A preparation method of Qifuyin compound granules

Abstract

The present invention belongs to the technical field of Chinese medicine preparations, and specifically relates to a Qifuyin compound granule, a preparation method and a quality control method thereof. The granule is composed of seven medicines, namely, ginseng, Rehmannia glutinosa, Chinese angelica, stir-fried Atractylodes macrocephala, roasted licorice, Chinese date seeds and processed Polygala tenuifolia; the preparation method of the granule is mainly to adopt different extraction methods for different raw materials, to enclose volatile oil with β-cyclodextrin, to concentrate the filtrate extracted by classification, to add inclusion compound and auxiliary materials, to mix and granulate to obtain the finished product; the present invention optimizes and screens the preparation process parameters of the granule, and also performs thin layer identification on ginseng, Rehmannia glutinosa, roasted licorice, Chinese angelica and processed Polygala tenuifolia in the quality control method, and performs quantitative detection on ginsenosides and 3,6′-dierucyl sucrose. Compared with traditional decoctions, the granules prepared by the present invention retain the effective ingredients such as volatile oils and ginsenosides to the maximum extent, improve the efficacy, save Chinese medicine resources, standardize the quality standards, and are suitable for industrial production.

Description

Technical Field

[0001] The invention belongs to the technical field of traditional Chinese medicine preparations, and particularly relates to a Qifuyin compound granule, a preparation method and a quality control method thereof. Background Art

[0002] Qi Fu Yin comes from "Jing Yue Quan Shu" and is composed of 7 herbs: ginseng, rehmannia root, angelica root, stir-fried atractylodes, roasted licorice, spinach seed and processed polygala. It is mainly used to treat dementia caused by deficiency of qi and blood in the five internal organs, especially the heart and spleen. It has the effects of replenishing qi and blood, calming the heart and strengthening the spleen, improving intelligence and tranquilizing the mind.

[0003] However, the traditional decoction of Qifuyin has a poor taste, a large dosage, and is difficult to control the dosage, which is not conducive to the treatment of many diseases. The active ingredients of the medicine may volatilize during the decoction process, thereby reducing the clinical efficacy. In addition, the time of taking the medicine is relatively particular, and the process of boiling the medicine is also relatively cumbersome. Compared with traditional decoctions, granules are stable in nature, easy to transport, carry, and store, and the production process of granules is simple and easy to achieve mechanized production. There is no quality standard for Qifuyin granules at present. Therefore, it is very necessary to establish the quality control standard of Qifuyin granules to make the preparation method of Qifuyin granules standardized, reasonable, and suitable for large-scale production. Summary of the invention

[0004] In view of the above problems, the present invention aims to provide a method for preparing Qifuyin compound granules and to perform quality control on the whole process, so as to establish the quality control standard of Qifuyin granules.

[0005] To achieve the above object, the present invention adopts the following technical solution:

[0006] A Qifuyin compound granule is prepared by extracting raw materials in the granule according to different active ingredients; wherein, in parts by weight, the raw materials include 300 parts of ginseng, 450 parts of prepared rehmannia root, 450 parts of angelica root, 250 parts of stir-fried atractylodes, 150 parts of roasted liquorice root, 300 parts of spiny jujube seeds, and 250 parts of processed polygala root;

[0007] The specific steps include:

[0008] (1) preparing each raw material drug according to the weight portion;

[0009] (2) adding ethanol to ginseng, heating under reflux for extraction, combining the filtrate, concentrating to obtain ginsenoside extract, and setting aside the ginseng residue;

[0010] (3) Take angelica and stir-fried atractylodes, add water and extract by steam distillation to obtain volatile oil and water extract, mix the volatile oil with ethanol, add β-cyclodextrin and water for inclusion, and dry to obtain inclusion compound, and use the angelica residue, stir-fried atractylodes residue and water extract for later use;

[0011] (4) mixing the ginseng residue, angelica residue and stir-fried atractylodes residue with the remaining raw materials, decocting with water, filtering, combining the filtrate with the ginsenoside extract and the water extract, and concentrating to obtain a clear paste; allowing the clear paste to stand, centrifuging, and concentrating to obtain an extract;

[0012] (5) Dry the extract, add the inclusion compound, grind to obtain dry extract powder, add auxiliary materials, mix well, granulate to obtain the finished product.

[0013] The above technical solution is preferably:

[0014] The unit prescription composition of the raw material medicine is preferably: 300g of ginseng, 450g of Rehmannia root, 450g of Angelica sinensis, 250g of stir-fried Atractylodes macrocephala, 150g of roasted Licorice root, 300g of Ziziphus jujuba seeds, and 250g of processed Polygala tenuifolia.

[0015] Preferably, in step (2), the selected ethanol is 50-70% by volume of medicinal ethanol, the weight ratio of the ginseng raw material to the ethanol is 1:3-18, and the extraction is carried out by heating and reflux for 1-3 times, each time for 1-2 hours; more preferably, the extraction is carried out twice, the first time 60% by volume ethanol accounting for 10 times the weight of the ginseng is added, and the second time 60% by volume ethanol accounting for 8 times the weight of the ginseng is added, and each extraction takes 1.5 hours.

[0016] Preferably, in step (2), the concentration is performed until each 1 mL of concentrated solution contains 0.5 to 1 g of the raw medicinal material.

[0017] Preferably, in step (2), the concentration conditions are a temperature of 55° C. and a pressure of 100 bar.

[0018] Preferably, in step (3), the weight of water is 6 to 10 times the total weight of Angelica sinensis and stir-fried Atractylodes macrocephala, and the steam distillation is performed for 3 to 7 hours; more preferably, the weight of water is 8 times the total weight of Angelica sinensis and stir-fried Atractylodes macrocephala, and the steam distillation is performed for 5 hours.

[0019] Preferably, in step (3), during inclusion, the volatile oil and ethanol are mixed in a volume ratio of 1:1, the inclusion method is a colloid mill method, the amount ratio of the volatile oil, water and β-cyclodextrin is 1mL:18-84mL:6-12g, and the inclusion time is 10-30min; more preferably, the amount ratio of the volatile oil, water and β-cyclodextrin is 1mL:27mL:9g, and the inclusion time is 20min.

[0020] Preferably, in step (4), the decoction is performed by adding water accounting for 3 to 18 times the total weight of all the raw materials in this step, and decocting for 1 to 3 times, each time for 1 to 2 hours; more preferably, the decoction is performed twice, with water accounting for 10 times the total weight of the raw materials being added for the first time and water accounting for 8 times the total weight of the raw materials being added for the second time, and each decoction is performed for 2 hours.

[0021] Preferably, in step (4), the concentration conditions for obtaining the clear paste are a temperature of 65°C and a pressure of 80 bar; the relative density of the clear paste is 1.02 to 1.08 (60±5°C); more preferably, the relative density of the clear paste is 1.02 to 1.06 (60±5°C).

