Granulation process and evaluation index of sugar-free granules capable of clearing away heat and toxic materials and eliminating dampness

Abstract

The invention discloses a granulation process and evaluation indexes of sugar-free granules for clearing away heat and toxic materials and eliminating dampness, and belongs to the technical field of medicine preparation, the granulation process comprises the following steps: weighing raw medicinal materials according to a prescription for clearing away heat and toxic materials and eliminating dampness, heating, refluxing and extracting for 1-3 times, adding 6-8 times of water each time, respectively extracting for 1-2 hours, filtering, merging filtrates, and concentrating at 80 DEG C under normal pressure until the relative density is 1.28-1.32, thereby obtaining the sugar-free granules for clearing away heat and toxic materials and eliminating dampness. The preparation method comprises the following steps: performing vacuum drying to obtain dry extract, crushing the dry extract into fine powder capable of passing through a 80-mesh sieve, then adding auxiliary materials with the same proportion as the fine powder, uniformly mixing, adding ethanol with the mass fraction of 60-80% to prepare a soft material, passing through a 12-mesh sieve, extruding to prepare wet granules, drying at 60 DEG C for 1.5 hours, and finishing the granules to obtain the traditional Chinese medicine composition. According to the invention, the rutin transfer rate and the dry extract rate are taken as evaluation indexes, the optimal extraction process of the sugar-free granules for clearing away heat and toxic materials and eliminating dampness is screened by an orthogonal test method, and the sugar-free granules for clearing away heat and toxic materials and eliminating dampness are successfully prepared by screening optimal forming process parameters by using a single factor analysis method and taking the forming rate, the dissolubility and the hygroscopicity as evaluation indexes.

Description

Technical Field

[0001] The invention relates to the technical field of medicine preparation, and in particular to a granulation process and evaluation index of sugar-free heat-clearing, detoxifying and dehumidifying granules. Background Art

[0002] The prescription for clearing heat, detoxifying and removing dampness is an empirical prescription that has been clinically used for many years in the First Affiliated Hospital of Guangzhou University of Chinese Medicine. It is mainly composed of medicinal ingredients such as Sophora japonica seeds, kapok flowers, chrysanthemums, and honeysuckle. It has the functions of clearing heat, detoxifying, removing water and dampness. It is currently mainly used in the clinical treatment of dysentery, enteritis, eczema, sores, red and swollen throat, red and painful eyes, etc. Kapok in the prescription has the effects of clearing heat, removing dampness, detoxifying and stopping bleeding. Its main ingredients are volatile oils and flavonoids, and it is mainly used to treat enteritis, diarrhea, bacterial dysentery and other diseases; Sophora japonica is good at clearing heat and cooling blood. "Compendium of Materia Medica" says that it "enters the liver and large intestine blood and cools blood, and treats wind-heat red eyes, red and white diarrhea, five hemorrhoids, intestinal wind, vomiting and metrorrhagia, and other blood". Its main active ingredient is rutin, which has the functions of stopping bleeding, anti-oxidation, and anti-inflammatory; Honeysuckle has the effects of clearing heat and detoxifying, cooling blood, and stopping diarrhea, and is often used for hot toxic bloody diarrhea and watery diarrhea; Imperata root can clear the heat of blood without hurting dryness; Mulberry leaf, "Compendium of Materia Medica" says that it "clears the lungs and drains the stomach, cools blood and dries dampness"; Chrysanthemum clears heat and detoxifies; Prunella vulgaris is good at clearing liver fire, dispersing knots and reducing swelling. All the medicines are used together to play the role of clearing heat and detoxification, removing dampness and promoting diuresis to stop diarrhea.

[0003] In order to make it easier for patients to carry and take, the Qingrejieduqushi recipe is prepared as a traditional Chinese medicine granule, with sucrose as an added excipient. However, excessive intake of high sugar can easily induce a variety of diseases, such as diabetes, cardiovascular disease, cancer, etc. Experts recommend that the intake of added sugar should be less than 25g per day. Therefore, for the health of more patients and to remove the medication restrictions for patients with metabolic diseases, it is planned to modify the original Qingrejieduqushi granules into sugar-free Qingrejieduqushi granules. However, the preparation method and evaluation indicators for sugar-free Qingrejieduqushi granules are still imperfect, which will lead to inaccurate drug testing results, thereby leading to uncontrollable drug quality. At the same time, the current evaluation indicators do not have an inspection of the excipient content and a measurement item for the content of the intrinsic quality indicators of the drug, and the quality of the product cannot be guaranteed during production.

[0004] In view of this, the inventor conducted in-depth research on this demand, and thus the present case was produced. Summary of the invention

[0005] In order to overcome the problem that the preparation method and evaluation index of the original heat-clearing, detoxifying and dampness-removing granules in the prior art are not perfect after being modified into sugar-free heat-clearing, detoxifying and dampness-removing granules, the present invention provides a granulation process and evaluation index of sugar-free heat-clearing, detoxifying and dampness-removing granules, takes rutin transfer rate and dry paste rate as evaluation indexes, and uses orthogonal experiment method as a method to screen the best extraction process of sugar-free heat-clearing, detoxifying and dampness-removing granules, and uses single factor analysis method, takes molding rate, solubility and hygroscopicity as evaluation indexes, screens the best molding process parameters, and successfully prepares sugar-free heat-clearing, detoxifying and dampness-removing granules.

