pressure in a rotary evaporator. The changes in the dry paste rate and the contents of the index components after concentration at 90℃ for 2, 4, and 6 hours were examined.

[0190] 3. Experimental Results 3.1 Considerations regarding concentration temperatures are shown in Table 10.

[0191] JPEG2025531477000013.jpg78170

[0192] The results in Table 10 show that: (1) Dry paste ratio: Different concentration temperatures did not significantly affect the dry paste ratio of Choling granules (RSD% was 1.46).

[0193] (2) Index components: Different concentration temperatures significantly affected the content of 23-acetylalisol B in Choling Granules (RSD% was 7.04), but not 23-acetylalisol C (RSD% was 1.87), and not the content of 23-acetylalisol B + C (RSD% was 3.89). This indicates that 23-acetylalisol B in Choling Granules is temperature sensitive.

[0194] The experimental results showed that the concentration temperature range was 70-95℃, as this temperature did not significantly affect the dry paste rate and index components of Choling granules.

[0195] 3.2 Considerations for concentration time are shown in Table 11.

[0196] JPEG2025531477000014.jpg82170

[0197] The results in Table 11 show that: (1) Dry paste ratio: Different concentration times did not significantly affect the dry paste ratio of Choling Granules (RSD% was 4.39).

[0198] Concentration for 2, 4, and 6 h had no significant effect on the content of 23-acetylalisol B in Choling Granules (RSD% was 3.18), a significant effect on 23-acetylalisol C (RSD% was 6.77), and no significant effect on the content of 23-acetylalisol B + C (RSD% was 3.50), indicating that 23-acetylalisol C is sensitive to concentration time.

[0199] The experimental results showed that concentration at 90°C for 6 hours did not affect the dry paste rate or indicator components of Zoliang granules, indicating that the Zoliang granule extract could be concentrated for less than 6 hours under reduced pressure concentration conditions.

[0200] 4. Experimental Conclusion The dry paste ratio and the contents of 23-acetylalisol B and 23-acetylalisol C were used as evaluation indicators to determine the effects of different concentration temperatures and times on Zhongling granules. The concentration temperature ranged from 70 to 95°C, and the concentration time did not exceed 6 hours.

[0201] Example 4 100L extraction tank experiment To further verify the applicability of the extraction and concentration process parameters in the modernized production facility, a small-scale test verification of three batches was conducted using a 100L extraction tank.

[0202] 1. Experimental process 1.1 Soaking, extraction 1.4490 kg of Zhu Ling, 1.4490 kg of Poria Cocos, 1.4490 kg of Zelkova, 1.4490 kg of Talc, and 1.4490 kg of Er Jiao were weighed out. The talc was placed in a decoction bag and the top was tied in the appropriate position. Except for Er Jiao, the remaining four herbs were placed in a 100L extraction tank, and 10 times the amount of water as the pieces were added. The pieces were soaked for 50 minutes, then heated and extracted for 45 minutes to release the medicinal liquid. The second time, 8 times the amount of water as the pieces were added, and the pieces were soaked for 30 minutes to release the medicinal liquid.

[0203] 1.2 Post-treatment of the extract (purification) The extracts were mixed, centrifuged in a centrifuge, and then filtered through a plate and frame filter.

[0204] 1.3 Concentration The precisely filtered chemical solution was poured into a 100L single-effect concentrator and concentrated at a vapor pressure of 0.01-0.03 MPa, a vacuum pressure of 0.02-0.04 MPa, and a temperature of 80-90°C. The chemical solution's condition needed to be continuously monitored during concentration. The concentration was continued until the relative density reached 1.004-1.007 (at 20°C). The molten glue was then sucked in, and concentration continued until the relative density reached 1.12-1.18 (at 20°C).

[0205] 2. Test Results Three consecutive batches of Zhuling Granules were extracted and tested in a 100L extraction tank. The matching of the main equipment for extraction, purification, concentration, etc. was high, and all key process parameters were within the control range, and the entire testing process was smooth. The dry paste rate of the three batches of Zhuling Granule concentrate prepared all met the standards of the Zhuling Tang reference sample, and the index component contents were higher than those of the reference sample with small differences (the dry paste rate of the three batches of finished products, the RSD% of the contents of 23-acetylalisol B and 23-acetylalisol C were less than 5). The results are shown in Table 12.

