阅读说明：本技术 白虎汤颗粒的制备工艺及质量标准研究 (Preparation process and quality standard research of Baihu decoction granules ) 是由 闫雪生 于蓓蓓 王平 刘瑾 孙丹丹 生立嵩 李富欣 王文静 杜立遥 于 2019-10-23 设计创作，主要内容包括：本发明属于中药制剂技术领域,具体涉及一种白虎汤颗粒的制备方法及质量标准的检测方法。提供了一种白虎汤颗粒的制备方法,以CaSO<Sub>4</Sub>·2H<Sub>2</Sub>0保留率、新芒果苷保留率、芒果苷保留率、除杂率为考查指标,确定了所述制备方法,该制备方法制备的白虎汤颗粒质量稳定,有效成分利用率高；本发明建立了白虎汤颗粒的质量控制方法,对制剂中的知母和炙甘草进行薄层色谱鉴别,对制剂中的知母和甘草进行含量测定,建立的方法操作简便,专属性强,结果准确,可用于白虎汤颗粒的质量控制。(The invention belongs to the technical field of traditional Chinese medicine preparations, and particularly relates to a preparation method of a Baihu decoction granule and a quality standard detection method. A method for preparing BAIHUTANG granule with CaSO is provided 4 ·2H 2 0 retention rate, new mangiferin retention rate, mangiferin retention rate and impurity removal rate are considered indexes, the preparation method is determined, the quality of the prepared white tiger soup particles is stable, and the utilization rate of active ingredients is high; the invention establishes a quality control method of the Baihu decoction granules, carries out thin-layer chromatography identification on rhizoma anemarrhenae and radix glycyrrhizae preparata in the preparation, carries out content measurement on the rhizoma anemarrhenae and the radix glycyrrhizae preparata in the preparation, has simple and convenient operation, strong specificity and accurate result, and can be used for the quality control of the Baihu decoction granules.)

1. The preparation method of the white tiger soup particles is characterized by comprising the following steps of:

(1) weighing gypsum, rhizoma anemarrhenae, radix Glycyrrhizae Preparata and semen oryzae Sativae according to the formula ratio, adding 8 times of water in the 1 st time, soaking for 0.5h, decocting, adding 6 times of water in the 2 nd and 3 rd times of decoction respectively, decocting for 0.5h in the three times, filtering with gauze, mixing the filtrates, and concentrating until the relative density is 1.02;

(2) adding a carboxymethyl chitosan solution into the concentrated filtrate obtained in the step (1) at 60 ℃, stirring for 20 min, standing for 12 h, centrifuging, filtering, and concentrating the filtrate to obtain a thick paste with the relative density of 1.10-1.12;

(3) adding adjuvant A, drying under reduced pressure, pulverizing into fine powder, adding adjuvant B, adding 88% ethanol to make soft mass, granulating, drying, grading, and packaging to obtain BAIHUTONG granule.

2. The preparation method according to claim 1, wherein the mass ratio of the gypsum, the rhizoma anemarrhenae, the radix glycyrrhizae preparata and the polished round-grained rice in the step (1) is 50: 18: 6: 9.

3. the preparation method according to claim 1, wherein the carboxymethyl chitosan solution of step (2) has a mass concentration of 0.5%; the volume ratio of the concentrated filtrate to the carboxymethyl chitosan solution is 1: 5.

4. The preparation method according to claim 1, wherein the auxiliary materials A and B in the step (3) constitute a total auxiliary material; the mass ratio of the thick paste in the step (2) to the total auxiliary materials is 1: 0.3; wherein the mass ratio of the auxiliary material A to the auxiliary material B in the step (3) is 2: 1; the ratio of dextrin to soluble starch in the auxiliary materials is 3: 2.

5. The method for detecting quality standards of BAIHUTANG granule as claimed in any one of claims 1-4, comprising: qualitatively identifying rhizoma anemarrhenae and radix Glycyrrhizae Preparata by TLC, and determining the content of rhizoma anemarrhenae and radix Glycyrrhizae by HPLC.

