Quality control method of traditional Chinese medicine composition preparation

Abstract

The invention relates to the technical field of analysis, and particularly discloses a quality control method of a traditional Chinese medicine composition preparation, the traditional Chinese medicine composition preparation is prepared from radix scutellariae, radix bupleuri, rheum officinale, fructus aurantii, herba artemisiae scopariae, polygonum cuspidatum, gardenia, radix paeoniae alba, radix aucklandiae and other bulk drugs, and the quality control method comprises an identification method and a content determination method. The quality control method provided by the invention has good specificity, separation degree, precision, accuracy, stability, durability and repeatability, and can effectively control the product quality of the traditional Chinese medicine composition preparation.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of analytical techniques, in particular to a quality control method of a traditional Chinese medicine composition preparation. BACKGROUND

[0002] The traditional Chinese medicine composition is composed of Huangqi, Chaihu, Dahuang, Jisheng, Yincheng, Huzhang, Zhizi, Jinqiancao, Baishao, Jiangbanxia, Shengjiang, etc. The combination of all the medicines can resolve the heat of Shao Yang and clear the heat of Yang Ming, and has the effects of clearing heat and dampness, soothing liver and gallbladder, and relieving stomach pain.

[0003] With the rapid development of traditional Chinese medicine, the types of traditional Chinese medicine used in clinical practice are increasing, and the determination and identification of the content of traditional Chinese medicine components have become an important work. Accurate determination of the content of components can ensure the quality and efficacy of traditional Chinese medicine, and improve the safety of drug use.

[0004] Although there are reports that thin layer chromatography and high performance liquid chromatography can be used to determine the active ingredients of Chaihu, Jisheng, Baishao, Zhizi and other medicinal materials, there is no literature or report disclosing the identification and content determination method of multiple components in the traditional Chinese medicine composition preparation.

[0005] The pharmaceutical composition contains dozens or even hundreds of bioactive components. For a listed drug, how to comprehensively control the quality of the traditional Chinese medicine composition and ensure the safety of drug use is an urgent problem to be solved. SUMMARY

[0006] The purpose of the present application is to provide a quality control method of a traditional Chinese medicine composition preparation, which includes identification method and content determination method, and can effectively and quickly comprehensively control the quality of the traditional Chinese medicine composition preparation.

[0007] In order to achieve the above-mentioned purpose of the application, the following technical solutions are adopted:

[0008] A quality control method of a traditional Chinese medicine composition preparation, the raw materials of the traditional Chinese medicine composition preparation include the following components in weight parts: Huangqi 69-205 parts, Chaihu 69-205 parts, Dahuang 52-155 parts, Jisheng 69-205 parts, Yincheng 69-205 parts, Huzhang 86-255 parts, Zhizi 69-205 parts, Jinqiancao 171-515 parts, Baishao 69-205 parts, Muxiang 69-205 parts, Jiangbanxia 52-155 parts, and Shengjiang 17-52 parts,

[0009] The quality control method of the traditional Chinese medicine composition preparation includes identification method and content determination method,

[0010] The identification method includes at least one of the following (1) to (4) identification methods:

[0011] (1) with the reference material of Radix Bupleuri as the control, thin layer chromatography with chloroform-methanol-water (30:8:1, by volume) as the developing agent;

[0012] (2) with the reference materials of Fructus Gardeniae and Radix Paeoniae Alba as the control, thin layer chromatography with chloroform-ethyl acetate-methanol-ammonia (4:4:5:1, by volume) as the developing agent;

[0013] (3) with the reference material of Radix Aucklandiae as the control, thin layer chromatography with petroleum ether (30-60℃)-ethyl formate-ethyl formate upper layer solution (15:5:1, by volume) as the developing agent;

[0014] (4) high performance liquid chromatography with naringin reference substance as the control, acetonitrile-0.1% phosphoric acid solution (18:82) as the mobile phase, and detection wavelength of 283 nm;

[0015] The content determination method includes the content determination method of baicalin in Radix Scophulariae and the content determination method of emodin in Radix et Rhizoma Rhei and Polygonum Cuspidatum.

[0016] Further, the identification method of item (1) includes the following steps: taking the traditional Chinese medicine composition preparation, extracting with methanol, removing impurities with chloroform, extracting with water-saturated n-butanol, combining the n-butanol layers, evaporating to dryness, dissolving the residue with methanol to obtain a test sample solution; taking the reference material of Radix Bupleuri, extracting with methanol under ultrasonic, filtering, and taking the filtrate as a reference material solution; taking the test sample solution and the reference material solution, spotting them on the same silica gel G thin layer plate, developing with chloroform-methanol-water as the developing agent, taking out, drying, spraying with 2% p-dimethylaminobenzaldehyde 40% sulfuric acid ethanol solution, and heating until the spots are clear; in the test sample chromatogram, the same color spots appear at the positions corresponding to the reference material chromatogram.

[0017] The identification method of item (2) includes the following steps:

[0018] Taking the traditional Chinese medicine composition preparation, extracting with methanol under ultrasonic, filtering, and taking the filtrate as a test sample solution; taking the reference material of Fructus Gardeniae, extracting with methanol under heating reflux, filtering, evaporating the filtrate to dryness, dissolving the residue with methanol to obtain a reference material solution of Fructus Gardeniae; taking the reference material of Radix Paeoniae Alba, extracting with methanol under ultrasonic, filtering, evaporating the filtrate to dryness, dissolving the residue with methanol to obtain a reference material solution of Radix Paeoniae Alba; taking the test sample solution and the two reference material solutions, spotting them on the same silica gel G thin layer plate, developing with chloroform-ethyl acetate-methanol-ammonia (4:4:5:1, by volume) as the developing agent, taking out, drying, spraying with 1% vanillin sulfuric acid solution, and heating until the spots are clear; in the test sample chromatogram, the same color spots appear at the positions corresponding to the reference material chromatograms of Fructus Gardeniae and Radix Paeoniae Alba.

[0019] The identification method of item (3) includes the following steps:

[0020] Take the Radix Aucklandiae control drug, add chloroform, ultrasonic treatment, filter, the filtrate as the control drug solution, take the test solution under (2) and the Radix Aucklandiae control drug solution, respectively, point on the same silica gel G thin layer plate, with volume ratio 15:5:1 of petroleum ether (30-60℃)-ethyl formate-methanol upper solution as developing agent, develop, take out, air dry, spray with 1% vanillin sulfuric acid solution, heat to clear spots, the test sample chromatogram, in the corresponding position with the control drug chromatogram, shows the same color spots;

[0021] The identification method of (4) includes the following steps:

[0022] Take the traditional Chinese medicine composition preparation, add methanol ultrasonic, filter, evaporate the filtrate, dissolve the residue with water, remove impurities with ethyl acetate, then extract with water-saturated n-butanol, combine the n-butanol layers, evaporate, dissolve the residue in methanol, filter, and the filtrate is the test sample solution; additionally, take the appropriate naringin control product, add methanol to prepare a solution containing 80 μg per 1 ml, which is the control sample solution, use acetonitrile-0.1% phosphoric acid solution (18:82) as the mobile phase; the detection wavelength is 283 nm; the column temperature is 25℃; precisely take 10 μl of the control sample solution and the test sample solution respectively, inject into the high performance liquid chromatograph, and the test sample chromatogram should present a chromatographic peak consistent with the retention time of the control sample chromatographic peak.

[0023] Further, the identification method of (1) includes the following steps:

[0024] For the thin layer identification of Bupleurum in the preparation, take the traditional Chinese medicine composition preparation, weigh 2-4 g, add 10-50 ml of methanol, ultrasonic, filter, evaporate the filtrate, dissolve the residue with water, extract 1-5 times with chloroform, discard the chloroform liquid, then extract 1-5 times with water-saturated n-butanol, combine the n-butanol layers, wash with 3% sodium hydroxide solution, evaporate the n-butanol liquid, dissolve the residue in methanol, which is the test sample solution; additionally, take 0.1-1 g of Bupleurum control drug powder, add 10-50 ml of methanol, ultrasonic treatment, filter, concentrate the filtrate, which is the control drug solution, take the control drug solution and the test sample solution, respectively, point on the same silica gel G thin layer plate, with volume ratio 30:8:1 of chloroform-methanol-water as developing agent, develop, take out, air dry, spray with 2% p-dimethylaminobenzaldehyde 40% sulfuric acid ethanol solution, heat to clear spots, the test sample chromatogram, in the corresponding position with the control drug chromatogram, shows the same color spots;

[0025] The identification method of (2) includes the following steps:

[0026] For the preparation of Radix Paeoniae Alba, Gardenia thin layer identification, take the traditional Chinese medicine composition preparation, 2-4 g, add methanol 10-50 ml, ultrasonic, filter, the filtrate as test solution; another gardenia control drug 0.1-0.8 g, add methanol 10-50 ml, reflux, filter, the filtrate is evaporated to dryness, the residue is dissolved with methanol, as the gardenia control drug solution; again take Radix Paeoniae Alba control drug 0.1-0.8 g, add methanol 10-50 ml, ultrasonic treatment, filter, the filtrate is evaporated to dryness, the residue is dissolved with methanol, as Radix Paeoniae Alba control drug solution, take the test solution and gardenia, Radix Paeoniae Alba two control drug solution, respectively, on the same silica gel G thin layer plate, with volume ratio of 4:4:5:1 chloroform-ethyl acetate-methanol-ammonia as developing agent, development, take out, dry, spray with 1% vanillin sulfuric acid solution, heated to the spots color clear, the test chromatogram, in the corresponding position with gardenia control drug and Radix Paeoniae Alba control drug chromatography, show the same color spots;

[0027] The identification method of item (3) comprises the following steps:

[0028] Take the control drug of Aucklandia 0.1-0.3 g, add chloroform 2-10 ml, ultrasonic, filter, the filtrate as control drug solution, take the test solution and control drug solution under item (2), respectively, on the same silica gel G thin layer plate, with volume ratio of 15:5:1 petroleum ether (30-60℃)-ethyl formate-methanol upper solution as developing agent, development, take out, dry, spray with 1% vanillin sulfuric acid solution, heated to the spots color clear, the test chromatogram, in the corresponding position with control drug chromatography, show the same color spots;

[0029] The identification method of item (4) comprises the following steps:

[0030] Take the traditional Chinese medicine composition preparation, 0.5-2 g, add methanol ultrasonic, filter, the filtrate is evaporated to dryness, the residue is dissolved with water, shaken with ethyl acetate, discard the ethyl acetate liquid, then shaken with water saturated n-butanol, combine the n-butanol layer, evaporate to dryness, the residue is dissolved with methanol, transfer to a 25 ml volumetric flask, dilute to the mark with methanol, shake well, filter, the filtrate as test solution; another naringin control 0.1-0.8 g, add methanol to prepare a solution containing 80 μg per 1 ml, as the control solution, with octadecylsilane bonded silica gel as the filler; with acetonitrile-0.1% phosphoric acid solution (18:82) as mobile phase; detection wavelength is 283 nm; column temperature is 25℃; respectively, 10 μl of the control solution and the test solution are precisely taken and injected into the high performance liquid chromatograph. The test chromatogram should present a chromatographic peak consistent with the retention time of the control chromatographic peak.

[0031] Further, the content determination method of Scutellaria includes the following steps:

[0032] Preparation of the control solution: accurately weigh the baicalin control sample, add 70% methanol to prepare the control solution;

[0033] Preparation of the test solution: accurately weigh the traditional Chinese medicine composition preparation, add 70% ethanol, ultrasonic, filter, and then filter the solution to obtain the test solution;

[0034] HPLC detection: the chromatographic column is C18, the mobile phase is composed of methanol and 0.1% phosphoric acid solution with a volume ratio of 45:55, the detection wavelength is 275-285 nm, the column temperature is 20-30℃, and the flow rate is 0.8-1.2 ml / min;

[0035] Determination method: accurately take 10 μl of the control solution and the test solution respectively, inject into the high performance liquid chromatograph, and then determine to obtain the content of baicalin.

[0036] Further, the content determination method of the baicalin comprises the following steps:

[0037] Preparation of the control solution: take the baicalin control sample, accurately weigh, add 70% methanol to prepare a solution containing 20 μg per 1 ml, and shake to obtain the control solution;

[0038] Preparation of the test solution: accurately weigh 0.1 g of the content of the traditional Chinese medicine composition preparation, accurately add 25 ml of 70% ethanol, filter, take 2 ml of the filtrate, dilute to the scale with 70% ethanol, shake, filter, and then take the filtrate to obtain the test solution;

[0039] HPLC detection: the chromatographic column is C18, the mobile phase is composed of methanol and 0.1% phosphoric acid solution with a volume ratio of 45:55, the detection wavelength is 275-285 nm, the column temperature is 20-30℃, and the flow rate is 0.8-1.2 ml / min;

[0040] Determination method: accurately take 10 μl of the control solution and the test solution respectively, inject into the high performance liquid chromatograph, and then determine to obtain the content of baicalin.