[0006] In order to achieve the above-mentioned purpose, the present invention provides the following technical solutions:

[0007] The invention discloses a granulation process of sugar-free heat-clearing, detoxifying and dehumidifying granules. The raw medicinal materials are weighed according to the heat-clearing, detoxifying and dehumidifying prescription, and the medicinal materials are heated and refluxed for extraction for 1 to 3 times, and 6 to 8 times of water is added each time, and the extraction is performed for 1 to 2 hours respectively and filtered, and the filtrate is combined, concentrated at 80°C and normal pressure to a relative density of 1.28 to 1.32, vacuum dried to form a dry extract, and crushed into fine powder that can pass through an 80-mesh sieve, and then auxiliary materials with the same proportion as the fine powder are added, mixed evenly, and 60 to 80% of ethanol by mass fraction is added to prepare a soft material, which is squeezed through a 12-mesh sieve to prepare wet granules, and dried at 60°C for 1.5 hours, and granulated to obtain the obtained granules. The raw medicinal materials of the sugar-free heat-clearing, detoxifying and dehumidifying granules include kapok flowers, honeysuckle flowers, chrysanthemum flowers, raw rehmannia roots, liquorice, Imperata roots, mulberry leaves, Sophora japonica seeds, Prunella vulgaris and Pueraria lobata.

[0008] Preferably, the auxiliary material includes any one of dextrin, soluble starch and mannitol.

[0009] Preferably, the raw medicinal materials are weighed according to the prescription of clearing heat, detoxifying and removing dampness, and the medicinal materials are heated and refluxed for extraction 3 times, and 8 times the amount of water is added each time, and the extraction is performed for 1 hour and filtered, the filtrate is combined, and concentrated at 80°C under normal pressure to a relative density of 1.28-1.32, vacuum dried to a dry extract and crushed into a fine powder that can pass through an 80-mesh sieve, and then mannitol, an auxiliary material, is added in the same proportion as the fine powder, mixed evenly, and 70% by mass ethanol is added to prepare a soft material, which is squeezed through a 12-mesh sieve to prepare wet granules, dried at 60°C for 1.5 hours, and granulated to obtain the product.

[0010] Preferably, the raw medicinal materials of the sugar-free heat-clearing, detoxifying and dehumidifying granules include 35-40 parts of kapok flowers, 1-5 parts of honeysuckle, 20-25 parts of chrysanthemums, 5-10 parts of rehmannia roots, 1-5 parts of licorice, 5-10 parts of Imperata roots, 1-5 parts of mulberry leaves, 10-15 parts of sophora japonica seeds, 10-15 parts of selfheal and 5-10 parts of kudzu vines.

[0011] The present invention also provides evaluation indicators for the above-mentioned sugar-free heat-clearing, detoxifying and dehumidifying granules, including: using an orthogonal test method, taking rutin transfer rate and dry paste rate as evaluation indicators, to screen the extraction process of the sugar-free heat-clearing, detoxifying and dehumidifying granules; using a single factor analysis method, taking molding rate, solubility and hygroscopicity as evaluation indicators, to screen the molding process parameters of the sugar-free heat-clearing, detoxifying and dehumidifying granules.

[0012] Preferably, the orthogonal test method is used to take the comprehensive index of rutin transfer rate and dry extract rate as the investigation index, and the amount of water added (A), extraction time (B), and number of extractions (C) are selected as the investigation factors. 4 )Orthogonal experimental design is used to arrange the levels of experimental factors.

[0013] Preferably, the rutin transfer rate detection method is as follows:

[0014] Chromatographic conditions: Kromasil 100-5-C18 (250×4.6 mm, 5 μm) was used as the chromatographic column, 0.1% phosphoric acid aqueous solution was used as the mobile phase A, acetonitrile was used as the mobile phase B, gradient elution, flow rate was 1.0 mL / min, column temperature was 25°C, detection wavelength was 358 nm, and injection was 10 μL;

[0015] Preparation of reference solution: Accurately weigh 1.00 mg of rutin reference substance, dissolve it in methanol, and dilute it to a 10 ml volumetric flask to obtain a 0.10 mg / mL rutin reference solution.

[0016] Preparation of test solution: weigh the single medicinal material according to the amount of medicinal material in the prescription for clearing heat, detoxifying and removing dampness, add 8 times the amount of water to extract twice, each time for 1.5 hours, combine the extracts, concentrate to 250mL at normal pressure, and the crude drug content is 0.468g / mL ， Accurately pipette 1 mL of the extract into a 25 mL volumetric flask, dilute to the mark with methanol, shake well, centrifuge at 4000 r / min for 5 min, and filter the supernatant through a 0.22 μm filter membrane to obtain the final product.

[0017] Preparation of negative control solution lacking Sophora japonica seeds: weigh out the remaining single herbs except Sophora japonica seeds according to 1 / 5 of the amount of herbs in the prescription for clearing away heat, detoxifying and removing dampness, and then prepare a negative control solution according to the preparation method of the test solution, and set aside;

[0018] Detection method: Take the test sample, reference sample, and negative control sample solution lacking Sophora japonica seeds for measurement, and inject them into liquid chromatography to obtain the result.

[0019] Preferably, the dry paste rate detection method is as follows:

[0020] Preparation of concentrated extract: weigh the single medicinal material according to the amount of medicinal material in the prescription for clearing heat, detoxifying and removing dampness, add 8 times the amount of water to extract twice, each time for 1.5 hours, combine the extracts, concentrate to 250mL at normal pressure, and the crude drug content is 0.468g / mL;

[0021] Detection method: Accurately pipette 25 mL of concentrated extract into an evaporating dish with constant weight, evaporate to dryness in a water bath, transfer to a vacuum drying oven at 105°C and dry to constant weight, cool in a drying oven for 30 minutes, then weigh and calculate the dry cream rate. The formula is as follows: Dry cream rate (%) = (WV / 25×W t )×100%;

[0022] W is the weight of dry extract in 25 mL of concentrate, V is the fixed volume, W t The weight of medicinal materials.