[0206] JPEG2025531477000015.jpg87170

[0207] Example 5 Pilot-scale production testing The production process route and main parameters of the Zhuling Granules formulation were basically determined through small-scale optimization. To further verify the rationality of the production equipment and process route flow, three batches of pilot test verification were carried out in a 500L extraction tank, specifically as follows:

[0208] 1. Testing process 1.1 Soaking, extraction 7.452 kg of Zhu Ling, 7.452 kg of Poria Cocos, 7.452 kg of Zelkova, and 7.452 kg of talc were weighed out. The talc was placed in a decoction bag and the top was tied at the appropriate position. The bag was then placed in a 500L extraction tank, and 10 times the amount of water as the pieces were added. The pieces were soaked for 50 minutes, then heated and extracted for 45 minutes to release the medicinal liquid. The second time, 8 times the amount of water as the pieces were added, and the pieces were soaked for 30 minutes to release the medicinal liquid.

[0209] 1.2 Post-treatment of the extract (purification) Each drug solution was centrifuged in a centrifuge and then precisely filtered through a plate and frame filter.

[0210] 1.3 Concentration The chemical solution was poured into a vacuum concentrator, and the temperature was controlled at 70-90°C, the vacuum at 0.03-0.06 MPa, and the vapor pressure at 0.09 MPa or less. The boiling state and temperature were constantly monitored, and the vacuum pressure and vapor pressure were adjusted until the relative density of the chemical solution reached 1.004-1.007 (20°C), and it was ready for use.

[0211] 1.4 Aglue dissolution, auxiliary material dissolution 7.452 kg of eriolytic gelatin was weighed out and placed in a double boiler. Water approximately 2 to 6 times the amount of eriolytic gelatin was added, and the mixture was heated to a boil and stirred constantly until it was dissolved into a uniform eriolytic gelatin solution. The upper layer of bubbles and impurities were removed, and the mixture was passed through a 100-mesh sieve for preparation for use.

[0212] 3.000 kg of auxiliary materials (40% of the dry extract) were weighed out, the auxiliary materials were placed in a stainless steel barrel, and water approximately 2 to 6 times the amount of the auxiliary materials was added, stirred to completely dissolve, and the mixture was ready for use.

[0213] 1.5 Mixing, concentration The concentrate, glue and auxiliary liquid were sucked into a single-effect concentrator and continued to be concentrated until a thick paste with a relative density of 1.12-1.16 (20°C) was formed, which was then poured into a rotating barrel, weighed and prepared for use.

[0214] 1.6 Drying The mixed concentrate was poured into a liquid storage tank and heated to 75-85°C. A vacuum belt dryer was turned on to dry and mill the powder. The parameters were as follows: the material feed temperature was 75-85°C, the material feed rate was 2.0-3.5 L / h, the crawler travel speed was 150±50 mm / min, the swing arm angle was 12-25°, the material distribution motor speed was 100 r / min, the first, second, third, and fourth heating temperatures were 95±5°C, 96±5°C, 96±5°C, and 30±5°C (cooling), respectively. The cutter was turned on for 10-20 seconds per cycle, and the vacuum pressure was 0.097-0.1 MPa. The rotating barrel collected the powder, sealed it, and prepared it for use.

[0215] 1.7 Granulation The dry powder was weighed and transferred to a granulator. The granulator's press roll pressure was adjusted to 12-20 MPa, the screw feeder rotation speed was adjusted to 20-40 r / min vertically, 70-120 r / min horizontally, and 4-10 r / min pressure roller rotation speed. The sieve mesh was set to 14 mesh. The equipment status and granule properties were observed during operation, and the granulation parameters were adjusted accordingly. The powder was added as needed, and the granules were collected. The vibrating sieve was turned on, and the granules were placed in the hopper. The hopper valve was adjusted so that the material feeding speed was such that the fine powder could be completely separated from the fine granules. The granules were then placed in the rotating barrel for use. The fine powder was then re-granulated.

[0216] 2. Test Results Three batches of Choling Tang preparation were tested consecutively on a pilot scale. The process route was reasonable, and the matching of major equipment such as extraction, centrifugation, microfiltration, concentration, drying, and granulation was high. All key process parameters were within the control range, and the entire testing process was smooth. The dry paste rate and index components of the three batches of Choling Tang preparation produced all conformed to the Choling Tang reference sample standards, with small differences (the dry paste rate, 23-acetylalisol B and 23-acetylalisol C, and characteristic polypeptide (donkey-derived polypeptide A1, A2) contents of the three batches of products had RSD% of less than 5). The results are shown in Table 13.

[0217] JPEG2025531477000016.jpg103170

[0218] Comparative Example 1 100L extraction tank (standard sample method) test

[0219] Based on the requirement that the quality standard of Choling-Tang preparations be basically consistent with the reference sample, we attempted to directly use the preparation method of the reference sample to extract the preparation, in order to ensure that the quality of the preparation was consistent with the reference sample.