[0041] The traditional Chinese medicine composition preparation contains 3.5 mg-14.1 mg of baicalin calculated as baicalin (C 21 H 18 O 11 ) per pill.

[0042] Further, the content determination method of the baicalin comprises the following steps:

[0043] Preparation of the control solution: take the baicalin control sample, accurately weigh, add 70% methanol to prepare a solution containing 20 μg per 1 ml, and shake to obtain the control solution;

[0044] Preparation of the test solution: accurately weigh 0.1 g of the content of the traditional Chinese medicine composition preparation, accurately add 25 ml of 70% ethanol, filter, take 2 ml of the filtrate, dilute to the scale with 70% ethanol, shake, filter, and then take the filtrate to obtain the test solution;

[0045] HPLC detection: the chromatographic column is C18, the mobile phase is composed of acetonitrile, methanol and 0.10% phosphoric acid solution with a volume ratio of 41-43:23:34-36, the detection wavelength is 252-256nm, the column temperature is 25-35℃, and the flow rate is 0.8-1.2ml / min;

[0046] Determination method: 10ul of the control solution and the test solution are precisely taken respectively, injected into the liquid chromatograph, and determined, and then the content of emodin in the test solution is calculated.

[0047] Further, the content determination method of the rhubarb and giant knotweed comprises the following steps:

[0048] Preparation of the control solution: emodin control sample is precisely weighed, methanol is added to prepare a solution containing 37ug per 1ml, and then the solution is obtained after shaking;

[0049] Preparation of the test solution: the content of the traditional Chinese medicine composition preparation is finely ground, 0.5g is precisely weighed, 10% hydrochloric acid methanol solution is precisely added, the weight is determined, 80℃ water bath heating reflux is carried out for 30 minutes, the weight is determined again, the weight loss is made up with methanol, and then the solution is obtained after shaking, filtering, precisely taking 4ml of the filtrate, placing in a 10ml volumetric flask, precisely adding 2% sodium hydroxide solution 2ml, diluting to the mark with methanol, shaking, filtering, and then the filtrate is obtained;

[0050] HPLC detection: the chromatographic column is C18, the mobile phase is composed of acetonitrile, methanol and 0.10% phosphoric acid solution with a volume ratio of 41-43:23:34-36, the detection wavelength is 252-256nm, the column temperature is 25-35℃, and the flow rate is 0.8-1.2ml / min;

[0051] Determination method: 10ul of the control solution and the test solution are precisely taken respectively, injected into the liquid chromatograph, and determined, and then the content of emodin in the test solution is calculated.

[0052] The traditional Chinese medicine composition preparation in the application contains 0.8mg-3.2mg of rhubarb and giant knotweed in terms of emodin (C 15 H 10 O5).

[0053] Further, the traditional Chinese medicine composition preparation in the application is in the form of capsules, tablets, pills, oral liquids, granules or powders.

[0054] Further, the active ingredients of the traditional Chinese medicine composition are prepared by the following steps:

[0055] A, ginger pinnate leaf is weighed, crushed, and passed through a 100 mesh sieve for standby use;

[0056] B. Weigh the Fructus Aurantii and ginger, add 5-9 times the amount of water, and extract the volatile oil for 8-12 hours. Collect and separate the volatile oil. Distill the water solution and reserve it for later use;

[0057] C. Weigh the Radix Scrophulariae, Radix Paeoniae Alba, Herba Artemisiae Scopariae, and Fructus Gardeniae, and decoct them in water for 2-4 times. Extract for 1-3 hours in the first time, and extract for 1-3 hours in the second, third, and fourth times, respectively. Add 7-10 times the amount of water each time. Combine the extraction liquid with the volatile oil extraction liquid obtained in step B, and concentrate under reduced pressure to the extract with a relative density of 1.25±0.05 (60°C). Reserve it for later use;

[0058] D. Weigh the Radix Bupleuri, Radix Aucklandiae, Radix et Rhizoma Rhei, Polygonum cuspidatum, and Herba Lysimachiae, and extract them in 60-80% ethanol for 2-4 times. Add 10-14 times the amount of extraction liquid in the first time, and extract for 2-4 hours. Add 8-12 times the amount of extraction liquid in the second and third times, and extract for 1-3 hours each time. Filter the extraction liquid, and concentrate it to a clear extract. Combine it with the water extraction extract obtained in step C, mix well, dry, and pulverize. Reserve it for later use;

[0059] The fine powder obtained in step A, the volatile oil obtained in step B, and the dry powder obtained in step D collectively constitute the active component of the traditional Chinese medicine composition.

[0060] Further, the preparation process of the capsule is as follows:

[0061] A. Weigh the Rhizoma Pinelliae, pulverize it, pass it through a 100-mesh sieve, and reserve it for later use;

[0062] B. Weigh the Fructus Aurantii and ginger, add 5-9 times the amount of water, and extract the volatile oil for 8-12 hours. Collect and separate the volatile oil. Distill the water solution and reserve it for later use;

[0063] C. Weigh the Radix Scrophulariae, Radix Paeoniae Alba, Herba Artemisiae Scopariae, and Fructus Gardeniae, and decoct them in water for 2-3 times. Extract for 1-3 hours in the first time, and extract for 1-3 hours in the second and third times, respectively. Add 7-10 times the amount of water each time. Filter the extraction liquid, and combine it with the volatile oil extraction liquid obtained in step B. Concentrate under reduced pressure to the extract with a relative density of 1.25±0.05 (60°C). Reserve it for later use;

[0064] D. Weigh the Radix Bupleuri, Radix Aucklandiae, Radix et Rhizoma Rhei, Polygonum cuspidatum, and Herba Lysimachiae, and extract them in 60-80% ethanol for 2-4 times. Add 10-14 times the amount of extraction liquid in the first time, and extract for 2-3 hours. Add 8-12 times the amount of extraction liquid in the second and third times, and extract for 1-3 hours each time. Filter the extraction liquid, and concentrate it to a clear extract. Combine it with the water extraction extract obtained in step C, mix well, dry, and pulverize. Reserve it for later use;

[0065] E. Mix the fine powder obtained in step A and the extract powder obtained in step D, mix well, dry, and mix with the volatile oil obtained in step B. Fill it into capsules to obtain the product.

[0066] Preferably, the preparation process of the capsule is as follows:

[0067] A. Take the pinellia ternate, crush, pass through a 100 mesh sieve, and reserve;

[0068] B. Take the fructus aurantii and raw ginger, add 6-8 times the amount of water, and extract the volatile oil for 9-11 hours. Collect and separate the volatile oil. Distill the water solution, and reserve;

[0069] C. Take the radix scrophulariae, radix paeoniae alba, herba artemisiae, and fructus gardeniae, and decoct with water for 2-3 times. Extract for 2 hours the first time, and extract for 1.5 hours the second and third times. Add 8-10 times the amount of water each time. Combine the extract with the volatile oil extract from step B, and concentrate under reduced pressure to an extract with a relative density of 1.25±0.05 (60°C). Reserve;

[0070] D. Take the radix bupleuri, radix saussureae lappae, radix et rhizoma rhei, rhizoma polygoni cuspidati, and herba lycopodii, and extract with 60-80% ethanol for 3 times. Add 11-13 times the amount of ethanol the first time and extract for 2-3 hours. Add 9-11 times the amount of ethanol the second and third times and extract for 1-3 hours each time. Filter the extract, and concentrate to a clear extract. Combine the extract with the water extract from step C, mix, dry, and pulverize. Reserve.

[0071] E. Mix the fine powder from step A and the dry extract powder from step D. Mix the volatile oil from step B with silica, mix, and fill into capsules to obtain the product.

[0072] Further, the raw materials of the traditional Chinese medicine composition preparation include the following components in the following amounts by weight: radix scrophulariae 90-150 parts, radix bupleuri 90-150 parts, radix et rhizoma rhei 80-120 parts, fructus aurantii 90-150 parts, herba artemisiae 90-150 parts, rhizoma polygoni cuspidati 120-200 parts, fructus gardeniae 90-150 parts, herba lycopodii 250-400 parts, radix paeoniae alba 90-150 parts, radix saussureae lappae 90-150 parts, pinellia ternate 80-120 parts, and raw ginger 25-40 parts.

[0073] Further, the raw materials of the traditional Chinese medicine composition preparation include the following components in the following amounts by weight: radix scrophulariae 90-150 parts, radix bupleuri 90-150 parts, radix et rhizoma rhei 80-120 parts, fructus aurantii 90-150 parts, herba artemisiae 90-150 parts, rhizoma polygoni cuspidati 120-200 parts, fructus gardeniae 90-150 parts, herba lycopodii 250-400 parts, radix paeoniae alba 90-150 parts, radix saussureae lappae 90-150 parts, pinellia ternate 80-120 parts, and raw ginger 25-40 parts.

[0074] Further, the raw materials of the traditional Chinese medicine composition preparation include the following components in the following amounts by weight: radix scrophulariae 90-150 parts, radix bupleuri 90-150 parts, radix et rhizoma rhei 80-120 parts, fructus aurantii 90-150 parts, herba artemisiae 90-150 parts, rhizoma polygoni cuspidati 120-200 parts, fructus gardeniae 90-150 parts, herba lycopodii 250-400 parts, radix paeoniae alba 90-150 parts, radix saussureae lappae 90-150 parts, pinellia ternate 80-120 parts, and raw ginger 25-40 parts.

[0075] Further, the raw materials of the traditional Chinese medicine composition preparation include the following components in weight parts: 150 parts of Huangqi, 70 parts of Chaihu, 90 parts of Dahuang, 90 parts of Zhike, 150 parts of Yincheng, 100 parts of Huzhang, 110 parts of Zhizi, 500 parts of Jinqiancao, 80 parts of Baishao, 200 parts of Muxiang, 110 parts of Jiangbanxia, and 20 parts of Shengjiang.

[0076] The quality control method of the traditional Chinese medicine composition preparation protected by the application can effectively and quickly identify the corresponding traditional Chinese medicine ingredients in the traditional Chinese medicine composition preparation of the application by using thin layer chromatography, with clear spots, good separation degree, and good repeatability, with naringin as a control and by using high performance liquid chromatography, with good separation degree and good repeatability. Meanwhile, the application establishes a baicalin and emodin high performance liquid content determination method for Huangqi and Dahuang and Huzhang in the traditional Chinese medicine composition preparation, with good specificity, good precision, good repeatability, good accuracy, good stability, good durability, and common use with the thin layer chromatography, effectively and quickly comprehensively controlling the quality of the traditional Chinese medicine composition preparation. BRIEF DESCRIPTION OF DRAWINGS

[0077] Figure 1 is a thin layer chromatogram (daylight) of Chaihu thin layer identification in the application;

[0078] Figure 2 is a thin layer chromatogram (ultraviolet) of Chaihu thin layer identification in the application;

[0079] Figure 3 is a thin layer chromatogram of Baishao and Zhizi thin layer identification in the application;

[0080] Figure 4 is a thin layer chromatogram of Muxiang thin layer identification in the application;

[0081] Figure 5 is a specificity chromatogram of Huangqi in the specificity test in the application;

[0082] Figure 6 is a standard curve of baicalin in the linearity investigation in the application;

[0083] Figure 7 is a baicalin control chromatogram in the application;

[0084] Figure 8 is a specificity chromatogram of Dahuang and Huzhang in the specificity test in the application;

[0085] Figure 9 is a standard curve of emodin in the linearity investigation in the application;

[0086] Figure 10 Reference chromatogram of emodin in the present application. DETAILED DESCRIPTION

[0087] In order to make the objectives, technical solutions and advantages of the present application clearer, the present application will be further described in detail below with examples. It should be understood that the specific examples described herein are only used to explain the present application and not to limit the present application.

[0088] Identification

[0089] The present application adopts thin layer chromatography identification method to establish the identification method of Bupleurum, Gardenia, Radix Paeoniae Alba and Radix Aucklandiae, and adopts liquid chromatography method to establish the identification method of naringin in Fructus Aurantii. The identification methods are continuously optimized and improved during the clinical test. The capsule prepared according to Example 1 is subjected to identification and content determination test, and the thin layer chromatography identification results of each medicine in the prescription are as follows:

[0090] 1. Materials

[0091] Traditional Chinese medicine composition preparation: the medicine capsule prepared according to Example 1, Shijiazhuang Yiling Pharmaceutical Co., Ltd., batch numbers 20080601, 20080602 and 20080603.

[0092] 2. Identification test

[0093] (1) Thin layer identification of Bupleurum:

[0094] The preparation method of the test solution was once "2g of the product was taken, 25ml of methanol was added, ultrasonic treatment was carried out for 20 minutes, filtration was carried out, the filtrate was evaporated to dryness, 25ml of water was added to dissolve the residue, 25ml of chloroform was extracted twice, 25ml of water-saturated n-butanol was extracted once, the n-butanol layer was evaporated to dryness, 5ml of methanol was added to dissolve the residue", the lower layer solution of chloroform-ethyl acetate-methanol-water (15:40:22:10) placed overnight below 10℃ was used as the developing agent, 2% p-dimethylaminobenzaldehyde 40% sulfuric acid ethanol solution was sprayed, heating was carried out at 60℃, and the results showed that there were no clear spots in the test sample chromatogram and the control medicine chromatogram under daylight and fluorescence. Again, ethyl acetate-ethanol-water (8:2:1), chloroform-methanol-water (13:7:2) lower layer solution placed overnight below 10℃ was used as the developing agent, and the results were observed under daylight and fluorescence, and there were no clear spots in the corresponding positions of the test sample chromatogram and the control medicine chromatogram, and a satisfactory thin layer chromatography method was not established.