[0022] Preferably, in step (4), when the centrifugal force of the centrifuge is lower than 9395×g, the medicinal solution becomes turbid and a small amount of sediment is left at the bottom of the container after being placed. Therefore, the centrifugal force needs to be maintained above 9395×g, that is, the centrifugal force of the centrifugation is 4175×g to 16700×g; more preferably, the centrifugal force of the centrifugation is 9395×g.

[0023] Preferably, in step (4), the concentration conditions for obtaining the extract are a temperature of 65°C and a pressure of 80 bar; the relative density of the extract is 1.22 to 1.30 (60±5°C); more preferably, the relative density of the extract is 1.22 to 1.28 (60±5°C).

[0024] Preferably, in step (4), the standing is standing at room temperature for 12 hours.

[0025] Preferably, in step (5), the drying method is vacuum belt drying, the feeding speed of the cloth motor is set to 2.0L / H, the cloth rod angle is 15 degrees, the crawler speed is 100mm / min, and the temperatures of the four stages are 80℃~95℃, 80℃~90℃, 75℃~85℃, and 25℃ respectively; more preferably, the temperatures of the four stages are 95℃, 90℃, 85℃, and 25℃ respectively.

[0026] Preferably, in step (5), the auxiliary materials include a filler and a lubricant; wherein the filler is selected from white dextrin, maltodextrin or soluble starch, and the lubricant is magnesium stearate; more preferably, the filler is soluble starch, and the lubricant is magnesium stearate accounting for 0.5% of the total mass of the granules.

[0027] Preferably, in step (5), the granulation method is dry granulation, the water content during granulation is controlled below 8%, and the environmental humidity during granulation, packaging and storage is controlled below 50.79%.

[0028] Preferably, the content of each bag of the finished product is 20g, wherein ginsenoside Rg1 (C 42 H 72 O 14 )、ginsenoside Re(C 48 H 82 O 18 )、ginsenoside Rb1(C 54 H 92 O 23) total content of at least 17mg / bag, 3,6′-biserucyl sucrose (C 36 H 46 O 17 ) content is at least 5.5 mg / bag.

[0029] The beneficial technical effects of the present invention are:

[0030] (1) Using different solvents to extract different active ingredients in the prescription, extracting the active ingredients in the prescription as completely as possible, thereby improving the efficacy and saving Chinese medicine resources;

[0031] (2) After the volatile oils extracted from angelica and stir-fried atractylodes by distillation are encapsulated with β-cyclodextrin, the volatile oils are isolated from the external environment, which maximizes the retention of the volatile oil content and its effective substances, thereby ensuring that its efficacy is fully exerted and greatly increasing the stability of the drug;

[0032] (3) The refined sample reduces the oral dose, and the granules are easy to carry, have a long shelf life, and can improve patient compliance;

[0033] (4) The present invention optimizes and screens the preparation process parameters of the granules, and provides a quality control method for the Qifuyin compound granules, which ensures the stability, uniformity and controllability of the product quality. In addition, the quality control method also has the advantages of strong operability and good repeatability.

[0034] Obviously, according to the above contents of the present invention, in accordance with common technical knowledge and customary means in the art, without departing from the above basic technical ideas of the present invention, other various forms of modification, replacement or change may be made.

[0035] The above contents of the present invention are further described in detail below through specific implementation methods in the form of examples. However, this should not be understood as the scope of the above subject matter of the present invention being limited to the following examples. All technologies realized based on the above contents of the present invention belong to the scope of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0036] Figure 1 This is the technical route for preparing the Qifuyin granules of the present invention;

[0037] Figure 2 This is the thin layer identification chromatogram of ginseng, where 1 is the negative control, 3 is the control herbal medicine, 2, 4, and 5 are Qifuyin Granules A, B, and C respectively;

[0038] Figure 3 This is the TLC identification chromatogram of Rehmannia glutinosa, where 1 is the negative control, 3 is the control herbal medicine, 2, 4, and 5 are Qifuyin Granules A, B, and C respectively;

[0039] Figure 4 This is the thin layer identification chromatogram of Angelica sinensis, where 1 is the negative control, 3 is the control herbal medicine, 2, 4, and 5 are Qifuyin Granules A, B, and C respectively;

[0040] Figure 5 This is the TLC identification chromatogram of Zhigancao, where 1 is the negative control, 3 is the control herbal material, 2, 4, and 5 are Qifuyin Granules A, B, and C respectively;

[0041] Figure 6 This is the thin layer identification chromatogram of processed Polygala tenuifolia, where 2 is the negative control, 4 is the reference medicinal material, and 1, 3, and 5 are Qi Fu Yin Granules A, B, and C respectively. DETAILED DESCRIPTION

[0042] In order to better illustrate the purpose, technical solutions and advantages of the present invention, the present invention will be further described below in conjunction with the accompanying drawings and specific embodiments.

[0043] Example 1

[0044] A Qifuyin compound granule, the raw material composition includes, by weight:

[0045] 300g ginseng, 450g Rehmannia root, 450g Chinese angelica root, 250g stir-fried Atractylodes macrocephala, 150g roasted Licorice root, 300g Ziziphus jujuba seeds, 250g processed Polygala tenuifolia;

[0046] The specific steps are as follows:

[0047] (1) preparing each raw material drug according to the weight portion;

[0048] (2) adding 60% ethanol to ginseng and heating under reflux twice, adding ethanol 10 times the weight of ginseng for the first time and 8 times the weight of ginseng for the second time, extracting for 1.5 hours each time, filtering, recovering ethanol, and concentrating at a temperature of 55° C. and a pressure of 100 bar to obtain a ginsenoside concentrate containing 0.5 to 1 g of raw medicinal material per 1 mL for use, and the ginseng residue for use;

[0049] (3) Take angelica and stir-fried atractylodes, add water that is 8 times the total weight of angelica and stir-fried atractylodes, and steam distill for 5 hours to obtain volatile oil and water extract, mix the volatile oil with ethanol in a volume ratio of 1:1, add β-cyclodextrin and water for inclusion, wherein the amount ratio of volatile oil, water and β-cyclodextrin is 1mL:27mL:9g, use a colloid mill method, grind and include for 20 minutes, dry to obtain the inclusion compound for later use, and use the water extract, angelica residue and stir-fried atractylodes residue for later use;

[0050] (4) Mixing the ginseng residue, angelica residue and stir-fried Atractylodes residue with the remaining raw materials, adding water and decocting twice, adding water accounting for 10 times the total weight of all the raw materials in step (4) for the first time, and adding water accounting for 8 times the total weight of all the raw materials for the second time, decocting for 2 hours each time, filtering, combining the filtrate with the ginsenoside concentrate and the water extract, and concentrating to a clear paste (60±5°C) with a relative density of 1.02 to 1.06 at a temperature of 65°C and a pressure of 80 bar; leaving the obtained clear paste to stand at room temperature for 12 hours, centrifuging at 9395×g, and concentrating to an extract (60±5°C) with a relative density of 1.22 to 1.28 at a temperature of 65°C and a pressure of 80 bar;

[0051] (5) The extract was subjected to vacuum belt drying, and the feeding speed of the distribution motor was set to 2.0 L / H, the distribution rod angle was set to 15 degrees, the crawler speed was set to 100 mm / min, and the temperatures of the four drying stages were 95°C, 90°C, 85°C, and 25°C, respectively. The inclusion compound was added and ground to obtain a dry paste powder; soluble starch and magnesium stearate accounting for 0.5% of the total mass of the granules were added to the dry paste powder, mixed well, and dry granulated to obtain 1000 g of the finished product, which was packaged in 20 g bags, and a total of 50 bags of Qifuyin granules were obtained.

