A method for testing sifenable capsules

Abstract

The present application relates to the technical field of drug efficacy detection, and particularly relates to a test method of Suguanning capsules. The present application adds the detection standards of Isatis indigotica Fort, Siphonostegia chinensis Maxim, Lonicera japonica and paracetamol and Bupleurum chinense to the original standard of Suguanning capsules, carries out thin layer chromatography on Isatis indigotica Fort, Siphonostegia chinensis Maxim and Lonicera japonica, and carries out the detection of specificity, linear relationship, precision, stability, reproducibility and sample recovery rate on paracetamol and Bupleurum chinense. The thin layer chromatography reproducibility and specificity of Bupleurum chinense and Lonicera japonica are good, and the negative has no interference, so the quality standard text is included. The thin layer chromatography specificity of Siphonostegia chinensis Maxim and Isatis indigotica Fort is poor, so the quality standard text is not included, and the detection standard of Suguanning capsules is improved.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of drug efficacy detection, and particularly relates to a test method of Sugenning capsules. BACKGROUND

[0002] The Sugenning capsule is a compound preparation composed of 7 medicines, i.e., Herba Swertiae Mileensis, Radix Bupleuri, Folium Isatidis, Flos Lonicerae Japonicae, Bovine Bezoar, acetaminophen and chlorpheniramine maleate, and is produced by Jilin Zhongyan Pharmaceutical Co., Ltd. According to the theory of traditional Chinese medicine and the principle of new drug naming, the Sugenning capsule can also be named as Goutoni Zhuanggu Tablet. The raw material composition of the Sugenning capsule specifically includes: Herba Swertiae Mileensis 320 g, Radix Bupleuri 320 g, Folium Isatidis 240 g, Flos Lonicerae Japonicae 240 g, Bovine Bezoar 8 g, acetaminophen 100 g and chlorpheniramine maleate 1.2 g.

[0003] The four medicines of Herba Swertiae Mileensis, Radix Bupleuri, Folium Isatidis and Flos Lonicerae Japonicae are decocted with water twice, the first time for 3 hours and the second time for 2 hours. The decoction is filtered, concentrated to a thick paste under reduced pressure, dried and pulverized into fine powder. The powder is mixed with Bovine Bezoar powder, acetaminophen and chlorpheniramine maleate, and then mixed with 75% ethanol. The mixture is dried and pulverized again to obtain gray-brown powder, which is the content of the Sugenning capsule. The powder is filled into capsules to obtain the Sugenning capsule.

[0004] The original detection standard of the Sugenning capsule only has two items, one is thin layer identification of Bovine Bezoar and chlorpheniramine maleate, and the other is determination of the content of acetaminophen by ultraviolet-visible spectrophotometry. The detection standard is low. In order to better control the product quality, it is urgent to improve the standard of the product. SUMMARY

[0005] The technical problem to be solved by the present application is to provide a test method of the Sugenning capsule to improve the quality detection standard of the Sugenning capsule by using a liquid chromatograph.

[0006] To solve the above technical problem, the technical scheme adopted by the present application is as follows:

[0007] S1, taking the content of the Sugenning capsule to prepare a test solution, a control solution and a negative control solution;

[0008] S2, specificity detection: 5 μL of the test solution, the control solution, the negative control solution and the methanol solution obtained in S1 are injected into a liquid chromatograph to obtain the solution chromatogram corresponding to each solution;

[0009] S3, linear relationship detection: different volumes of the test solution were taken, then methanol was added for constant volume, different mass concentrations of the solution were prepared, the liquid chromatogram of the prepared solution with different mass concentrations was determined, the peak area was taken as the vertical coordinate, the mass concentration was taken as the horizontal coordinate for linear regression and standard curve drawing;

[0010] S4, precision detection: 5 μL of the control solution prepared in S1 was taken, and the liquid chromatogram was determined by continuous injection for 6 times;

[0011] S5, stability detection: 5 μL of the test solution and the control solution obtained in S1 were taken, and the liquid chromatogram was determined at different times after preparation;

[0012] S6, reproducibility detection: 0.1 g of the contents of the Sugenning capsules was taken for 6 times, and the test solution was prepared, and the liquid chromatogram was determined;

[0013] S7, sample addition recovery rate detection: 0.05 g of 9 samples of the contents of the Sugenning capsules was taken, 0.5, 1 and 1.5 times of the control substance were added to the samples respectively, and the liquid chromatogram was determined.

[0014] According to the test method of the Sugenning capsule provided by the application, when the acetaminophen in the Sugenning capsule is determined, S1 further comprises the following steps:

[0015] S101, the finely ground contents of the Sugenning capsule were dissolved in methanol, ultrasonic treatment was carried out, the solution was cooled to room temperature, and then filtered after shaking, the obtained filtrate was constant volume to 50 mL in a volumetric flask, 1 mL of the solution in the 50 mL volumetric flask was taken, and then constant volume to 10 mL in a volumetric flask, and then filtered after shaking again to obtain the filtrate, that is, the acetaminophen test solution;

[0016] S102, the acetaminophen control substance was taken and constant volume to 10 mL in a volumetric flask with methanol to prepare the acetaminophen control substance stock solution, and the acetaminophen control substance stock solution was taken and constant volume to 5 mL in a volumetric flask with methanol to prepare the acetaminophen control solution;

[0017] S103, the contents of the Sugenning capsule without acetaminophen were taken, and the acetaminophen negative control solution was prepared according to the method described in S101.

[0018] According to the test method of the Sugenning capsule provided by the application:

[0019] S111, 5.0 g of the content of the Sujingning capsule is taken, added into 40 mL of methanol, dissolved, and then filtered after ultrasonic treatment for 30 min. The filtrate is evaporated to dryness, and the obtained residue is dissolved with 15 mL of NaOH test solution in several portions. The solution is extracted with 40 mL of water-saturated n-butanol solution, and the extracted n-butanol solution is washed with 100 mL of n-butanol-saturated ammonia test solution. After the n-butanol-saturated ammonia washing solution is discarded, the n-butanol solution is washed with 50 mL of n-butanol-saturated water solution for two times. The n-butanol solution is taken, evaporated to dryness, and then added into a 2 mL volumetric flask with methanol, shaken, filtered through a microporous filter membrane with a pore size of 0.45 μm, and then the filtrate is taken, i.e. the Bupleurum test sample solution is obtained;

[0020] S112, 10.02 mg, 10.04 mg and 10.07 mg of the Bupleurum saikosaponin a, Bupleurum saikosaponin b1 and Bupleurum saikosaponin b2 reference substances are taken, added into a 10 mL volumetric flask with methanol, and then a Bupleurum saikosaponin a, Bupleurum saikosaponin b1 and Bupleurum saikosaponin b2 reference substance stock solution with a mass concentration of 0.9820 mg / mL, 0.9839 mg / mL and 0.9869 mg / mL respectively is prepared. 1 mL of each of the above Bupleurum saikosaponin a, Bupleurum saikosaponin b1 and Bupleurum saikosaponin b2 reference substance stock solution is taken, added into a 10 mL volumetric flask with methanol, and then a Bupleurum saikosaponin reference substance mixed solution is prepared.

