CN115337333A

CN115337333A - Preparation method of classical famous prescription abalone shell powder preparation

Info

Publication number: CN115337333A
Application number: CN202210969641.2A
Authority: CN
Inventors: 贾晓斌; 封亮; 方占民; 黄俊明; 侯金才; 郭勇; 刘磊
Original assignee: Jingjinji Lianchuang Drug Research Beijing Co ltd; Shineway Pharmaceutical Zhangjiakou Co ltd; Shenwei Pharmaceutical Group Co Ltd
Current assignee: Jingjinji Lianchuang Drug Research Beijing Co ltd; Shineway Pharmaceutical Zhangjiakou Co ltd; Shenwei Pharmaceutical Group Co Ltd
Priority date: 2022-08-12
Filing date: 2022-08-12
Publication date: 2022-11-15
Anticipated expiration: 2042-08-12
Also published as: CN115337333B

Abstract

The invention provides a preparation method of a classical name prescription abalone shell powder preparation, belonging to the technical field of medicine preparation, wherein the preparation method comprises the following steps of preparing abalone shell, chrysanthemum, cassia seed, notopterygium root and liquorice, the preparation process comprises a preparation process and a forming process, the preparation process sequentially comprises a processing process, a crushing and mixing process, a sterilization process and a drying process, and a medicine intermediate is obtained after the drying process; the molding process is to prepare the drug intermediate into a preparation; the sterilization process is to perform sterilization by adopting circulating steam. The preparation process of the invention well retains the effective components in the traditional Chinese medicine, the index components of the abalone shell powder, namely chlorogenic acid and glycyrrhizic acid, have less loss, the preparation conforms to the pharmacopoeia standard, and the effectiveness and the safety of the abalone shell powder preparation are ensured.

Description

Preparation method of classical famous prescription abalone shell powder preparation

Technical Field

The invention relates to a preparation method of a traditional Chinese medicine, in particular to a preparation method of a classical famous formula abalone shell powder preparation, belonging to the technical field of medicine preparation.

Background

The classical name prescription has wide application in modern clinic, has good curative effect foundation, becomes an important source for the research and development of new drugs in the world at present, and has important significance for developing and preparing the abalone shell powder on a large scale as the classical name prescription. The abalone shell powder is prepared from abalone shell, notopterygium root (rhizoma phragmitis removed), cassia seed and chrysanthemum, and licorice root (processed and water cut) and half of licorice root (processed and water cut) respectively, and the abalone shell powder is prepared into powder for two coins for each dose, wherein the powder is prepared by using water for one dose and decocting for six minutes, and is prepared into powder for warm taking after eating and lying. Has the main effects of calming the liver and suppressing yang, clearing liver and improving vision, and is mainly used for treating liver meridian heat accumulation type anterior uveitis, filiform keratitis, herpes simplex keratitis, viral keratitis and the like.

In order to prepare the classical well-known formula abalone shell powder into a safe, effective, stable and uniform modern preparation, some key problems in the traditional Chinese medicine preparation need to be solved urgently. For example, in the preparation process of the traditional Chinese medicine, because the processing process parameters are not specific, the loss of active ingredients in the medicinal materials is large, the difference between each batch of decoction pieces obtained after processing is large, and finally the quality of the finished product is unstable; and the abalone shell powder is used as the raw traditional Chinese medicine powder, so that the microbial limit is generally over-standard. In addition, the efficacy of the medicine is provided by the effective components of each medicine, and in the prior art, the index components of the abalone shell powder, namely chlorogenic acid and glycyrrhizic acid, are usually greatly lost in the preparation process, so that the efficacy is influenced to a certain extent.

Disclosure of Invention

Aiming at the problems, the invention provides a preparation method of a abalone shell powder preparation, which reduces the loss of chlorogenic acid and glycyrrhizic acid as index components, improves the safety of the preparation and improves the stability of the active ingredients of the preparation.

In order to achieve the purpose, the technical scheme of the invention is a preparation method of a classical famous prescription abalone shell powder preparation, the abalone shell powder preparation uses medicinal materials including abalone shell, chrysanthemum, cassia seed, notopterygium root and liquorice, and comprises a preparation process and a forming process, wherein the preparation process sequentially comprises a processing process, a crushing and mixing process, a sterilizing process and a drying process, and a medicine intermediate is obtained after the drying process; the molding process is to prepare the drug intermediate into a preparation; the sterilization process is to perform sterilization by adopting circulating steam.

Concretely, the five medicinal materials of the abalone shell, the notopterygium root, the cassia seed, the chrysanthemum and the liquorice are processed through a processing procedure to obtain corresponding decoction pieces;

through the crushing and mixing process, abalone shell, notopterygium root, cassia seed, chrysanthemum and liquorice decoction pieces are crushed into fine powder and sieved, and then the powder obtained by crushing and sieving each decoction piece is uniformly mixed according to a certain proportion;

sterilizing the uniformly mixed powder by adopting a flowing steam method to obtain an intermediate through a sterilization process;

removing excessive water in the sterile powder through a drying process to prepare a dried intermediate;

adding adjuvants into the dried intermediate by molding process to obtain solid oral preparation.

In the invention, the abalone shell medicinal material is shell of Haliotis disc abalone hannai ino of abalone family animal, the Notopterygium root medicinal material is dried root and rhizome of Notopterygium root Notopterygium incisum Ting ex H.T.Chang of Umbelliferae family, the Cassia seed medicinal material is mature fruit of Cassia obtusifolia L.of Leguminosae family, and the Chrysanthemum medicinal material is dried head-shaped inflorescence of Chrysanthemum morifolium Ramat of Compositae family; the Glycyrrhrizae radix is dried root and rhizome of Glycyrrhiza uralensis Fisch of Leguminosae.