6. The detection method according to claim 5, wherein the qualitative identification of Anemarrhena asphodeloides and prepared licorice root is carried out by the following method:

A. weighing 2 g of BAIHU decoction, grinding, adding 10 mL of methanol, respectively, and performing ultrasonic treatment for 30 min to obtain supernatant as sample solution; precisely weighing mangiferin and new mangiferin reference substances respectively, and adding methanol to obtain solutions with mangiferin concentration of 0.25 mg/mL-1 and new mangiferin concentration of 0.23 mg/mL-1 as reference substance solutions; weighing 0.5 g of rhizoma anemarrhenae powder, and preparing into rhizoma anemarrhenae reference solution by the same method as the test solution; weighing other prescription medicinal materials except rhizoma anemarrhenae according to the proportion of the white tiger decoction particles, preparing rhizoma anemarrhenae deficiency negative particles according to the preparation process of the particles, and preparing rhizoma anemarrhenae deficiency negative control solution according to the preparation method of the test solution; performing test according to thin-layer chromatography (2015 edition of Chinese pharmacopoeia, Tonghe 0502), respectively sucking 5 μ L of mangiferin control solution, neomangiferin control solution, rhizoma anemarrhenae lacking negative control solution, and test solution, respectively dropping on the same polyamide film, developing with ethanol-water as developing agent at volume ratio of 1: 1, taking out, air drying, and inspecting under ultraviolet lamp 365 nm to obtain test sample, rhizoma anemarrhenae control solution chromatogram and control chromatogram at corresponding positions, wherein the fluorescent spots of the same color are displayed, and the rhizoma anemarrhenae lacking negative particles have no spots;

B. weighing 15 g of BAIHU decoction granule, adding diethyl ether 40 mL, heating and refluxing for 1 h, filtering, discarding ether solution, adding methanol 30 mL into the residue, heating and refluxing for 1 h, filtering, evaporating the filtrate to dryness, adding water 40 mL into the residue to dissolve, extracting with n-butanol for 3 times, 20 mL each time, mixing n-butanol solutions, washing with water for 3 times, discarding water solution, evaporating n-butanol solution to dryness, adding methanol 5 mL into the residue to dissolve, and using as sample solution; taking liquiritin reference substance, adding methanol to prepare solution containing 2 mg per 1mL as reference substance solution; weighing other prescription medicinal materials except Glycyrrhrizae radix according to the proportion of BAIHUTANG granule, preparing into negative granule without Glycyrrhrizae radix according to the preparation process of the granule, and preparing into negative control solution without Glycyrrhrizae radix according to the preparation method of the sample solution; performing thin-layer chromatography (2015 version of Chinese pharmacopoeia, general 0502 of the fourth ministry of pharmacopoeia of China), sucking 5 μ L of a licorice-lacking negative control solution, a liquiritin control solution and three batches of test solution, respectively dropping the solution on a same silica gel G thin-layer plate made of 1% sodium hydroxide solution, developing by taking ethyl acetate-formic acid-glacial acetic acid-water as a developing agent with the volume ratio of 15: 1: 2, taking out, drying in the air, uniformly spraying 10% sulfuric acid ethanol solution for color development, heating at 105 ℃ for 5-10 min, and inspecting under an ultraviolet lamp (365 nm), wherein fluorescent spots with the same color are formed on the positions corresponding to the color spectrums of the three batches of test solution and the control solution, and spots do not exist at the positions where the licorice-lacking negative particles.