[0021] S113, the content of the Sujingning capsule without Bupleurum is taken, and a Bupleurum negative reference solution is prepared according to the preparation method of the Bupleurum test sample solution.

[0022] According to the test method of the Sujingning capsule provided by the application, when the paracetamol in the Sujingning capsule is detected, the liquid chromatography detection conditions are as follows: an Agilent 1260 type high performance liquid chromatograph is used, the chromatographic column is selected to be an Agilent Eclipse SB-C18 chromatographic column with a size of 4.6*250 mm and 5 μm, the column temperature is set to be 25 ℃, the detection wavelength is set to be 240 nm, the flow rate is set to be 1.0 mL / min, the injection amount is set to be 5 μL, and the mobile phase is set to be a water:methanol solution with a ratio of 80:20.

[0023] When the Bupleurum in the Sujingning capsule is detected, the liquid chromatography detection conditions are as follows: an Agilent 1260 type high performance liquid chromatograph is used, the chromatographic column is selected to be an Agilent Eclipse SB-C18 chromatographic column with a size of 4.6*250 mm and 5 μm, the column temperature is set to be 30 ℃, the detection wavelength is set to be 210 nm, the flow rate is set to be 1.0 mL / min, the injection amount is set to be 10 μL, and the mobile phase is set to be a water:acetonitrile solution with a ratio of 66:34.

[0024] The content of the Sujianning capsule in S101 is 0.1 g; and the ultrasonic conditions are: ultrasonic treatment for 30 min under the conditions of a power of 150 W and a frequency of 40 KHz.

[0025] The content of the Sujianning capsule in S102 is 10.01 mg, the mass concentration of the control sample stock solution is 0.9910 mg / mL, 0.35 mL of the control sample stock solution is taken, and the mass concentration of the control sample solution is 0.0694 mg / mL.

[0026] The content of the Sujianning capsule in S3 is 0.5-3.5 mL, the concentration of the control sample solution is 0.9900 mg / mL when detecting acetaminophen in the Sujianning capsule, and the concentration of the control sample solution is 1.9619 mg / mL when detecting acetaminophen in the Sujianning capsule.

[0027] The mass concentration of the control sample solution in S4 is 0.0693 mg / mL when the control sample solution is an acetaminophen control sample solution, and the mass concentration of the control sample solution in S4 is 0.3956 mg / mL when the control sample solution is a radix bupleuri control sample solution.

[0028] The different time periods in S5 include 0 h, 2 h, 4 h, 6 h, 8 h, 10 h, 12 h, 18 h, 24 h, 28 h, 32 h and 36 h.

[0029] Compared with the prior art, the Sujianning capsule detection method has the following advantages:

[0030] The Sujianning capsule detection method adds the detection standards of Isatidis Folium, Sphagneticolas Rehbra-rioides, Lonicera Japonica, acetaminophen and radix bupleuri to the original standard of the Sujianning capsule, carries out thin layer chromatography on Isatidis Folium, Sphagneticolas Rehbra-rioides and Lonicera Japonica, and carries out the detection of specificity, linear relationship, precision, stability, reproducibility and sample recovery rate on radix bupleuri and acetaminophen, so that the reproducibility and specificity of the thin layer chromatography of radix bupleuri and Lonicera Japonica are good, the negative is not interfered, and the quality standard text is included. The specificity of the thin layer chromatography of Sphagneticolas Rehbra-rioides and Isatidis Folium is poor, so the quality standard text is not included, and the detection standard of the Sujianning capsule is improved. BRIEF DESCRIPTION OF DRAWINGS

[0031] Figure 1 is a chromatogram of the acetaminophen sample solution in Example 5;

[0032] Figure 2 is a chromatogram of the acetaminophen control sample solution in Example 5;

[0033] Figure 3 is a chromatogram of the acetaminophen negative control solution in Example 5;

[0034] Figure 4 is a chromatogram of the methanol solution in Example 5;

[0035] Figure 5 is a standard curve plot for acetaminophen in Example 5;

[0036] Figure 6 is a chromatogram of saikosaponin a, saikosaponin bl, and saikosaponin b2 in Example 7 with a mobile phase ratio of water:acetonitrile of 66:34;

[0037] Figure 7 is a chromatogram of the Bupleurum test solution in Example 7 with a mobile phase ratio of water:acetonitrile of 66:34;

[0038] Figure 8 is a chromatogram of the Bupleurum negative control solution in Example 7 with a mobile phase ratio of water:acetonitrile of 66:34;

[0039] Figure 9 is a chromatogram of the Bupleurum saikosaponin bl test solution in Example 7 with a mobile phase ratio of water:acetonitrile of 60:40;

[0040] Figure 10 is a chromatogram of the Bupleurum saikosaponin bl control solution in Example 7 with a mobile phase ratio of water:acetonitrile of 60:40;

[0041] Figure 11 is a chromatogram of the Bupleurum saikosaponin bl test solution in Example 7 with a mobile phase ratio of water:acetonitrile of 64.5:35.5;

[0042] Figure 12 is a chromatogram of the Bupleurum saikosaponin bl control solution in Example 7 with a mobile phase ratio of water:acetonitrile of 64.5:35.5;

[0043] Figure 13 is a chromatogram of the Bupleurum saikosaponin bl test solution in Example 7 with a mobile phase ratio of 0.05% formic acid in water:acetonitrile of 64:36;

[0044] Figure 14 is a chromatogram of the Bupleurum saikosaponin bl control solution in Example 7 with a mobile phase ratio of 0.05% formic acid in water:acetonitrile of 64:36;

[0045] Figure 15 is a chromatogram of the Bupleurum saikosaponin bl test solution in Example 7 with a mobile phase ratio of 0.1% phosphoric acid in water:acetonitrile of 64:36;

[0046] Figure 16 is the chromatogram of the reference solution of saikosaponin b1 in Example 7 when the proportion of the mobile phase of 0.1% phosphoric acid aqueous solution: acetonitrile is 64:36;

[0047] Figure 17 is the chromatogram of the test solution of saikosaponin b1 in Example 7;

[0048] Figure 18 is the chromatogram of the reference solution of saikosaponin b1 in Example 7;

[0049] Figure 19 is the chromatogram of the negative control solution of saikosaponin b1 in Example 7;

[0050] Figure 20 is the chromatogram of the methanol solution in Example 7;

[0051] Figure 21 is the standard curve of saikosaponin b1 in Example 7. DETAILED DESCRIPTION

[0052] Example 1

[0053] In this example, the Sugenning capsule is a compound preparation composed of 320 g of Glechoma longituda L, 320 g of Bupleurum chinense DC, 240 g of Indigolfera, 240 g of Honeysuckle, 8 g of ox gall, 100 g of acetaminophen and 1.2 g of chlorpheniramine maleate.

[0054] This example provides a method for identifying Bupleurum chinense DC in Sugenning capsules.