Further, the processing procedure comprises the following specific steps:

taking a cassia seed medicinal material, washing with clear water to remove most of impurities, placing on a Chinese medicinal material sorting table, removing adhered stubborn impurities with a brush, washing with clear water twice, drying in the sun, and appropriately crushing;

taking a chrysanthemum medicinal material, placing the chrysanthemum medicinal material on a Chinese medicinal material sorting table, and manually removing non-medicinal parts such as branch tips, calyx and the like and impurities such as dust, silt, soil blocks and the like;

placing semen Cassiae on Chinese medicinal material sorting table, and manually removing non-medicinal parts such as branch tip and calyx, and impurities such as dust, silt, and soil;

placing Notopterygii rhizoma on a Chinese medicinal material sorting table, washing with water, and oven drying below 60 deg.C;

the liquorice medicinal material is taken and placed on a Chinese medicinal material sorting table, impurities are manually removed, ash and scraps, non-medicinal parts and the like are sieved, and the size strips are separated into two grades. Washing selected Glycyrrhrizae radix with water, soaking in a moistening tank with water 3-5 cm higher than Glycyrrhrizae radix at 35 deg.C for 3 hr. Regulating the thickness of cut herbs to 2-4mm by using a beveling type herbal medicine cutting machine, drying the cut liquorice slices in an oven at the spreading thickness of 20-30 mm, setting the drying temperature at 60 ℃ and the drying time for 5 hours.

Further, the pulverizing and mixing process comprises the following steps:

respectively putting processed Concha Haliotidis decoction pieces, notopterygii rhizoma decoction pieces, semen Cassiae decoction pieces, flos Chrysanthemi decoction pieces, and Glycyrrhrizae radix decoction pieces into a pulverizing device, pulverizing into fine powder, and sieving;

according to the following steps: 2:2:2:1, taking the five-flavor decoction pieces, crushing and sieving the powder, and uniformly mixing the powder in a mixing mechanical device to obtain mixed powder.

The crushing device comprises a firewood field type crusher, a universal pulverizer, a ball mill and a dust-absorbing micro crusher.

Further, the sterilization process is to perform sterilization by using normal pressure circulating steam.

Further, the sterilization process adopts flowing steam sterilization at the normal pressure of 100 ℃ for 30min.

The sterilization process comprises the steps of filling the mixed powder into a sterile bag, placing the sterile bag into a circulating steam sterilization device, and introducing steam for sterilization;

taking out the powder after the circulation steam sterilization, and drying the powder in a drying oven provided with a sterile filtering device at the temperature of 65 ℃ for 12 hours; after drying, a pharmaceutical intermediate is obtained.

The above molding process comprises directly packaging the intermediate into powder.

The molding process can also be that the medicine intermediate is taken according to a certain proportion, added with auxiliary materials, mixed evenly and then subpackaged in a capsule shell to prepare capsules. The auxiliary materials comprise magnesium stearate, sodium dodecyl sulfate and pregelatinized starch.

The forming process can also be that the drug intermediate is taken according to a certain proportion, added with auxiliary materials and uniformly mixed, pressed into slices with required hardness by a dry extrusion granulator, crushed into granules by a granulator and subpackaged into granules.

The preparation method of the classical famous prescription abalone shell powder preparation has the beneficial effects that:

the preparation process of the invention well retains the effective components in the traditional Chinese medicine, the index components of the abalone shell powder, namely chlorogenic acid and glycyrrhizic acid, have less loss, the preparation conforms to the pharmacopoeia standard, and the effectiveness and the safety of the abalone shell powder preparation are ensured.

In the invention, various parameters of the processing procedure of the liquorice in the preparation of the abalone shell powder preparation are optimized, so that a stable and reliable processing method of the liquorice is obtained; in the sterilization process, the sterilization is carried out by adopting a normal-pressure flowing steam method, and the preferred steam flowing time is 30 minutes. The contents of chlorogenic acid and glycyrrhizic acid are not changed before and after sterilization, and the microbial limit after sterilization meets the pharmacopeia standard, thereby ensuring the effectiveness and safety of the abalone shell powder preparation.

Compared with the traditional preparation process, the flow-through steam sterilization process can retain the content of the heat stability components to a greater extent; meanwhile, the content of liquiritin and glycyrrhizic acid in the circulating steam sample is almost the same as that in the freeze-dried sample and is similar to that of the preparation raw materials. The sample obtained by the abalone shell steam-dispersing process has more component quantity and index component content than the sample obtained by the traditional preparation process, so unexpected excellent experimental results are obtained.

The abalone shell powder prepared by the method meets the requirements of modern preparations, well retains the active ingredients in the traditional Chinese medicine, can be produced in a large scale, has uniform and stable quality, and is convenient to carry and store. The Concha Haliotidis powder preparation has microorganism limit in accordance with pharmacopeia standard, and each effective component can be well retained, and can be used for effectively treating liver meridian heat accumulation type anterior uveitis, filamentary keratitis, herpes simplex keratitis and viral keratitis.

Drawings

The present invention will be described in further detail with reference to the accompanying drawings and specific embodiments.