7. The detection method according to claim 6, wherein the method A further comprises the following steps:

weighing 1 g of BAIHU decoction, grinding, adding 30% acetone 10 mL, ultrasonic treating for 30 min, and collecting supernatant as sample solution; accurately weighing timosaponin BII reference substance, adding 30% acetone to obtain solution with concentration of 0.56 mg/mL-1 as reference solution; weighing other prescription medicinal materials except rhizoma anemarrhenae according to the prescription proportion, preparing rhizoma anemarrhenae deficiency negative granules according to the preparation process of the granules, and preparing rhizoma anemarrhenae deficiency negative control solution according to the preparation method of the test solution; performing thin layer chromatography (2015 edition of Chinese pharmacopoeia, general rule of the fourth part 0502), respectively sucking 5 μ L of a control solution, a rhizoma anemarrhenae deficiency negative control solution and a test solution, respectively dropping on the same silica gel G thin layer plate, developing with n-butanol-glacial acetic acid-water as a developing agent at a volume ratio of 4: 1:5, taking out, air drying, spraying a 5% vanillin sulfuric acid ethanol test solution, heating at 105 ℃ until the color development of the spots is clear, and as a result, the spots with the same color appear on the corresponding positions of the chromatogram of the test solution and the control solution, and no spots exist in the positions of rhizoma anemarrhenae deficiency negative particles.

8. The detection method according to claim 5, wherein the HPLC method for detecting the content of rhizoma anemarrhenae and radix Glycyrrhizae comprises:

(1) chromatographic conditions are as follows: lichrospher C18 (5 μm, 4.6 mm. times.250 mm) high performance liquid chromatography column; mobile phase: acetonitrile A, 25 mmol.L-1 potassium dihydrogen phosphate B, gradient elution (0-2 min, 98 percent B, 2-8 min, 98-85 percent B, 8-15min, 85-78 percent B, 15-20 min, 78-72 percent B, 20-30 min, 72-50 percent B), column temperature of 30 ℃, flow rate of 1 mL.min-1, detection wavelength of 257 nm and sample injection amount of 10 mu L;

(2) preparation of mixed control solution: weighing 4 reference substances of neomangiferin, mangiferin, liquiritin and monoammonium glycyrrhizinate, dissolving with methanol, fixing the volume, and preparing into a mixed reference substance solution, wherein the concentration of neomangiferin is 26.22 mug.mL < -1 >, the concentration of mangiferin is 20.38 mug.mL < -1 >, the concentration of liquiritin is 26.46 mug.mL < -1 >, the concentration of monoammonium glycyrrhizinate is 30.67 mug.mL < -1 >, and filtering with a 0.45 mu m microporous membrane to obtain a subsequent filtrate;

(3) preparation of a test solution: grinding BAIHU decoction granule, collecting 1 g, adding 25 mL of methanol, weighing, performing ultrasonic treatment at 250W and 40KHz frequency for 30 min, cooling, weighing, supplementing the weight loss with methanol, shaking, filtering, and collecting the filtrate;

(4) preparation of negative control solution

Weighing other medicinal materials except rhizoma anemarrhenae and radix Glycyrrhizae Preparata, and preparing into negative control solution without rhizoma anemarrhenae and radix Glycyrrhizae Preparata according to the preparation method of BAIHU decoction granule and the preparation method of test solution, wherein the negative control solution has no color spectrum peak at the position corresponding to the color spectrum peak of the control solution, which indicates that Gypsum Fibrosum and semen oryzae Sativae in the prescription has no interference to measurement;

(5) respectively sucking 10 μ L of the mixed reference solution, the test solution and the negative reference solution, injecting into a high performance liquid chromatograph, and detecting at 257 nm according to the above chromatographic conditions, wherein the mixed reference solution and the test solution show absorption peaks in the same retention time, and no chromatographic peak appears in the negative reference of rhizoma anemarrhenae and radix Glycyrrhizae;

(6) the determination method comprises the following steps: respectively sucking 10 μ L of the mixed reference solution, and injecting into high performance liquid chromatograph, wherein each 1 g of the product contains rhizoma anemarrhenae based on the content of neomangiferin and mangiferin, the content of neomangiferin should not be less than 0.4 mg, and the content of mangiferin should not be less than 0.5 mg; the product contains Glycyrrhrizae radix in each 1 g, and calculated by content of glycyrrhizin and monoammonium glycyrrhizinate, the content of glycyrrhizin should not be less than 0.3 mg, and the content of monoammonium glycyrrhizinate should be 0.3 mg.

Technical Field

The invention belongs to the technical field of traditional Chinese medicine preparations, and particularly relates to a preparation method of a Baihu decoction granule and a quality standard detection method.