[0055] Take the content of Sugenning capsules, 3 g in total, and dissolve in 30 mL of methanol. Under the condition of power 150 W and frequency 40 KHz, ultrasonic for 30 min, then filter to obtain the filtrate. The obtained filtrate is evaporated to dryness, and the residue is dissolved with 15 mL of NaOH solution to obtain the dissolved solution. The dissolved solution is extracted with 20 mL of water-saturated n-butanol solution for 2 times. The two extracted solutions are combined to obtain the n-butanol solution, which is washed with 100 mL of ammonia-saturated n-butanol washing solution, and then washed with 50 mL of water-saturated n-butanol washing solution for 2 times to obtain the washed n-butanol solution, which is evaporated to dryness. The residue is dissolved with 1 mL of anhydrous ethanol to obtain the test solution of Bupleurum chinense DC;

[0056] Take the content of Sugenning capsules without Bupleurum chinense DC and prepare the negative control solution of Bupleurum chinense DC according to the above method;

[0057] Take saikosaponin a and saikosaponin b1 respectively, and add methanol solution to obtain saikosaponin a control solution and saikosaponin b1 control solution with a mass concentration of 1 mg / mL respectively;

[0058] Saikosaponin a and saikosaponin b1 are commercially available from Shanghai Yuan Ye Biological Technology Co., Ltd.

[0059] Thin layer chromatography identification: 5 μL of the above Bupleurum test solution, Bupleurum negative control solution, saikosaponin a control solution and saikosaponin b1 control solution are taken and spotted on the same silica gel G thin layer plate, and a chloroform / methanol / water solution with a proportion of 30:10:1 is used as a developing agent for development, then the silica gel G thin layer plate is taken out and dried, 1 mL of sulfuric acid ethanol is diluted to 10 mL of sulfuric acid ethanol solution, then 2% of p-dimethylaminobenzaldehyde by mass is added and sprayed on the dried silica gel G thin layer plate, and the plate is heated in a water bath at a temperature of 105°C until the spots are clearly colored, then the heated silica gel G thin layer plate is placed under a UV light lamp with a wavelength of 365 nm for inspection, and the corresponding thin layer chromatogram is obtained.

[0060] In the thin layer chromatogram of the Bupleurum test solution, the same color spots are shown at positions corresponding to the thin layer chromatograms of the saikosaponin a control solution and the saikosaponin b1 control solution, and the thin layer chromatogram of the Bupleurum negative control solution has no interference, and has good specificity;

[0061] To detect the stability of the chromatogram, the chromatogram is developed under normal temperature and humidity, high temperature conditions of 40°C, high humidity conditions of relative humidity 90%, low temperature conditions of 4°C, and using silica gel from different manufacturers, and the specificity is good; 5 batches of samples are taken, and the Bupleurum is identified by the method, and the reproducibility is good.

[0062] Example 2

[0063] The content of the Sugenning capsule used in this example is the same as that of Example 1.

[0064] This example provides a method for identifying honeysuckle in Sugenning capsules.

[0065] 1 g of the content of the Sugenning capsule is dissolved in 10 mL of water, ultrasonic is performed at a power of 150 W and a frequency of 40 KHz for 30 min, and then filtration is performed to obtain a filtrate, the filtrate is extracted twice with 20 mL of ethyl acetate, the ethyl acetate solutions after the two extractions are combined, and the solvent is evaporated by water bath heating at a temperature of 105°C, and the obtained residue is dissolved in 1 mL of methanol as a honeysuckle test solution;

[0066] Take 0.2 g of Honeysuckle and 1 g of the content of Sujingning capsules without Honeysuckle, respectively, and prepare Honeysuckle reference solution and Honeysuckle negative reference solution according to the above method;

[0067] Thin layer chromatography identification: take 10 μL of Honeysuckle test solution, Honeysuckle reference solution and Honeysuckle negative reference solution, respectively, and spot them on the same silica gel G thin layer plate at appropriate intervals. Use the upper solution of butyl acetate / formic acid / methanol / water solution (14:5:4:5) as the developing agent for development. Then take out the silica gel G thin layer plate and dry it, and then place it under UV light at a wavelength of 365 nm for observation. The corresponding thin layer chromatogram is obtained.

[0068] In the thin layer chromatogram of Honeysuckle test solution, the same color spots appear at the corresponding position of the thin layer chromatogram of Honeysuckle reference solution, and the thin layer chromatogram of Honeysuckle negative reference solution has no interference, showing good specificity.

[0069] To test the stability of thin layer chromatography, the specificity is good under the conditions of normal temperature and humidity, high temperature of 40℃, high humidity of 90% relative humidity, low temperature of 4℃ and use of silica gel from different manufacturers. Five batches of samples were taken to identify Honeysuckle using this method, and the reproducibility was good.

[0070] Example 3

[0071] The content of Sujingning capsules used in this example is the same as that of Example 1.

[0072] This example provides three methods for identifying Gegen in Sujingning capsules.

[0073] The first method for identifying Gegen in Sujingning capsules is as follows:

[0074] Take 5 g of the content of Sujingning capsules and dissolve it in 20 mL of cyclohexane. Ultrasonic for 30 min under the condition of power 150 W and frequency 40 KHz, and then filter to obtain the filtrate. Concentrate the filtrate to 1 mL as Gegen test solution 1.

[0075] Take 1 g of Gegen medicinal material and prepare Gegen reference medicinal solution 1 according to the above method.

[0076] Take Gegenin ABBA and add it to cyclohexane to prepare Gegen reference solution 1 with a mass concentration of 1 mg / mL.

[0077] Take 1 g of the content of Sujingning capsules without Gegen and prepare Gegen negative reference solution 1 according to the above method.

[0078] Thin layer chromatography identification: 4 μL of each of the above Asarum sieboldii Miq. test solution 1 and Asarum sieboldii Miq. negative control solution 1, 5 μL of each of Asarum sieboldii Miq. control medicinal material solution 1 and Asarum sieboldii Miq. control solution 1 were taken and spotted on the same silica gel G thin layer plate at proper intervals, and developed with n-hexane / trichloromethane / methanol (30:15:1) as developing agent, then the silica gel G thin layer plate was taken out and dried, sprayed with dilute ethanol solution containing 0.3% fast blue BB salt, and placed at 40°C for 1 hour for color development, and the corresponding thin layer chromatogram was obtained;

[0079] In the thin layer chromatogram of Asarum sieboldii Miq. test solution 1, no corresponding spot was present at the position corresponding to the thin layer chromatogram of Asarum sieboldii Miq. control medicinal material solution 1 and Asarum sieboldii Miq. control solution 1.