[0095] The preparation method of the test sample solution provided by the present application is used to take the traditional Chinese medicine composition preparation, take 3g, add methanol 20ml, ultrasonic treatment for 20 minutes, filter, evaporate the filtrate, add water 25ml to dissolve, shake extraction with chloroform for 3 times, 25ml each time, discard the chloroform liquid, then shake extraction with water-saturated n-butanol for 2 times (15ml, 25ml), combine the n-butanol layer, wash with 3% sodium hydroxide solution for 3 times, 30ml each time, evaporate the n-butanol liquid, add methanol 2ml to dissolve, and take the residue as the test sample solution; another 0.5g of Radix Bupleuri control medicinal material powder is added with methanol 20ml, ultrasonic treatment for 10 minutes, filtration, and the filtrate is concentrated to 5ml as the control medicinal material solution, 5ul of the control medicinal material solution and 10ul of the test sample solution are taken and spotted on the same silica gel G thin layer plate, with chloroform-methanol-water (30:8:1) as the developing agent, sprayed with 2% p-dimethylaminobenzaldehyde 40% sulfuric acid ethanol solution, heated at 60°C until the spots develop clear color, and observed under daylight and ultraviolet light (365nm), the test sample chromatogram and the control medicinal material chromatogram show clear main spots at corresponding positions, and the negative sample has no interference, and the specificity is strong (TLC spectrum as shown in Figure 1 and Figure 2 , wherein, 1 is the Radix Bupleuri negative sample; 2 is the control medicinal material; 3-5 are the test samples).

[0096] (2) Thin layer chromatography identification of Radix Paeoniae Alba and Gardenia:

[0097] The preparation method of the test sample solution provided by the present application is used to take the traditional Chinese medicine composition preparation, take 3g, add methanol 20ml, ultrasonic treatment for 20 minutes, filter, evaporate the filtrate, add water 25ml to dissolve, shake extraction with chloroform for 3 times, 25ml each time, discard the chloroform liquid, then shake extraction with water-saturated n-butanol for 2 times (15ml, 25ml), combine the n-butanol layer, wash with 3% sodium hydroxide solution for 3 times, 30ml each time, evaporate the n-butanol liquid, add methanol 2ml to dissolve, and take the residue as the test sample solution; another 0.5g of Radix Bupleuri control medicinal material powder is added with methanol 20ml, ultrasonic treatment for 10 minutes, filtration, and the filtrate is concentrated to 5ml as the control medicinal material solution, 5ul of the control medicinal material solution and 10ul of the test sample solution are taken and spotted on the same silica gel G thin layer plate, with chloroform-methanol-water (30:8:1) as the developing agent, sprayed with 2% p-dimethylaminobenzaldehyde 40% sulfuric acid ethanol solution, heated at 60°C until the spots develop clear color, and observed under daylight and ultraviolet light (365nm), the test sample chromatogram and the control medicinal material chromatogram show clear main spots at corresponding positions, and the negative sample has no interference, and the specificity is strong (TLC spectrum as shown in

[0098] The preparation method of the test sample solution provided by the present application is used to take the traditional Chinese medicine composition preparation, take 3g, add methanol 20ml, ultrasonic treatment for 20 minutes, filter, evaporate the filtrate, add water 25ml to dissolve, shake extraction with chloroform for 3 times, 25ml each time, discard the chloroform liquid, then shake extraction with water-saturated n-butanol for 2 times (15ml, 25ml), combine the n-butanol layer, wash with 3% sodium hydroxide solution for 3 times, 30ml each time, evaporate the n-butanol liquid, add methanol 2ml to dissolve, and take the residue as the test sample solution; another 0.5g of Radix Bupleuri control medicinal material powder is added with methanol 20ml, ultrasonic treatment for 10 minutes, filtration, and the filtrate is concentrated to 5ml as the control medicinal material solution, 5ul of the control medicinal material solution and 10ul of the test sample solution are taken and spotted on the same silica gel G thin layer plate, with chloroform-methanol-water (30:8:1) as the developing agent, sprayed with 2% p-dimethylaminobenzaldehyde 40% sulfuric acid ethanol solution, heated at 60°C until the spots develop clear color, and observed under daylight and ultraviolet light (365nm), the test sample chromatogram and the control medicinal material chromatogram show clear main spots at corresponding positions, and the negative sample has no interference, and the specificity is strong (TLC spectrum as shown in

[0099] The white peony root and gardenia were used as control drugs, and "3 g of the product was added to 20 ml of methanol, ultrasonic treatment was performed for 20 minutes, and then filtration was performed, and the filtrate was used as the test sample solution". 5 μl of the above test sample solution and the solution of the above two control drugs were spotted on the same silica gel G thin layer plate, chloroform-ethyl acetate-methanol-ammonia (4:4:5:1) was used as the developing agent, 1% vanillin sulfuric acid solution was sprayed, heating was performed at 105°C until the spots were clearly colored, and good thin layer chromatography effect was obtained. The white peony root and gardenia negatives had no interference. The chromatography system can simultaneously identify the white peony root and gardenia, and the method is simple and easy to operate (TLC atlas is shown in Figure 3 , wherein, sample 1 is the white peony root negative; 2 is the white peony root control drug; 3-5 are the test samples; 6 is the gardenia control drug; 7 is the gardenia negative.

[0100] (3) Thin layer chromatography identification of costus:

[0101] The control drug of costus was used as a control, and "3 g of the product was added to 20 ml of methanol, ultrasonic treatment was performed for 20 minutes, and then filtration was performed, and the filtrate was used as the test sample solution". Trichloromethane-cyclohexane (5:1) was used as the developing agent, and the results were observed under ultraviolet light (365 nm). No clear spots were obtained. Then 1% vanillin sulfuric acid solution was sprayed, and the spots were clearly colored by hot air blowing. The test sample chromatogram and the control drug chromatogram showed a blue spot at the corresponding position, but the Rf value was low.

[0102] The upper solution of petroleum ether (30-60°C)-ethyl formate-formic acid (15:5:1) was used as the developing agent, 1% vanillin sulfuric acid solution was sprayed, and the spots were clearly colored by hot air blowing. The test sample chromatogram and the control drug chromatogram showed a same color spot at the corresponding position, and the negative had no interference. The above experiments show that the chromatography system uses the control drug of costus as a control, has strong specificity, and the spots are clear (TLC atlas is shown in Figure 4 , wherein, sample 1 is the costus negative; 2 is the control drug; 3-5 are the test samples.

[0103] (4) Liquid chromatography identification of bitter orange. The control drug of bitter orange was used as a control, and "3 g of the product was added to 20 ml of methanol, ultrasonic treatment was performed for 20 minutes, and then filtration was performed, and the filtrate was used as the test sample solution". Cyclohexane-trichloromethane-ethyl acetate (4:1:1) was used as the developing agent, 10% sulfuric acid ethanol solution was sprayed, heating was performed at 105°C until the spots were clearly colored, and observation was performed under ultraviolet light (365 nm). Under this condition, good test effect was obtained, and the negative had no interference. However, during the stability investigation, it was found that the reproducibility was poor. Therefore, the method was deleted from the text in consideration of the universality of the method.

[0104] The sample extracted by ultrasonic in methanol was used as the test sample, and naringin and neohesperidin were used as the reference substances. Gradient elution was attempted using acetonitrile-0.1% phosphoric acid solution and methanol-0.1% phosphoric acid solution, but no good separation effect was obtained, and the negative sample was interfered. The mobile phase was adjusted to methanol-1.5% glacial acetic acid solution for isocratic elution, but no good chromatographic effect was obtained.

[0105] The test sample processing method was adjusted to "take 1 g of the product, add 25 ml of methanol, ultrasonic treatment (power 250 W, frequency 40 kHz) for 30 minutes, filter, evaporate the filtrate to dryness, add 15 ml of water to dissolve, extract twice with ethyl acetate, 20 ml each time, discard the ethyl acetate liquid, then extract twice with water-saturated n-butanol, 20 ml each time, combine the n-butanol layers, evaporate to dryness, dissolve the residue in methanol and dilute to 25 ml" as the test sample solution. The chromatographic conditions were adjusted to "octadecylsilane-bonded silica gel as the filler; acetonitrile-0.1% phosphoric acid solution (18:82) as the mobile phase; detection wavelength 283 nm; column temperature 25°C". Under these chromatographic conditions, the negative sample had no interference, but neohesperidin in the test sample did not reach baseline separation. Therefore, only the detection method using naringin as the reference was included in the main text.

[0106] (5) The identification method of Jinyinchen was studied. Jinyinchen

Himalayan wormwood (Mianyinchen)

[0107] Chlorogenic acid reference substance and Jinyinchen reference drug were used as the reference, and the Jinyinchen reference drug solution, test sample solution and negative blank solution were prepared by ultrasonic extraction in methanol. Gradient elution was attempted using acetonitrile-0.1% phosphoric acid solution and methanol-0.1% phosphoric acid solution. The results showed that the chromatographic peaks corresponding to the chlorogenic acid reference substance and the Jinyinchen reference drug could be detected in the test sample chromatogram, but the negative sample was interfered. An effective liquid chromatography identification method could not be established, so it was not included in the main text.

[0108] (6) TLC identification of Rhizoma Pinelliae Preparata:

[0109] Take the product 2 g, add methanol 30 ml, ultrasonic treatment for 20 minutes, filter, evaporate the filtrate, add water 30 ml to the residue, make it dissolve; extracted with diethyl ether 2 times, 20 ml each time, discard the diethyl ether layer, the water layer is extracted with water saturated n-butanol 2 times, 20 ml each time, combine the n-butanol layer, washed with 1% sodium hydroxide solution 2 times, 20 ml each time, separate the n-butanol layer, evaporate to dryness, add methanol 1 ml to the residue, make it dissolve, as the test solution. Take another Pinellia ternate control drug 1 g, add methanol 20 ml, ultrasonic treatment for 30 minutes, filter, evaporate the filtrate to dryness, add methanol 1 ml to the residue, make it dissolve, as the control drug solution. Take 3-8 μl of the test solution and 5 μl of the control drug solution, respectively, and point them on the same silica gel G thin layer plate, with cyclohexane-ethyl acetate-formic acid (5:1:0.1) as the developing agent, develop, take out, dry, spray with phosphomolybdic acid test solution, heat at 105°C until the spots develop clear color. In the test chromatogram, a clear blue-black main spot appears at the corresponding position of the control drug chromatogram, but the negative interference is not used.

[0110] (7) TLC identification of Herba Lysimachiae:

[0111] Take the product 2 g, add methanol 20 ml, ultrasonic treatment for 20 minutes, filter, evaporate the filtrate to dryness, add water 15 ml to the residue, make it dissolve; extracted with diethyl ether 2 times, 15 ml each time, combine the diethyl ether layer, evaporate to dryness, add methanol 2 ml to the residue, make it dissolve as the test solution, take another Lysimachia clethroides control drug 0.3 g, add ethanol 20 ml, heat reflux for 30 minutes, filter, evaporate the filtrate to dryness, add ethanol 1 ml to the residue, make it dissolve, as the control drug solution. With toluene-ethyl acetate-formic acid (10:8:1) as the developing agent, develop, take out, dry, spray with 3% aluminum chloride ethanol solution, heat at 105°C until the spots develop clear color, observe under ultraviolet light (365 nm). In the test chromatogram, a red fluorescent spot appears at the corresponding position of the control drug chromatogram, but the spot is not clear.

[0112] Take the product 2 g, add methanol 25 ml, ultrasonic treatment for 20 minutes, filter, evaporate the filtrate to dryness, add water 25 ml to the residue, make it dissolve, extract with chloroform 2 times, 25 ml each time; the water layer is extracted with water saturated n-butanol 25 ml once, evaporate the n-butanol layer to dryness, add methanol 5 ml to the residue, make it dissolve, as the test solution; with Lysimachia clethroides control drug as the control, toluene-ethyl acetate-formic acid (10:3:1), chloroform-ethyl acetate-methanol-water (15:40:22:10) lower layer solution after standing overnight below 10°C as the developing agent, spray with 10% sulfuric acid ethanol solution, heat at 105°C until the spots develop clear color, observe under ultraviolet light (365 nm), the result is not satisfactory.

[0113] The mobile phase was chloroform-methanol-water (30:8:1), and the sample was sprayed with 10% sulfuric acid in ethanol and heated to 105°C until the spots were clearly colored. Under ultraviolet light (365 nm), the test sample chromatogram showed a fluorescent spot of the same color as the corresponding spot in the control drug chromatogram, but the negative control was interfered, so it was not used.