Background

The Jing Fang is the ancestor of the medical prescriptions, which refers to a prescription recorded in the treatise on the exogenous febrile diseases (classified as the treatise on exogenous febrile diseases and the general lack of supply) by Zhang Zhongjing in the Han Dynasty, and has the characteristics and advantages of "general, simple, cheap and effective". Wherein the Baihu decoction is issued from Shanghai treatise on Shanghai, is a typical prescription for treating yang-Ming-Qi-branch heat excess syndrome, is composed of 4 medicinal materials including gypsum, rhizoma anemarrhenae, radix Glycyrrhizae Preparata and semen oryzae Sativae, and has the effects of clearing heat and promoting fluid production.

As a classic famous prescription in Shanghai treatise on Cold-induced diseases, the white tiger decoction mainly focuses on the influence of different compatibility combinations on the dissolution of chemical components and the clinical application of the white tiger decoction, and the white tiger decoction is prepared into a proper dosage form, so that the aim of new use of ancient prescriptions is achieved.

Currently, there is little doctor in applying the menstruation prescription in modern clinical treatment, which causes the current situation: firstly, the medicine is not used, because the user needs to be familiar with and master the mutual compatibility and dosage of the medicines on the basis of the basic theory of the traditional Chinese medicine, and the experience of clinical practice is realized, so that the medicine has stronger practical and empirical properties; the menstruation is dare not to use, the menstruation side has good effect but is not a universal side, the correct application has good effect on treating diseases, if the menstruation side is misused, the menstruation side has no effect, and the menstruation side can also hurt patients, so the menstruation side is not used as the menstruation side is responsible for the disease; most prescriptions are less in flavor and have economic benefits than traditional prescriptions, especially for the first reason. Due to the rapid progress and development of modern scientific technology and modern medicine, the clinical symptoms of the current are greatly different from the ancient times, and the exertion of the clinical effects of the menstruation recipe is greatly impacted and challenged, so that the basic material research and the clinical practice research of the menstruation recipe are carried out by combining the gist of the original Zhang Zhongjing theory and the current science and technology, and more reference values can be laid for the further development and improvement of the menstruation recipe.

Disclosure of Invention

In order to overcome the defects of the prior art, the invention aims to provide a preparation method of the white tiger decoction particles and a detection method of quality standards, and the quality of the white tiger decoction particles is ensured by using the detection method.

In order to realize the purpose, the invention adopts the following technical scheme:

the preparation method of the white tiger decoction particles comprises the following steps:

(1) weighing gypsum, rhizoma anemarrhenae, radix Glycyrrhizae Preparata and semen oryzae Sativae according to the formula ratio, adding 8 times of water in the 1 st time, soaking for 0.5h, decocting, adding 6 times of water in the 2 nd and 3 rd times of decoction respectively, decocting for 0.5h in the three times, filtering with gauze, mixing the filtrates, and concentrating until the relative density is 1.02;

(2) adding a carboxymethyl chitosan solution into the concentrated filtrate obtained in the step (1) at 60 ℃, stirring for 20 min, standing for 12 h, centrifuging, filtering, and concentrating the filtrate to obtain a thick paste with the relative density of 1.10-1.12;

(3) adding adjuvant A, drying under reduced pressure, pulverizing into fine powder, adding adjuvant B, adding 88% ethanol to make soft mass, granulating, drying, grading, and packaging to obtain BAIHUTONG granule.

Preferably, the mass ratio of the gypsum, the rhizoma anemarrhenae, the radix glycyrrhizae preparata and the polished round-grained rice in the step (1) is 50: 18: 6: 9.

preferably, the mass concentration of the carboxymethyl chitosan solution in the step (2) is 0.5%; the volume ratio of the concentrated filtrate to the carboxymethyl chitosan solution is 1: 5.

Preferably, the auxiliary materials A and B in the step (3) form a total auxiliary material; the mass ratio of the thick paste in the step (2) to the total auxiliary materials is 1: 0.3; wherein the mass ratio of the auxiliary material A to the auxiliary material B in the step (3) is 2: 1; the ratio of dextrin to soluble starch in the auxiliary materials is 3: 2.