[0023] Preferably, the forming rate is the total weight of the weighed and dried particles, and the particles that can pass through the No. 1 sieve but not through the No. 5 sieve are screened as qualified particles, and the forming rate is calculated by weighing, forming rate (%) = qualified particle mass / total particle mass × 100%.

[0024] Preferably, the solubility is as follows: take 2.50 g of qualified particles, add 50 mL of hot water, stir for 5 min, centrifuge the solution at 3000 r / min for 3 min, discard the supernatant, vacuum dry the bottom precipitate at 105°C to a constant mass, weigh and calculate the solubility rate, solubility rate = (particle mass - precipitate mass) / particle mass.

[0025] Preferably, the hygroscopicity is prepared by placing a desiccator with a supersaturated ammonium chloride solution at the bottom at 25°C for 24 hours to create a high humidity environment, accurately weighing 2.00 g of sample particles, spreading them in a weighing bottle of constant mass, placing them in a high humidity desiccator, and absorbing moisture at a constant temperature (T=25°C) for 24 hours, weighing the weight of the particles after moisture absorption, and moisture absorption rate=(mass of particles after moisture absorption-mass of sampled particles) / mass of sampled particles.

[0026] The beneficial effects produced by the technical solution of the present invention are as follows:

[0027] (1) This application uses rutin transfer rate and dry extract rate as the evaluation indicators of the extraction process, and adopts L9(3 4 ) The optimal water extraction process of the sugar-free Qingrejiedu Qushi granules was selected by orthogonal experiment, and an HPLC method for determining the rutin content in Qingrejiedu Qushi granules was established. The optimal extraction process was: adding 8 times the amount of water, heating reflux extraction 3 times, each extraction for 1 hour. After multiple verifications, it was proved that the process has good repeatability, stability and reliability, and can be used as the extraction process of the sugar-free Qingrejiedu Qushi granules.

[0028] (2) This application selected soluble starch, dextrin, and mannitol as auxiliary materials, and used particle size, solubility, and molding rate as comprehensive indicators combined with moisture absorption rate to investigate the molding process of sugar-free heat-clearing, detoxifying, and dehumidifying granules, and determined that 70% ethanol was the best for preparing soft materials. Subsequently, the three composite ratios were used to screen granule auxiliary materials. The final comprehensive results showed that the sugar-free granules prepared by mannitol alone as an auxiliary material had the best comprehensive performance, a high molding rate, good solubility, uniform particle size, and low moisture absorption rate, which can improve the moisture absorption and easy agglomeration of traditional Chinese medicine preparations and achieve the effect of rapid release. In addition, mannitol tastes slightly sweet, and can also regulate the bitterness of traditional Chinese medicine to a certain extent, increasing patient compliance. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0030] Figure 1 This is the HPLC chromatogram of the rutin reference solution in Experiment 3.1 of this application;

[0031] Figure 2 The HPLC chromatogram of the negative reference solution lacking Sophora japonica seeds in Experiment 3.1 of this application;

[0032] Figure 3 This is the HPLC chromatogram of the test solution in Experiment 3.1 of this application;

[0033] Figure 4 This is a linear relationship diagram of the peak areas of rutin reference solution at 6 concentrations in Experiment 3.2 of this application. DETAILED DESCRIPTION

[0034] In order to make the purpose, technical scheme and advantages of the embodiments of the present invention clearer, the technical scheme in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are part of the embodiments of the present invention, rather than all the embodiments. Therefore, the following detailed description of the embodiments provided by the present invention is not intended to limit the scope of the claimed invention, but merely represents the selected embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0035] The present invention is further described in detail below in conjunction with specific embodiments:

[0036] Example 1

[0037] The invention discloses a granulation process of sugar-free heat-clearing, detoxifying and dehumidifying granules. The raw medicinal materials are weighed according to a heat-clearing, detoxifying and dehumidifying prescription, and the medicinal materials are heated and refluxed for extraction once, and 6 times the amount of water is added each time, and the extraction is performed for 1 hour and filtered respectively, and the filtrate is combined, concentrated at 80°C under normal pressure to a relative density of 1.28-1.32, vacuum dried to obtain a dry extract, and crushed into fine powder that can pass through a No. 5 sieve, and then auxiliary materials with the same proportion as the fine powder are added, mixed evenly, 60% ethanol is added to prepare a soft material, and wet granules are prepared by extrusion through a 12-mesh sieve, and dried at 60°C for 1.5 hours, and granulated to obtain the sugar-free heat-clearing, detoxifying and dehumidifying granules.

[0038] Example 2

[0039] The invention discloses a granulation process of sugar-free heat-clearing, detoxifying and dehumidifying granules. The raw medicinal materials are weighed according to a heat-clearing, detoxifying and dehumidifying prescription, and the medicinal materials are heated and refluxed for extraction twice, and 8 times of water is added each time, and the extraction is respectively performed for 1 hour and filtered, and the filtrate is combined, concentrated at 80°C under normal pressure to a relative density of 1.28-1.32, vacuum dried to obtain a dry extract, and crushed into fine powder that can pass through a No. 5 sieve, and then auxiliary materials with the same proportion as the fine powder are added, mixed evenly, 60% ethanol is added to prepare a soft material, and wet granules are prepared by extrusion through a 12-mesh sieve, and dried at 60°C for 1.5 hours, and granulated to obtain the sugar-free heat-clearing, detoxifying and dehumidifying granules.