FIG. 1 is a graph of the cumulative dissolution of various chlorogenic acids in the artificial intestinal juice of example 1;

FIG. 2 is a graph showing the cumulative dissolution of glycyrrhizic acid from each sample of the artificial intestinal juice according to example 1;

FIG. 3 is a graph of the cumulative dissolution of various chlorogenic acids in artificial gastric juice of example 1;

FIG. 4 is a graph showing the cumulative dissolution rate of glycyrrhizic acid in each sample of artificial gastric juice in example 1;

FIG. 5 is a graph of the cumulative dissolution of various chlorogenic acids in the artificial intestinal juice of example 2;

FIG. 6 is a graph showing the cumulative dissolution of glycyrrhizic acid from each sample of the artificial intestinal juice of example 2;

FIG. 7 is a graph of the cumulative dissolution of various chlorogenic acids in artificial gastric fluid of example 2;

FIG. 8 is a graph showing the cumulative dissolution rate of glycyrrhizic acid in each sample of artificial gastric juice in example 2;

FIG. 9 is a graph of the cumulative dissolution of various chlorogenic acids in the artificial intestinal juice of example 3;

FIG. 10 is a graph showing the cumulative dissolution rate of glycyrrhizic acid in each sample of the artificial intestinal juice according to example 3;

FIG. 11 is a graph of the cumulative dissolution of various chlorogenic acids in artificial gastric fluid of example 3;

FIG. 12 is a graph showing the cumulative dissolution rate of glycyrrhizic acid in each sample of artificial gastric juice in example 3;

FIG. 13 is a graph of the results of mass spectrometry analysis of lyophilized powder samples of Concha Haliotidis in example 4;

FIG. 14 is a chart showing the results of mass spectrometry of the dispersed steam sample of Cassia tora in example 4

Fig. 15 is a route chart of the preparation process of the abalone shell powder preparation of the present invention.

Detailed Description

The technical solution in the embodiments of the present invention is clearly and completely described below with reference to the drawings in the embodiments of the present invention. In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present invention, but the present invention may be practiced in other ways than those specifically described and will be readily apparent to those of ordinary skill in the art without departing from the spirit of the present invention, and therefore the present invention is not limited to the specific embodiments disclosed below.

A preparation method of a classic and famous prescription abalone shell powder preparation, the abalone shell powder preparation uses the medicinal materials including abalone shell, chrysanthemum, cassia seed, notopterygium root and licorice, includes the preparation process and the forming process, the preparation process includes the processing procedure, the crushing and mixing procedure, the sterilization procedure and the drying procedure in turn, the medicine intermediate is obtained after the drying procedure; the molding process is to prepare the drug intermediate into a preparation; the sterilization process is to perform sterilization by adopting circulating steam.

(1) Processing five medicinal materials of abalone shell, notopterygium root, cassia seed, chrysanthemum and liquorice by a processing procedure to obtain corresponding decoction pieces; specifically, the method comprises the following steps:

taking a cassia tora medicinal material, washing with clear water to remove most impurities, placing on a Chinese medicinal material sorting table, removing adhered stubborn impurities by using a brush, washing twice with clear water, drying in the sun, and crushing properly;

the liquorice is taken and placed on a Chinese medicinal material sorting table, impurities are manually removed, ash and scraps, non-medicinal parts and the like are screened, and the size strips are separated into two grades. Washing selected liquorice with water, putting the liquorice in a moistening pool, adding water to the water to be more than 3-5 cm of the liquorice, setting the water temperature to be 35 ℃, and soaking for 3 hours. Regulating the thickness of cut herbs to 2-4mm by using a beveling type herbal medicine cutting machine, drying the cut liquorice slices in an oven at the spreading thickness of 20-30 mm, setting the drying temperature at 60 ℃ and the drying time for 5 hours.

(2) Through the crushing and mixing process, abalone shell, notopterygium root, cassia seed, chrysanthemum and liquorice decoction pieces are crushed into fine powder and sieved, and then the powder obtained by crushing and sieving each decoction piece is uniformly mixed according to a certain proportion; specifically, the method comprises the following steps:

respectively putting processed Concha Haliotidis decoction pieces, notopterygii rhizoma decoction pieces, semen Cassiae decoction pieces, flos Chrysanthemi decoction pieces, and Glycyrrhrizae radix decoction pieces into a pulverizing device, pulverizing into fine powder, and sieving; according to the following steps: 2:2:2:1, taking the five-flavor decoction pieces, crushing and sieving the powder, and uniformly mixing the powder in a mixing mechanical device to obtain mixed powder. The crushing device comprises a firewood type crusher, a universal pulverizer, a ball mill and a dust collection micro-crusher.

(3) Sterilizing the uniformly mixed powder by adopting a flowing steam method to obtain an intermediate through a sterilization process; (4) excess water in the sterile powder is removed by a drying step to produce a dried intermediate.

Preferably, the sterilization process is performed by using normal pressure flowing steam, and more preferably, the sterilization process is performed by using normal pressure flowing steam at 100 ℃ for 30min.

The sterilization process comprises the steps of filling the mixed powder into a sterile bag, placing the sterile bag into a circulating steam sterilization device, and introducing steam for sterilization; taking out the powder after the circulation steam sterilization, and drying the powder in a drying oven provided with a sterile filtering device at the temperature of 65 ℃ for 12 hours; after drying, a pharmaceutical intermediate is obtained.

(5) Adding adjuvants into the dried intermediate by molding process to obtain solid oral preparation.

The forming process can also be that the drug intermediate is taken according to a certain proportion, added with auxiliary materials and uniformly mixed, pressed into slices with required hardness by a dry extrusion granulator, then crushed into granules by a granulator, and the granules are subpackaged into granules.