[0114] (8) After obtaining the clinical batch, the identification method of ginger was studied using 6-gingerol as a control.

[0115] The test sample solution was prepared by adding 4 g of the product to 20 ml of ethyl acetate and ultrasonically treating for 10 minutes, filtering, and evaporating the filtrate to dryness. The residue was dissolved in 1 ml of ethyl acetate. The dry ginger control drug material was prepared in the same way as the test sample solution. The 6-gingerol control was prepared by adding ethyl acetate to obtain a solution containing about 0.5 mg per 1 ml. The three solutions were spotted on the same silica gel G plate using petroleum ether (60-90°C)-chloroform-ethyl acetate (2:1:1), petroleum ether (60-90°C)-chloroform-ethyl acetate (3:0.5:1), and petroleum ether (60-90°C)-ethyl acetate (3:1) as the developing agents. After development, the plate was sprayed with phosphomolybdic acid solution and heated to 105°C until the spots were clearly colored. The results were not ideal for thin layer chromatography. The developing agents were adjusted to petroleum ether (60-90°C)-chloroform-ethyl acetate (7:2:1) and cyclohexane-chloroform-ethyl acetate (9:2:1). After development, the plate was sprayed with phosphomolybdic acid solution and heated to 105°C until the spots were clearly colored. The spots were clear and well separated, but the negative control was interfered, so a mature thin layer chromatography identification method was not established and was not included in the text.

[0116] (9) The volatile oil thin layer identification method was studied. After obtaining the clinical batch, the identification method of volatile components was studied using the control drug material of Fructus Aurantii and the control of limonene. The research content is as follows:

[0117] "Take 10 g of the product, place it in a round-bottom flask, add 200 ml of water, connect a volatile oil tester, connect a reflux condenser, heat to boiling, and keep it slightly boiling for 2 hours, cool, and take the n-hexane liquid" as the test solution; take the Citrus aurantium L. control drug material, and prepare a control drug material solution by the same method; then take the limonene control, add n-hexane to prepare a control solution containing about 0.7 mg per ml. Take the above-mentioned control solution, control drug material solution and test solution, and spot them on a silica gel GF254 thin layer plate, use "n-hexane-ethyl acetate (12:1), n-hexane-ethyl acetate (6:1), n-hexane-ethyl acetate (3:1), n-hexane-ethyl acetate-formic acid (6:1:0.1), petroleum ether (60-90°C)-ethyl acetate (6:1), petroleum ether (60-90°C)-butyl acetate (6:1)" as the developing agent, develop, and observe under a UV light (254 nm), under a UV light (365 nm), or spray with 5% vanillin sulfuric acid ethanol solution, 10% sulfuric acid ethanol solution, 10% phosphomolybdic acid ethanol solution, 5% ferric trichloride ethanol solution, heat at 105°C until the spots develop clearly, and all do not achieve ideal thin layer effects.

[0118] Then use "petroleum ether (60-90°C)-ethyl acetate-ammonia water (4:1:0.2)" as the developing agent, use the Citrus aurantium L. control drug material and limonene control as controls, spray with 10% phosphomolybdic acid ethanol solution, heat at 105°C until the spots develop clearly, and achieve a good thin layer effect, but the negative is interfered, a mature thin layer chromatography identification method is not established, and therefore is not included in the text.

[0119] (10) The corresponding content determination methods for Scutellaria baicalensis Georgi and Rheum officinale Baill and Polygonum cuspidatum Sieb.et Zucc. in the preparation have been established, and therefore the thin layer identification research is not performed.

[0120] Since the traditional Chinese medicine composition preparation is a large compound preparation, the active ingredients therein are complex and interact with each other in numerous ways, the identification method adopted by the present application can accurately and effectively identify the components of multiple medicinal materials in the traditional Chinese medicine composition preparation, the identification test solution of Gardenia jasminoides Ellis, Paeonia lactiflora Pall. and Saussurea lappa Clarke adopts the same treatment method, time and steps are saved, the specificity is extremely strong, very accurate results are achieved, the product quality of the traditional Chinese medicine composition preparation is effectively controlled, the safety and reliability of the drug are improved, and the safety of the people using the drug is ensured.

[0121] Content determination

[0122] 1. Scutellaria baicalensis Georgi content determination

[0123] In order to effectively control the quality of the finished product, the effective component of the Huangqi in the water extract part of the prescription is selected as the content determination index, and a content determination method of the Huangqi in the traditional Chinese medicine composition preparation is established by using high performance liquid chromatography. The test result shows that the method has strong specificity, good reproducibility, short analysis time, stable and reliable result, and can effectively control the quality of the traditional Chinese medicine composition preparation.

[0124] 1.1 Test material, instrument and reagent

[0125] The traditional Chinese medicine composition preparation is a medicine capsule prepared according to the preparation method in the embodiment 2, which is provided by Shijiazhuang Yiling Pharmaceutical Co., Ltd., and the batch number is 20080101;

[0126] The Huangqi reference substance is purchased from the China Institute for Drug Control, and the batch number is 110715-200212;

[0127] The high performance liquid chromatograph (Agilent 1100 series) is provided with a VWD ultraviolet detector.

[0128] The chromatographic column is filled with octadecylsilane bonded silica gel.

[0129] The methanol used for the mobile phase is chromatographic pure, and the rest of the reagents and reagents are analytical pure, and the water is purified water.

[0130] 1.2 Selection of mobile phase

[0131] The methanol-0.1% phosphoric acid solution (45:55) is used as the mobile phase, the chromatographic peak separation degree is better, and the peak shape is symmetrical.

[0132] 1.3 Chromatographic conditions

[0133] The chromatographic column is filled with octadecylsilane bonded silica gel.

[0134] The mobile phase is methanol-0.1% phosphoric acid solution (45:55).

[0135] The flow rate is 1.0 ml / min.

[0136] The detection wavelength is 280 nm.

[0137] The theoretical plate number is not less than 2500 according to the Huangqi peak.

[0138] 1.4 Preparation of reference solution

[0139] The Huangqi reference substance is accurately weighed, 70% ethanol is added to prepare a solution containing 20.7 μg per 1 ml, and then shaken to obtain the solution.

[0140] 1.5 Investigation on the preparation method of the test sample

[0141] ① Selection of extraction solvent:

[0142] Take the contents of the product, fine, take 0.1 g, accurately weighed, add 70% methanol, 70% ethanol ultrasonic extraction for 15 minutes, the results are shown in Table 1.

[0143] Table 1 extraction solvent on the determination of baicalin content

[0144]

[0145] From the test results can be seen, respectively, with 70% methanol and 70% ethanol as the extraction solvent for the extraction of baicalin, determination, content has no significant difference, but 70% ethanol toxicity small, cheap, so select 70% ethanol as the extraction solvent.

[0146] ② ultrasonic time inspection:

[0147] Take the contents of the product, fine, take 0.1 g, accurately weighed, add 70% ethanol ultrasonic extraction, ultrasonic time was investigated, the results are shown in Table 2.

[0148] Table 2 ultrasonic time on the determination of baicalin content

[0149]

[0150] From the test results can be seen, ultrasonic time on the determination of baicalin content little effect, ultrasonic 5 minutes can be fully extracted baicalin, but taking into account the differences between different ultrasonic instruments, to ensure that the baicalin extraction is complete, so select ultrasonic time for 10 minutes.

[0151] 1.6 specificity test

[0152] Take the medicine in the prescription except Huangqi, according to the preparation process to obtain negative sample, according to the processing method of test solution, get negative sample solution. Take baicalin reference substance solution, negative sample solution and test solution, according to the selected determination conditions for detection, the results of no interference, good specificity (baicalin reference substance, negative test sample, test sample chromatogram see Figure 5 ).

[0153] 1.7 linear investigation

[0154] Accurately take the reference solution (20.68 μg / ml) 4, 8, 12, 16, 20, 25 μl, respectively, into the liquid chromatograph, the determination of peak area, with peak area integral value as the ordinate, the sample size as the abscissa, draw the standard curve. The results show that: baicalin sample size in the range of 82.72 ng ~ 517.00 ng shows a good linear relationship, the regression equation is: y = -2.2693 + 3.2683x, r = 0.99999 (standard curve see Figure 6The measurement results are shown in Table 3.

[0155] Table 3 Standard Curve

[0156]

[0157] 1.8 Determination of purity of reference standard:

[0158] 15 μl of a 0.12925 mg / mL baicalin reference solution was injected into the liquid chromatograph, and the absorbance value was determined. The purity of baicalin was determined to be 98.1% by the normalization method (chromatogram shown). Figure 7 ).

[0159] 1.9 Precision Test

[0160] Take 10 μl of the reference solution and the test solution and inject each solution 5 times. Measure the peak area and the results are shown in Table 4.

[0161] Table 4 Precision Test Results

[0162]

[0163] The test results show that the RSDs of the measured values ​​of the reference solution and the test solution were 0.81% and 0.44%, respectively, indicating that the instrument has good precision.

[0164] 1.10 Stability Test

[0165] Take samples of the reference solution and the test solution, inject 10 μl of each sample at regular intervals, and measure the peak area. The results are shown in Table 5.

[0166] Table 5. Stability Test Results

[0167]

[0168]

[0169] The results above show that the detection values ​​of the reference solution and the test solution placed at room temperature are stable within 24 hours, with RSD values ​​of 0.79% and 0.48%, respectively.

[0170] 1.11 Repeatability Test:

[0171] Nine samples from the same batch were taken, and three portions of each sample (high, medium, and low) were prepared according to the preparation method for the test solution. The test results are shown in Table 6.

[0172] Table 6 Results of Repeatability Tests

[0173]

[0174] From the above results, the repeatability of the test sample preparation and determination method for the content of Baicalin is good.

[0175] 1.12 Recovery test:

[0176] The same batch of sample was taken in 9 portions, divided into three groups, and 25 ml of high, medium and low concentration Baicalin control 70% ethanol solution was added respectively, ultrasonic extraction was carried out, and the recovery rate was calculated. The results are shown in Table 7 (the content of Baicalin in the sample is 22.90 mg / g).

[0177] Table 7 Recovery test results

[0178]

[0179] From the above results, the average recovery rate in the Baicalin spiking recovery test of the product is 100.07%, and the RSD of the determination results is 2.48%, indicating that the prepared Baicalin content determination method has good accuracy.

[0180] 1.13 Effect of column temperature on Baicalin content determination: The same liquid flow rate was used to detect the same test sample at different column temperatures, and the content change was observed, as shown in Table 8.

[0181] Table 8 Effect of column temperature on Baicalin content determination

[0182]

[0183] From the determination results, the column temperature has no significant effect on the content determination results when the column temperature is between 20℃ and 30℃, and the RSD is 1.62%.

[0184] 1.14 Effect of different chromatographic columns on Baicalin content determination: Under the same liquid phase condition parameters, different chromatographic columns were replaced to investigate the effect of chromatographic columns on Baicalin content determination, and the test results are shown in Table 9.

[0185] Table 9 Effect of chromatographic columns on Baicalin content determination

[0186]

[0187] From the test results, replacing the chromatographic column has little effect on the Baicalin content determination results.

[0188] 1.15 Effect of detection wavelength on Baicalin content determination: Under the same liquid phase condition parameters, the detection wavelength was adjusted to investigate the effect of detection wavelength on Baicalin content determination, and the test results are shown in Table 10.

[0189] Table 10 Effect of detection wavelength on Baicalin content determination

[0190]

[0191] From the determination results, it can be seen that the detection wavelength has no great influence on the content determination results at 275nm-285nm, and the RSD is 0.45%.

[0192] 1.16 Determination of Baicalin: Three batches of capsules were determined by the developed content determination method, and the determination results are shown in Table 11.

[0193] Table 11 Determination results of baicalin content

[0194]

[0195] The baicalin in 6 batches of large production samples of the product was determined by the developed content determination method, and the determination results are shown in Table 12.

[0196] Table 12 Determination results of baicalin content in large production samples

[0197]

[0198] 1.17 Determination of the content limit of baicalin in the product

[0199] From the production of the second phase of the clinical sample, 6 batches of large production samples were prepared. Considering that there are many factors affecting the content, combined with the content determination data and the degree of dispersion of the 6 batches of large production samples, the content limit of baicalin in the product was developed by using the average content of the 6 batches of large production samples ±60%.

[0200] The average content of baicalin in the six batches of samples is 8.8mg per particle, and the upper limit of 60% is 14.1mg per particle, and the lower limit of 60% is 3.5mg per particle, so the content limit of baicalin is 3.5mg-14.1mg per particle. 21 H 18 O 11 ) should be 3.5mg-14.1mg per particle.

[0201] 2. Determination of emodin content:

[0202] In order to effectively control the quality of the finished product, the effective component emodin in rhubarb and giant knotweed in alcohol extraction part of prescription is selected as the content determination index, and a high performance liquid chromatography method is adopted to establish the content determination method of emodin in the traditional Chinese medicine composition preparation. The test results show that the method has strong specificity, good reproducibility, short analysis time, stable and reliable results, and can effectively control the quality of the traditional Chinese medicine composition preparation.