[0052] According to the above steps, three batches of samples were prepared for use, and were respectively named Qifuyin Granules A, B, and C.

[0053] Example 2 Extraction process study

[0054] 1Study on Ginseng Extraction Technology

[0055] According to the screening results of the process route, the alcohol extraction effects of ginsenoside Rg1+Re content and ginsenoside Rb1 content are better than water extraction, so ginseng is determined to be extracted with alcohol. The alcohol extraction process of ginseng was studied. The orthogonal experiment was used to screen the ethanol concentration, total alcohol addition amount, extraction time and number of extractions. The results showed that these four factors had no significant effect on the extraction rate of the three index components of ginseng. However, considering that the ginseng residue after alcohol extraction still needs to be extracted with water, reasonable adjustments were made from the perspective of actual production, and the optimal extraction process was determined to be extracted twice with 60% medicinal ethanol, with the amount of alcohol added being 10 times and 8 times the weight of ginseng, respectively, and extracted for 1.5 hours respectively.

[0056] 2 Study on the extraction technology of volatile oil from Angelica sinensis and stir-fried Atractylodes macrocephala

[0057] In order to select the best extraction process of volatile oil from Angelica sinensis and Atractylodes macrocephala, an orthogonal experiment was designed to investigate the main influencing factors of water addition (times) and extraction time (h). 450g of Angelica sinensis and 250g of Atractylodes macrocephala were weighed, a total of 9 portions, and the experiment was carried out. The content of ligustilide and volatile oil was determined by high performance liquid chromatography. The factor levels are shown in Table 1, and the experimental results and data analysis are shown in Table 2, Table 3, and Table 4.

[0058] Table 1. Levels of factors for extracting volatile oil from Angelica sinensis and stir-fried Atractylodes macrocephala

[0059]

[0060] Table 2 Results of the experiment on extracting volatile oil from Angelica sinensis and Atractylodes macrocephala

[0061]

[0062]

[0063] Table 3 Analysis of variance of volatile oil content

[0064]

[0065] Table 4 Analysis of variance table of ligustilide content

[0066]

[0067] According to the results, the influence of the two factors on the extraction of volatile oil is B>A, that is, the extraction time has the greatest influence on the extraction of volatile oil, followed by the amount of water added. The results of variance analysis show that the extraction time (B) has a significant effect on the extraction of oil (mL) and ligustilide content (mg / g angelica) of angelica and stir-fried atractylodes (P<0.05). According to the results of variance analysis, considering the actual situation such as production time, the optimal process conditions for oil extraction are determined to be A2B2. Verification experiments were carried out according to the preferred extraction process conditions. The results showed that the RSD values ​​of the extracted volatile oil amount and ligustilide content were less than 3%. The volatile oil extraction process is stable, reasonable and feasible. That is, the extraction process is that the amount of water added is 8 times the total weight of angelica and stir-fried atractylodes, and the steam distillation extraction is 5 hours.

[0068] 3 Study on the inclusion technology of volatile oil β-cyclodextrin

[0069] 3.1 Study on the inclusion method of volatile oil β-cyclodextrin

[0070] If the volatile oil extracted from Angelica sinensis and Atractylodes macrocephala is directly sprayed into the particles, it is easy to volatilize and affect the quality of the preparation; if it is encapsulated with β-cyclodextrin, the oxidation deterioration and volatilization loss during production and storage are reduced, and the stability of the drug is greatly increased, thereby ensuring the efficacy of the drug. After the volatile oil is encapsulated, the irritation and bad smell are reduced, and it is also convenient for preparation. An experiment was conducted to determine the encapsulation method of volatile oil and β-cyclodextrin. Weigh 90g of β-cyclodextrin respectively, slowly add 10mL of volatile oil (dissolved in the same volume of ethanol), use saturated aqueous solution method, colloid mill method and ultrasonic method for encapsulation, and test the encapsulation rate and yield of the three methods respectively. The experimental results are shown in Table 5.

[0071] Table 5 Inclusion rate (%) and yield (%) of different inclusion methods

[0072]

[0073] From the results in Table 5, it can be seen that, with inclusion yield (%) and inclusion rate (%) as evaluation indicators, the colloid milling method is the best among the three inclusion methods, so the volatile oil is included by the colloid milling method.

[0074] 3.2 Study on the process conditions of β-cyclodextrin inclusion of volatile oil

[0075] In order to select the best technology for inclusion of volatile oil of Angelica sinensis and Atractylodes macrocephala, an orthogonal experiment was designed and L9(3 4 ) orthogonal table, the ratio of volatile oil to β-cyclodextrin, the ratio of β-cyclodextrin to water, and the inclusion time (min) were investigated. 10mL of volatile oil (dissolved in the same volume of ethanol) was measured, a total of 9 portions, and orthogonal experiments were carried out. The factor levels are shown in Table 6, and the experimental results and data analysis are shown in Tables 7 and 8.

[0076] Table 6 β-cyclodextrin inclusion factor level table of volatile oil

[0077]

[0078] Table 7 β-cyclodextrin inclusion volatile oil experimental results

[0079]

[0080] Table 8 Analysis of variance of β-cyclodextrin inclusion volatile oil

[0081]

[0082] According to the results, the order of influence of each factor on the experimental results is A>C>B, that is, the ratio of oil to β-cyclodextrin has the greatest influence on the inclusion of volatile oil. It can be seen from the variance analysis table in Table 8 that the ratio of oil to β-cyclodextrin has a significant effect on the inclusion of volatile oil, and the inclusion time and the ratio of β-cyclodextrin to water have no significant effect on the inclusion of volatile oil. The intuitive analysis results show that the optimal inclusion process conditions are A2B1C2. The verification experiment was carried out according to the preferred extraction process conditions. The results showed that the RSD values ​​of the inclusion rate and yield were less than 3%, and the extraction process was stable, reasonable and feasible. That is, the inclusion process is that the ratio of volatile oil to β-cyclodextrin is 1:9, the ratio of β-cyclodextrin to water is 1:3 (the dosage ratio of volatile oil, water and β-cyclodextrin is 1mL:27mL:9g), and the inclusion time is 20min.