The abalone shell powder preparation can detect the contents of chlorogenic acid and glycyrrhizic acid as index components under the following chromatographic conditions.

Chromatographic conditions are as follows: octadecylsilane chemically bonded silica is used as a filling agent; gradient elution is carried out by taking acetonitrile-0.1 percent phosphoric acid aqueous solution as a mobile phase, and the flow rate is 1 ml/min; the detection wavelength is 242 nanometers; the column temperature was 30 ℃.

Example 1: preparation process investigation of classical famous prescription abalone shell powder

1 method of experiment

Preparation of a raw powder sample: weighing 7.46g of mixed powder according to the fine powder of each medicinal material prepared in the steps (1) and (2) in the method.

Preparation of a lyophilized sample: weighing 7.46g of mixed powder according to the method, placing the mixed powder into a decocting earthen pot, adding 300mL of pure water, decocting with strong fire at 150 ℃ (1400W) till boiling, decocting with slow fire at 100 ℃ for 30min while keeping the liquid medicine slightly boiling, placing the liquid medicine at 180mL, pre-freezing in a refrigerator at-80 ℃ for 3h, and drying in a freeze dryer for 48h after completely freezing to obtain the concha haliotidis freeze-dried sample.

The invention-preparation of a flow-through steam sample: according to the method, the fine powder of each medicinal material prepared after the steps (1) and (2) is prepared, 7.46g of mixed powder (ten times daily dosage) is weighed, the mixed powder is placed in a circulating steam device (the device is a conventional device capable of introducing steam, and is not particularly limited), steam is introduced for 30min at normal temperature and normal pressure (the water temperature is about 100 ℃), and then the mixed powder is transferred to a 60 ℃ oven to be dried for 12h, so that the traditional Chinese medicine composition is obtained.

Each of the samples referred to in the following examples was prepared in accordance with the above-described method.

Dissolution test method: weighing 7.46g of the above raw powder sample, lyophilized sample, and circulating steam sample, taking 900mL of artificial gastric juice and 900mL of artificial intestinal juice as dissolution medium, sampling at rotation speed of 100 r.min-1 at 15, 30, 45, 60, 75, 100, and 120min respectively, detecting chlorogenic acid and glycyrrhizic acid content at each sampling point, and calculating cumulative dissolution rate according to dissolution rate and release rate determination method (second method of 0931).

1.1 high performance liquid detection conditions of the sample: 1. gradient elution condition a (see table 1): octadecylsilane chemically bonded silica is used as a filling agent; gradient elution with acetonitrile (a) -0.1% aqueous phosphoric acid solution (B) as mobile phase, as shown in the table below; flow rate 1.0 ml/min; the detection wavelength is 242 nanometers; the column temperature was 30 ℃. The number of chlorogenic acid peaks in theoretical plate should be not less than 8000, and calculated according to liquiritin peak should be not less than 5000.

TABLE 1 high Performance liquid gradient elution conditions

2 Experimental materials and apparatus

2.1 materials of the experiment

Homemade raw powder of abalone shell powder (batch No. 20220525), homemade freeze-dried sample of abalone shell powder (batch No. 20220521), homemade steam sample of abalone shell powder (batch No. 20220526), phosphoric acid (chromatographic grade, batch No. 179831, 500 mL/bottle, fisher), water (purified water, tianjin Waha food Co., ltd.), acetonitrile (chromatographic grade, batch No. 241341,4L/bottle, fisher)

2.2 Experimental facilities

Dissolution apparatus (708-DS, agilent technologies, ltd.), electric heating constant temperature air-blast drying oven (DHG-9240A, shanghai essence macro experimental facilities, ltd.), high performance liquid chromatograph (Waters 2695, wate technologies, ltd.), electronic balance (FA 1004B, shanghai hel pu international trade, ltd.), decocting earthenware pot (DTL 20A10, nano-liya kitchenware, ltd.), freeze dryer (Alpha 2-4LSCbasic, CHRIST, germany), homemade circulating steam equipment, and the like.

3 results of the experiment

The results of the cumulative dissolution test of three samples of the abalone shell powder in the artificial intestinal juice and the artificial gastric juice are shown in figures 1 to 4.

4 analysis of results

Analyzing the experimental results, comparing the cumulative dissolution rates of the index components chlorogenic acid and glycyrrhizic acid in the artificial intestinal juice and the artificial gastric juice of the original powder sample, the freeze-dried sample and the circulating steam sample, and finding that the cumulative dissolution rates of the chlorogenic acid and the glycyrrhizic acid in the artificial intestinal juice and the artificial gastric juice of the circulating steam sample are both at the highest values, and the difference ratio is large, so that the feasibility of preparing the abalone shell powder preparation by a circulating steam method can be preliminarily determined.

Example 2: inspection of classical famous-style abalone shell powder on steam temperature of steam circulation process

1 Experimental method

1.1 preparation of circulating steam samples with different steam-passing temperatures: and taking three parts of 74.6g (ten times of daily dose) of the concha haliotidis powder (the mixed powder) and placing the three parts in self-made circulating steam equipment, ordinary high-pressure sterilization equipment and special high-pressure sterilization equipment respectively, introducing steam for 30min at normal temperature and normal pressure (the water temperature is about 90 ℃), introducing steam for 30min at normal temperature and normal pressure (the water temperature is about 100 ℃), introducing steam for 30min at pressurized heating temperature (the water temperature is about 110 ℃), introducing steam for 30min at heated pressurized heating (the water temperature is about 120 ℃), and then transferring the mixture to a 60 ℃ oven to be dried for 12h to obtain the traditional Chinese medicine.