[0203] 2.1 Materials, instruments and reagents

[0204] Emodin reference substance: purchased from China Institute for Drug Control, batch number: 0756-200211;

[0205] Traditional Chinese medicine composition preparation: a medicine capsule prepared according to Example 2, provided by Shijiazhuang Hill Pharmaceutical Co., Ltd., batch number: 20080101;

[0206] High performance liquid chromatograph (Agilent 1100 series) VWD ultraviolet detector;

[0207] Chromatographic column: packed with octadecylsilane bonded silica gel;

[0208] Acetonitrile and methanol used for mobile phase were chromatographic pure, and the rest of reagents and reagents were analytical pure, and water was purified water.

[0209] 2.2 Mobile phase selection: acetonitrile-methanol-0.1% phosphoric acid solution (42:23:35) was used as the mobile phase, and the chromatographic peak separation was better, and the peak shape was symmetrical.

[0210] 2.3 Chromatographic conditions:

[0211] Chromatographic column: packed with octadecylsilane bonded silica gel;

[0212] Mobile phase: acetonitrile-methanol-0.1% phosphoric acid solution (42:23:35);

[0213] Flow rate: 1.0 ml / min;

[0214] Detection wavelength: 254 nm;

[0215] Theoretical plate number: should not be less than 3000 according to the emodin peak.

[0216] 2.4 Preparation of reference solution: accurately weigh a certain amount of emodin reference substance, add methanol to prepare a solution containing 37 μg per 1 ml, and shake well to obtain.

[0217] 2.5 Preparation of test solution:

[0218] Investigation of reflux time:

[0219] Take the content, grind finely, accurately weigh 0.5 g, accurately weigh, add 10% hydrochloric acid methanol solution, reflux in 80°C water bath, and investigate the reflux time, and the investigation results are shown in Table 13:

[0220] Table 13 Effect of reflux time on determination of emodin content

[0221]

[0222]

[0223] From the test results can be seen, heating reflux 20 min when emodin has been extracted completely, in order to ensure sufficient extraction, so selected reflux time of 30 minutes.

[0224] 2.6 specificity test:

[0225] Take the medicine in the prescription except for rhubarb, giant knotweed, according to the preparation process to obtain negative sample, according to the processing method of test solution, get negative sample solution. Take the control sample solution, negative test sample solution and test sample solution, according to the selected detection conditions for detection, the results of negative without interference (negative sample, emodin control sample, sample chromatogram see Figure 8 ).

[0226] 2.7 linear relationship:

[0227] Precisely take the control sample solution (0.0369mg / ml) 2, 4, 6, 8, 10, 20 μl, respectively, into the liquid chromatograph, the determination of peak area, with peak area integral value as the ordinate, the injection amount of as the abscissa, draw the standard curve. The results show that: emodin injection amount in the range of 73.8ng~738ng shows good linear relationship, the regression equation is: y=-3.8926+3.6720x, r=1.0000 (standard curve see Figure 9 ). The determination results are shown in table 14.

[0228] Table 14 standard curve

[0229]

[0230] 2.8 control sample purity determination:

[0231] Take 20 μl of emodin control sample solution with a concentration of 0.1107mg / ml, inject into the liquid chromatograph to determine the absorption value, and determine the purity of emodin by normalization method is 99.6% (chromatogram see Figure 10 )

[0232] 2.9 precision test:

[0233] Take the control sample solution and test sample solution, each injection 6 times, 10 μl each time, determine the peak area, the detection results are shown in table 15.

[0234] Table 15 precision test results

[0235]

[0236] From the test results can be seen, the detection value of control sample solution and test sample solution RSD is 0.39% and 0.12% respectively, indicating that the instrument precision is good.

[0237] 2.10 stability test:

[0238] The control sample solution and the test sample solution were taken every certain time, 10 μl each time, the peak area was determined, and the test results are shown in Table 16.

[0239] Table 16 Stability test results

[0240]

[0241] From the above results, it can be seen that the control sample solution and the test sample solution placed at room temperature are stable within 20 hours, and the RSD values are 1.35% and 0.89%, respectively.

[0242] 2.11 Reproducibility test:

[0243] Nine samples from the same batch were taken, and high, medium and low sample amounts were taken, three samples for each sample amount. Each test sample solution was prepared according to the preparation method of the test sample solution, and the test results are shown in Table 17.

[0244] Table 17 Reproducibility test results

[0245]

[0246] From the above results, it can be seen that the test sample preparation and determination method for the determination of emodin content in the product is good in reproducibility.

[0247] 2.12 Recovery test:

[0248] The recovery test was performed by adding 25 ml of 10% emodin hydrochloric acid methanol solution of high, medium and low concentrations to three groups of nine samples from the same batch, respectively, and then performing reflux extraction and determination. The recovery rate was calculated, and the results are shown in Table 18 (the emodin content in the sample was 4.02 mg / g).

[0249] Table 18 Recovery determination results

[0250]

[0251]

[0252] From the above results, it can be seen that the average recovery rate of emodin in the sample recovery test is 98.06%, and the RSD of the determination results is 2.22%, indicating that the emodin content determination method is good in accuracy.

[0253] 2.13 Effect of different chromatographic columns on determination of emodin content: Under the same liquid chromatography conditions, different chromatographic columns were replaced to investigate the effect of chromatographic columns on the determination results of emodin content, and the test results are shown in Table 19.

[0254] Table 19 Effect of different chromatographic columns on determination of emodin content

[0255]

[0256] From the test results, it can be seen that the replacement of chromatographic column has less effect on the determination results of emodin content.

[0257] 2.14 Effect of mobile phase on determination of emodin content:

[0258] Under the same liquid phase condition parameters, the proportion of mobile phase was adjusted to investigate the effect of different proportions of mobile phase on the determination results of emodin content, and the test results are shown in Table 20.

[0259] Table 20 Effect of mobile phase on determination of emodin content

[0260]

[0261]

[0262] From the determination results, it can be seen that when the proportion of mobile phase is slightly adjusted, it has no great effect on the determination results, and the RSD is 0.79%.

[0263] 2.15 Effect of column temperature on determination of emodin content: Under the same liquid phase condition parameters, the column temperature was changed to investigate the effect of different column temperatures on the determination results of emodin content, and the test results are shown in Table 21.

[0264] Table 21 Effect of column temperature on determination of emodin content

[0265]

[0266] From the determination results, it can be seen that when the column temperature is changed, it has no great effect on the determination results, and the RSD is 0.25%.

[0267] 2.16 Effect of detection wavelength on determination of emodin content: Under the same liquid phase condition parameters, the detection wavelength was slightly changed to investigate the effect of different detection wavelengths on the determination results of emodin content, and the test results are shown in Table 22.

[0268] Table 22 Effect of detection wavelength on determination of emodin content

[0269]

[0270] From the determination results, it can be seen that when the detection wavelength is slightly adjusted, it has no great effect on the determination results, and the RSD is 0.38%.

[0271] 2.17 Determination of emodin: The emodin content in three batches of finished products was determined by the developed emodin content determination method, and the determination results are shown in Table 23.

[0272] Table 23 emodin content determination results

[0273]

[0274] The emodin content of 6 batches of large production samples of the product was determined using the developed content determination method, and the determination results are shown in Table 24.

[0275] Table 24 emodin content determination results of large production samples

[0276]

[0277] 2.18 Determination of the emodin content limit in the product

[0278] From the start of the second phase of clinical sample production, the preparation of 6 batches of large production samples was completed. Considering the many factors that affect the content, combined with the content determination data and the degree of dispersion of the 6 batches of large production samples, the emodin content limit in the product was developed in the form of the average content of the 6 batches of large production samples ± 60%.

[0279] The average content of emodin in the six batches of samples was 2.0 mg / tablet, the upper limit was 3.2 mg / tablet, and the lower limit was 0.8 mg / tablet, so the emodin content limit of each tablet of Radix et Rhizoma Rhei and Polygonum cuspidatum was 0.8 mg-3.2 mg. 15 H 10 O5) should be 0.8 mg-3.2 mg.

[0280] Example 1

[0281] Identification and content determination method of traditional Chinese medicine composition capsules

[0282] The raw material formula is: Huangqi 137g, Chaihu 137g, Dahuang 103g, Zhicheng 137g, Yincheng 137g, Huzhang 171g, Zhizi 137g, Jinqiancao 342g, Baishao 137g, Muxiang 137g, Jiangbanxia 103g, Shengjiang 34g.

[0283] Preparation process:

[0284] A, Jiangbanxia was weighed, crushed, and passed through a 100 mesh sieve for use;

[0285] B, Zhicheng and Shengjiang were weighed, 7 times the amount of water was added, and the volatile oil was extracted for 10 hours, and the volatile oil was collected and separated; the water solution after distillation was reserved;

[0286] C, Huangqi, Baishao, Yincheng, and Zhizi were weighed, and water was added for decoction 3 times, the first extraction was 2 hours, and the second and third extractions were 1.5 hours each time, 9 times the amount of water was added each time, the extraction liquid was filtered, combined with the volatile oil water extraction liquid obtained in step B, and concentrated under reduced pressure to a relative density of 1.25±0.05 (60°C) of the extract, which was reserved;

[0287] D, take the Bupleurum, costus, rhubarb, giant knotweed, money grass, 70% ethanol extraction 3 times, the first time to add 12 times the amount of extraction 2.5 hours, the second, the third time to add 10 times each for 2 hours, the filtrate, concentrated into a clear paste, with the water extract of step C paste combined, mix well, dry, crush for use;

[0288] E, the fine powder of step A, step D dry paste powder, mix well, step B volatile oil with silica adsorption volatile oil mix well, capsule, get.

[0289] Identification:

[0290] Bupleurum thin layer identification, take the contents of this product, fine, take 3g, add methanol 20ml, ultrasonic treatment 20 minutes, filter, filter dry, add water 25ml to dissolve, with chloroform shake extraction 3 times, 25ml each time, discard the chloroform liquid, then use water saturated n-butanol shake extraction 2 times (15ml, 25ml), combined n-butanol layer, washed with 3% sodium hydroxide solution 3 times, 30ml each time, take the n-butanol liquid dry, the residue is dissolved in 2ml of methanol as test solution; another 0.5g of Bupleurum control drug powder, add methanol 20ml, ultrasonic treatment 10 minutes, filter, filter concentrate to 5ml, as control drug solution, according to the thin layer chromatography test, 5ul of control drug solution, 10ul of test solution, respectively, on the same silica gel G thin layer plate, with volume ratio of 30:8:1 of chloroform-methanol-water as developing agent, development, take out, dry, spray with 2% p-dimethylaminobenzaldehyde 40% sulfuric acid ethanol solution, heated to 60 DEG C, until the color spots clear, in the light and ultraviolet light (365nm) under the observation, the test chromatogram, in the corresponding position of the control drug chromatogram, showed the same color spots.

[0291] Thin-layer identification of Baishao and Zhizi: take the contents of the product, grind finely, take 3 g, add methanol 20 ml, ultrasonic treat for 20 minutes, filter, take the filtrate as the test solution; take Zhizi control drug 0.5 g, add methanol 20 ml, heat reflux for 30 minutes, filter, evaporate the filtrate to dryness, add methanol 2 ml to dissolve, as Zhizi control drug solution; take Baishao control drug 0.5 g, add methanol 20 ml, ultrasonic treat for 20 minutes, filter, evaporate the filtrate to dryness, add methanol 2 ml to dissolve, as Baishao control drug solution, according to the test of thin-layer chromatography, take 5 μl of the test solution and the above two control drug solutions respectively, and point them on the same silica gel G thin-layer plate, with trichloromethane-ethyl acetate-methanol-ammonia water (4:4:5:1) as developing agent, develop, take out, dry, spray with 1% vanillin sulfuric acid solution, heat at 105 ℃ until the spots develop clearly, the test solution chromatogram shows the same color spots as the Zhizi control drug and Baishao control drug chromatograms at the corresponding positions.

[0292] Thin-layer identification of Muxiang: take Muxiang control drug 0.2 g, add trichloromethane 5 ml, ultrasonic treat for 30 minutes, filter, the filtrate as the control drug solution, according to the test of thin-layer chromatography, take 5 μl of the test solution in Artemisia capillaris thin-layer identification and the above control drug solution respectively, and point them on the same silica gel G thin-layer plate, with petroleum ether (30-60 ℃)-ethyl formate-methanol (15:5:1) as developing agent, develop, take out, dry, spray with 1% vanillin sulfuric acid solution, blow with hot air until the spots develop clearly, the test solution chromatogram shows the same color spots as the control drug chromatogram at the corresponding positions.