[0220] The standard sample was prepared as follows: 13.8 g of Choling Root, Poria Root, Zelkova, and talc were added to 800 mL of water, decoction was added, and the mixture was filtered to obtain approximately 400 mL of decoction. 13.8 g of Erjiao was added and dissolved in hot water, and the mixture was concentrated under reduced pressure to approximately 200 mL and freeze-dried to obtain the standard Choling Tang sample.

[0221] The preparation method of the reference sample is characterized by the fact that it is not soaked, the amount of water added is 14.5 times, and it is extracted once.

[0222] Following the standard sample preparation method, the formulation was extracted directly into a 100L extraction tank. The applicability of the method was evaluated based on the dry paste rate of the extract and the content indexes of 23-acetylalisol B and 23-acetylalisol C. The experimental procedure was as follows:

[0223] 1. Testing process 1.1 Extraction Weigh out 1.035 kg of Poria cocos, 1.035 kg of Poria cocos, 1.035 kg of Zelkova, and 1.035 kg of talc. Place the talc into a decoction bag and tie the top at the appropriate position. Place the bag in a 100L extraction tank, add 14.5 times the amount of water as the pieces, and extract for 45 minutes to release the liquid.

[0224] 1.2 Purification The extract was centrifuged in a centrifuge and then filtered through a plate and frame filter.

[0225] 1.3 Concentration The chemical solution after microfiltration was poured into a 100L single-effect concentrator and concentrated at a vapor pressure of 0.01-0.03 MPa, a vacuum pressure of 0.02-0.04 MPa, and a temperature of 80-90°C. The condition of the chemical solution had to be continuously monitored during concentration. The solution was concentrated until the relative density reached 1.004 (at 20°C). The molten glue was then sucked in, and concentration continued until the relative density reached 1.042 (at 20°C).

[0226] 2. Detection results Experiments were conducted using the reference sample method directly in a 100L extraction tank. The dry paste rate of the prepared Zhongling granule concentrate was within the range of the reference sample, and the content of indicator components was lower than that of the reference sample. Furthermore, compared to the determined Zhongling granule process (batch numbers: ZLKLXS2203011-ZLKLXS2203013), the dry paste rate was 7.8% lower. The results are shown in Table 14. Table 14 indicates that the extraction rate of the reference sample method used in modern processes is low, and it cannot meet the quality standard requirements of the reference sample. Therefore, research and determination of the process applicable to modern production equipment is necessary to ensure that the Zhongling granules to be prepared meet the standards of the reference sample for Zhongling Tang.

[0227] JPEG2025531477000017.jpg98170

[0228] The method for preparing Zhuling Granules, a modern formulation of the classical prescription Zhuling Tang provided by the present invention, includes the firing of decoction pieces, soaking, extraction, extract treatment (purification), concentration, drying, granulation, etc. The preparation method provided by the present invention has a stable and controllable production process, and can prepare granules that basically meet the quality standards of the "reference sample of Zhuling Tang prepared according to the principle of 'respecting the ancients'." Furthermore, compared to traditional decoctions, the Zhuling Tang formulation has the advantages of good stability, portability, and ease of administration.

[0229] The present invention provides a method for firing the decoction pieces used in preparing Zhuling Tang solid preparations and a quality standard for the decoction pieces, thereby ensuring stable and uniform quality of the raw materials and controlling the quality of the preparation from the source.

[0230] This invention provides an extraction method for Choling Tang formulations. Compared with traditional Chinese herbal preparations, which require excessive extraction and high energy consumption to maximize extraction rates (extracting for 1-2 hours each time often results in the extraction of impurities such as fiber and starch, without increasing the extraction of active ingredients, and the destruction of some components during prolonged heating), the extraction method proposed by this invention aims to produce granules that essentially match the quality standards of the reference sample while simultaneously compensating for losses in the refining, concentration, and drying processes. The extraction method determines the soaking time, amount of water, number of extractions, and extraction time of the extraction process, ensuring that the quality of the formulation essentially matches the chemical composition of traditional decoctions and reducing production costs.

[0231] The entire post-extraction process of this invention is carried out under low temperature conditions (below 100°C), and the purification process uses physical methods such as centrifugation and plate and frame filtration to remove impurities, the concentration process uses reduced pressure concentration, and the drying process uses vacuum belt drying. The low temperature is more advantageous for protecting and preserving the ingredients, and avoiding the destruction of the substance ingredients due to high temperatures. This minimizes the destruction during the industrialized preparation production process and ensures the consistency between traditional decoctions and modern preparations.