The detection method for the quality standard of the Baihu decoction granules comprises the following steps: qualitatively identifying rhizoma anemarrhenae and radix Glycyrrhizae Preparata by TLC, and determining the content of rhizoma anemarrhenae and radix Glycyrrhizae by HPLC.

Further, the qualitative identification of rhizoma anemarrhenae and radix glycyrrhizae preparata is carried out by the following method:

A. weighing 2 g of BAIHU decoction, grinding, adding 10 mL of methanol, respectively, and performing ultrasonic treatment for 30 min to obtain supernatant as sample solution; precisely weighing mangiferin and new mangiferin reference substances respectively, adding methanol to obtain mangiferin with concentration of 0.25 mg/mL^-1The concentration of the new mangiferin is 0.23 mg/mL^-1The solution of (4) as a control solution; weighing 0.5 g of rhizoma anemarrhenae powder, and preparing into rhizoma anemarrhenae reference solution by the same method as the test solution; weighing other prescription medicinal materials except rhizoma anemarrhenae according to the proportion of the white tiger decoction particles, preparing rhizoma anemarrhenae deficiency negative particles according to the preparation process of the particles, and preparing rhizoma anemarrhenae deficiency negative control solution according to the preparation method of the test solution; performing thin layer chromatography (2015 version of the fourth general rule 0502 of Chinese pharmacopoeia)Respectively dropping 5 μ L of mangiferin reference solution, neomangiferin reference solution, rhizoma anemarrhenae deficiency negative reference solution, and test solution on the same polyamide film, developing with ethanol-water as developing agent at volume ratio of 1: 1, taking out, air drying, and inspecting under ultraviolet lamp 365 nm to obtain test sample, rhizoma anemarrhenae reference solution, and reference solution chromatogram with the same color fluorescent spots, and no rhizoma anemarrhenae deficiency negative particles;

B. weighing 15 g of BAIHU decoction granule, adding diethyl ether 40 mL, heating and refluxing for 1 h, filtering, discarding ether solution, adding methanol 30 mL into the residue, heating and refluxing for 1 h, filtering, evaporating the filtrate to dryness, adding water 40 mL into the residue to dissolve, extracting with n-butanol for 3 times, 20 mL each time, mixing n-butanol solutions, washing with water for 3 times, discarding water solution, evaporating n-butanol solution to dryness, adding methanol 5 mL into the residue to dissolve, and using as sample solution; taking liquiritin reference substance, adding methanol to prepare solution containing 2 mg per 1mL as reference substance solution; weighing other prescription medicinal materials except Glycyrrhrizae radix according to the proportion of BAIHUTANG granule, preparing into negative granule without Glycyrrhrizae radix according to the preparation process of the granule, and preparing into negative control solution without Glycyrrhrizae radix according to the preparation method of the sample solution; performing thin-layer chromatography (2015 version of Chinese pharmacopoeia, general 0502 of the fourth ministry of pharmacopoeia of China), sucking 5 μ L of a licorice-lacking negative control solution, a liquiritin control solution and three batches of test solution, respectively dropping the solution on a same silica gel G thin-layer plate made of 1% sodium hydroxide solution, developing by taking ethyl acetate-formic acid-glacial acetic acid-water as a developing agent with the volume ratio of 15: 1: 2, taking out, drying in the air, uniformly spraying 10% sulfuric acid ethanol solution for color development, heating at 105 ℃ for 5-10 min, and inspecting under an ultraviolet lamp (365 nm), wherein fluorescent spots with the same color are formed on the positions corresponding to the color spectrums of the three batches of test solution and the control solution, and spots do not exist at the positions where the licorice-lacking negative particles.