[0083] 4 Study on the water extraction technology of seven medicinal materials including Rehmannia glutinosa

[0084] In order to select the best decoction process of ginseng residue, angelica residue, stir-fried atractylodes residue and remaining raw materials, an orthogonal experiment was designed and L9(3 4 ) orthogonal table, the total amount of water added (times), total extraction time (h), extraction times (times) three main influencing factors were investigated. Weigh 150g of ginseng, 225g of angelica, 125g of stir-fried atractylodes, 225g of prepared rehmannia, 75g of roasted licorice, 150g of spiny jujube seeds, and 125g of processed polygala in 9 portions. Ginseng was extracted with alcohol according to the preferred process, angelica and stir-fried atractylodes were extracted according to the preferred process, and the above three kinds of drug residues and the rest of the medicinal materials were extracted according to the above L9 (3 4 ) orthogonal table was used for the experiment, and the obtained filtrate was combined with ginsenoside extract and water extract and concentrated to 5.0 kg. The content of 3,6′-dicerucyl sucrose was determined by high performance liquid chromatography. The factor levels are shown in Table 9, and the experimental results and data analysis are shown in Tables 10 and 11.

[0085] Table 9 Water extraction factor levels of seven medicinal materials including Radix Rehmanniae Preparata

[0086]

[0087] Table 10 Water extraction test results of seven medicinal materials such as Radix Rehmanniae Preparata

[0088]

[0089] Table 11 Analysis of variance of water extraction of seven medicinal materials including Radix Rehmanniae Preparata

[0090]

[0091] According to the results in Tables 10 and 11, the order of influence of each factor on the test results is C>A>B, and the number of extractions has a significant effect on the decoction effect (P<0.05). The intuitive analysis results show that the optimal extraction conditions are A3B3C3. In view of the fact that the oil-extracting drug residue has been extracted for 5 hours, the actual conditions such as the extraction solvent cost, energy cost, labor cost and subsequent concentration cost consumed in the production process are comprehensively considered for reasonable adjustment, and the optimal extraction process is determined to be A3B3C2. The verification experiment was carried out according to the preferred extraction process conditions. The results showed that the RSD values ​​of the paste yield and the content of 3,6′-dicorinoyl sucrose were less than 5%, and the extraction process was stable, reasonable and feasible. That is, the extraction process is to add water and decoct twice, the first time adding water accounting for 10 times the total weight of the raw material, the second time adding water accounting for 8 times the total weight of the raw material, and each extraction takes 2 hours.

[0092] Example 3 Purification process study

[0093] 1. Study on relative density of clear paste

[0094] In order to select the process parameters for the relative density of the clear paste, a single factor test was conducted on the relative density of the clear paste. Weigh 750g of ginseng, 1125g of Rehmannia glutinosa, 1125g of Angelica sinensis, 625g of stir-fried Atractylodes macrocephala, 375g of roasted Licorice root, 750g of Semen Ziziphi Spinosae, and 625g of processed Polygala tenuifolia. Extract according to the above preferred process conditions, divide the total extract into 4 parts (1 is 0.5 prescription volume, 2 is 0.5 prescription volume, 3 is 0.75 prescription volume, and 4 is 0.75 prescription volume), concentrate to a certain density, stand and centrifuge. Determine the total weight before and after centrifugation, and use high performance liquid chromatography to determine the content of ginsenoside Rg1+Re+Rb1 and 3,6′-dimercaptoyl sucrose. The data results are shown in Table 12.

[0095] Table 12 Data results before and after centrifugation

[0096]

[0097] According to the analysis of the experimental results, when the relative density of the clear paste is 1.02-1.06, the weight (g), dry matter content (%), total ginsenoside content (mg), and 3,6′-dicercosylsucrose content (mg) before and after centrifugation do not change much. When the relative density of the clear paste is 1.08, the evaluation indicators decrease significantly. Therefore, the relative density of the clear paste is selected to be 1.02-1.06.

[0098] 2. Study on relative density of extract

[0099] In order to select the relative density process parameters of the extract, a single factor test was conducted on the relative density of the extract. Weigh 3.3kg of ginseng, 4.95kg of Rehmannia glutinosa, 4.95kg of Angelica sinensis, 2.75kg of stir-fried Atractylodes macrocephala, 1.65kg of roasted Licorice root, 3.3kg of Ziziphus jujuba seeds, and 2.75kg of processed Polygala tenuifolia. According to the above preferred conditions, the experiment was carried out, and when it was concentrated to 60.5kg, a clear paste (sampling and testing) was obtained, and the clear paste was divided into 5 portions (1 for 1.6 prescription amounts, 2 for 1.6 prescription amounts, 3 for 2.4 prescription amounts, 4 for 2.4 prescription amounts, and 5 for 3 prescription amounts), allowed to stand, centrifuged, and concentrated to relative densities of 1.22, 1.24, 1.26, 1.28, and 1.30, respectively. The extract was dried by vacuum belt drying, and the four-stage drying temperatures were 95°C, 90°C, 85°C, and 25°C. The total weight before and after centrifugation was measured, and the content of ginsenoside Rg1+Re+Rb1 and 3,6′-dicerinoylsucrose was determined by high performance liquid chromatography. The experimental results are shown in Table 13.

[0100] Table 13 Extract density screening results

[0101]

[0102] During the experiment, the sample extract of batch number 5 (relative density of the extract is 1.30) could not be dried because of its high relative density and difficulty in loading. As can be seen from Table 13, when the relative density of the extract is 1.22-1.28, the amount of ginsenoside Rg1+Re+Rb1 (mg) and 3,6′-dicercosyl sucrose (mg) did not change much before and after drying, and the transfer rate was greater than 88%, and the moisture content of the dry extract powder met the requirements of granules (<8%). Therefore, the relative density of the extract is determined to be 1.22-1.28.

[0103] 3 Drying process research

[0104] In actual production, the drying methods are mainly atmospheric pressure drying and reduced pressure drying. Atmospheric pressure drying takes a long time and has high temperature, which will cause losses to heat-unstable ingredients, and the product quality is difficult to control. Reduced pressure drying is carried out under closed and vacuum conditions. The main advantages are short drying time, thorough drying, loose products and easy crushing. In order to prevent the effective ingredients from being damaged by heat for a long time, combined with actual production conditions, this experiment uses vacuum belt drying, and the vacuum degree is controlled at 2200~2400pa.

[0105] 3.1 Preliminary selection of drying temperature

[0106] In order to select the drying temperature process parameters, the drying temperature was investigated. The liquid medicine of batch number 4 in the process research of the relative density of the above-mentioned clear paste was combined and pumped into a rotary evaporator for concentration. Concentrate to a relative density of 1.25, take 95g (2 portions) of each, and put them into a vacuum drying oven for drying. Set 85℃ and 95℃ respectively. The content of ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1 and 3,6′-dicorinoylsucrose were determined by high performance liquid chromatography. The experimental results are shown in Table 14.

[0107] Table 14 Drying temperature preliminary results

[0108]

[0109] The results showed that drying temperature below 95°C had little effect on ginsenosides and 3,6′-dicerinoylsucrose.

[0110] 3.2 Drying temperature screening study

[0111] In order to select the drying process parameters, a single factor test was conducted on different heating parameters. Weigh 1.8kg of ginseng, 2.7kg of Rehmannia root, 2.7kg of Angelica sinensis, 1.5kg of stir-fried Atractylodes macrocephala, 0.9kg of roasted Licorice root, 1.8kg of Ziziphus jujuba seeds, and 1.5kg of processed Polygala tenuifolia. Extract according to the optimal conditions of the above experiment. When the relative density is concentrated to 1.05, a clear paste is obtained. Let it stand, centrifuge, and concentrate to a relative density of 1.24 to obtain an extract weighing 10.20kg. Divide the extract into 3 parts for drying. Determine the moisture content (%) of the dry paste powder and observe the drying condition of the material. The heating parameters are shown in Table 15, and the experimental results are shown in Table 16.