[0080] Second identification method of Asarum sieboldii Miq. in Sujingning capsules:

[0081] 5 g of the content of Sujingning capsules was dissolved in 50 mL of water, and ultrasonic treatment was carried out at a power of 150 W and a frequency of 40 KHz for 30 min, and filtration was carried out twice to obtain a filtrate, and then the filtrate was acidified with HCl to a pH value of 2.5, and then placed in a separatory funnel, and extracted twice with 15 mL of diethyl ether, and the diethyl ether liquid obtained after the two extractions was combined, and then the solvent was evaporated by heating in a water bath at a temperature of 105°C to obtain a residue, and then the residue was dissolved in dichloromethane to obtain Asarum sieboldii Miq. test solution 2;

[0082] The content of Sujingning capsules without Asarum sieboldii Miq. was taken, and Asarum sieboldii Miq. negative control solution 2 was prepared according to the above method;

[0083] 0.5 g of Asarum sieboldii Miq. control medicinal material was taken, and Asarum sieboldii Miq. control medicinal material solution 2 was prepared according to the above method;

[0084] Asarum sieboldii Miq. ABBA was taken and dissolved in dichloromethane to obtain Asarum sieboldii Miq. control solution 2 with a mass concentration of 0.9820 mg / mL;

[0085] Thin layer chromatography identification: 8 μL of each of the above Asarum sieboldii Miq. test solution 2, Asarum sieboldii Miq. negative control solution 2, Asarum sieboldii Miq. control medicinal material solution 2 and Asarum sieboldii Miq. control solution 2 was taken and spotted on the same silica gel G thin layer plate at proper intervals, and developed with dichloromethane / acetone / glacial acetic acid solution (16:4:0.3) as developing agent, then the silica gel G thin layer plate was taken out and dried, sprayed with 10% phosphomolybdic acid ethanol solution, and then sprayed with 0.4% NaOH solution, and placed in sunlight for observation, and the corresponding thin layer chromatogram was obtained;

[0086] No obvious spot corresponding to the thin layer chromatogram of Asarum sieboldii Miq. control solution 2 was present in the thin layer chromatogram of Asarum sieboldii Miq. test solution 2;

[0087] Third identification method of Asarum sieboldii Miq. in Sujingning capsules:

[0088] Take 5g of the contents of Suganning capsules and dissolve them in methanol. Acidify with HCl to pH 2.5, then extract with ether three times. Combine the extracts obtained from the three extractions and evaporate the solvent by heating in a water bath at 105℃. Dissolve the residue in 2mL of chloroform to prepare the Dryopteris crassirhizoma test solution 3.

[0089] Take the contents of the Suganning capsules (without Dryopteris crassirhizoma) and prepare Dryopteris crassirhizoma negative control solution 3 according to the above method;

[0090] Take 0.5g of Dryopteris crassirhizoma reference material and prepare Dryopteris crassirhizoma reference material solution 3 according to the above method;

[0091] Prepare a reference solution of Dryopteris crassirhizoma with a mass concentration of 1.0212 mg / mL by adding ABBA of Dryopteris crassirhizoma into chloroform.

[0092] Thin-layer chromatography identification: Take 8 μL each of the above-mentioned Dryopteris crassirhizoma test solution 3, Dryopteris crassirhizoma negative control solution 3, Dryopteris crassirhizoma reference medicinal material solution 3 and Dryopteris crassirhizoma reference solution 3, and spot them at appropriate intervals on the same silica gel G thin-layer plate. Develop the plate with a dichloromethane / acetone / glacial acetic acid solution in a ratio of 16:4:0.3. After that, remove the G thin-layer plate and let it dry. Spray it with a 10% concentration of phosphomolybdic acid ethanol solution, and then spray it with a 0.4% concentration of NaOH solution. Examine it under sunlight.

[0093] The thin-layer chromatogram of Dryopteris crassirhizoma test solution 3 did not show obvious spots, which corresponded to the thin-layer chromatogram of Dryopteris crassirhizoma reference solution 3.

[0094] Example 4

[0095] The contents of the Suganning capsules used in this embodiment are the same as those in Embodiment 1.

[0096] This embodiment provides three methods for identifying Isatis tinctoria leaves in Suganning capsules.

[0097] The first method for identifying Isatis tinctoria leaves in Suganning capsules:

[0098] Take 5g of the contents of Suganning capsules and dissolve them in 20mL of ether. Sonicate for 20min at 150W and 40KHz, then filter to obtain the filtrate. Evaporate the solvent by heating in a water bath at 105℃. Dissolve the residue in 0.5mL of chloroform to obtain Daqingye test solution 1.

[0099] Take the contents of the Suganning capsules (without Isatis tinctoria leaves) and prepare Isatis tinctoria leaf negative control solution 1 according to the above method;

[0100] Take 0.5 g of Isatis indigotica Fort. control drug, and prepare Isatis indigotica Fort. control drug solution 1 according to the above method;

[0101] Take 0.1 mg of Indigotin reference substance and 0.1 mg of Indirubin reference substance, and add them into 1 mL of chloroform to prepare Isatis indigotica Fort. reference substance solution 1;

[0102] Thin-layer chromatography identification: take 8 μL of each of the above-mentioned Isatis indigotica Fort. test sample solution 1, Isatis indigotica Fort. negative control substance solution 1, Isatis indigotica Fort. control drug solution 1 and Isatis indigotica Fort. reference substance solution 1, and spot them on the same silica gel G thin-layer plate at appropriate intervals, develop them with a developing agent of toluene / ethyl acetate in a ratio of 3:1, then take out the silica gel G thin-layer plate and dry it, develop it again in the above-mentioned developing agent, take out the silica gel G thin-layer plate and dry it, and then place it under sunlight for inspection;

[0103] No obvious spots corresponding to the thin-layer chromatogram of Isatis indigotica Fort. control drug solution 1 and Isatis indigotica Fort. reference substance solution 1 are found in the thin-layer chromatogram of Isatis indigotica Fort. test sample solution 1;

[0104] Second identification method for Isatis indigotica Fort. in Sugen Ning capsules:

[0105] Take 5 g of Sugen Ning capsule content, dissolve it in 20 mL of chloroform, heat it to reflux for 1 h, filter it, and then concentrate the filtrate to 1 mL to obtain Isatis indigotica Fort. test sample solution 2;

[0106] Take Sugen Ning capsule content without Isatis indigotica Fort. and Isatis indigotica Fort. control drug, and prepare Isatis indigotica Fort. negative test sample solution 2 and Isatis indigotica Fort. control drug solution 2, respectively, according to the above method;

[0107] Take Indigotin reference substance and Indirubin reference substance, and add them into chloroform to obtain Isatis indigotica Fort. reference substance solution 2 with a mass concentration of 1 mg / mL;

[0108] Thin-layer chromatography identification: take 5 μL of each of the above-mentioned Isatis indigotica Fort. test sample solution 2 and Isatis indigotica Fort. negative test sample solution 2, and take 8 μL of each of Isatis indigotica Fort. control drug solution 2 and Isatis indigotica Fort. reference substance solution 2, and spot them on the same silica gel G thin-layer plate at appropriate intervals, develop them with a developing agent of cyclohexane / chloroform / acetone solution in a ratio of 5:4:2, then take out the silica gel G thin-layer plate and dry it, and place it under sunlight for inspection;

[0109] No obvious spots corresponding to the thin-layer chromatogram of Isatis indigotica Fort. control drug solution 2 and Isatis indigotica Fort. reference substance solution 2 are found in the thin-layer chromatogram of Isatis indigotica Fort. test sample solution 2;

[0110] Third identification method for Isatis indigotica Fort. in Sugen Ning capsules:

[0111] Take 10 g of the contents of the Sujingning capsules and dissolve in 50 mL of chloroform. Heat to reflux for 30 h, then filter. Concentrate the filtrate to 10 mL, add 100-200 mesh activated alumina, 12 g, stir well, then evaporate to dryness. Dry load the resulting dry powder onto a column of activated alumina, 100-200 mesh, 6 g, and 15 cm in diameter. Elute with 45 mL of chloroform, then discard the first 13 mL of eluate. Collect the pink eluate, 10 mL, and evaporate to dryness. Dissolve the residue in 0.5 mL of chloroform to obtain the test solution 3 of Isatis indigotica Fort;

[0112] Take the contents of the Sujingning capsules without Isatis indigotica Fort and the control drug of Isatis indigotica Fort, and prepare the negative test solution 3 of Isatis indigotica Fort and the control drug solution 3 of Isatis indigotica Fort, respectively, according to the above method.