[0293] Identification of Jisheng: take the contents of the product, grind finely, take 1 g, add methanol 25 ml, ultrasonic treat for 30 minutes, filter, evaporate the filtrate to dryness, add water 15 ml to dissolve, shake with ethyl acetate to extract twice, 20 ml each time, discard the ethyl acetate liquid, shake with water-saturated n-butanol to extract twice, 20 ml each time, combine the n-butanol layers, evaporate to dryness, dissolve the residue in methanol, transfer to a 25 ml volumetric flask, dilute to the mark with methanol, shake well, filter, the filtrate as the test solution. Take naringin control substance, add methanol to prepare a solution containing 80 μg per 1 ml, as the control solution. According to the test of high performance liquid chromatography (Chinese Pharmacopoeia 2020 edition general 0512), use octadecylsilane-bonded silica gel as the filler; use acetonitrile-0.1% phosphoric acid solution (18:82) as the mobile phase; the detection wavelength is 283 nm; the column temperature is 25 ℃; the theoretical plate number should not be less than 3000 calculated by naringin peak. Precisely take 10 μl of the control solution and the test solution respectively, inject into the high performance liquid chromatograph, the test solution chromatogram should show a chromatographic peak consistent with the retention time of the control solution chromatographic peak.

[0294] Content determination:

[0295] The content determination method of Scutellariae Radix includes the following steps:

[0296] Preparation of the control solution: take the baicalin control product, accurately weigh, add 70% methanol to prepare a solution containing 20 μg per 1 ml, shake well, and obtain;

[0297] Preparation of the test solution: take the content of the traditional Chinese medicine composition preparation, grind finely, take 0.1 g, accurately weigh, place in a conical flask with a plug, accurately add 70% ethanol 25 ml, weigh, ultrasonic treat for 10 minutes, cool, weigh again, make up the weight loss with 70% ethanol, shake well, filter, accurately take 2 ml of the filtrate, place in a 10 ml volumetric flask, dilute to the mark with 70% ethanol, shake well, filter, take the filtrate, and obtain;

[0298] HPLC detection: the chromatographic column is C18 (Phenomenex Gemini 5 μm C 18 4.6x150mm), the mobile phase is composed of methanol and 0.1% phosphoric acid solution with a volume ratio of 45:55, the detection wavelength is 280 nm, the column temperature is 25°C, and the flow rate is 1.0 ml / min;

[0299] Determination method: accurately take 10 μl of the control solution and the test solution respectively, inject into the high performance liquid chromatograph, determine, and obtain.

[0300] The content determination method of Rhubarb and Giant Knotweed includes the following steps:

[0301] Preparation of the control solution: take the emodin control product, accurately weigh, add methanol to prepare a solution containing 37 μg per 1 ml, shake well, and obtain;

[0302] Preparation of the test solution: take the content of the traditional Chinese medicine composition preparation, grind finely, take 0.5 g, accurately weigh, place in a conical flask with a plug, accurately add 10% hydrochloric acid methanol solution 25 ml, weigh, heat in a water bath at 80°C for 30 minutes, cool, weigh again, make up the weight loss with methanol, shake well, filter, accurately take 4 ml of the filtrate, place in a 10 ml volumetric flask, accurately add 2% sodium hydroxide solution 2 ml, dilute to the mark with methanol, shake well, filter, take the filtrate, and obtain;

[0303] HPLC detection: the chromatographic column is C18 (Agilent SB-C 18 5 μm C18 4.6x150mm), the mobile phase is composed of acetonitrile, methanol and 0.10% phosphoric acid solution with a volume ratio of 42:23:35, the detection wavelength is 254 nm, the column temperature is 30°C, and the flow rate is 1.0 ml / min;

[0304] Determination method: 10 μl of the control solution and the sample solution were respectively injected into the liquid chromatograph, and then determination was carried out, and thus the content was obtained.

[0305] Example 2

[0306] Identification and content determination method of the traditional Chinese medicine composition capsule

[0307] The raw material formula is: Huangqi 128g, Chaihu 128g, Dahuang 120g, Zhike 128g, Yincheng 128g, Huzhang 200g, Zhizi 150g, Jinqiancao 250g, Baishao 128g, Muxiang 128g, Jiangbanxia 90g, Shengjiang 40g.

[0308] Preparation process:

[0309] A. Jiangbanxia is weighed, crushed, and passed through a 100-mesh sieve for standby use;

[0310] B. Zhike and Shengjiang are weighed, 9 times the amount of water is added, and volatile oil is extracted for 12 hours; the water solution after distillation is reserved;

[0311] C. Huangqi, Baishao, Yincheng, and Zhizi are weighed, and water is added for decoction for 14 times; the first extraction is carried out for 13 hours, and the second and third extractions are carried out for 11 hours each time, 110 times the amount of water is added each time, the extraction liquid is filtered, combined with the volatile oil water extraction liquid obtained in step B, and concentrated under reduced pressure to the extract with a relative density of 1.25±0.05 (60℃), which is reserved;

[0312] D. Chaihu, Muxiang, Dahuang, Huzhang, and Jinqiancao are weighed, and 80% ethanol is added for extraction for 3 times; 12 times the amount of solvent is added for the first extraction for 2 hours, and 10 times the amount of solvent is added for the second and third extractions for 2 hours each time, the extraction liquid is filtered, concentrated into a clear extract, combined with the water extraction extract obtained in step C, mixed, dried, and crushed for standby use;

[0313] E. The fine powder obtained in step A and the dry extract powder obtained in step D are mixed, dried, mixed with the volatile oil obtained in step B, and then filled into capsules to obtain the product.

[0314] The identification and content determination method are the same as those in Example 1.

[0315] Example 3

[0316] Identification and content determination method of the traditional Chinese medicine composition tablet

[0317] Raw material formula:

[0318] Huangqi 69g, Chaihu 205g, Dahuang 52g, Zhike 205g, Yincheng 69g, Huzhang 255g, Zhizi 69g, Jinqiancao 515g, Baishao 69g, Muxiang 205g, Banxia 52g, and Shengjiang 17-52g.

[0319] Preparation process:

[0320] A. Take Pinellia ternate, crush, pass through a 100-mesh sieve, and reserve;

[0321] B. Take Fructus Aurantii and Rhizoma Zingiberis Recens, add 5 times the amount of water, extract volatile oil for 10 hours, collect and separate the volatile oil; distill the water solution, and reserve;

[0322] C. Take Radix Scrophulariae, Radix Paeoniae Alba, Herba Artemisiae Scopariae, and Fructus Gardeniae, add water, decoct 4 times, extract for 2 hours the first time, extract for 1 hour each of the second and third times, add 10 times the amount of water each time, combine the extract with the volatile oil extract obtained in step B, concentrate under reduced pressure to an extract with a relative density of 1.25±0.05 (60°C), and reserve;

[0323] D. Take Radix Bupleuri, Radix Aucklandiae, Radix et Rhizoma Rhei, Rhizoma Polygoni Cuspidati, and Herba Lysimachiae, add 60% ethanol, extract 3 times, add 10 times the amount of extraction for 3 hours the first time, add 8 times the amount of extraction for 2 hours each of the second and third times, filter the extract, concentrate to a clear extract, combine with the water extract obtained in step C, mix well, dry, and pulverize to reserve;

[0324] E. Mix the fine powder obtained in step A and the extract powder obtained in step D, mix well, dry, mix with the volatile oil obtained in step B, and prepare into tablets according to the conventional method to obtain the product.

[0325] Identification:

[0326] Thin-layer identification of Radix Bupleuri: take the contents of the product, finely grind, take 3 g, add methanol 20 ml, ultrasonically treat for 20 minutes, filter, evaporate the filtrate to dryness, dissolve the residue in water 25 ml, extract 3 times with chloroform 25 ml each time, discard the chloroform liquid, extract 2 times with water-saturated butanol 15 ml and 25 ml, wash the butanol layer 3 times with 3% sodium hydroxide solution 30 ml each time, evaporate the butanol liquid to dryness, dissolve the residue in methanol 2 ml to obtain a test solution; take another 0.5 g of Radix Bupleuri control material powder, add methanol 20 ml, ultrasonically treat for 10 minutes, filter, concentrate the filtrate to 5 ml to obtain a control material solution, test by thin-layer chromatography, take 5 μl of the control material solution and 10 μl of the test solution, spot on the same silica gel G thin-layer plate, develop with chloroform-methanol-water (30:8:1 by volume), take out, dry, spray with 2% p-dimethylaminobenzaldehyde in 40% sulfuric acid ethanol solution, heat at 60°C until the spots develop clearly, and observe under daylight and ultraviolet light (365 nm), the same colored spots appear in the test chromatogram at the positions corresponding to the control chromatogram.

[0327] Thin-layer identification of Baishao and Zhizi: take the contents of the product, grind finely, take 3 g, add methanol 20 ml, ultrasonic treat for 20 minutes, filter, take the filtrate as the test solution; take Zhizi control drug 0.5 g, add methanol 20 ml, heat reflux for 30 minutes, filter, evaporate the filtrate to dryness, add methanol 2 ml to dissolve, as Zhizi control drug solution; take Baishao control drug 0.5 g, add methanol 20 ml, ultrasonic treat for 20 minutes, filter, evaporate the filtrate to dryness, add methanol 2 ml to dissolve, as Baishao control drug solution, according to the test of thin-layer chromatography, take 5 μl of the test solution and the above two control drug solutions respectively, and point them on the same silica gel G thin-layer plate, with trichloromethane-ethyl acetate-methanol-ammonia water (4:4:5:1) as developing agent, develop, take out, dry, spray with 1% vanillin sulfuric acid solution, heat at 105 ℃ until the spots develop clearly. In the test solution chromatogram, there are spots of the same color at the corresponding positions of Zhizi control drug and Baishao control drug chromatogram.

[0328] Thin-layer identification of Muxiang: take Muxiang control drug 0.2 g, add trichloromethane 5 ml, ultrasonic treat for 30 minutes, filter, take the filtrate as the control drug solution, according to the test of thin-layer chromatography, take 5 μl of the test solution in Artemisia capillaris thin-layer identification and the above control drug solution respectively, and point them on the same silica gel G thin-layer plate, with petroleum ether (30-60 ℃)-ethyl formate-methanol (15:5:1) as developing agent, develop, take out, dry, spray with 1% vanillin sulfuric acid solution, blow with hot air until the spots develop clearly. In the test solution chromatogram, there are spots of the same color at the corresponding positions of the control drug chromatogram.

[0329] Identification of Zhike: take the contents of the product, grind finely, take 1 g, add methanol 25 ml, ultrasonic treat for 30 minutes, filter, evaporate the filtrate to dryness, add water 15 ml to dissolve, shake with ethyl acetate to extract twice, 20 ml each time, discard the ethyl acetate liquid, shake with water-saturated n-butanol to extract twice, 20 ml each time, combine the n-butanol layers, evaporate to dryness, dissolve the residue in methanol, transfer to a 25 ml volumetric flask, dilute to the mark with methanol, shake well, filter, take the filtrate as the test solution. Take naringin control substance, add methanol to prepare a solution containing 80 μg per 1 ml, as the control solution. According to the test of high performance liquid chromatography (Chinese Pharmacopoeia 2020 edition general 0512), use octadecylsilane-bonded silica gel as the filler; use acetonitrile-0.1% phosphoric acid solution (18:82) as the mobile phase; the detection wavelength is 283 nm; the column temperature is 25 ℃; the theoretical plate number should not be less than 3000 calculated by naringin peak. Precisely take 10 μl of the control solution and the test solution respectively, inject into the high performance liquid chromatograph. In the test solution chromatogram, there should be a chromatographic peak consistent with the retention time of the control solution chromatogram peak.

[0330] Content determination:

[0331] The content determination method of Scutellariae Radix comprises the following steps:

[0332] Preparation of the control solution: take the baicalin control sample, accurately weigh, add 70% methanol to prepare a solution containing 20 μg per 1 ml, shake well, and obtain;

[0333] Preparation of the test solution: take the content of the traditional Chinese medicine composition preparation, grind finely, take 0.1 g, accurately weigh, place in a conical flask with a plug, accurately add 70% ethanol 25 ml, weigh, ultrasonic treat for 10 minutes, cool, weigh again, make up the weight loss with 70% ethanol, shake well, filter, accurately take 2 ml of the filtrate, place in a 10 ml volumetric flask, dilute to the mark with 70% ethanol, shake well, filter, take the filtrate, and obtain;

[0334] HPLC detection: the chromatographic column is C18 (Symmetry shield RP18 5 μm 4.6*150 mm), the mobile phase is composed of methanol and 0.1% phosphoric acid solution with a volume ratio of 45:55, the detection wavelength is 275 nm, the column temperature is 30°C, and the flow rate is 1.0 ml / min; TM

[0335] Determination method: accurately take 10 μl of the control solution and the test solution respectively, inject into the high performance liquid chromatograph, determine, and obtain.