[0040] Example 3

[0041] The invention discloses a granulation process of sugar-free heat-clearing, detoxifying and dehumidifying granules. The raw medicinal materials are weighed according to a heat-clearing, detoxifying and dehumidifying prescription, and the medicinal materials are heated and refluxed for extraction for 3 times, and 8 times of water is added each time, and the extraction is performed for 1 hour and filtered respectively, and the filtrate is combined, concentrated at 80 DEG C and normal pressure to a relative density of 1.28-1.32, vacuum dried to obtain a dry extract, and crushed into fine powder that can pass through a No. 5 sieve, and then auxiliary materials with the same proportion as the fine powder are added, mixed evenly, 60% ethanol is added to prepare a soft material, and wet granules are prepared by extrusion through a 12-mesh sieve, and dried at 60 DEG C for 1.5 hours, and granulated to obtain the sugar-free heat-clearing, detoxifying and dehumidifying granules.

[0042] Example 4

[0043] The invention discloses a granulation process of sugar-free heat-clearing, detoxifying and dehumidifying granules. The raw medicinal materials are weighed according to a heat-clearing, detoxifying and dehumidifying prescription, and the medicinal materials are heated and refluxed for extraction for 3 times, and 10 times the amount of water is added each time, and the extraction is performed for 2 hours and filtered respectively, and the filtrate is combined, concentrated at 80°C under normal pressure to a relative density of 1.28-1.32, vacuum dried to obtain a dry extract, and crushed into fine powder that can pass through a No. 5 sieve, and then auxiliary materials with the same proportion as the fine powder are added, mixed evenly, 60% ethanol is added to prepare a soft material, and wet granules are prepared by extrusion through a 12-mesh sieve, and dried at 60°C for 1.5 hours, and granulated to obtain the sugar-free heat-clearing, detoxifying and dehumidifying granules.

[0044] This application uses rutin transfer rate and dry paste rate as evaluation indicators, and orthogonal test method as a method to screen the best extraction process of sugar-free heat-clearing, detoxifying and dehumidifying granules. It also uses single factor analysis method to screen the best molding process parameters with molding rate, solubility and hygroscopicity as evaluation indicators, and successfully prepares sugar-free heat-clearing, detoxifying and dehumidifying granules. It provides more data support for the subsequent development of our hospital's sugar-free heat-clearing, detoxifying and dehumidifying prescription and promotes the healthy development of traditional Chinese medicine preparations. The following experiments were conducted:

[0045] 1. Materials and instruments

[0046] 1.1 Instrument

[0047] Agilent 1260 InfinityⅡ high performance liquid chromatograph (Agilent Technologies Co., Ltd.); XA-105 electronic balance (Mettler Toledo, Switzerland, 1 / 100,000); XPR36 electronic balance (Mettler Toledo, Switzerland, 1 / 1000); LS2200C SCS electronic balance (Shanghai Tianmei Balance Instrument Co., Ltd., 1 / 10000); electric blast drying oven (Shanghai Yiheng Scientific Instrument Co., Ltd.), electric heating jacket (Tianjin Test Instrument Co., Ltd.); SHZ-D(Ⅲ) circulating water vacuum pump (Gongyi Yuhua Instrument Co., Ltd.).

[0048] 1.2 Medicinal materials and reagents

[0049] The experimental samples included kapok (batch number: 2307001, origin in Guangdong), honeysuckle (batch number: 2307001, origin in Henan), chrysanthemum (batch number: 2307001, origin in Zhejiang), raw rehmannia root (batch number: 2307001, origin in Henan), licorice (batch number: 2307001, origin in Gansu), Imperata root (batch number: 2307001, origin in Guangdong), mulberry leaf (batch number: 2307001, origin in Guangdong), and chrysanthemum root (batch number: 2307001, origin in Zhejiang). 1, origin Guangxi), Sophora japonica (batch number: 2307001, origin Jiangsu), Prunella vulgaris (batch number: 2307001, origin Hubei), Pueraria lobata (2306001, origin Guangxi) were purchased from Lingnan Chinese Medicine Pieces Co., Ltd. and verified by Huang Yuechun, chief pharmacist of the First Affiliated Hospital of Guangzhou University of Chinese Medicine, and all met the relevant provisions of the 2020 edition of the "Chinese Pharmacopoeia (Part I)" under the items of various medicinal materials and pieces. Rutin reference substance (batch number 10080-202012, content 92.2%) was purchased from China Food and Drug Inspection Institute. Dextrin (Dongguan Dongyue Glucose Factory Co., Ltd.), soluble starch (Xiangtan County Starch Products Co., Ltd.), mannitol (Qingdao Mingyue Seaweed Group Co., Ltd.), methanol (chromatographic grade, Merck, Germany), acetonitrile (chromatographic grade, Merck, Germany), ammonium chloride (analytical grade, Tianjin Damao Chemical Reagent Factory), phosphoric acid (analytical grade, Tianjin Fuyu Fine Chemical Co., Ltd.), and water was double distilled water.