[0113] Take the control of indigirin and dissolve in chloroform to obtain the control solution 3 of Isatis indigotica Fort, with a mass concentration of 0.1 mg / mL.

[0114] Thin layer chromatography identification: take 5 μL of each of the test solution 3 of Isatis indigotica Fort, the negative test solution 3 of Isatis indigotica Fort, the control drug solution 3 of Isatis indigotica Fort, and the control solution 3 of Isatis indigotica Fort, and spot them on the same silica gel G thin layer plate at appropriate intervals. Develop with a benzene / acetone solution in a ratio of 4:1. Then take out the silica gel G thin layer plate, dry it, and observe it under daylight.

[0115] The thin layer chromatogram of the test solution 3 of Isatis indigotica Fort does not show obvious spots corresponding to the thin layer chromatograms of the control drug solution 3 of Isatis indigotica Fort and the control solution 3 of Isatis indigotica Fort.

[0116] Example 5

[0117] The contents of the Sujingning capsules used in this example are the same as those in Example 1.

[0118] This example provides a method for determining the content of acetaminophen in Sujingning capsules.

[0119] Take 0.1 g of the finely ground contents of the Sujingning capsules and dissolve in methanol in a conical flask. Seal the flask, then weigh it. Then ultrasonically treat it for 30 min at a power of 150 W and a frequency of 40 KHz. After the solution cools to room temperature, shake it well, then filter it. Take 1 mL of the filtrate, then dilute it to a volume of 10 mL in a volumetric flask. Shake it well again, then filter it to obtain the filtrate, which is the test solution of acetaminophen.

[0120] Take 10.01 mg of paracetamol control substance with a purity of 99% and add methanol to a constant volume of 10 mL in a volumetric flask to prepare a paracetamol control substance stock solution with a mass concentration of 0.9910 mg / mL. Take 0.35 mL of the paracetamol control substance stock solution and add methanol to a constant volume of 5 mL in a volumetric flask to prepare a paracetamol control substance solution with a mass concentration of 0.0694 mg / mL.

[0121] Take the contents of the Sujianning capsules without paracetamol and prepare a paracetamol negative control substance solution according to the method for preparing the test sample solution.

[0122] Selection of chromatographic conditions: Agilent 1260 high performance liquid chromatograph was used, the chromatographic column was an Agilent Eclipse SB-C18 chromatographic column with a size of 4.6 x 250 mm and 5 μm, the column temperature was set to 25°C, the detection wavelength was set to 240 nm, the flow rate was set to 1.0 mL·min -1 ; the injection volume was set to 5 μL, and the mobile phase was set to a water:methanol solution with a ratio of 80:20.

[0123] Determination of specificity:

[0124] Take 5 μL of each of the above paracetamol test sample solution, paracetamol control substance solution, paracetamol negative control substance solution, and methanol solution and inject them into the high performance liquid chromatograph to obtain the chromatograms corresponding to each solution.

[0125] Determination of linear relationship:

[0126] Take 3 mL, 2 mL, 1 mL, 0.7 mL, 0.5 mL, and 0.1 mL of the paracetamol solution with a mass concentration of 0.9900 mg / mL, respectively, and then add methanol to a constant volume of 10 mL in a volumetric flask to prepare paracetamol solutions with mass concentrations of 0.2970 mg / mL, 0.1980 mg / mL, 0.0990 mg / mL, 0.0693 mg / mL, 0.0495 mg / mL, and 0.0099 mg / mL, respectively.

[0127] Determine the peak area under the chromatographic conditions for the determination of paracetamol content in Sujianning capsules, and the data details are shown in Table 1. The standard curve was plotted with the peak area (Y) as the vertical coordinate and the mass concentration (X) as the horizontal coordinate, and the standard curve is shown in Figure 5 The regression equation is Y = 24093X + 78.103, and R = 0.9996.

[0128] Table 1 Relationship between paracetamol concentration and peak area

[0129] mass concentration / mg mL -1 ]] Peak area 0.2970 7199.500 0.1980 4913.000 0.0990 2452.550 0.0693 1658.100 0.0495 1370.500 0.0099 286.830

[0130] Thus, it is determined that the concentration of paracetamol control sample is linearly correlated with the peak area when the mass concentration of paracetamol control sample is in the range of 0.0099 mg / mL to 0.2970 mg / mL;

[0131] Determination of precision:

[0132] 5 μL of paracetamol control sample solution with a mass concentration of 0.0693 mg / mL is precisely taken, and the sample is continuously injected for 6 times, the peak area is recorded, and the relative standard deviation RSD is calculated;

[0133] The RSD of the peak area integral value of the paracetamol peak control sample solution is 1.48%, indicating that the precision of the instrument is good, and the results are shown in Table 2 below,

[0134] Table 2 RSD of peak area integral value of paracetamol peak control sample solution

[0135]

[0136]

[0137] Stability test:

[0138] 5 μL of paracetamol test sample solution and paracetamol control sample solution are taken, and the peak area is determined according to the method for determining the content of paracetamol in Speeding Ning capsules at 0 h, 2 h, 4 h, 6 h, 8 h, 10 h, 12 h, 18 h, 24 h, 28 h, 32 h and 36 h after preparation, respectively. The results show that the RSD of the peak area of the paracetamol control sample solution is 0.57%, and the RSD of the peak area of paracetamol in the paracetamol test sample solution is 0.59%, as shown in Table 3 below; indicating that paracetamol in the paracetamol test sample solution and the paracetamol control sample solution is stable at room temperature within 36 h;

[0139] Table 3 Stability test table of paracetamol test sample solution and its control sample solution

[0140] Time / h Reference solution Test solution 0 1563.500 1404.900 2 1555.600 1406.800 4 1569.400 1391.000 6 1568.900 1406.200 8 1565.900 1396.700 10 1558.400 1402.400 12 1558.700 1404.300 18 1578.100 1414.200 24 1576.100 1393.000 28 1577.700 1416.600 32 1580.500 1404.800 36 1579.800 1415.600 STDEV 9.015 8.264 AVERAGE 1569.383 1404.708 RSD 0.57％ 0.59％

[0141] Reproducibility test:

[0142] 6 portions of 0.1 g of Speeding Ning capsule content are taken, and paracetamol test sample solution is prepared according to the method for determining the content of paracetamol in Speeding Ning capsules, and the sample is injected under the same chromatographic conditions for determination. The results show that the content of paracetamol is 0.3001 g / g, and the RSD is 0.45%, as shown in Table 4, indicating that the method has good reproducibility;