1.2 dissolution test method: weighing the above flowing steam sample (90 deg.C), flowing steam sample (100 deg.C), flowing steam sample (110 deg.C), and flowing steam sample (120 deg.C) 7.46g respectively, and measuring with dissolution rate and release rate (second method of 0931), using 900mL artificial gastric juice and 900mL artificial intestinal juice as dissolution medium, and rotating at 100 r.min ^－1 Sampling is carried out at 15 min, 30min, 45min, 60min, 75 min, 100 min and 120min respectively, the contents of chlorogenic acid and glycyrrhizic acid at each sampling point are detected respectively, and the cumulative dissolution rate is calculated.

1.3 high performance liquid detection conditions of the sample: 1. gradient elution condition a (same table 1): octadecylsilane chemically bonded silica is used as a filling agent; gradient elution with acetonitrile (a) -0.1% aqueous phosphoric acid solution (B) as mobile phase, as shown in the table below; flow rate 1.0 ml/min; the detection wavelength is 242 nanometers; the column temperature was 30 ℃. The theoretical plate number of chlorogenic acid peak should be not less than 8000, and calculated according to glycyrrhizin peak should be not less than 5000.

1.4 detection method of microorganism:

the preparation of the test solution takes 10g of the medicine powder before the circulation steam sterilization and the medicine powder after the circulation steam sterilization obtained by different circulation steam temperatures respectively, adds the sterilized tryptone soy peptone liquid culture medium to 100ml, leads the concentration to be 1, and prepares the test solution for standby, and each condition is divided into three groups in parallel.

Microorganism counting the test solution is sequentially diluted by 10 times to 1 10000, 1ml of the test solution is sucked from 1. Meanwhile, 1ml of blank diluent (physiological saline) was respectively sucked and placed in 2 dishes as blank controls, and 46 ℃ agar medium was immediately poured into the dishes until the mixture solidified. The above 2 parallel samples were used for detecting total aerobic bacteria (by using sterilized trypticase soy agar medium-TSA plate, incubation temperature 33 ℃ C., and incubation time 5 d) and total mold and yeast (by using Sasa dextrose agar medium-SDA plate, incubation temperature 25 ℃ C., and incubation time 7 d), respectively, and the results were observed. The equipment and materials used in the microbiological detection experiments are shown in the following table:

TABLE 2 Experimental Equipment and materials

2 Experimental materials and Equipment

2.1 materials of the experiment

Self-made abalone shell steam sample-90 ℃ (batch number 20220601-1), abalone shell steam sample-100 ℃ (batch number 20220601-2), self-made abalone shell steam sample-110 ℃ (batch number 20220601-3), self-made abalone shell steam sample-120 ℃ (batch number 20220601-4) phosphoric acid (chromatographic grade, batch number 179831, 500 mL/bottle, fisher), water (purified water, tianjin Waha food Co., ltd.), acetonitrile (chromatographic grade, batch number 241341,4L/bottle, fisher)

2.2 Experimental facilities

Dissolution apparatus (708-DS, agilent technologies, inc.), electric heating constant temperature air-blast drying oven (DHG-9240A, shanghai Jing Macro laboratory instruments, inc.), high performance liquid chromatograph (Waters e2695, watt science and technology, inc., USA), electronic balance (FA 1004B, shanghai Help International trade, inc.), common high-pressure sterilization apparatus (JC-STSX 50L digital display automatic control type, qingdao Chuangju environmental protection group, inc.), special high-pressure sterilization apparatus (HT 2000FJ, shanghai Huo Tong laboratory instruments, inc.), self-made circulation steam equipment, etc.

3 results of the experiment

See fig. 5-8, and the following table:

TABLE 3 microbial content of samples obtained before and after steam sterilization by different steam temperatures for Concha Haliotidis powder

TABLE 4 chlorogenic acid and glycyrrhizic acid content of samples obtained before and after steam sterilization by different steam temperatures of Concha Haliotidis powder

4 analysis of results

Analyzing the dissolution experiment results, comparing the cumulative dissolution rates of index components chlorogenic acid and glycyrrhizic acid in the artificial intestinal juice and the artificial gastric juice at-90 ℃ of a circulating steam sample, -100 ℃ of the circulating steam sample, -110 ℃ of the circulating steam sample and-120 ℃ of the circulating steam sample, and finding that the cumulative dissolution rates of the chlorogenic acid and the glycyrrhizic acid in the artificial intestinal juice and the artificial gastric juice of the circulating steam sample at-100 ℃ are both at the highest values, and the difference ratio is larger, so that the content of the chlorogenic acid and the glycyrrhizic acid is presumably reduced due to the increase of the steam temperature; analyzing the detection result of the microorganisms, and finding that the content of the microorganisms of the medicinal powder corresponding to the four different steam-introducing temperatures before sterilization is higher than the limit standard, and the content of the microorganisms of the medicinal powder obtained after sterilization at the four different steam-introducing temperatures meets the limit standard except for the steam sample of-90 ℃; the detection results of the index components are analyzed, the content of the index components of the samples corresponding to the four different steam introducing temperatures is reduced along with the increase of the steam introducing temperatures, the experimental results of the content are consistent with those of a dissolution experiment, and the adverse effect on the content of the index components caused by the increase of the temperatures is further proved.

To sum up, on the premise of ensuring that the content of microorganisms meets the limit standard, the temperature of the circulating steam should be selected to be a temperature which has little influence on the content of the index components chlorogenic acid and glycyrrhizic acid, so that the temperature used by the circulating steam is finally determined to be 100 ℃.