[0050] 2. Methods

[0051] 2.1 Chromatographic conditions

[0052] Chromatographic column: Kromasil 100-5-C18 (250×4.6 mm, 5 μm); mobile phase A is 0.1% phosphoric acid aqueous solution, phase B is acetonitrile, gradient elution (see Table 1); flow rate 1.0 mL / min; column temperature: 25°C; detection wavelength 358 nm; injection volume 10 μL.

[0053] Table 1 Gradient elution program

[0054]

[0055] 2.2 Preparation of reference solution

[0056] Accurately weigh 1.00 mg of rutin reference substance, dissolve it in methanol, and dilute it to volume in a 10 mL volumetric flask to obtain a rutin reference substance solution with a content of 0.10 mg / mL.

[0057] 2.3 Preparation of test solution

[0058] According to the amount of medicinal materials in the prescription for clearing heat, detoxifying and removing dampness, weigh the single medicinal materials such as kapok, chrysanthemum, and sophora japonica, add 8 times the amount of water, extract twice for 1.5 hours, combine the extracts, and concentrate to 250mL at normal pressure. Accurately pipette 1mL of the extract into a 25mL volumetric flask, add methanol to dilute to the scale, shake well, centrifuge at 4000r / min for 5min, and filter the supernatant through a 0.22μm filter membrane to obtain the product.

[0059] 2.4 Preparation of negative control solution lacking Sophora japonica

[0060] According to 1 / 5 of the amount of medicinal materials in the prescription for clearing heat, detoxifying and removing dampness, weigh out the remaining single medicinal materials except Sophora japonica seeds, and then prepare the negative control solution according to the method under 2.3 and set aside.

[0061] 2.5 Determination of dry paste rate

[0062] Accurately pipette 25 mL of concentrated extract into an evaporating dish with constant weight, evaporate to dryness in a water bath, transfer to a vacuum drying oven at 105°C and dry to constant weight, cool in a drying oven for 30 minutes, then weigh and calculate the dry paste rate using the following formula:

[0063] Dry paste rate (%) = (WV / 25×W t )×100%

[0064] W is the weight of dry extract in 25 mL of concentrate, V is the fixed volume, W t The weight of medicinal materials.

[0065] 3. Methodological investigation and content determination

[0066] 3.1 Specificity experiment

[0067] Take 10 μL of each of the test sample, reference sample, and negative control sample solution lacking Sophora japonica seeds and inject them for determination. Figures 1 to 3 As shown, at the corresponding retention time position of the rutin reference substance, the test solution has the same absorption peak, and the negative reference solution lacking Sophora japonica has no obvious absorption peak, indicating that the negative has no interference and the method has good specificity.

[0068] 3.2 Linear Relationship

[0069] Take an appropriate amount of rutin reference substance and prepare 0.422 mg / mL and 0.310 mg / mL rutin reference substance solutions. Take the 0.422 mg / mL rutin reference substance solution and dilute it in multiple proportions to 0.211 mg / mL, 0.106 mg / mL, 0.053 mg / mL, and 0.026 mg / mL rutin standard solution. Inject and measure the chromatograms of the above 6 concentrations of rutin reference substance solutions according to the chromatographic conditions under 2.1. Figure 4 As shown in the figure, the standard curve is drawn with the peak area as the ordinate and the standard concentration as the abscissa, and the regression equation is obtained: y = 14886x-22.325, R 2 =1, indicating a good linear relationship in the range of 0.026-0.422 mg / mL.

[0070] 3.3 Precision test

[0071] Take one portion of the rutin reference solution under Item 2.2, and inject it six times continuously according to the chromatographic conditions under Item 2.1. Record the chromatogram and calculate the RSD value of the rutin peak area to be 0.17% (see Table 2), indicating that the instrument has good precision.

[0072] Table 2 Precision test

[0073]

[0074]

[0075] 3.4 Repeatability test

[0076] Six samples were prepared according to the method in 2.3 and measured by chromatographic strip injection in 2.1. The RSD value of the rutin peak area was 1.3% (see Table 3), indicating that the method had good repeatability.

[0077] Table 3 Repeatability test

[0078]

[0079] 3.5 Stability test

[0080] The test solution under 2.3 was sampled at 0, 2, 4, 8, 12, and 24 hours, and the peak area of ​​rutin was determined and the RSD was calculated. The results showed that the RSD value of the peak area of ​​rutin was 1.2% (see Table 4), indicating that the test solution had good stability within 24 hours.

[0081] Table 4 Stability test

[0082]

[0083] 3.6 Sample recovery test

[0084] Accurately draw 6 portions of extract with known content, add rutin reference substance approximately 1:1 according to the rutin content of the extract, prepare the sample solution according to the method under 2.3, inject according to the chromatographic conditions under 2.1, measure the absorbance value and calculate the rutin content according to the regression equation to obtain the sample recovery. The average sample recovery of rutin is 103.39%, and the RSD value is 2.47% (see Table 5), indicating that the method has good accuracy.

[0085] Table 5 Sample recovery test

[0086]

[0087] 3.7 Orthogonal test

[0088] The orthogonal test method was used to take the comprehensive index of rutin transfer rate and dry extract rate as the investigation index, and the amount of water added (A), extraction time (B), and extraction times (C) were selected as the investigation factors. 4 ) Orthogonal experimental design was used to arrange the experimental factor levels (see Table 6). The test solution was prepared using the method in 2.2, and the rutin content was determined using the chromatographic conditions in 2.1.