[0143] Table 3 Paracetamol reproducibility test data table

[0144] Test No. Sample weight / g Acetaminophen content g / g 1 0.10004 0.2978 2 0.10005 0.2997 3 0.10002 0.2999 4 0.10007 0.3017 5 0.10006 0.3009 6 0.10002 0.3008 STDEV / 0.001354 AVERAGE / 0.3001 RSD / 0.45％

[0145] Add sample recovery rate test:

[0146] Take 0.05 g of Sujingning capsule content sample with 0.3001 g / g of acetaminophen content, 0.5, 1 and 1.5 times the amount of acetaminophen control added to the sample, respectively, according to the chromatographic conditions of the determination method of acetaminophen content in Sujingning capsules, record the peak area, calculate the recovery rate, the results are shown in Table 5, according to Table 5, the recovery rate meets the requirements;

[0147] Table 5 Acetaminophen addition recovery data

[0148]

[0149]

[0150] Content verification of acetaminophen in Sujingning capsules:

[0151] In order to eliminate the error of test results caused by instrument difference, the existing Sujingning capsule samples were determined by using instrument Agilent 1260, different operators and ZXRBAX Eclipse Plus C18 chromatographic column under the same system conditions, the determination results are shown in Table 6, the acetaminophen content determination value is basically consistent, which can exclude the error caused by instrument, personnel and chromatographic column change, and prove that the acetaminophen is stable in nature;

[0152] Table 6 Content verification of acetaminophen

[0153] Test name Acetaminophen (g / g) Routine conditions 0.3001 Different instruments 0.2967 Different operators 0.2922 Different chromatographic columns 0.2909

[0154] Content of acetaminophen in Sujingning capsule sample:

[0155] Prepare the acetaminophen test solution according to the above method, take 200201, 210101, 201201 and 200301 Sujingning capsules of different batch numbers, calculate the content of acetaminophen therein, the results are shown in Table 7.

[0156] Table 7 Acetaminophen content in different batches of Sujingning capsules

[0157]

[0158]

[0159] Example 6

[0160] The content of the Sujingning capsule used in this example is the same as that of Example 1.

[0161] This example provides a method for determining the content of Bupleurum in Sujingning capsules.

[0162] Take 5.0 g of finely ground Sujingning capsule content and dissolve it in 40 mL of methanol. After ultrasonic treatment for 30 min, filter the solution. Evaporate the filtrate to dryness. Dissolve the residue obtained in 15 mL of NaOH test solution in portions. Shake the solution with 40 mL of water-saturated n-butanol solution. Wash the extracted n-butanol solution with 100 mL of n-butanol-saturated ammonia solution. Discard the n-butanol-saturated ammonia wash solution. Then wash the solution twice with 50 mL of n-butanol-saturated water solution. Take the n-butanol solution, evaporate it to dryness, and add methanol to make up the volume to 2 mL in a volumetric flask. Shake well, filter through a microporous filter membrane with a pore size of 0.45 μm, and take the filtrate to obtain the Bupleurum test solution.

[0163] Take 10.02 mg, 10.04 mg, and 10.07 mg of Bupleurum saikosaponin a, Bupleurum saikosaponin b1, and Bupleurum saikosaponin b2 reference substances with a purity of 98% and add methanol to make up the volume to 10 mL in a volumetric flask to prepare Bupleurum saikosaponin a, Bupleurum saikosaponin b1, and Bupleurum saikosaponin b2 reference substance stock solutions with mass concentrations of 0.9820 mg / mL, 0.9839 mg / mL, and 0.9869 mg / mL, respectively. Take 1 mL of each of the above Bupleurum saikosaponin a, Bupleurum saikosaponin b1, and Bupleurum saikosaponin b2 reference substance stock solutions and add methanol to make up the volume to 10 mL in a volumetric flask to prepare a Bupleurum saikosaponin reference mixture solution.

[0164] Take Sujingning capsule content without Bupleurum and prepare a Bupleurum negative control solution according to the preparation method of the Bupleurum test solution.

[0165] Take 10 μL of each of the Bupleurum reference mixture solution, the Bupleurum test solution, and the Bupleurum negative control solution and inject them into the HPLC for determination.

[0166] Selection of chromatographic conditions: use an Agilent 1260 type high-performance liquid chromatograph, select an Agilent Eclipse SB-C18 chromatographic column with a size of 4.6 x 250 mm and a particle size of 5 μm, set the column temperature to 30°C, set the detection wavelength to 210 nm, set the flow rate to 1.0 mL·min -1 ; set the injection volume to 10 μL, and set the mobile phase to a water:acetonitrile solution with a ratio of 66:34.

[0167] Under this determination method, the results show that the saikosaponin a, saikosaponin b1, saikosaponin b2 control solution and the bupleurum test solution have common peaks at the same retention time, but the bupleurum negative control solution and the saikosaponin a and saikosaponin b2 control solution have interference, so the saikosaponin b1 is selected as the suitable control, and because the retention time of the saikosaponin b1 is long, the proportion of the mobile phase is adjusted to shorten the retention time of the saikosaponin b1;

[0168] The proportion of the mobile phase of the water: acetonitrile solution is changed from 66:34 to 60:40, 10 μL of the saikosaponin b1 control solution and the bupleurum test solution are injected into the HPLC according to the chromatographic conditions under this method for determination, the bupleurum test solution and the bupleurum control solution have multiple peaks at the same retention time, and the tailing exists, so the proportion of the mobile phase is adjusted again, and the tailing is adjusted by adding acid;

[0169] The injection amount is changed to 5 μL, and the mobile phase is changed from the water: acetonitrile solution with the proportion of 60:40 to the 0.04% concentration acetic acid water solution: acetonitrile with the proportion of 64.5:35.5, the bupleurum test solution and the bupleurum control solution have common peaks at the same retention time, but the tailing still exists;

[0170] The injection amount is changed to 5 μL, and the mobile phase is changed from the 0.04% concentration acetic acid water solution: acetonitrile solution with the proportion of 64.5:35.5 to the 0.05% concentration formic acid water solution: acetonitrile solution with the proportion of 64:36, the tailing of the bupleurum test solution still exists;

[0171] The injection amount is changed to 5 μL, and the mobile phase is changed from the 0.05% concentration formic acid water: acetonitrile solution with the proportion of 64:36 to the 0.1% concentration phosphoric acid water solution: acetonitrile solution with the proportion of 64:36, it is found that Figure 15 and Figure 16 The bupleurum test solution and the bupleurum control solution have common peaks at the same retention time, the tailing disappears, and the condition is stable;

[0172] Therefore, the chromatographic conditions for determining the content of the bupleurum are determined as follows: the Agilent 1260 high performance liquid chromatograph, the Agilent ZORBAX SB-C18 chromatographic column, the mobile phase is the 0.1% phosphoric acid water solution-acetonitrile solution with the proportion of 64:36, the detection wavelength is 210 nm, the column temperature is 25°C, the flow rate of the mobile phase is 1.0 mL / min, and the injection amount is 5 μL;

[0173] Table 8: Data table of the saikosaponin extraction method and content determination

[0174] Extraction method Bupleurum saikoside b1 / mg·g -1 ]] Refiux method 0.3654 Ultrasonic method (20 min) 0.2735 Ultrasonic method (30 min) 0.3487 Ultrasonic method (40 min) 0.3437

[0175] The above results show that the content of saikosaponin b1 extracted by reflux method and ultrasonic for 30 and 40 min has little difference, so the test sample extraction method of ultrasonic for 30 min which is convenient and environmentally friendly is used.