Example 3: inspection of classical famous-style abalone shell powder on steam-passing time of steam-passing process

1 method of experiment

1.1 preparation of circulation steam samples at different steam-passing time: placing 74.6g (ten times daily dose) of Concha Haliotidis powder in self-made circulating steam equipment, introducing steam at room temperature and normal pressure (water temperature about 100 deg.C) for 30min, 45min and 60min, and oven drying at 60 deg.C for 12 hr.

1.2 dissolution test methods: weighing 7.46g of the flowing steam sample (20 min), 25min, 30min, 45min and 60min respectively, and taking 900mL of artificial gastric juice and 900mL of artificial intestinal juice as dissolution media according to dissolution and release measurement method (second method of 0931), with rotation speed of 100 r.min ^－1 Sampling is carried out at 15 min, 30min, 45min, 60min, 75 min, 100 min and 120min respectively, the contents of chlorogenic acid and glycyrrhizic acid at each sampling point are detected respectively, and the cumulative dissolution rates are calculated.

1.4 detection method of microorganism:

And (3) counting microorganisms, taking the test solution, sequentially diluting the test solution to 1 10000 in an incremental manner by 10 times, and placing 1ml of the test solution in a flat dish, wherein 1. Meanwhile, 1ml of blank diluent (physiological saline) was respectively sucked and placed in 2 plates as blank control, and 46 ℃ agar medium was immediately poured into the plates until the mixture was solidified. The above 2 parallel samples were used for detecting total aerobic bacteria (by using sterilized trypticase soy agar medium-TSA plate, incubation temperature 33 ℃ C., and incubation time 5 d) and total mold and yeast (by using Sasa dextrose agar medium-SDA plate, incubation temperature 25 ℃ C., and incubation time 7 d), respectively, and the results were observed. The equipment and materials used in the microbiological assay are shown in the following table, which is also given in table 2.

2 Experimental materials and apparatus

2.1 Experimental materials

Self-made abalone shell steam sample-20 min (batch number 20220607-1), abalone shell steam sample-25 min (batch number 20220607-2), abalone shell steam sample-30 min (batch number 20220607-3), self-made abalone shell steam sample-45 min (batch number 20220607-4), self-made abalone shell steam sample-60 min (batch number 20220607-5) phosphoric acid (chromatographic grade, batch number 179831, 500 mL/bottle, fisher), water (purified water, tianjin Waha food Co., ltd.), acetonitrile (chromatographic grade, batch number 241341,4L/bottle, fisher)

2.2 Experimental facilities

Dissolution apparatus (708-DS, agilent technologies, ltd.), electric heating constant temperature air-blast drying oven (DHG-9240A, shanghai essence macro experimental facilities, ltd.), high performance liquid chromatograph (Waters e2695, wate technologies, ltd.), electronic balance (FA 1004B, shanghai hel pu international trade ltd.), homemade circulation steam equipment, and the like.

3 results of the experiment

See figures 9-12 and the following table.

TABLE 5 microbial content of samples obtained from different aeration times of Concha Haliotidis powder before and after aeration steam sterilization

TABLE 6 content of chlorogenic acid and glycyrrhizic acid in Concha Haliotidis powder before and after steam sterilization

4 analysis of results

Analyzing the dissolution experiment results, comparing the cumulative dissolution rates of index components chlorogenic acid and glycyrrhizic acid in the artificial intestinal juice and the artificial gastric juice for-20 min, -25min, -30min, -45min and-60 min of the circulating steam sample, and finding that the cumulative dissolution rates of chlorogenic acid and glycyrrhizic acid in the artificial intestinal juice and the artificial gastric juice for-30 min of the circulating steam sample are at the highest values and have larger difference proportion, so that the content of chlorogenic acid and glycyrrhizic acid is presumably reduced due to the prolonging of the steam-passing time; analyzing the microorganism detection result, finding that the microorganism contents of the medicinal powder corresponding to five different steam-introducing times before sterilization are all higher than the limit standard, and the microorganism contents of the medicinal powder obtained after five different steam-introducing times after sterilization all accord with the limit standard, but the microorganism contents of the samples obtained by a circulating steam sample of-20 min and a circulating steam sample of-25 min are higher, so the microorganism contents are abandoned due to poor sterilization effect; the detection results of the index components are analyzed, the content of the index components of the samples corresponding to the five different steam-through times is found to be reduced along with the prolonging of the steam-through time, the experimental result is consistent with the dissolution experiment, and the adverse effect on the content of the index components caused by the prolonging of the steam-through time is further proved.

In summary, on the premise of ensuring that the content of microorganisms meets the limit standard, the steam-through time of the circulating steam should be selected to have a value with small influence on the contents of the index components chlorogenic acid and glycyrrhizic acid, so that the steam-through time of the circulating steam is finally determined to be 30min.

Example 4: comparative analysis of samples obtained by different preparation processes of classical famous prescription abalone shell powder

1. Experimental methods

1.1 Analytical experimental method for UPLC-Q-TOF-MS mass spectrum

1.1.1 preparation of lyophilized sample test solution: taking 7.4g of the fine powder of the abalone shell, placing the fine powder in a decocting earthen pot, adding 300mL of water, heating the mixture to 150 ℃ with strong fire until the mixture is boiled, keeping the mixture slightly boiling at 100 ℃ with mild fire, decocting for 30min to obtain 180mL of a residue-liquid mixture, pre-freezing the mixture for 3h at-80 ℃, freeze-drying the mixture under the experimental conditions of-80 ℃,0.1mpa and 48h, dissolving 0.1g of freeze-dried sample in 10mL of absolute ethyl alcohol, performing ultrasonic treatment at 250 ℃ for 30min (w, 40kHZ), and passing through a membrane to obtain the traditional Chinese medicine.