[0336] The content determination method of Rheum officinale and Polygonum cuspidatum comprises the following steps:

[0337] Preparation of the control solution: take the emodin control sample, accurately weigh, add methanol to prepare a solution containing 37 μg per 1 ml, shake well, and obtain;

[0338] Preparation of the test solution: take the content of the traditional Chinese medicine composition preparation, grind finely, take 0.5 g, accurately weigh, place in a conical flask with a plug, accurately add 10% hydrochloric acid methanol solution 25 ml, weigh, heat in a water bath at 80°C for 30 minutes, cool, weigh again, make up the weight loss with methanol, shake well, filter, accurately take 4 ml of the filtrate, place in a 10 ml volumetric flask, accurately add 2% sodium hydroxide solution 2 ml, dilute to the mark with methanol, shake well, filter, take the filtrate, and obtain;

[0339] HPLC detection: the chromatographic column is C18 (CAPCELL PAK 5 μm C MG 4.6*150 mm), the mobile phase is composed of acetonitrile, methanol and 0.10% phosphoric acid solution with a volume ratio of 41:23:36, the detection wavelength is 256 nm, the column temperature is 25°C, and the flow rate is 1.0 ml / min; 18

[0340] ​​Determination method: precisely pipette 10 μl of the control solution and the sample solution respectively, inject into the liquid chromatograph, and determine, then obtain.

[0341] Example 4

[0342] Discrimination and content determination method of traditional Chinese medicine composition granules in the application

[0343] Raw material formula: Huangqi 150g, Chaihu 150g, Dahuang 80g, Zhike 150g, Yincheng 90g, Huzhang 200g, Zhizi 90g, Jinqiancao 400g, Baishao 90g, Muxiang 150g, Jiangbanxia 80g, Shengjiang 40g.

[0344] Preparation process:

[0345] A. Jiangbanxia is weighed, crushed, and passed through a 100-mesh sieve for standby use;

[0346] B. Zhike and Shengjiang are weighed, 9 times the amount of water is added, and volatile oil is extracted for 10 hours. The volatile oil is collected and separated. The water solution after distillation is standby used;

[0347] C. Huangqi, Baishao, Yincheng, and Zhizi are weighed, and water is added for decoction for 4 times. The first extraction is for 3 hours, and the second and third extractions are for 1 hour each time. 7 times the amount of water is added each time. The extraction liquid is filtered and combined with the volatile oil water extraction liquid obtained in step B. The extraction liquid is concentrated under reduced pressure to a relative density of 1.25±0.05 (60℃) of the extract, which is standby used;

[0348] D. Chaihu, Muxiang, Dahuang, Huzhang, and Jinqiancao are weighed, and 80% ethanol is added for extraction for 2 times. 10 times the amount of extraction is added for the first time for 2 hours. 10 times the amount of extraction is added for the second and third times for 1 hour each time. The extraction liquid is filtered and concentrated into a clear extract. The water extraction extract obtained in step C is combined, mixed, dried, and crushed for standby use;

[0349] E. The fine powder obtained in step A and the extract powder obtained in step D are mixed, dried, and mixed with the volatile oil obtained in step B. Granulation is performed, and the granules are finished, thereby obtaining the granules.

[0350] Discrimination:

[0351] Thin-layer identification of Bupleurum: take the content of the product, grind finely, take 3 g, add methanol 20 ml, ultrasonic treatment for 20 minutes, filter, evaporate the filtrate, add water 25 ml to dissolve the residue, extract with chloroform 3 times, 25 ml each time, discard the chloroform liquid, then extract with water-saturated n-butanol 2 times (15 ml, 25 ml), combine the n-butanol layer, wash with 3% sodium hydroxide solution 3 times, 30 ml each time, take the n-butanol liquid to evaporate, add methanol 2 ml to dissolve the residue as the test solution; take Bupleurum control drug material powder 0.5 g, add methanol 20 ml, ultrasonic treatment for 10 minutes, filter, concentrate the filtrate to 5 ml as the control drug solution, test by thin-layer chromatography, take 5 μl of the control drug solution and 10 μl of the test solution, point on the same silica gel G thin-layer plate, use chloroform-methanol-water (30:8:1 by volume) as the developing agent, develop, take out, dry, spray with 2% p-dimethylaminobenzaldehyde 40% sulfuric acid ethanol solution, heat at 60°C until the spots develop clearly, observe under daylight and ultraviolet light (365 nm), the same colored spots appear in the corresponding positions of the test chromatogram and the control drug chromatogram.

[0352] Thin-layer identification of Radix Paeoniae Alba and Gardenia: take the content of the product, grind finely, take 3 g, add methanol 20 ml, ultrasonic treatment for 20 minutes, filter, take the filtrate as the test solution; take Gardenia control drug material 0.5 g, add methanol 20 ml, heat reflux for 30 minutes, filter, evaporate the filtrate, add methanol 2 ml to dissolve the residue as the Gardenia control drug solution; take Radix Paeoniae Alba control drug material 0.5 g, add methanol 20 ml, ultrasonic treatment for 20 minutes, filter, evaporate the filtrate, add methanol 2 ml to dissolve the residue as the Radix Paeoniae Alba control drug solution, test by thin-layer chromatography, take 5 μl of the test solution and the two control drug solutions, point on the same silica gel G thin-layer plate, use chloroform-ethyl acetate-methanol-ammonia water (4:4:5:1 by volume) as the developing agent, develop, take out, dry, spray with 1% vanillin sulfuric acid solution, heat at 105°C until the spots develop clearly, the same colored spots appear in the corresponding positions of the test chromatogram and the Radix Paeoniae Alba and Gardenia control drug chromatograms.

[0353] Thin-layer identification of Radix Aucklandiae: take Radix Aucklandiae control drug material 0.2 g, add chloroform 5 ml, ultrasonic treatment for 30 minutes, filter, take the filtrate as the control drug solution, test by thin-layer chromatography, take 5 μl of the test solution in Artemisia capillaris thin-layer identification and the control drug solution, point on the same silica gel G thin-layer plate, use petroleum ether (30-60°C)-ethyl formate-formic acid upper layer solution (15:5:1 by volume) as the developing agent, develop, take out, dry, spray with 1% vanillin sulfuric acid solution, blow with hot air until the spots develop clearly, the same colored spots appear in the corresponding positions of the test chromatogram and the control drug chromatogram.

[0354] Identification of Fructus Aurantii: take the contents of the product, finely grind 1 g, add 25 ml of methanol, ultrasonic treat for 30 minutes, filter, evaporate the filtrate to dryness, add 15 ml of water to the residue to dissolve, extract with ethyl acetate twice, 20 ml each time, discard the ethyl acetate liquid, then extract with water-saturated n-butanol twice, 20 ml each time, combine the n-butanol layers, evaporate to dryness, dissolve the residue in methanol, transfer to a 25 ml volumetric flask, dilute to the mark with methanol, shake well, filter, and the filtrate is used as the test solution. Take an appropriate amount of naringin reference substance, add methanol to prepare a solution containing 80 μg per 1 ml, as the reference solution. According to the test of high performance liquid chromatography (Chinese Pharmacopoeia 2020 edition general 0512), use octadecylsilane-bonded silica gel as the filler; use acetonitrile-0.1% phosphoric acid solution (18:82) as the mobile phase; the detection wavelength is 283 nm; the column temperature is 25°C; the theoretical plate number calculated by naringin peak should not be less than 3000. Precisely take 10 μl of the reference solution and the test solution respectively, inject into the high performance liquid chromatograph, and in the test solution chromatogram, there should be a chromatographic peak consistent with the retention time of the reference solution chromatogram peak.

[0355] Content determination:

[0356] The content determination method of Scutellariae Radix includes the following steps:

[0357] Preparation of reference solution: take baicalin reference substance, accurately weigh, add 70% methanol to prepare a solution containing 20 μg per 1 ml, shake well, and obtain;

[0358] Preparation of test solution: take the contents of the traditional Chinese medicine composition preparation, finely grind, take 0.1 g, accurately weigh, place in a conical flask with a plug, accurately add 25 ml of 70% ethanol, weigh, ultrasonic treat for 10 minutes, cool, re-weigh, make up the weight loss with 70% ethanol, shake well, filter, accurately take 2 ml of the filtrate, place in a 10 ml volumetric flask, dilute to the mark with 70% ethanol, shake well, filter, and take the filtrate, which is obtained.

[0359] HPLC detection: the chromatographic column is C18 (Waters sunfire 5 μm C 18 4.6×150 mm), the mobile phase is composed of methanol and 0.1% phosphoric acid solution with a volume ratio of 45:55, the detection wavelength is 285 nm, the column temperature is 20°C, and the flow rate is 1.0 ml / min;

[0360] Determination method: accurately take 10 μl of the reference solution and the test solution respectively, inject into the high performance liquid chromatograph, and determine, which is obtained.

[0361] The content determination method of Rheum officinale and Polygonum cuspidatum includes the following steps:

[0362] Preparation of control solution: precisely take emodin control sample, add methanol to prepare a solution containing 37 μg per 1 ml, shake well, and you get it;

[0363] Preparation of test solution: take the content of traditional Chinese medicine composition preparation, grind, take 0.5 g, precisely weigh, put into a conical flask with a plug, precisely add 10% hydrochloric acid methanol solution 25 ml, weigh, heat at 80℃ water bath for 30 minutes, cool, weigh again, make up the weight loss with methanol, shake well, filter, precisely take 4 ml of the filtrate, put into a 10 ml volumetric flask, precisely add 2% sodium hydroxide solution 2 ml, dilute to the mark with methanol, shake well, filter, take the filtrate, and you get it;

[0364] HPLC detection: the chromatographic column is C18 (Waters sufire TM 5μm C 18 4.6×150mm), the mobile phase is composed of acetonitrile, methanol and 0.10% phosphoric acid solution with a volume ratio of 43:23:34, the detection wavelength is 252 nm, the column temperature is 35℃, and the flow rate is 1.0 ml / min;

[0365] Determination method: precisely take 10 μl of control solution and test solution respectively, inject into the liquid chromatograph, determine, and you get it.

[0366] The above only describes the preferred embodiments of the present application and is not used to limit the present application. Any modification, equivalent replacement or improvement made within the spirit and principle of the present application shall be included in the protection scope of the present application.

Claims

1. A method for quality control of a traditional Chinese medicine composition preparation, characterized in that, The raw materials of the traditional Chinese medicine composition preparation include the following components in parts by weight: Scutellaria baicalensis 69-205 parts, Bupleurum chinense 69-205 parts, Rheum palmatum 52-155 parts, Citrus aurantium 69-205 parts, Artemisia capillaris 69-205 parts, Polygonum cuspidatum 86-255 parts, Gardenia jasminoides 69-205 parts, Lysimachia christinae 171-515 parts, Paeonia lactiflora 69-205 parts, Aucklandia lappa 69-205 parts, Pinellia ternata 52-155 parts, and Zingiber officinale 17-52 parts. The quality control methods for the traditional Chinese medicine composition preparations include identification methods and content determination methods. The identification method includes at least one of the following identification methods (1) to (4): (1) Thin-layer chromatography with Bupleurum chinense as reference material and chloroform-methanol-water in a volume ratio of 30:8:1 as the developing solvent; (2) Thin-layer chromatography with gardenia and white peony as reference materials and chloroform-ethyl acetate-methanol-ammonia water in a volume ratio of 4:4:5:1 as the developing solvent; (3) Thin-layer chromatography with costus root as reference material and petroleum ether (30-60℃)-ethyl formate-formic acid upper layer solution with a volume ratio of 15:5:1 as developing solvent. (4) High-performance liquid chromatography with naringin as reference standard and acetonitrile-0.1% phosphoric acid solution (18:82) as mobile phase; detection wavelength 283 nm. The content determination method is to determine the content of baicalin and rhein using high performance liquid chromatography.