[0089] Table 6 Factor level table

[0090]

[0091] Using the comprehensive weighted scoring method, the weighted coefficient of rutin transfer rate was set to 0.6, and the weighted coefficient of dry paste rate was set to 0.4. The results of the orthogonal test are shown in Table 7. The influence of the three factors on the extraction process is C>A>B, that is, the number of extractions has the greatest impact on the test results, followed by the extraction time, and finally the amount of water added. The orthogonal analysis results in Table 8 show that only the results of the number of extractions among the three factors are statistically significant (P<0.05). Therefore, considering the production economy and time cost, the optimal process was finally determined to be 3 extractions, each for 1 hour, and 8 times the amount of water added.

[0092] Table 7 Orthogonal experimental design and results

[0093]

[0094] Note: Comprehensive score = [rutin transfer rate / maximum amount × 0.6 + dry paste rate / maximum amount × 0.4] × 100%.

[0095] Table 8 Results of variance analysis

[0096]

[0097] 3.8 Verification test

[0098] According to the prescription amount ratio, three portions of each single medicinal material such as Sophora japonica, Kapok, and Chrysanthemum were weighed, and extracted according to the above-selected best extraction process (8 times the amount of water, 3 extractions, 1 hour each time), and the average transfer rate of rutin was measured to be 48.55%, RSD was 1.3% (n = 3); the average dry extract rate was 32.74%, RSD was 0.69% (n = 3). The results show that the extraction efficiency of the preferred process is high and the results are stable, and the extraction method is feasible (see Table 9).

[0099] Table 9 Verification test results

[0100]

[0101] 4. Molding process research

[0102] 4.1 Screening of wetting agents and test conditions

[0103] In order to preliminarily screen the experimental conditions for the preparation of heat-clearing, detoxifying and dehumidifying granules, 70% ethanol was used as a wetting agent, the drug-excipient ratio was 1:1, the wet granules were dried at 60°C, and dried for 1.5 h. Dextrin was used as an excipient, and the difficulty of preparing granules with different types of wetting agents was preliminarily investigated. The results are shown in Table 10. The results show that the molding rate of 70% ethanol as a wetting agent is slightly lower than that of 60% ethanol, but the soft material is not easy to stick to the screen during the preparation process, the material loss is less, and the particle size is relatively uniform. Considering all factors, 70% ethanol was used as a wetting agent.

[0104] Table 10 Preparation of granules with different wetting agents

[0105]

[0106] 4.2 Preparation of particles

[0107] Weigh the prescribed amount of kapok, sophora japonica seeds, chrysanthemum and other medicinal materials, decoct according to the above-mentioned optimal extraction process, concentrate the decoction to a relative density of 1.28-1.32 (80°C), vacuum dry, grind the dry extract into fine powder that can pass through a No. 5 sieve, weigh the medicinal powder: auxiliary materials (1:1) and mix them evenly, add 70% ethanol to prepare a soft material, extrude through a 12-mesh sieve to prepare wet granules, dry at 60°C for 1.5 hours, and granulate to obtain the product.

[0108] 4.3 Screening of excipients for granules

[0109] Using dextrin, soluble starch and mannitol as auxiliary materials, granules were prepared according to the method under 4.2. The comprehensive score of appearance, forming rate and solubility was used as the evaluation index, and the weight coefficients were 30%, 40% and 30% respectively. The optimal auxiliary materials for preparing sugar-free heat-clearing, detoxifying and dehumidifying granules were comprehensively selected in combination with the anti-hygroscopicity of the granules.

[0110] 4.4 Evaluation index of particles

[0111] 4.4.1 Appearance

[0112] The appearance of the particles was evaluated using particle uniformity and color as evaluation indicators. The scoring criteria are shown in Table 11.

[0113] Table 11 Particle evaluation standard table

[0114]

[0115] 4.4.2 Qualification Rate

[0116] The total weight of the dried particles was weighed, and the particles that passed through the No. 1 sieve but not through the No. 5 sieve were screened as qualified particles. The molding rate was calculated by weighing, and the molding rate (%) = qualified particle mass / total particle mass × 100%.

[0117] 4.4.3 Solubility

[0118] Take 2.50 g of qualified particles, add 50 mL of hot water, stir for 5 min, centrifuge the solution at 3000 r / min for 3 min, discard the supernatant, vacuum dry the bottom precipitate at 105°C to a constant mass, weigh it and calculate the dissolution rate, dissolution rate = (particle mass - precipitate mass) / particle mass.

[0119] 4.4.4 Hygroscopicity

[0120] Place a desiccator with a supersaturated ammonium chloride solution at the bottom at 25°C for 24 hours to create a high humidity environment, accurately weigh 2.00g of sample particles, spread them flat in a weighing bottle with a constant mass, put them in a high humidity desiccator, and absorb moisture at a constant temperature (T = 25°C) for 24 hours, and weigh the weight of the particles after moisture absorption. Moisture absorption rate = (mass of particles after moisture absorption - mass of sampled particles) / mass of sampled particles.

[0121] 4.5 Results and Analysis

[0122] The comprehensive scores of appearance, forming rate and solubility were used as evaluation indicators, and the best excipients were selected in combination with the anti-hygroscopic properties of the particles. The results in Table 12 showed that the mannitol group had the highest comprehensive score, and its hygroscopic rate was lower than that of the other groups, with good anti-hygroscopic properties; followed by the dextrin group, which also had a high comprehensive score, but its anti-hygroscopic properties were relatively low. The comprehensive results showed that the sugar-free heat-clearing, detoxifying and dehumidifying granules prepared with mannitol as an excipient had the characteristics of uniform particle size, high forming rate, good solubility and stable anti-hygroscopic quality, and were the best excipients for preparing sugar-free heat-clearing, detoxifying and dehumidifying granules.