1.1.2 preparation of a test solution of a circulating steam sample: 74.6g of the fine powder of the abalone shell powder (ten times of daily dose) is taken and placed in a self-made flowing steam device, steam is introduced for 30min at normal temperature and normal pressure (the water temperature is about 100 ℃), then the mixture is transferred to a 60 ℃ oven to be dried for 12h, 0.1g of the dried sample is taken and dissolved in 10mL of absolute ethyl alcohol, ultrasonic treatment is carried out for 30min (250w, 40kHZ) at 37 ℃, and the membrane is filtered, thus obtaining the traditional Chinese medicine.

1.1.3 liquid chromatography conditions for mass spectrometry: a Waters I-class liquid chromatograph; the chromatographic column is an Acquity UPLC HSS T3 column (100 mm. Times.2.1mm, 1.8 μm); the mobile phase is 0.1 percent aqueous formic acid solution-acetonitrile; gradient elution: 0-0.5min, 5% acetonitrile; 0.5-10 min, 5-50% acetonitrile; 10-10.5min, 50-95% acetonitrile; 10.5-12min, 95% acetonitrile; 12-12.5min, 95% -5% acetonitrile; 12.5-14min, 5% acetonitrile; the volume flow is 0.4mL/min; the column temperature is 40 ℃; the sample size is 2 mu L;2D detection wavelength is 260 nm and 300nm; flow-through steam samples and freeze-dried samples were analyzed by UPLC-Q-TOF-MS.

1.1.4 Mass Spectrometry conditions: respectively carrying out MS E scanning in an electrospray ion source (ESI) positive ion mode and an electrospray ion source (ESI negative ion mode); low impact voltage 6V; the high collision voltage is 30-70V; collecting mass range m/z 50-1200; capillary voltage 3kV (ESI +) and 2.5kV (ESI-); the taper hole voltage is 40V; the temperature of desolventizing gas is 500 ℃; the volume flow of the desolventizing gas is 1000L/h; the scanning time is 14min; leucine Enkephalin (LE) calibrates mass number in real time with positive ion at m/z 556.277 1 and negative ion at m/z 554.2615.

1.2 index component content determination experimental method

1.2.1 preparation of lyophilized sample test solution: taking 7.4g of the fine powder of the abalone shell, placing the fine powder in a decocting earthen pot, adding 300mL of water, heating the mixture to 150 ℃ with strong fire until the mixture is boiled, keeping the mixture slightly boiling at 100 ℃ with mild fire, decocting for 30min to obtain 180mL of a residue-liquid mixture, pre-freezing the mixture for 3h at-80 ℃, freeze-drying the mixture under the experimental conditions of-80 ℃,0.1mpa and 48h, dissolving 0.1g of freeze-dried sample in 10mL of absolute ethyl alcohol, performing ultrasonic treatment at 250 ℃ for 30min (w, 40kHZ), and passing through a membrane to obtain the traditional Chinese medicine.

1.2.2 preparation of a flowing steam sample test solution: taking 74.6g of fine powder of the abalone shell (ten times of daily dose), placing the fine powder in a self-made circulating steam device, steaming for 30min at normal temperature and normal pressure (water temperature is about 100 ℃), then transferring the fine powder to a 60 ℃ oven to be dried for 12h, taking 0.1g of dried sample, dissolving the sample in 10mL of absolute ethyl alcohol, performing ultrasonic treatment for 30min at 37 ℃ (250w, 40kHZ), and passing through a membrane to obtain the traditional Chinese medicine.

1.2.3 liquid chromatography conditions: octadecylsilane chemically bonded silica is used as a filling agent; gradient elution with acetonitrile (a) -0.1% aqueous phosphoric acid solution (B) as mobile phase, as shown in the table below; flow rate 1.0 ml/min; the detection wavelength was 267 nm; the column temperature was 35 ℃. The number of theoretical plates should be not less than 8000 calculated according to isochlorogenic acid A peak, and should be not less than 5000 calculated according to liquiritin peak.

TABLE 7 high Performance liquid gradient elution conditions

Time (minutes)	Acetonitrile (%)	0.1% phosphoric acid water (%)
			0	3	97
10	15	85
			13	15	85
40	45	55
			45	60	40
50	90	10
			66	3	97
70	3	97

2 Experimental materials and apparatus

2.1 Experimental materials

The method comprises the steps of preparing a freeze-dried sample of the sea-ear shell powder (batch number 20220615-1), preparing a steam sample of the sea-ear shell powder (batch number 20220615-2), preparing phosphoric acid (chromatographic grade, batch number 179831, 500 mL/bottle, fisher), acetonitrile (chromatographic grade, batch number 241341,4L/bottle, fisher), preparing absolute ethyl alcohol (mass spectrum grade, TEDIA company, USA, batch number 20205273), and preparing water (purified water, tianjin Wahaha food Co., ltd.).