[0112] Table 15 Heating parameter design table

[0113]

[0114] Table 16 Material drying results

[0115]

[0116] From the results in Table 16, it can be seen that according to the moisture requirement of the granules (<8%), this study set the four-stage drying temperatures to 95°C, 90°C, 85°C, and 25°C. In actual production, the temperature parameters can be adjusted according to the specific equipment.

[0117] Example 4 Granulation and Forming Process Research

[0118] 1. Choice of granulation method

[0119] At present, the commonly used granulation technologies for Chinese medicine granules are mainly wet granulation and dry granulation. Dry granulation is a granulation process that uses a dry granulator to directly press the mixed raw material powder and then crush it into whole particles. Its advantage is that it can be granulated directly without adding wetting agents, drying particles, etc., and the process is simple and convenient. Because the dry matter proportion of Qifuyin granules is large (>80%) and the proportion of auxiliary materials is small, dry granulation is used for granulation.

[0120] 2 Granulation filling agent screening

[0121] Conventional excipients for Chinese medicine granules generally include lactose, dextrin, soluble starch, etc. Considering that this product is mainly used for middle-aged and elderly patients, sugar-containing excipients are not selected. In view of the characteristics of large paste output and strong hygroscopicity of this prescription, it is particularly important to select excipients with low hygroscopicity. Since white dextrin, soluble starch and maltodextrin have low hygroscopicity and good stability, these three excipients are investigated with molding rate, hygroscopicity, solubility and fluidity as evaluation indicators. The experimental results are shown in Table 17.

[0122] Table 17 Excipient screening results

[0123]

[0124] During the experiment, after being placed at 75% relative humidity for 80 hours, the moisture absorption rate of the dry paste powder was the highest, and the addition of different excipients could significantly reduce the hygroscopicity of the material. According to the data in the table, it can be seen that the comprehensive score of soluble starch is the highest, so soluble starch is selected as the molding excipient of this granule.

[0125] 3. Lubricant dosage screening

[0126] The commonly used lubricant in dry granulation is magnesium stearate, and the commonly used addition amount is 0.1% to 1.0%. In order to improve the molding rate, the amount of magnesium stearate was screened and the granulation condition was observed. The results are shown in Table 18.

[0127] Table 18 Experimental results

[0128]

[0129] It can be seen from Table 18 that when the addition amount of magnesium stearate is 0.5%, there is no sticking phenomenon and the one-time forming rate reaches 73.00%. Therefore, the addition amount of magnesium stearate is selected to be 0.5% of the total mass of the granules.

[0130] 4. Determination of critical relative humidity (CRH) of particles

[0131] 14 portions of sample granules prepared by the preferred preparation process are placed in a weighing bottle that has been dried to constant weight, with a thickness of about 2 mm, accurately weighed, and placed in constant temperature and humidity chambers with different humidity for 120 hours, and 2 portions are measured in parallel in each constant temperature and humidity chamber to calculate the moisture absorption rate. A curve is drawn with relative humidity (%) as the horizontal coordinate and moisture absorption rate (%) as the vertical coordinate, and tangents are drawn from the inflection points at both ends of the curve. The value corresponding to the intersection of the tangents on the horizontal coordinate is the critical relative humidity. According to the experimental results, the CRH of the granules is calculated to be 50.79%. Therefore, the environmental humidity of this product during granulation, packaging and storage needs to be controlled below 50.79%, thereby reducing the influence of moisture on the properties and stability of the drug.

[0132] In summary, in the granulation process, dry granulation was adopted, soluble starch was selected as a filler, magnesium stearate (0.5% of the total mass of the granules) was used as a lubricant, and the ambient humidity was controlled below 50.79% during granulation, packaging and storage. Test Example 1 Determination of the content of total ginsenosides and methodological verification

[0133] 1 Determination of the content of total ginsenosides

[0134] 1.1 Preparation of test solution

[0135] Take 4g of this product, grind it, weigh it accurately, put it in a stoppered conical flask, accurately add 50mL of 80% methanol by volume, stopper it, weigh it, treat it ultrasonically (power 450W, frequency 40kHz) for 30 minutes, let it cool, weigh it again, make up the lost weight with 80% methanol, shake it well, filter it, accurately measure 25mL of the filtrate, evaporate it to dryness, add 10mL of water to dissolve the residue, shake it with ethyl acetate for 3 times (30mL, 30mL, 20mL), discard the ethyl acetate solution, add 80mL of n-butanol to the aqueous solution, shake it well, wash it with ammonia test solution for 3 times, 30mL each time, combine the ammonia test solutions, shake it with water-saturated n-butanol for 2 times, 20mL each time, combine the n-butanol solutions, evaporate it to dryness, dissolve the residue with methanol, transfer it to a 10mL volumetric flask, add methanol to the scale, shake it well, filter it, and take the filtrate to obtain it.

[0136] 1.2 Preparation of reference solution

[0137] Take appropriate amounts of ginsenoside Rg1 reference substance, ginsenoside Re reference substance and ginsenoside Rb1 reference substance, weigh them accurately, add methanol to make a mixed solution containing ginsenoside Rg10.1 mg, ginsenoside Re 0.1 mg and ginsenoside Rb10.2 mg per 1 mL, shake well, and obtain.

[0138] 1.3 Chromatographic conditions

[0139] The chromatographic column is Agilent Poroshell 120EC-C18 (50mm×4.6mm, 2.7μm); acetonitrile is mobile phase A, water is mobile phase B, and gradient elution is performed according to the parameters in Table 19; the flow rate is 0.8mL / min; the detection wavelength is 203nm; 5μL of each of the above two solutions is accurately drawn, injected into the liquid chromatograph, and measured. The theoretical plate number calculated based on the total amount of ginsenosides should not be less than 6000.

[0140] Table 19 Ginseng gradient elution program

[0141]

[0142] 1.4 Content determination

[0143] The reference solution was used to locate the components in the test sample, and the content of total ginsenosides in three batches (A, B, and C) of samples was calculated by the external standard method, as shown in Table 20.

[0144] Table 20 Ginseng total saponin content determination results

[0145]

[0146] 2 Methodological verification of the determination of ginsenoside Rg1, ginsenoside Re and ginsenoside Rb1

[0147] 2.1 Exclusivity Investigation

[0148] Take a negative control without ginseng medicinal materials and prepare a negative control solution according to the preparation method of the test solution. Accurately pipette 5μL of each of the above three solutions and inject them into the liquid chromatograph for determination. The results show that the negative control has no interference and the method has good specificity.