[0176] Example 7

[0177] The test sample treatment method is determined as follows: an appropriate amount of the contents of Suguanning capsules is finely ground, 5.0 g of which is weighed, 40 mL of methanol is added, ultrasonic treatment is performed for 30 min, filtration is performed, the filtrate is evaporated to dryness, the residue is dissolved with 15 mL of sodium hydroxide test solution in portions, the solution is extracted with 40 mL of water-saturated n-butanol by shaking, the n-butanol liquid after extraction is washed with 100 mL of n-butanol-saturated ammonia test solution, the ammonia washing liquid is discarded, and the n-butanol liquid is washed with water saturated with n-butanol twice, each time with 50 mL, the n-butanol liquid is taken, evaporated to dryness, the residue is dissolved with methanol, and the volume is made up to 2 mL in a volumetric flask, shaken well, filtered through a 0.45 μm microporous filter, and the filtrate is taken, i.e. the test sample solution, is obtained.

[0178] Preparation of negative control solution: the medicinal materials except for Bupleurum chinense in the prescription are taken according to the prescription proportion, and the Bupleurum chinense negative control solution is prepared according to the method;

[0179] The Bupleurum chinense saikosaponin b1 control solution is prepared according to the preparation method of the test sample solution;

[0180] 5 μL of the above Bupleurum chinense saikosaponin b1 control solution, test sample solution, Bupleurum chinense negative control solution and methanol solvent are injected into the high performance liquid chromatograph, and the results are shown in Table 8. Figures 18-20 ;

[0181] Linear relationship detection:

[0182] 20.02 mg of Bupleurum chinense saikosaponin b1 control solution with a purity of 98% is weighed, and methanol is added to prepare a control solution with a concentration of 1.9619 mg / mL; 3.5 mL, 2 mL, 1.5 mL, 1 mL and 0.5 mL of the Bupleurum chinense saikosaponin b1 control solution with a mass concentration of 1.9619 mg / mL are taken, methanol is added to make the volume of the volumetric flask 5 mL, and Bupleurum chinense saikosaponin b1 control solutions with concentrations of 1.3733 mg / mL, 0.7847 mg / mL, 0.5885 mg / mL, 0.3923 mg / mL and 0.1961 mg / mL are prepared;

[0183] The peak area is determined under the chromatographic conditions for determination of the content of Bupleurum chinense saikosaponin b1 in Suguanning capsules, the peak area is taken as the vertical coordinate, and the mass concentration is taken as the horizontal coordinate, a standard curve is drawn, and linear regression is performed; the data details are shown in Table 9, and the standard curve graph is shown in Figure 21The linear relationship between the concentration of saikosaponin b1 reference substance and the peak area was determined when the mass concentration of saikosaponin b1 was in the range of 0.1961-1.9619 mg / mL, and the regression equation was Y=1460.2X+9.8946, R=0.9996;

[0184] Table 9 Linear relationship data table between the concentration of saikosaponin b1 reference substance and the peak area

[0185] Concentration (mg / ml) Peak area 1.9619 2855.500 1.3733 2046.800 0.7847 1168.200 0.5885 855.940 0.3923 554.750 0.1961 312.590

[0186] Precision test:

[0187] 5 μL of saikosaponin b1 reference substance solution with a mass concentration of 0.3956 mg / mL was successively injected for 6 times, the peak area was recorded, and the RSD was calculated, as shown in Table 10;

[0188] The RSD of the peak area integral value of saikosaponin b1 was 0.91%, indicating that the precision of the instrument was good;

[0189] Table 10 Precision test data table

[0190]

[0191]

[0192] Stability test:

[0193] 5 μL of the test sample solution and saikosaponin b1 reference substance solution were taken, respectively, and the peak area was determined according to the method for determining the content of acetaminophen in Sujianning capsules at 0 h, 1 h, 2 h, 4 h, 6 h, 8 h, 10 h, 12 h, 24 h, 34 h and 36 h after preparation. The RSD of the peak area of the reference substance solution was 1.30%, and the RSD of the peak area of saikosaponin b1 in the test sample solution was 1.73%, indicating that saikosaponin b1 in the test sample solution and the reference substance solution was stable at room temperature within 36 h;

[0194] Table 11 Stability test data

[0195] Time / h Peak area integration value of reference Peak area integration value of test 0h 1155.400 1011.000 1h 1155.500 1017.200 2h 1160.100 1019.200 4h 1157.400 1022.900 6h 1160.700 1022.300 8h 1157.100 1023.400 10h 1161.700 1023.300 12h 1162.200 1024.600 24h 1171.600 1047.100 34h 1198.600 1072.300 36h 1193.800 1046.100 STDEV 15.272 17.915 AVERAGE 1166.736 1029.945 RSD 1.30％ 1.73％

[0196] Reproducibility test:

[0197] 5 g of the contents of Sujianning capsules was weighed in total 6 portions, and the test sample solution was prepared according to the method for determining the content of Bupleurum in Sujianning capsules. The sample was injected and determined under the same chromatographic conditions, and the results were shown in Table 12. The content of saikosaponin b1 was 0.2674 mg / g, and the RSD was 1.50%, respectively, indicating that the method was good in reproducibility;

[0198] Table 12 Reproducibility test data table

[0199]

[0200] Add sample recovery rate test:

[0201] Take the known content of 0.2674 mg / g of the speed sense Ning capsule (batch number 201103) content sample 9, 2.5 g each, accurately weighed, respectively, with the sample of 0.5, 1, 1.5 times the amount of saikosaponin b1 control, according to the content determination method of saikosaponin b1 in speed sense Ning capsule under the chromatographic conditions of determination, record the peak area, calculate the recovery rate, the results are shown in table 13, the recovery rate meets the requirements;

[0202] Table 13 data table of adding sample recovery rate test

[0203]

[0204]

[0205] Content verification of saikosaponin b1 in speed sense Ning capsule:

[0206] In order to eliminate the error of test results caused by the difference of instrument, Agilent 1260 instrument, different operation and ZXRBAX Eclipse Plus C18 chromatographic column were used under the same system conditions, the content of known content of speed sense Ning capsule sample was determined in parallel, the results were shown in table 14;

[0207] Table 14 results of determination of saikosaponin b1 content under different conditions

[0208] Test name Bupleurum saikoside b1 / mg·g -1 ]] Routine conditions 0.2674 Different instruments 0.2693 Different operators 0.2679 Different chromatographic columns 0.2754

[0209] The content determination value was basically consistent, which could exclude the error caused by instrument, personnel and chromatographic column change, and proved that the saikosaponin b1 was stable in nature;

[0210] According to the content determination method of saikosaponin b1 in speed sense Ning capsule in the quality standard of the product, the sample solution was prepared, 3 batches of different speed sense Ning capsule with batch number of 201103, 200203 and 201001 and 5 batches of speed sense Ning capsule extract with batch number of 201204, 201205, 201206, 201207 and 201208 were taken, and the content was calculated, the results were shown in table 15;

[0211] Table 15 content of saikosaponin b1 in different batches of speed sense Ning capsule

[0212]

[0213]

[0214] The above merely describes preferred embodiments of the present application, and is not intended to limit the present application. Any simple modification, change and equivalent variation of the above embodiments according to the technical essence of the present application are still within the protection scope of the technical scheme of the present application.