2.2 Experimental facilities

Acquity UPLC H-Class ultra high performance liquid chromatograph, waters Xevo G2-XS QTof mass spectrometer (equipped with electrospray ionization: ESI), data processing software (MassLynx V4.2 and UNIFI, waters corporation, USA); an ultra pure water system (Elix Essential 5uv + milli-Q Reference, germany merck porcelain factory), a decocting earthenware pot (Kang Shu ceramics limited, jiang xi), a freeze dryer (Alpha 2-4LSCbasic type, germany CHRIST), an electric ceramic furnace (DTL 21B, jieyinda small electric appliance limited), a low speed centrifuge (TD 6.6, shanghai Lu Xiangyi centrifuge instrument limited), a universal high speed pulverizer (QE-100, zhejiang xie jiehu trade), an ultrasonic machine (KH-800 DE type, kunshan wound), an electronic balance (TD 10002A, tianjin horse balance base instrument), a high efficiency liquid chromatograph (Waters e2695, wott scientific and technology limited, usa), an electric heating constant temperature blast drying box (DHG-9240A, shanghai essence macro experimental facility limited, self-made by seiki), and circulating steam experimental facilities, etc.

3 results of the experiment

See fig. 13-14 and the following table.

TABLE 8 determination of content of index components in samples obtained by different preparation processes of Concha Haliotidis powder

Index component	Raw materials of preparation	Freeze-dried sample	Flow through steam sample
				Chlorogenic acid	0.57％	0.43％	0.55％
Liquiritin	0.95％	0.89％	0.88％
				Isochlorogenic acid A	0.80％	0.70％	0.78％
Glycyrrhizic acid	3.61％	3.38％	3.48％

4 analysis of results

Analyzing the mass spectrometry analysis experiment result, the abalone shell dispersed flowing steam sample contains 13 more components than the freeze-dried sample, and the reason is considered probably because part of volatile components are volatilized along with water vapor and part of unstable component structures are damaged due to heating in the preparation process of the freeze-dried sample, so that compared with the traditional preparation process, the flowing steam process can reserve the total number of the original components of the preparation to a greater extent, and plays a more key role in playing the drug effect; the analysis of the content determination results of the index components shows that the content of chlorogenic acid and isochlorogenic acid A in a dispersed flowing steam sample of the abalone shell is higher than that in a freeze-dried sample, and the content of the chlorogenic acid and isochlorogenic acid A in the flowing steam sample is similar to that of raw materials of a preparation, and the heat stability of the chlorogenic acid and isochlorogenic acid A is poor by looking up related documents, so that the conclusion can be drawn that the content of heat-stable components can be retained to a greater extent by a flowing steam process compared with the traditional preparation process; meanwhile, the contents of liquiritin and glycyrrhizic acid in the circulating steam sample are almost the same as those in the freeze-dried sample and are similar to those of preparation raw materials, so that the two preparation processes have no adverse effect on the content of relatively stable components. In summary, the abalone shell samples obtained by the steam-dispersing process have larger component amount and index component content than the samples obtained by the traditional preparation process, so unexpected excellent experimental results are obtained.

It should be apparent that the described embodiments are only some of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

Claims

1. A preparation method of a classic name prescription abalone shell powder preparation, the abalone shell powder preparation uses the medicinal materials including abalone shell, chrysanthemum, cassia seed, notopterygium root and licorice, and is characterized by comprising a preparation process and a forming process, wherein the preparation process sequentially comprises a processing procedure, a crushing and mixing procedure, a sterilization procedure and a drying procedure, and a medicine intermediate is obtained after the drying procedure; the molding process is to prepare the drug intermediate into a preparation; the sterilization process is to perform sterilization by adopting circulating steam.

2. The preparation method of the classic famous prescription abalone shell powder preparation as claimed in claim 1, wherein the processing steps are as follows:

taking a abalone shell medicinal material, removing impurities, cleaning, drying in the sun, and crushing to obtain abalone shell decoction pieces;

taking a chrysanthemum medicinal material, and removing non-medicinal parts and impurities to obtain chrysanthemum decoction pieces;

removing non-medicinal parts and impurities according to semen Cassiae medicinal material to obtain semen Cassiae decoction pieces;

cleaning and drying notopterygium root medicinal materials to obtain notopterygium root decoction pieces;

taking a licorice medicinal material, removing non-medicinal parts and impurities, cleaning, soaking, cutting, and drying to obtain licorice decoction pieces.

3. The method of claim 2, wherein the non-medicinal portion of the chrysanthemum comprises the branch and calyx of chrysanthemum; the non-medicinal part of semen Cassiae comprises branch tip and calyx of semen Cassiae; drying Notopterygii rhizoma at below 60 deg.C; soaking Glycyrrhrizae radix at 35 deg.C for 3 hr, cutting into pieces with thickness of 2-4mm, drying at 60 deg.C for 5 hr.

4. The method as claimed in claim 1, wherein the pulverizing and mixing step comprises the following steps:

the processed abalone shell powder is respectively put into a crushing device to be respectively crushed into fine powder, and then the fine powder is sieved and evenly mixed to obtain mixed powder.

5. The method of claim 4, wherein the weight ratio of the fine powder of concha haliotidis, the fine powder of chrysanthemum, the fine powder of cassia seed, the fine powder of notopterygium root and the fine powder of licorice in the mixed powder is 2:2:2:2: 1.

6. the method of claim 1, wherein the formulation comprises powders, capsules, or granules.

7. The method of claim 6, wherein the excipients used in the manufacturing process of the classical prescription abalone shell powder formulation include magnesium stearate, sodium lauryl sulfate, pregelatinized starch.

8. The process for preparing a classical and famous abalone shell powder formulation according to any one of claims 1-7, wherein the sterilization is carried out by flowing steam under normal pressure.

9. The method for preparing a classical and famous formula Concha Haliotidis powder preparation according to claim 8, wherein the sterilization process is performed by normal pressure sterilization with 100 deg.C flowing steam for 30min.