[0149] 2.2 System suitability test

[0150] Accurately pipette 5 μL of the reference solution, repeat the injection 6 times, measure and record the peak areas of ginsenoside Rg1, ginsenoside Re and ginsenoside Rb1, and calculate the RSD. The RSD of the retention time and peak area were less than 0.8%, indicating that the instrument had good precision.

[0151] 2.3 Investigation of linear relationship

[0152] Take appropriate amount of ginsenoside Rg1, ginsenoside Re and ginsenoside Rb1 reference substances, accurately weigh them, place them in a 50mL volumetric flask, dissolve them by ultrasonication with 80% volume fraction methanol, cool to room temperature, dilute to the scale with 80% volume fraction methanol, shake well to obtain ginsenoside stock solution, accurately measure 1.0mL, 2.0mL, 4.0mL, 6.0mL, 8.0mL and 10.0mL of the reference substance stock solution in sequence, place them in 6 10mL volumetric flasks respectively, dilute to the scale with 80% volume fraction methanol, to obtain a series of ginsenoside Rg1, ginsenoside Re and ginsenoside Rb1 linear solutions, filter them, and measure the filtrate in sequence under the above-mentioned chromatographic conditions, and record the peak area. The standard curve was drawn with the concentration of the reference solution as the horizontal axis (X) and the peak area as the vertical axis (Y), and the regression equation of ginsenoside Rg1 was obtained: Y = 39.7782x + 0.0404, r = 0.999; the regression equation of ginsenoside Re: Y = 33.5135x + 0.0377, r = 0.999; the regression equation of ginsenoside Rb1: Y = 28.8340x + 0.0721, r = 0.999. The results showed that the linear relationship was good in the range of ginsenoside Rg1 injection concentration of 0.02273-0.22734 mg / mL, ginsenoside Re injection concentration of 0.03025-0.3025 mg / mL, and ginsenoside Rb1 injection concentration of 0.04532-0.4532 mg / mL.

[0153] 2.4 Stability test

[0154] The same test solution was taken and measured at 0h, 4h, 8h, 12h, 16h, and 20h after preparation at room temperature. The results showed that the peak area RSD of the test solution was less than 3.1% after being placed at room temperature for 24 hours, indicating that the test solution had good stability within 24 hours at room temperature.

[0155] 2.5 Repeatability test

[0156] Take 6 portions of batch A sample, prepare 6 test solutions, inject and analyze them in sequence, determine the peak area, calculate the content and relative standard deviation. The results show that the RSD of the repeatability test of ginsenoside Rg1, ginsenoside Re and ginsenoside Rb1 is less than 1.9%, and the repeatability is good.

[0157] 2.6 Intermediate precision test

[0158] Different analysts used different instruments on different dates to prepare 6 test solutions in parallel, and 5 μL of the reference solution and the test solution were accurately aspirated and injected into the liquid chromatograph to record the peak areas of ginsenoside Rg1, ginsenoside Re and ginsenoside Rb1. The results showed that the intermediate precision RSD was less than 1.9%, proving that the intermediate precision was good.

[0159] 2.7 Accuracy test

[0160] Take 6 samples with known ginsenoside Rg1, ginsenoside Re and ginsenoside Rb1 contents (ginsenoside Rg1 content is 0.567 mg / g, ginsenoside Re content is 0.801 mg / g, ginsenoside Rb1 content is 1.244 mg / g), each 2 g (equivalent to 1 / 2 of the sample volume specified in the standard), accurately weigh, place in 6 conical flasks, and accurately add ginsenoside reference solution (C Rg1 =0.1164mg / mL, C Re =0.1525mg / mL, C Rb1 =0.2538mg / mL) 10mL, prepare the sample recovery rate sample solution according to the above test sample preparation method, accurately draw 5μL of the sample recovery rate sample solution and the reference solution, inject them into the high performance liquid chromatograph in turn, and calculate the recovery rate according to the external standard method based on the peak area measured. The results show that the recovery rate limit is within 85% to 110%, which shows that the accuracy of the method is good.

[0161] 2.8 Durability test

[0162] In order to investigate the influence of slight fluctuations in chromatographic conditions on the separation and content determination of ginsenoside Rg1, ginsenoside Re and ginsenoside Rb1, a durability experiment was conducted. 5 μL of reference solution and test solution were accurately aspirated and injected into the liquid chromatograph. The peak areas of ginsenoside Rg1, ginsenoside Re and ginsenoside Rb1 were recorded under different chromatographic conditions. The contents of ginsenoside Rg1, ginsenoside Re and ginsenoside Rb1 were calculated by external standard method. The results showed that under the conditions of different chromatographic columns, slight changes in column temperature and flow rate, the RSD of ginsenoside Rg1 content was 1.63%, the minimum separation was 2.08, the RSD of ginsenoside Re content was 1.83%, the minimum separation was 2.08, and the RSD of ginsenoside Rb1 content was 1.47%, the minimum separation was 1.79, which showed that the method had good durability.

[0163] Test Example 23: Determination of 6′-Dierucyl Sucrose Content and Methodological Verification

[0164] Determination of 13,6′-dicerucyl sucrose content

[0165] 1.1 Preparation of test solution

[0166] Take 1g of this product, grind it into powder, weigh it accurately, put it in a 50mL volumetric flask, add an appropriate amount of 70% methanol, ultrasonically treat it (power 200W, frequency 40kHz) for 30 minutes, cool it, add 70% methanol by volume to the scale, shake it well, filter it, and take the filtrate to obtain it.

[0167] 1.2 Preparation of reference solution

[0168] Take an appropriate amount of 3,6′-dicerinoylsucrose reference substance, weigh it accurately, and add methanol to prepare a reference substance solution containing 10 μg per 1 mL.

[0169] 1.3 Chromatographic conditions

[0170] Octadecylsilane bonded silica gel was used as the filler (column length 150 mm), acetonitrile-0.1% phosphoric acid aqueous solution with a volume ratio of 17:83 was used as the mobile phase; the detection wavelength was 329 nm. The theoretical plate number calculated based on the 3,6′-dicerucyl sucrose peak should not be less than 3000.

[0171] 1.4 Content determination

[0172] The reference solution was used to locate the components in the test sample, and the content of 3,6′-dicerucyl sucrose in three batches (A, B, and C) of samples was calculated by the external standard method, as shown in Table 21.

[0173] Table 21 Sample 3,6′-dicerucyl sucrose content determination results

[0174]

[0175] Methodological Validation of Determination of 23,6′-Dierucyl Sucrose Content

[0176] 2.1 Exclusivity Investigation

[0177] Take a negative control without processed Polygala tenuifolia and prepare a negative control solution according to the preparation method of the test solution. Accurately pipette 5 μL of each of the three solutions, inject into the liquid chromatograph, and measure according to the method. The results show that the negative control has no interference and the method has good specificity.

[0178] 2.2 System suitability test

[0179] Accurately pipette 10 μL of the reference solution, repeat the injection 6 times, measure and record the peak area of ​​3,6′-dicerucyl sucrose, and calculate the RSD. The results show that the instrument has good precision.