[0123] Table 12 Screening results of excipients for granules

[0124]

[0125] Note: Comprehensive score = [Appearance score / maximum amount × 0.3 + Forming rate / maximum amount × 0.4 + Solubility / maximum amount × 0.3] × 100%.

[0126] 5. Discussion

[0127] 5.1 Extraction process investigation

[0128] In this experiment, the rutin transfer rate and dry extract rate were used as the evaluation indicators of the extraction process. 4 ) The orthogonal test optimized the optimal water extraction process of sugar-free Qingrejiedu Qushi granules, and established an HPLC method for determining the content of rutin in Qingrejiedu Qushi granules. The optimal extraction process was: add 8 times the amount of water, heat reflux extraction 3 times, each extraction 1h. Three batches of samples were prepared again, and the process was verified to have good repeatability, stability and reliability, and can be used as the extraction process of sugar-free Qingrejiedu Qushi granules. The established HPLC method is fast, simple and convenient, and can be used for the follow-up study of sugar-free Qingrejiedu Qushi granules.

[0129] 5.2 Molding process investigation

[0130] Due to the diverse flavors and complex ingredients of traditional Chinese medicine preparations, the stability and safety of excipients should be considered in the selection of excipients. Common fillers for traditional Chinese medicine preparations include sucrose, dextrin, soluble starch, mannitol and lactose. However, lactose may cause gastrointestinal allergic reactions in some patients with intolerance. Therefore, soluble starch, dextrin and mannitol were finally selected as excipients for research in this experiment. Formability is an important factor in evaluating the yield and cost of particles. The weight coefficient is set to 40%, solubility and particle size are 30%. In addition, hygroscopicity has a great influence on the quality stability of particles. Therefore, this experiment uses soluble starch, dextrin and mannitol as excipients, and uses particle size, solubility and forming rate as comprehensive indicators combined with hygroscopicity to investigate the forming process of sugar-free heat-clearing, detoxifying and dehumidifying particles. During the investigation, it was found that the proportion of wetting agent has a great influence on the forming property of particles, so it was preliminarily screened. It was determined that 70% ethanol has the best effect in preparing soft materials. 70% ethanol was used as a wetting agent to further screen the best excipients for the granules. Mannitol, dextrin and soluble starch were used as excipients, and the three were compounded in a ratio to screen the excipients for the granules. The final comprehensive results showed that the sugar-free granules prepared with mannitol alone as an excipient had the best comprehensive performance, a high molding rate, good solubility, uniform particle size, and low moisture absorption rate, which could improve the moisture absorption and easy agglomeration of traditional Chinese medicine preparations and achieve the effect of rapid release; in addition, mannitol tasted slightly sweet, and could also adjust the bitterness of traditional Chinese medicine to a certain extent, and increase patient compliance. Therefore, mannitol was the best molding excipient for sugar-free heat-clearing, detoxifying and dehumidifying granules. The final molding process was to mix mannitol and dry paste powder in a ratio of 1:1, use 70% ethanol as a wetting agent, extrude through a 12-mesh sieve to prepare wet granules, and dry at 60℃ for 1.5h to prepare sugar-free heat-clearing, detoxifying and dehumidifying granules, which had high comprehensive indicators and were stable and reliable.

[0131] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A granulation process for a sugar-free heat-clearing, detoxifying and dehumidifying granule, characterized in that: According to the prescription of clearing away heat, detoxifying and removing dampness, the raw medicinal materials are weighed, and the medicinal materials are heated and refluxed for extraction 1 to 3 times, and 6 to 8 times the amount of water is added each time, and the extraction is performed for 1 to 2 hours respectively and filtered, and the filtrate is combined, concentrated at 80°C under normal pressure to a relative density of 1.28 to 1.32, vacuum dried to a dry extract, and crushed into a fine powder that can pass through an 80-mesh sieve, and then auxiliary materials with the same proportion as the fine powder are added, mixed evenly, and 60 to 80% by mass of ethanol are added to prepare a soft material, and wet granules are prepared by extrusion through a 12-mesh sieve, and dried at 60°C for 1.5 hours, and granulated to obtain; The raw medicinal materials of the sugar-free heat-clearing, detoxifying and dehumidifying granules include kapok, honeysuckle, chrysanthemum, raw rehmannia, liquorice, Imperata root, mulberry leaf, Sophora japonica seed, Prunella vulgaris and Pueraria lobata.

2. The granulation process of the sugar-free heat-clearing, detoxifying and dehumidifying granules according to claim 1 is characterized in that: The auxiliary material includes any one of dextrin, soluble starch and mannitol.

3. The granulation process of the sugar-free heat-clearing, detoxifying and dehumidifying granules according to claim 2 is characterized in that: Weigh the raw medicinal materials according to the prescription of clearing heat, detoxifying and removing dampness, heat the medicinal materials under reflux extraction for 3 times, add 8 times the amount of water each time, extract for 1 hour respectively and filter, combine the filtrate, concentrate at 80°C under normal pressure to a relative density of 1.28-1.32, vacuum dry to become a dry extract and grind into fine powder that can pass through an 80-mesh sieve, then add mannitol as an auxiliary material in the same proportion as the fine powder, mix well, add 70% by mass ethanol to prepare a soft material, extrude through a 12-mesh sieve to prepare wet granules, dry at 60°C for 1.5 hours, and granulate to obtain.