[0232] The present invention provides a process for concentrating Choling-to extract preparations, in which the concentration temperature is 70-90°C and the concentration time does not exceed 6 hours.

[0233] The present invention provides the types, dosages, and addition methods of auxiliary materials for Zhongling Tang solid preparations. By changing the traditional method of adding auxiliary materials for granules (adding them before granulation and mixing them evenly), the concentration process is started ahead of the time of adding the auxiliary materials, and the auxiliary materials are added during the concentration process. This not only solves the problems of clogging and material loss due to tape adhesion caused by the viscous medicinal solution of Zhongling Tang preparations (including gelatin, polysaccharides, etc.) during the drying process, but also solves the problem of uniform mixing of auxiliary materials and raw materials compared with the traditional method of adding auxiliary materials before granulation. Furthermore, since the paste yield of Zhongling granules is low, adding auxiliary materials at the concentration stage can relatively reduce the loss of dried paste during the drying process.

[0234] The drying method for the Choling Tang solid preparation provided by the present invention is a vacuum belt drying method, which has the advantages of low drying temperature, short drying time, little loss, and high efficiency, and is particularly suitable for Choling Tang preparation materials which are highly viscous and prone to aggregation.

[0235] This invention provides a research and evaluation method for the preparation of a solid formulation of Zhuling. The evaluation method uses the dry paste rate as the main evaluation index, supplemented by content measurement indexes in stages. The contents of 23-acetylalisol B and 23-acetylalisol C are used as secondary indexes in the extraction, purification, and concentration stages, and the contents of 23-acetylalisol B, 23-acetylalisol C, and characteristic polypeptides are used as secondary indexes in the drying and granulation stages. Changes in the indexes are tracked starting from the input of raw materials, and each process is systematically evaluated.

[0236] The present invention provides quality standards for Choling-Tang preparations, which stipulate the properties, identification, exudates, characteristic spectra, and detection methods for content measurement of Choling-Tang preparations, and also set upper and lower limits for content measurement.

[0237] The present invention provides a formulation quality control mode that uses a reference sample as a benchmark, and uses the dry paste rate + characteristic spectrum + index component content to comprehensively monitor the overall chemical component composition in the standard decoction, and uses the established multi-component control indexes in product stability studies to ensure the commercialization of finished pharmaceutical products.

[0238] This invention provides a research method based on the principle of transmission of quantity and quality from raw materials → process → intermediates → formulation. By determining the upper and lower limits of the contents of L-hydroxyproline, glycine, alanine, proline, donkey-derived polypeptide A1, and donkey-derived polypeptide A2 in the drug pieces, the quality of the formulation can be guaranteed, and a quality standard system for managing the entire process from medicinal materials, drug pieces, intermediate products, and solid formulations is being established.

[0239] The above are only preferred embodiments of the present invention, and those skilled in the art may make some improvements and modifications without departing from the principle of the present invention, and these improvements and modifications should also be considered within the protection scope.

Claims

1. A method for preparing Choreito, comprising the steps of: Soaking the four kinds of herbal medicines in water to obtain a prepreg liquid; performing a first boiling extraction on the prepreg liquid to obtain a first water extract and a dregs; Mixing the residue with water to perform a second boiling extraction to obtain a second water extract; performing a first concentration on the first aqueous extract to obtain a first concentrate; mixing the first concentrate and the second aqueous extract to perform a second concentration to obtain a second concentrate; mixing the second concentrate with an aqueous solution of Erjiao to obtain the Zhuling Tang; the four herbal medicines include Poria cocos, Poria cocos, Zelkova and talc, and during the soaking, the ratio of the mass of water to the total mass of the four herbal medicines is 14:1 or less; During the second boiling extraction, the ratio of the mass of water to the total mass of the four herbal medicines is 12:1 or less; A method for preparing Choling-to, characterized in that the temperatures of the first concentration and the second concentration are independently 90°C or lower.

2. During the soaking, the ratio of the mass of water to the total mass of the four herbal medicines is (8-14):1, and the soaking time is 40-60 minutes; 2. The method according to claim 1, wherein the first boiling extraction comprises a temperature-raising step by steam heating and a boiling and keeping warm step by steam heating, the keeping warming time of the boiling and keeping warm step is 40 to 90 minutes, the steam pressure of the boiling and keeping warm step is 0.1 MPa or less, and the steam pressure of the temperature-raising step is 0.2 MPa or less.