[0180] 2.3 Investigation of linear relationship

[0181] Take an appropriate amount of 3,6′-diesinapoylsucrose reference substance, accurately weigh it, use 70% volume fraction methanol to make a solution containing 0.2695 mg of 3,6′-diesinapoylsucrose per 1 mL, shake well, and use it as the 3,6′-diesinapoylsucrose reference substance stock solution. Accurately measure 0.1 mL, 0.2 mL, 0.4 mL, 0.6 mL, 0.8 mL, and 1.0 mL of the 3,6′-diesinapoylsucrose reference substance stock solution in sequence, place them in 6 10 mL volumetric flasks, respectively, dilute to the scale with 70% volume fraction methanol, and obtain a series of 3,6′-diesinapoylsucrose linear solutions. Filter, and measure in sequence under the above-mentioned chromatographic conditions, and record the peak areas. The standard curve was drawn with the concentration of the reference solution as the abscissa (X) and the peak area as the ordinate (Y). The regression equation of 3,6′-dicerinoylsucrose was obtained as follows: Y=25984.0391x+5157.6959, R 2 =0.9999. The experimental results show that the linear relationship of 3,6′-dicerucyl sucrose is good in the range of 3.42-34.20 μg / mL.

[0182] 2.4 Stability test

[0183] The same test solution was taken and measured at room temperature at 0h, 4h, 8h, 12h, 16h, 20h, and 24h after preparation. The results showed that the peak area RSD of the test solution was less than 0.4% when it was placed at room temperature for 24 hours, indicating that the test solution had good stability within 24 hours at room temperature.

[0184] 2.5 Repeatability test

[0185] Take 6 samples of batch A respectively, prepare 6 test solutions according to the test solution preparation method of Test Example 2, inject and analyze them in sequence, determine the peak area, calculate the content and RSD. The results show that the RSD of the repeatability test of 3,6′-dicerinoylsucrose is less than 1.3%, and the repeatability is good.

[0186] 2.6 Intermediate precision test

[0187] Different analysts used different instruments on different dates to prepare 6 test solutions in parallel, and accurately aspirated 10 μL of the reference solution and the test solution, respectively, and injected them into the liquid chromatograph to record the peak area of ​​3,6′-dicerucyl sucrose. The results showed that the intermediate precision RSD was less than 2.0%, proving that the intermediate precision was good.

[0188] 2.7 Accuracy test

[0189] Take 6 samples with known 3,6′-dicorinoyl sucrose content (3,6′-dicorinoyl sucrose content is 0.594 mg / g), each 0.5 g (equivalent to 1 / 2 of the sample sampling amount specified in the standard), accurately weigh, place in 6 50 mL volumetric flasks, accurately add 3,6′-dicorinoyl sucrose reference solution (0.3046 mg / mL) 1 mL, prepare the sample recovery rate sample solution according to the above test solution method, accurately draw 10 μL of the sample recovery rate sample solution and the reference solution, inject them into the high performance liquid chromatograph in turn, and determine the content according to the external standard method according to the peak area measured, and calculate the recovery rate. The results show that the recovery rate limit is within 85% to 110%, which shows that the accuracy of the method is good.

[0190] 2.8 Durability test

[0191] In order to investigate the influence of slight fluctuations in chromatographic conditions on the separation and content determination of 3,6′-dicansinoyl sucrose, a durability experiment was conducted. 10 μL of reference solution and test solution were accurately aspirated and injected into the liquid chromatograph. The sample was analyzed under different chromatographic conditions, the peak area of ​​3,6′-dicansinoyl sucrose was recorded, and the content of 3,6′-dicansinoyl sucrose was calculated by external standard method. The results showed that under different chromatographic columns, slight changes in column temperature and flow rate, the RSD of 3,6′-dicansinoyl sucrose content was 1.36%, and the minimum separation was 1.65, which showed that the method had good durability.

[0192] Test Example 3 Identification of Ginseng in Qifuyin Compound Granules

[0193] 1 Preparation of test solution

[0194] Prepare the test solution according to the test solution preparation method in Test Example 1.

[0195] 2. Preparation of control medicinal material solution

[0196] Take 0.5 g of ginseng reference medicinal material, grind it into powder, put it in a stoppered conical flask, add 50 mL of methanol, ultrasonically treat it for 30 minutes, filter, evaporate the filtrate to dryness, add 20 mL of water to dissolve the residue, shake and extract it with ethyl acetate twice, 30 mL each time, discard the ethyl acetate solution, shake and extract the aqueous solution with water-saturated n-butanol twice, 30 mL each time, combine the n-butanol solutions, wash it with ammonia test solution twice, 30 mL each time, discard the ammonia solution, evaporate the n-butanol solution, add 10 mL of methanol to dissolve the residue, and use it as the reference medicinal material solution.

[0197] 3. Preparation of negative control solution

[0198] Take a sample without ginseng medicinal materials and prepare a negative control solution according to the test solution preparation method of Test Example 1.

[0199] 4 Thin layer identification

[0200] Take 10 μL of each of the two control medicinal material solutions and spot them on the same silica gel G thin layer plate, use chloroform-methanol-water (7:3:0.5) as the developing agent, develop, take out, dry, spray with 10% sulfuric acid ethanol solution, heat at 105℃ until the spots are clearly colored, and examine under sunlight. In the chromatogram of the test sample, spots of the same color appear at the corresponding position of the chromatogram of the control medicinal material. The results are shown in Figure 2 .

[0201] Test Example 4 Identification of Radix Rehmanniae Preparata in Qifuyin Compound Granules

[0202] 1 Preparation of test solution

[0203] Take 6 g of this product, grind it into powder, put it in a stoppered conical flask, add 20 mL of water, ultrasonically treat it for 30 minutes, centrifuge it, take the supernatant, shake it with ethyl acetate twice, 20 mL each time, combine the ethyl acetate liquid, evaporate it to dryness, add 1 mL of methanol to dissolve the residue, and use it as the test solution.

[0204] 2. Preparation of control medicinal solution

[0205] Take 1 g of Rehmannia glutinosa control medicinal material, add 20 mL of water, heat and reflux for 30 minutes, cool, centrifuge, take the supernatant, shake and extract twice with ethyl acetate, 30 mL each time, combine the ethyl acetate solutions, evaporate to dryness, add 0.5 mL of methanol to the residue to dissolve it, and use it as the control medicinal material solution.

[0206] 3. Preparation of negative control solution

[0207] Take a sample without Rehmannia glutinosa medicinal material and prepare a negative control solution according to the method for preparing the test sample solution.

[0208] 4 Thin layer identification

[0209] Take 10 μL of each of the above two solutions and spot them on the same silica gel G thin layer plate, use xylene-ethyl acetate (1:1) as the developing solvent, develop, take out, dry, spray with dinitrophenylhydrazine ethanol test solution, and examine under sunlight. In the chromatogram of the test sample, spots of the same color appear at the corresponding position of the chromatogram of the control medicinal material. Figure 3 .