Further, the method A can also adopt the following method:

weighing 1 g of BAIHU decoction, grinding, adding 30% acetone 10 mL, ultrasonic treating for 30 min, and collecting supernatant as sample solution; accurately weighing timosaponin BII reference substance, adding 30% acetone to obtain a solution with a concentration of 0.56 mg/mL^-1The solution of (4) as a control solution; according to the positionWeighing other prescription medicinal materials except rhizoma anemarrhenae according to the proportion, preparing rhizoma anemarrhenae deficiency negative granules according to the preparation process of the granules, and preparing rhizoma anemarrhenae deficiency negative control solution according to the preparation method of the test solution; performing thin layer chromatography (2015 edition of Chinese pharmacopoeia, general rule of the fourth part 0502), respectively sucking 5 μ L of a control solution, a rhizoma anemarrhenae deficiency negative control solution and a test solution, respectively dropping on the same silica gel G thin layer plate, developing with n-butanol-glacial acetic acid-water as a developing agent at a volume ratio of 4: 1:5, taking out, air drying, spraying a 5% vanillin sulfuric acid ethanol test solution, heating at 105 ℃ until the spots are clearly developed, and as a result, the spots with the same color are developed on the positions corresponding to the chromatogram of the test solution and the control solution, and no spots are formed in the positions of rhizoma anemarrhenae deficiency negative particles.

Further, the method for determining the content of rhizoma anemarrhenae and radix glycyrrhizae by using the HPLC method comprises the following steps:

(1) chromatographic conditions are as follows: lichrospher C18 (5 μm, 4.6 mm. times.250 mm) high performance liquid chromatography column; mobile phase: acetonitrile-B25 mmol. L^-1Potassium dihydrogen phosphate, gradient eluting (0-2 min, 98% B; 2-8 min, 98% -85% B; 8-15min, 85% -78% B; 15-20 min, 78% -72% B; 20-30 min, 72% -50% B), column temperature 30 deg.C, flow rate 1 mL/min^-1The detection wavelength is 257 nm, and the sample injection amount is 10 mu L;

(2) preparation of mixed control solution: weighing 4 kinds of reference substances including neomangiferin, mangiferin, liquiritin and monoammonium glycyrrhizinate, dissolving with methanol, and diluting to desired volume to obtain mixed reference solution, wherein the concentration of neomangiferin is 26.22 μ g/mL^-1The mangiferin concentration is 20.38 mug.mL^-1The concentration of liquiritin is 26.46 mug. multidot.mL^-1The concentration of the monoammonium glycyrrhizinate is 30.67 mu g/mL^-1Filtering with 0.45 μm microporous membrane, and collecting the filtrate.

(3) Preparation of a test solution: grinding BAIHU decoction granule, collecting 1 g, adding 25 mL of methanol, weighing, performing ultrasonic treatment at 250W and 40KHz frequency for 30 min, cooling, weighing, supplementing the weight loss with methanol, shaking, filtering, and collecting the filtrate;

(4) preparation of negative control solution

Weighing other medicinal materials except rhizoma anemarrhenae and radix Glycyrrhizae Preparata, and preparing into negative control solution without rhizoma anemarrhenae and radix Glycyrrhizae Preparata according to the preparation method of BAIHU decoction granule and the preparation method of test solution, wherein the negative control solution has no color spectrum peak at the position corresponding to the color spectrum peak of the control solution, which indicates that Gypsum Fibrosum and semen oryzae Sativae in the prescription has no interference to measurement;

(5) respectively sucking 10 μ L of the mixed reference solution, the test solution and the negative reference solution, injecting into a high performance liquid chromatograph, and detecting at 257 nm according to the above chromatographic conditions, wherein the mixed reference solution and the test solution show absorption peaks in the same retention time, and no chromatographic peak appears in the negative reference of rhizoma anemarrhenae and radix Glycyrrhizae;

(6) the determination method comprises the following steps: respectively sucking 10 μ L of the mixed reference solution, and injecting into high performance liquid chromatograph, wherein each 1 g of the product contains rhizoma anemarrhenae based on the content of neomangiferin and mangiferin, the content of neomangiferin should not be less than 0.4 mg, and the content of mangiferin should not be less than 0.5 mg; the product contains Glycyrrhrizae radix in each 1 g, and calculated by content of glycyrrhizin and monoammonium glycyrrhizinate, the content of glycyrrhizin should not be less than 0.3 mg, and the content of monoammonium glycyrrhizinate should be 0.3 mg.