Claims

1. A method of testing Sengenol capsules, characterized in that, The Sujin Ning capsule is a compound preparation composed of 320 g of Herba Lycopi, 320 g of Radix Bupleuri, 240 g of Folium Isatidis, 240 g of Flos Lonicerae, 8 g of Bovis Calculus, 100 g of acetaminophen and 1.2 g of chlorpheniramine maleate, and comprises the following steps: S1, taking the content of the Sujin Ning capsule for preparing the test sample solution, the control sample solution and the negative control sample solution; including determining the acetaminophen in the Sujin Ning capsule, S1 further comprises the following steps: S101, taking the finely ground content of the Sujin Ning capsule and adding it into methanol for dissolution, ultrasonic treatment, after the solution is cooled to room temperature, shake well and filter, the obtained filtrate is diluted to 50 mL in a volumetric flask, 1 mL of the solution in the 50 mL volumetric flask is taken, and then diluted to 10 mL in a volumetric flask, shake well again and filter to obtain the filtrate, which is the acetaminophen test sample solution; S102, taking the acetaminophen control sample and adding it into methanol to prepare an acetaminophen control sample stock solution in a 10 mL volumetric flask, and then taking 1 mL of the acetaminophen control sample stock solution and adding it into methanol to prepare an acetaminophen control sample solution in a 5 mL volumetric flask; S103, taking the content of the Sujin Ning capsule without acetaminophen and preparing the acetaminophen negative control sample solution according to the steps in S101; including determining the Radix Bupleuri in the Sujin Ning capsule, S1 further comprises the following steps: S111, taking 5.0 g of the finely ground content of the Sujin Ning capsule and adding it into 40 mL of methanol for dissolution, ultrasonic treatment for 30 min and then filtering, the filtrate is evaporated to dryness, the obtained residue is dissolved with 15 mL of NaOH test solution, the solution is extracted with 40 mL of water-saturated n-butanol solution, the extracted n-butanol solution is washed with 100 mL of n-butanol-saturated ammonia test solution, the n-butanol-saturated ammonia washing solution is discarded, and then the n-butanol solution is washed with 50 mL of n-butanol-saturated water solution for two times, the n-butanol solution is taken, evaporated to dryness, and the obtained residue is diluted to 2 mL in a volumetric flask with methanol, shaken well, filtered through a microporous filter membrane with a pore size of 0.45 μm, and the filtrate is obtained, which is the Radix Bupleuri test sample solution; S112, taking 10.02 mg, 10.04 mg and 10.07 mg of saikosaponin a, saikosaponin b1 and saikosaponin b2 control samples respectively, adding them into methanol to prepare saikosaponin a, saikosaponin b1 and saikosaponin b2 control sample stock solutions with mass concentrations of 0.9820 mg / mL, 0.9839 mg / mL and 0.9869 mg / mL respectively in 10 mL volumetric flasks, and then taking 1 mL of each of the saikosaponin a, saikosaponin b1 and saikosaponin b2 control sample stock solutions and adding them into methanol to prepare a saikosaponin control sample mixed solution in a 10 mL volumetric flask; S113, taking the content of the Sujin Ning capsule without Radix Bupleuri and preparing the Radix Bupleuri negative control sample solution according to the preparation method of the Radix Bupleuri test sample solution; S2, specificity detection: taking 5 μL of each of the test sample solution, the control sample solution, the negative control sample solution and the methanol solution in S1 and injecting them into a liquid chromatograph to obtain the solution chromatograms corresponding to each solution; S3, linear relationship detection: different volumes of test solution were taken, then methanol was added for constant volume, different mass concentrations of solution were prepared, the liquid chromatograms of different mass concentrations of solution prepared were determined, the peak area was taken as the vertical coordinate, the mass concentration was taken as the horizontal coordinate for linear regression and standard curve was drawn; S4, precision detection: 5 μL of the control solution prepared in S1 was taken, 6 times of continuous injection were performed, and the liquid chromatograms were determined; S5, stability detection: 5 μL of the test solution and the control solution obtained in S1 were taken, and the liquid chromatograms were determined at different times after preparation; S6, reproducibility detection: 0.1 g of the contents of Sujingning capsules was taken for 6 times, and the test solution was prepared, and the liquid chromatograms were determined; S7, sample addition recovery rate detection: 0.05 g of 9 samples of Sujingning capsule contents was taken, 0.5, 1 and 1.5 times of the control substance was added to the sample, and the liquid chromatograms were determined; The liquid chromatography detection conditions of paracetamol in Sujingning capsules were as follows: Agilent 1260 type high performance liquid chromatograph was used, the size of Agilent Eclipse SB-C18 chromatographic column was 4.6*250 mm, 5 μm, the column temperature was set to 25 ℃, the detection wavelength was set to 240 nm, the flow rate was set to 1.0 mL / min; the injection amount was set to 5 μL, and the mobile phase was set to water:methanol solution, the ratio was 80:20; The liquid chromatography detection conditions of Bupleurum in Sujingning capsules were as follows: Agilent 1260 high performance liquid chromatograph, Agilent ZORBAX SB-C18 chromatographic column, the mobile phase was 0.1% phosphoric acid aqueous solution-acetonitrile solution with a ratio of 64:36, the detection wavelength was 210 nm, the column temperature was 25 ℃, the flow rate of the mobile phase was 1.0 mL / min, and the injection amount was 5 μL.

2. A method of testing Sufentanil Capsules as claimed in claim 1, wherein, The mass of the contents of Sujingning capsules in S101 was 0.1 g; the ultrasonic conditions were as follows: ultrasonic treatment for 30 min under the conditions of power 150 W and frequency 40 KHz.

3. The method of claim 1, wherein the Sufentanil Capsule is Sufentanil Capsule. The mass of the control substance in S102 was 10.01 mg, the mass concentration of the control substance stock solution was 0.9910 mg / mL, 0.35 mL of the control substance stock solution was taken, and the mass concentration of the control solution was 0.0694 mg / mL.

4. The method of claim 1, wherein the Sufentanil Capsule is Sufentanil Capsule. The range of different volumes in S3 was 0.5-3.5 mL, when paracetamol in Sujingning capsules was detected, the concentration of the control solution was 0.9900 mg / mL.

5. The method of claim 1, wherein the Sufentanil Capsule is Sufentanil Capsule. The mass concentration of the control solution when the control substance solution was paracetamol control substance solution in S4 was 0.0693 mg / mL; the mass concentration of the control solution when the control substance solution was Bupleurum control substance solution in S4 was 0.3956 mg / mL.

6. The method of claim 1, wherein the Sufentanil Capsule is Sufentanil Capsule. The different times in S5 included 0 h, 2 h, 4 h, 6 h, 8 h, 10 h, 12 h, 18 h, 24 h, 28 h, 32 h and 36 h.

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