2. The quality control method as described in claim 1, characterized in that, The identification method described in item (1) includes the following steps: Take the traditional Chinese medicine composition preparation, extract with methanol, remove impurities with chloroform, and then extract with water-saturated n-butanol. Combine the n-butanol layers, evaporate to dryness, and dissolve the residue in methanol to obtain the test solution. Separately, take Bupleurum reference material, add methanol, sonicate, and filter. Use the filtrate as the reference material solution. Take the reference material solution and the test solution and spot them separately on the same silica gel G thin-layer plate. Develop with chloroform-methanol-water as the developing solvent. Remove, air dry, spray with 2% p-dimethylaminobenzaldehyde in 40% sulfuric acid ethanol solution, and heat until the spots are clearly visible. In the chromatogram of the test sample, spots of the same color appear at the corresponding positions as in the chromatogram of the reference material. The identification method described in item (2) includes the following steps: Take a traditional Chinese medicine composition preparation, add methanol, sonicate, filter, and take the filtrate as the test solution; take gardenia reference material, add methanol, heat and reflux to extract, filter, evaporate the filtrate to dryness, dissolve the residue in methanol to obtain the gardenia reference material solution; take white peony reference material, add methanol, sonicate, filter, evaporate the filtrate to dryness, dissolve the residue in methanol to obtain the white peony reference material solution; take the test solution and the above two reference material solutions, spot them separately on the same silica gel G thin layer plate, use chloroform-ethyl acetate-methanol-ammonia water in a volume ratio of 4:4:5:1 as the developing solvent, develop, remove and dry, spray with 1% vanillin sulfuric acid solution, heat until the spots are clearly visible, the test sample chromatogram shows spots of the same color at the corresponding positions as the gardenia reference material and white peony reference material chromatograms; The identification method described in item (3) includes the following steps: Take the reference medicinal material of Costus root, add chloroform, sonicate, filter, and use the filtrate as the reference medicinal material solution. Take the test solution under item (2) and the Costus root reference medicinal material solution, and spot them on the same silica gel G thin layer plate. Use the upper layer solution of petroleum ether (30-60℃)-ethyl formate-formic acid with a volume ratio of 15:5:1 as the developing solvent, develop, take out, dry, spray with 1% vanillin sulfuric acid solution, and heat until the spots are clearly visible. In the chromatogram of the test sample, spots of the same color appear at the corresponding positions as in the chromatogram of the reference medicinal material. The identification method described in item (4) includes the following steps: Take the traditional Chinese medicine composition preparation, add methanol, sonicate, filter, evaporate the filtrate to dryness, dissolve the residue in water, remove impurities with ethyl acetate, and then extract with water-saturated n-butanol. Combine the n-butanol layers, evaporate to dryness, dissolve the residue in methanol, filter, and use the filtrate as the test solution. Separately, take an appropriate amount of naringin reference standard, add methanol to prepare a solution containing 80 μg per ml, and use it as the reference solution. Use acetonitrile-0.1% phosphoric acid solution (18:82) as the mobile phase; the detection wavelength is 283 nm; the column temperature is 25℃; accurately pipette 10 μl of the reference solution and the test solution respectively, and inject them into the high performance liquid chromatograph. The chromatogram of the test solution should show a chromatographic peak with the same retention time as the chromatographic peak of the reference standard.

3. The quality control method as described in claim 2, characterized in that, The identification method described in item (1) includes the following steps: Take 2-4g of the traditional Chinese medicine composition preparation, add 10-50ml of methanol, sonicate, filter, evaporate the filtrate to dryness, dissolve the residue in water, extract with chloroform 1-5 times, discard the chloroform solution, then extract with water-saturated n-butanol 1-5 times, combine the n-butanol layers, wash with 3% sodium hydroxide solution, evaporate the n-butanol solution to dryness, dissolve the residue in methanol, and use this as the test solution; separately take 0.1-1g of Bupleurum chinense reference herb powder, add 10-50ml of methanol... 50 ml, sonicated, filtered, and the filtrate concentrated to serve as the reference medicinal material solution. The reference medicinal material solution and the test solution were applied separately to the same silica gel G thin-layer plate. The plate was developed using a chloroform-methanol-water mixture with a volume ratio of 30:8:

1. The plate was removed, dried, and sprayed with a 2% p-dimethylaminobenzaldehyde solution in 40% sulfuric acid ethanol. The plate was heated until the spots were clearly visible. In the chromatogram of the test sample, spots of the same color appeared at the corresponding positions as in the chromatogram of the reference medicinal material. The identification method described in item (2) includes the following steps: Take 2-4g of the traditional Chinese medicine composition preparation, add 10-50ml of methanol, sonicate, filter, and use the filtrate as the test solution; separately take 0.1-0.8g of gardenia reference material, add 10-50ml of methanol, heat under reflux, filter, evaporate the filtrate to dryness, dissolve the residue in methanol, and use the gardenia reference material solution; then take 0.1-0.8g of white peony reference material, add 10-50ml of methanol, sonicate, filter, evaporate the filtrate to dryness, dissolve the residue in methanol, and use the white peony reference material solution. Apply the test solution and the gardenia and white peony reference material solutions separately to the same silica gel G thin-layer plate, develop with a 4:4:5:1 volume ratio of chloroform-ethyl acetate-methanol-ammonia water as the developing solvent, remove, air dry, spray with 1% vanillin sulfuric acid solution, and heat until the spots are clearly visible. In the chromatogram of the test sample, spots of the same color appear at the corresponding positions as in the chromatograms of gardenia and white peony reference materials. The identification method described in item (3) includes the following steps: Take 0.1-0.3g of Costus root reference material, add 2-10ml of chloroform, sonicate, filter, and use the filtrate as the reference material solution. Take the test solution and Costus root reference material solution from item (2) and spot them on the same silica gel G thin layer plate. Use the upper layer of petroleum ether (30-60℃)-ethyl formate-formic acid with a volume ratio of 15:5:1 as the developing solvent, develop, remove, air dry, spray with 1% vanillin sulfuric acid solution, and heat until the spots are clearly visible. In the chromatogram of the test sample, spots of the same color appear at the corresponding positions as in the chromatogram of the reference material. The identification method described in item (4) includes the following steps: Weigh 0.5-2g of the traditional Chinese medicine composition preparation, add methanol, sonicate, filter, evaporate the filtrate to dryness, dissolve the residue in water, extract with ethyl acetate by shaking, discard the ethyl acetate solution, and then extract with water-saturated n-butanol by shaking. Combine the n-butanol layers, evaporate to dryness, dissolve the residue in methanol, transfer to a 25ml volumetric flask, dilute to the mark with methanol, shake well, filter, and use the filtrate as the test solution; separately, take an appropriate amount of naringin reference standard, add methanol to prepare a solution containing 80μg per ml, as the reference solution, use octadecylsilane-bonded silica gel as the stationary phase; use acetonitrile-0.1% phosphoric acid solution (18:82) as the mobile phase; the detection wavelength is 283nm; the column temperature is 25℃; accurately pipette 10μl of the reference solution and the test solution respectively, inject them into the high performance liquid chromatograph, the chromatogram of the test solution should show a chromatographic peak with the same retention time as the chromatographic peak of the reference standard.

4. The quality control method as described in claim 1, characterized in that, The method for determining the content of Scutellaria baicalensis includes the following steps: Preparation of reference solution: Accurately weigh baicalin reference standard and add 70% methanol to prepare reference solution; Preparation of the test solution: Accurately weigh the traditional Chinese medicine composition preparation, add 70% ethanol, sonicate, filter, and measure the filtrate to obtain the test solution; HPLC detection: The chromatographic column was C18, the mobile phase consisted of methanol and 0.1% phosphoric acid solution in a volume ratio of 45:55, the detection wavelength was 275-285 nm, the column temperature was 20-30℃, and the flow rate was 0.8-1.2 ml / min. Determination method: Accurately pipette 10 μl of the reference solution and the test solution into the high performance liquid chromatograph and determine the result.

5. The quality control method as described in claim 4, characterized in that, The method for determining the content of Scutellaria baicalensis includes the following steps: Preparation of reference solution: Accurately weigh baicalin reference standard, add 70% methanol to prepare a solution containing 20 μg per ml, shake well, and the solution is ready; Preparation of the test solution: Weigh 0.1g of the contents of the traditional Chinese medicine composition preparation accurately, add 25ml of 70% ethanol accurately, filter, take 2ml of the filtrate, dilute with 70% ethanol to the mark, shake well, filter, and take the filtrate to obtain the test solution; HPLC detection: The chromatographic column was C18, the mobile phase consisted of methanol and 0.1% phosphoric acid solution in a volume ratio of 45:55, the detection wavelength was 275-285 nm, the column temperature was 20-30℃, and the flow rate was 0.8-1.2 ml / min. Determination method: Accurately pipette 10 μl of the reference solution and the test solution into the high performance liquid chromatograph and determine the result.

6. The quality control method as described in claim 1, characterized in that, The method for determining the content of rhubarb and Japanese knotweed includes the following steps: Preparation of reference solution: Take rhein reference standard, add methanol and shake well to obtain the solution; Preparation of the test solution: Take the contents of the traditional Chinese medicine composition preparation, add 10% hydrochloric acid methanol solution, heat to reflux, filter, add 2% sodium hydroxide solution to the filtrate, and dilute with methanol to obtain the test solution; HPLC detection: The chromatographic column was C18, and the mobile phase consisted of acetonitrile, methanol and 0.10% phosphoric acid solution in a volume ratio of 41-43:23:34-36. The detection wavelength was 252-256 nm, the column temperature was 25-35℃, and the flow rate was 0.8-1.2 ml / min. Determination method: Accurately pipette 10 μl of the reference solution and the test solution into the liquid chromatograph and determine the result.

7. The quality control method as described in claim 6, characterized in that, The method for determining the content of rhubarb and Japanese knotweed includes the following steps: Preparation of reference solution: Weigh the rhein reference standard accurately, add methanol to prepare a solution containing 37 μg per ml, shake well, and the solution is ready. Preparation of the test solution: Take the contents of the traditional Chinese medicine composition preparation, grind it into a fine powder, take 0.5g, accurately weigh it, accurately add 25ml of 10% hydrochloric acid methanol solution, weigh it, heat it in an 80℃ water bath under reflux for 30 minutes, cool it, weigh it again, make up the lost weight with methanol, shake it well, filter it, accurately measure 4ml of the filtrate, put it in a 10ml volumetric flask, accurately add 2ml of 2% sodium hydroxide solution, dilute it with methanol to the mark, shake it well, filter it, and take the filtrate to obtain the test solution; HPLC detection: The chromatographic column was C18, and the mobile phase consisted of acetonitrile, methanol and 0.10% phosphoric acid solution in a volume ratio of 41-43:23:34-36. The detection wavelength was 252-256 nm, the column temperature was 25-35℃, and the flow rate was 0.8-1.2 ml / min. Determination method: Accurately pipette 10 μl of the reference solution and the test solution into the liquid chromatograph and determine the result.

8. The quality control method according to any one of claims 1 to 7, characterized in that, The dosage form of the traditional Chinese medicine composition is capsule, tablet, pill, oral liquid, granule or powder.

9. The quality control method as described in claim 8, characterized in that, The preparation process of capsules is as follows: A. Weigh out ginger and Pinellia ternata, crush them, and pass them through a 100-mesh sieve for later use; B. Weigh out the bitter orange peel and fresh ginger, add 6-8 times the amount of water, extract the volatile oil for 9-11 hours, collect and separate the volatile oil; the distilled aqueous solution is for later use. C. Weigh out Scutellaria baicalensis, Paeonia lactiflora, Artemisia capillaris, and Gardenia jasminoides. Add water and decoct 2-3 times. The first extraction takes 2 hours, and the second and third extractions take 1.5 hours each time. Add 8-10 times the amount of water each time. Filter the extract and combine it with the volatile oil-water extract obtained in step B. Concentrate under reduced pressure to a relative density of 1.25±0.05 (60℃) to obtain an extract for later use. D. Weigh out Bupleurum, Aucklandia, Rhubarb, Polygonum cuspidatum, and Lysimachia christinae. Extract with 60-80% ethanol three times. For the first extraction, add 11-13 times the amount of ethanol and extract for 2-3 hours. For the second and third extractions, add 9-11 times the amount of ethanol and extract for 1-3 hours each. Filter the extract, concentrate it into a clear paste, combine it with the water extract obtained in step C, mix well, dry, and pulverize for later use. E. Mix the fine powder obtained in step A and the dry powder obtained in step D, dry them, adsorb the volatile oil obtained in step B with silica, mix them, and then fill them into capsules to obtain the final product.

10. The quality control method according to any one of claims 1-7, characterized in that, The raw materials of the traditional Chinese medicine composition preparation include the following components in parts by weight: Scutellaria baicalensis 128 parts, Bupleurum chinense 128 parts, Rheum palmatum 120 parts, Citrus aurantium 128 parts, Artemisia capillaris 128 parts, Polygonum cuspidatum 200 parts, Gardenia jasminoides 150 parts, Lysimachia christinae 250 parts, Paeonia lactiflora 128 parts, Aucklandia lappa 128 parts, Pinellia ternata 90 parts and fresh ginger 40 parts.

11. The quality control method according to any one of claims 1-7, characterized in that, The raw materials of the traditional Chinese medicine composition preparation include the following components in parts by weight: 137 parts of Scutellaria baicalensis, 137 parts of Bupleurum chinense, 103 parts of Rheum palmatum, 137 parts of Citrus aurantium, 137 parts of Artemisia capillaris, 171 parts of Polygonum cuspidatum, 137 parts of Gardenia jasminoides, 342 parts of Lysimachia christinae, 137 parts of Paeonia lactiflora, 137 parts of Aucklandia lappa, 103 parts of Pinellia ternata, and 34 parts of Zingiber officinale.

12. The quality control method according to any one of claims 1-7, characterized in that, The raw materials of the traditional Chinese medicine composition preparation include the following components in parts by weight: 150 parts of Scutellaria baicalensis, 70 parts of Bupleurum chinense, 90 parts of Rheum palmatum, 90 parts of Citrus aurantium, 150 parts of Artemisia capillaris, 100 parts of Polygonum cuspidatum, 110 parts of Gardenia jasminoides, 500 parts of Lysimachia christinae, 80 parts of Paeonia lactiflora, 200 parts of Aucklandia lappa, 110 parts of Pinellia ternata, and 20 parts of fresh ginger.

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