[0210] Test Example 5 Identification of Angelica sinensis in Qifuyin Compound Granules

[0211] 1 Preparation of test solution

[0212] Take 5 g of this product, grind it into powder, put it in a stoppered conical flask, add 20 mL of water, ultrasonically treat for 30 minutes, centrifuge, take the supernatant, shake and extract twice with ether, 30 mL each time, combine the ether solution (keep the water solution for later use), recover the solvent to dryness, add 1 mL of cyclohexane to the residue to dissolve it, and use it as the test solution.

[0213] 2. Preparation of control medicinal material solution

[0214] Take 0.5 g of Angelica sinensis reference medicinal material, place it in a stoppered conical flask, add 20 mL of methanol, ultrasonically treat for 30 minutes, filter, evaporate the filtrate to dryness, add 2 mL of methanol to the residue to dissolve it, and use it as the reference medicinal material solution.

[0215] 3. Preparation of negative control solution

[0216] Take a sample without Angelica sinensis and prepare a negative control solution according to the test solution preparation method.

[0217] 4 Thin layer identification

[0218] 2 μL of the test solution and 1 μL of the control drug solution were taken and spotted on the same silica gel G thin layer plate, respectively. Petroleum ether (60℃~90℃)-ethyl acetate (2:1) was used as the developing agent. The plate was developed, taken out, dried, and examined under ultraviolet light (365nm). In the chromatogram of the test sample, a fluorescent spot of the same color appeared at the corresponding position of the chromatogram of the control drug. Figure 4 Test Example 6 Identification of Radix Glycyrrhizae Radix in Qifuyin Compound Granules

[0219] 1 Preparation of test solution

[0220] Take the reserved water solution in the preparation of the test solution in Experimental Example 5, add ethyl acetate and shake to extract twice, 30 mL each time, combine the ethyl acetate liquid, evaporate to dryness, add 1 mL of methanol to dissolve the residue, and use it as the test solution.

[0221] 2. Preparation of control medicinal material solution

[0222] Take 0.5 g of licorice control medicinal material, add 50 mL of methanol, heat and reflux for 30 minutes, filter, evaporate the filtrate to dryness, add 1 mL of methanol to the residue to dissolve it, and use it as the control medicinal material solution.

[0223] 3. Preparation of negative control solution

[0224] Take a sample without licorice herbs and prepare a negative control solution according to the test solution preparation method.

[0225] 4 Thin layer identification

[0226] 5 μL of the test solution and 2 μL of the control drug solution were taken and spotted on the same silica gel G thin layer plate, respectively. Ethyl acetate-formic acid-glacial acetic acid-water (15:1:1:2) was used as the developing agent. After development, the plate was taken out, dried, sprayed with 10% sulfuric acid ethanol solution, heated at 60°C until the spots were clearly colored, and then placed under sunlight for inspection. In the chromatogram of the test sample, spots of the same color appeared at the corresponding positions of the chromatogram of the control drug. Figure 5 .

[0227] Test Example 7 Identification of processed Polygala tenuifolia in Qifuyin compound granules

[0228] 1 Preparation of test solution

[0229] Take 3 g of this product, grind it into powder, put it in a stoppered conical flask, add 40 mL of 70% methanol, ultrasonically treat it for 30 minutes, filter it, evaporate the filtrate to dryness, add 20 mL of 10% sodium hydroxide solution to the residue to dissolve it, heat it under reflux for 1.5 hours, cool it, adjust the pH value to 4-5 with hydrochloric acid, shake it with ether for 2 times, 20 mL each time, discard the ether solution, shake the aqueous solution with ethyl acetate for 2 times, 20 mL each time, combine the ethyl acetate solutions, evaporate them to dryness, add 2 mL of methanol to the residue to dissolve it, and use it as the test solution.

[0230] 2. Preparation of control solution

[0231] Take the Polygala tenuifolia saponin reference substance, add methanol to make a 1mL solution containing 1.5mg, as the reference substance solution.

[0232] 3. Preparation of negative control solution

[0233] Take a sample without Polygala tenuifolia and prepare a negative control solution according to the method for preparing the test solution.

[0234] 4 Thin layer identification

[0235] Take 5 μL of each of the above two solutions and spot them on the same silica gel G thin layer plate, use ethyl acetate-glacial acetic acid-water (8:1:1) as the developing agent, develop, take out, dry, spray with 10% sulfuric acid ethanol solution, heat at 105℃ until the spots are clearly colored, and examine under sunlight. In the chromatogram of the test sample, spots of the same color appear at the corresponding position of the chromatogram of the reference sample. Figure 6 .

Claims

1. A method for preparing Qifuyin compound granules, characterized in that: The specific steps are as follows: (1) Prepare the raw materials according to weight. The raw materials are composed of the following components by weight: 300 g of ginseng, 450 g of Rehmannia root, 450 g of Angelica sinensis, 250 g of stir-fried Atractylodes macrocephala, 150 g of roasted Licorice root, 300 g of Ziziphus jujuba seeds, and 250 g of processed Polygala tenuifolia. (2) Add 60% ethanol to ginseng and heat under reflux twice. The first time, add ethanol that is 10 times the weight of ginseng, and the second time, add ethanol that is 8 times the weight of ginseng. Each extraction takes 1.5 hours. Filter and recover ethanol. Concentrate at 55 °C and 100 bar to obtain a ginsenoside concentrate containing 0.5 to 1 g of raw medicinal material per 1 mL for later use. The ginseng residue is also set aside for later use. (3) Take angelica and stir-fried atractylodes, add water that is 8 times the total weight of angelica and stir-fried atractylodes, and steam distill for 5 hours to obtain volatile oil and water extract. Mix the volatile oil with ethanol in a volume ratio of 1:1, add β-cyclodextrin and water for inclusion, wherein the amount ratio of volatile oil, water and β-cyclodextrin is 1 mL:27 mL:9 g. Use the colloid mill method, grind and include for 20 minutes, dry to obtain the inclusion compound for later use, and set aside the water extract, angelica residue and stir-fried atractylodes residue for later use; (4) Mixing the ginseng residue, angelica residue and stir-fried Atractylodes residue with the remaining raw materials, decocting twice with water, adding water accounting for 10 times the total weight of all the raw materials in step (4) for the first time, and adding water accounting for 8 times the total weight of all the raw materials for the second time, decocting for 2 hours each time, filtering, combining the filtrate with the ginsenoside concentrate and the water extract, and concentrating to a clear paste with a relative density of 1.02-1.06 at a temperature of 65°C and a pressure of 80 bar; leaving the obtained clear paste to stand at room temperature for 12 hours, centrifuging at 9395 × g, and concentrating to an extract with a relative density of 1.22-1.28 at a temperature of 65°C and a pressure of 80 bar; (5) The extract is subjected to vacuum belt drying, the feeding speed of the distribution motor is set to 2.0 L / H, the distribution rod angle is set to 15 degrees, the crawler speed is set to 100 mm / min, and the temperatures of the four drying stages are respectively 95°C, 90°C, 85°C, and 25°C. The inclusion compound is added and crushed to obtain a dry paste powder; soluble starch and magnesium stearate accounting for 0.5% of the total mass of the granules are added to the dry paste powder, the mixture is mixed, and dry granulation is performed to obtain a finished product.

Images