4. The granulation process of the sugar-free heat-clearing, detoxifying and dehumidifying granules according to claim 1 is characterized in that: The raw medicinal materials of the sugar-free heat-clearing, detoxifying and dampness-removing granules include 35-40 parts of kapok flowers, 1-5 parts of honeysuckle flowers, 20-25 parts of chrysanthemums, 5-10 parts of raw rehmannia roots, 1-5 parts of licorice, 5-10 parts of Imperata roots, 1-5 parts of mulberry leaves, 10-15 parts of Sophora japonica seeds, 10-15 parts of Prunella vulgaris and 5-10 parts of Pueraria lobata.

5. An evaluation index for the sugar-free heat-clearing, detoxifying and dehumidifying granules according to any one of claims 1 to 4, characterized in that: include: Orthogonal test method was used to screen the extraction process of sugar-free Qingrejieduqushi granules with rutin transfer rate and dry extract rate as evaluation indicators. The single factor analysis method was used to screen the molding process parameters of the sugar-free heat-clearing, detoxifying and dehumidifying granules with molding rate, solubility and hygroscopicity as evaluation indicators.

6. The evaluation index of the sugar-free heat-clearing, detoxifying and dehumidifying granules according to claim 5 is characterized in that: The orthogonal test method was used to take the comprehensive index of rutin transfer rate and dry extract rate as the investigation index, and the amount of water added (A), extraction time (B), and extraction times (C) were selected as the investigation factors. 4 )Orthogonal experimental design is used to arrange the levels of experimental factors.

7. The evaluation index of the sugar-free heat-clearing, detoxifying and dehumidifying granules according to claim 6 is characterized in that: The rutin transfer rate detection method is as follows: Chromatographic conditions: Kromasil 100-5-C18 (250×4.6 mm, 5 μm) was used as the chromatographic column, 0.1% phosphoric acid aqueous solution was used as the mobile phase A, acetonitrile was used as the mobile phase B, gradient elution, flow rate was 1.0 mL / min, column temperature was 25°C, detection wavelength was 358 nm, and injection was 10 μL; Preparation of reference solution: Accurately weigh 1.00 mg of rutin reference substance, dissolve it in methanol, and dilute it to a 10 ml volumetric flask to obtain a 0.10 mg / mL rutin reference solution. Preparation of test solution: weigh the single medicinal material according to the amount of medicinal material in the prescription for clearing heat, detoxifying and removing dampness, add 8 times the amount of water to extract twice, each time for 1.5 hours, combine the extracts, concentrate to 250mL at normal pressure, and the crude drug content is 0.468g / mL ， Accurately pipette 1 mL of the extract into a 25 mL volumetric flask, dilute to the mark with methanol, shake well, centrifuge at 4000 r / min for 5 min, and filter the supernatant through a 0.22 μm filter membrane to obtain the final product. Preparation of negative control solution lacking Sophora japonica seeds: weigh out the remaining single herbs except Sophora japonica seeds according to 1 / 5 of the amount of herbs in the prescription for clearing away heat, detoxifying and removing dampness, and then prepare a negative control solution according to the preparation method of the test solution, and set aside; Detection method: Take the test sample, reference sample, and negative control sample solution lacking Sophora japonica seeds for measurement, and inject them into liquid chromatography to obtain the result.

8. The evaluation index of the sugar-free heat-clearing, detoxifying and dehumidifying granules according to claim 6 is characterized in that: The dry paste rate detection method is as follows: Preparation of concentrated extract: weigh the single medicinal material according to the amount of medicinal material in the prescription for clearing heat, detoxifying and removing dampness, add 8 times the amount of water to extract twice, each time for 1.5 hours, combine the extracts, concentrate to 250mL at normal pressure, and the crude drug content is 0.468g / mL; Detection method: Accurately pipette 25 mL of concentrated extract into an evaporating dish with constant weight, evaporate to dryness in a water bath, transfer to a vacuum drying oven at 105°C and dry to constant weight, cool in a drying oven for 30 minutes, then weigh and calculate the dry cream rate. The formula is as follows: Dry cream rate (%) = (WV / 25×W t )×100%; W is the weight of dry extract in 25 mL of concentrate, V is the fixed volume, W t The weight of medicinal materials.

9. The evaluation index of the sugar-free heat-clearing, detoxifying and dehumidifying granules according to claim 5 is characterized in that: The forming rate is the total weight of the weighed and dried particles, and the particles that can pass through the No. 1 sieve but not through the No. 5 sieve are qualified particles. The forming rate is calculated by weighing, and the forming rate (%) = qualified particle mass / total particle mass × 100%.

10. The evaluation index of the sugar-free heat-clearing, detoxifying and dehumidifying granules according to claim 5 is characterized in that: The solubility is as follows: take 2.50 g of qualified particles, add 50 mL of hot water, stir for 5 min, centrifuge the solution at 3000 r / min for 3 min, discard the supernatant, vacuum dry the bottom precipitate at 105°C to a constant mass, weigh it and calculate the solubility rate, solubility rate = (particle mass - precipitate mass) / particle mass; The hygroscopicity is prepared by placing a desiccator with a supersaturated ammonium chloride solution at the bottom at 25°C for 24 hours to create a high humidity environment, accurately weighing 2.00 g of sample particles, spreading them in a weighing bottle of constant mass, placing them in a high humidity desiccator, and absorbing moisture at a constant temperature (T = 25°C) for 24 hours, weighing the weight of the particles after moisture absorption, and moisture absorption rate = (mass of particles after moisture absorption - mass of sampled particles) / mass of sampled particles.

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