Advantageous effects

(1) The invention provides a preparation method of white tiger soup particles, which is determined by taking CaSO 4.2H 20 retention rate, neomangiferin retention rate, mangiferin retention rate and impurity removal rate as examination indexes.

(2) The invention establishes a quality control method of the Baihu decoction granules, carries out thin-layer chromatography identification on rhizoma anemarrhenae and radix glycyrrhizae preparata in the preparation, carries out content measurement on the rhizoma anemarrhenae and the radix glycyrrhizae preparata in the preparation, has simple and convenient operation, strong specificity and accurate result, and can be used for the quality control of the Baihu decoction granules.

Drawings

FIG. 1 is a process for preparing BAIHUTANG granule;

FIG. 2 is a TLC chart of Zhimu and Zhigan Cao; wherein, FIG. 2A is a TLC diagram of mangiferin and new mangiferin, FIG. 2A-1 is a TLC diagram of mangiferin reference, FIG. 2A-2 is a TLC diagram of new mangiferin reference, FIG. 2A-3 is a TLC diagram of mangiferin and new mangiferin as reference medicinal material of rhizoma anemarrhenae, and FIG. 2A-4 is a TLC diagram of mangiferin and new mangiferin as negative granules lacking rhizoma anemarrhenae; FIG. 2A-5, 2A-6, 2A-7 are TLC images of three batches of Baihu decoction granules to be tested for mangiferin and neomangiferin; wherein FIG. 2B is a TLC chart of timosaponin BII, FIG. 2B-1 is a reference sample of timosaponin BII, FIG. 2B-2 is a TLC chart of timosaponin BII lacking rhizoma anemarrhenae negative granules, and FIGS. 2B-3, 2B-4, and 2B 5 are TLC charts of timosaponin BII to be tested from three batches of BAIHUTANG granules; FIG. 2C is a TLC image of Glycyrrhiza glabra, FIG. 2C-1 lacks Glycyrrhiza glabra negative particles; FIG. 2C-2 glycyrrhizin control; fig. 2C-3, fig. 2C-4, and fig. 2C-5 show three batches of Baihu decoction granules to be tested.

FIG. 3 is an HPLC chart of 4 mixed control (A), anemarrhena negative control (B), and test sample (C);

FIG. 4 is a HPLC chart of a control, wherein 1 is neomangiferin, 2 is mangiferin, 3 is liquiritin, and 4 is monoammonium glycyrrhizinate;

FIG. 5 is finger print of BAIHUGUO granule, wherein 6 is neomangiferin, 8 is mangiferin, 12 is liquiritin, and 18 is monoammonium glycyrrhizinate;

FIG. 6 shows the finger prints of 10 batches of BAIHUTANG granule samples, wherein S1-S10 is 10 batches of BAIHUTANG granule samples, and R is the control.

Detailed Description

The present invention is further illustrated by the following examples, which should be construed as merely illustrative and not limitative.

The following examples used starting materials and reagents, respectively: gypsum Fibrosum (Shanxi, batch No. 170502), rhizoma anemarrhenae (Hebei, batch No. 161101), radix Glycyrrhizae Preparata (Gansu, batch No. 170201), all of which are purchased from Shandong Baiweitang Chinese medicinal decoction pieces Limited, and semen oryzae Sativae is commercially available from Ningpo rice of Jinlongyu. Rhizoma anemarrhenae as reference material (batch No. 121070-. Mangiferin reference substance (batch number: B20837-20 mg, not less than 98%), neomangiferin reference substance (batch number: B21397-20 mg, not less than 98%), liquiritin reference substance (batch number: B20414-20 mg, not less than 98%), monoammonium glycyrrhizinate reference substance (batch number: B20418-20 mg, not less than 98%), and all the reference substances are purchased from Shanghai-sourced leaf Biotechnology Co., Ltd. Monopotassium phosphate (liquid phase, 100 g of chemical reagent of Kemiou, Tianjin, Inc.), acetonitrile and methanol for the liquid phase, Wahaha purified water for the liquid phase, and analytically pure other reagents.