3. 2. The method of claim 1, wherein the ratio of the mass of water to the total mass of the four herbal medicines during the second boiling extraction is (6-12):1, the second boiling extraction includes a steam heating temperature raising step and a steam heating boiling warm keeping step, the boiling warm keeping time is 25-60 minutes, the steam pressure during the boiling warm keeping step is 0.1 MPa or less, and the steam pressure during the temperature raising step is 0.2 MPa or less.

4. 2. The method according to claim 1, wherein the mass ratio of the Poria Cocos, Poria Cocos, Zeaxanthin and talc is (0.7-1.3):(0.7-1.3):(0.7-1.3):(0.7-1.3).

5. the first concentration and the second concentration are independently carried out under steam heating conditions; The preparation method according to claim 1, wherein the temperatures of the first condensation and the second condensation are independently 70 to 90°C, the vacuum pressures of the first condensation and the second condensation are independently 0.03 to 0.06 MPa, the incubation times of the first condensation and the second condensation are independently 6 hours or less, and the vapor pressures of the first condensation and the second condensation are independently 0.01 to 0.05 MPa.

6. The method according to claim 1, wherein the weight ratio of the gelatin to water in the gelatin aqueous solution is 1:(2-6).

7. A method for preparing a polygala extract, comprising: The method comprises concentrating the Choling Tang prepared by the method according to any one of claims 1 to 6 to obtain the Choling extract; The method for preparing a Chorizo ​​extract is characterized in that the temperature of the concentration is 90°C or less, and the relative density of the Chorizo ​​extract at 20°C is 1.12 to 1.

16.

8. The concentration is carried out under steam heating conditions; The method according to claim 7, wherein the temperature of the concentration is 70 to 90°C, the vacuum pressure of the concentration is 0.03 to 0.06 MPa, the incubation time of the concentration is not more than 6 hours, and the vapor pressure of the concentration is 0.01 to 0.05 MPa.

9. A method for preparing a solid formulation of Choling including a dry powder or granules, comprising: The method for preparing the dry powder comprises: a step of mixing the Choling Tang prepared by the method according to any one of claims 1 to 6 with an aqueous solution of auxiliary materials, and then concentrating the mixture to obtain a thick paste; and vacuum drying the mixed thick paste to obtain the dried Chorizome powder. The mass ratio of the auxiliary materials to the Zhuling Tang is (1-7):50, and the mass of the Zhuling Tang is calculated as the total mass of Zhuling, Poria, Zeaxanthus, Talc and Erjiao; The method for preparing the granules comprises dry granulating the dried powder of Choling to obtain the Choling granules.

10. 10. The method according to claim 9, wherein the auxiliary material is at least one selected from the group consisting of dextrin, lactose, and soluble starch.

11. 10. The method according to claim 9, wherein the concentration is carried out under steam heating conditions, the temperature of the concentration is 70-90°C, the vacuum pressure of the concentration is 0.03-0.06 MPa, the incubation time of the concentration is not more than 6 hours, and the heating steam pressure of the concentration is 0.01-0.05 MPa.

12. 12. The method according to claim 9 or 11, wherein the relative density of the mixed thick paste at 20°C is 1.12 to 1.

16.

13. The vacuum drying is vacuum belt drying, and the operating parameters of the vacuum belt drying include a material supply temperature of 75 to 85°C, a material supply rate of 2.0 to 3.5 L / h, a crawler travel speed of 150±50 mm / min, a material distribution motor rotation speed of 100 r / min, a swing arm angle of 12 to 25°, heating temperature zones of a first zone of 95±5°C, a second zone of 96±5°C, and a third zone of 96±5°C, in that order, a vacuum degree of 0.097 to 0.1 MPa, a cutter speed of 10 to 20 seconds / cycle, and a cooling temperature of 30±5°C; 10. The method according to claim 9, wherein the dry granulation is carried out in a granulator, and the operating parameters of the dry granulation include: a pressure of a press roll of the granulator is 12-20 MPa; a rotation speed of the press roll of the granulator is 4-10 r / min; a vertical rotation speed of a screw feeder of the granulator is 25-40 r / min; a horizontal rotation speed of the screw feeder of the granulator is 70-120 r / min; and a mesh number of a sieve is 14 mesh.

14. The dry paste ratio of the Choling solid preparation is 18-23%, the total content of 23-acetylalisol B and 23-acetylalisol C in the Choling solid preparation is 0.011-0.020 wt%, and the donkey-derived polypeptide A in the Choling solid preparation is １ and donkey-derived polypeptide A ２ 14. A solid preparation of Poria cocos prepared by the method according to any one of claims 9 to 13, characterized in that the total content of

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