Preparation process and quality detection method of Shexiang Xintongning dropping pills

Abstract

The present invention discloses a preparation process and a quality detection method for Shexiang Xintongning Pills, belonging to the technical field of traditional Chinese medicine preparation. By determining the extraction method of medicinal materials, extraction solvent, extraction time, dropping parameters of the pills, etc., the present invention can better extract the active ingredients from the medicinal materials, improve the purity, dissolution rate and extraction rate of the active ingredients, and further improve the utilization rate of the medicinal materials and the absorption rate of the active ingredients in the medicinal materials. The established content determination methods for ginsenoside Rg1, tetrahydropalmatine, cinnamic acid and ferulic acid in the present invention have high precision, strong reproducibility, strong specificity, high stability, accurate results, and can ensure the quality stability of the components in the pills. The established TLC identification methods for Corydalis Rhizoma, Chuanxiong Rhizoma, Ginseng Radix et Rhizoma, Styrax, and Borneolum Syntheticum in the present invention have strong specificity, clear chromatographic spots, and no interference from negative controls.

Description

Technical Field

[0001] The present invention relates to a preparation process and a quality detection method of Shexiang Xintongning Pills, belonging to the technical field of traditional Chinese medicine preparation. Background Art

[0002] Shexiang Xintongning Tablets are composed of six herbs, namely Corydalis Yanhusuo processed with vinegar, Moschus, Styrax, Chuanxiong Rhizoma, Ginseng Radix et Rhizoma, and Borneolum Syntheticum, and have the effects of promoting qi and opening orifices, activating blood circulation to remove stasis, and dredging collaterals to relieve pain. Clinically, it is used for chest pain, chest tightness, distending pain in both flanks, shortness of breath, palpitation and other symptoms caused by angina pectoris of coronary heart disease with qi stagnation and blood stasis type. The existing dosage form of the Shexiang Xintongning formula is tablets. After oral administration, tablets generally need to be disintegrated and dissolved before being absorbed, and the drug effect is relatively slow, so it cannot be used for first aid of angina pectoris. Moreover, tablets can only be taken orally and cannot be sublingually administered, and the active ingredients of the drug effect cannot be directly absorbed into the blood to exert the curative effect. In addition, due to the slow dissolution rate of the drug in tablets, its bioavailability is relatively low.

[0003] Therefore, the present invention aims to provide a preparation method of Shexiang Xintongning Pills to solve the above problems. Among the raw materials of traditional Chinese herbs in the Shexiang Xintongning formula, the main active ingredients of Corydalis Yanhusuo are alkaloid components, the main active ingredients of Ginseng Radix et Rhizoma are saponin components, and the main active ingredients of Chuanxiong Rhizoma are organic acid components. The properties of the three active ingredients vary greatly. Therefore, the extraction and purification processes of the herbs have a great influence on the transfer rate of different active ingredients during the preparation of pills, which directly affects the drug effect of the pills.

[0004] In addition, although a quality control method for Shexiang Xintongning Tablets has been established, there is no relevant quality detection method for Shexiang Xintongning Pills.

[0005] It should be noted that the above content belongs to the technical cognition scope of the inventor and does not necessarily constitute the prior art. Summary of the Invention

[0006] The present invention provides a preparation process and a quality detection method of Shexiang Xintongning Pills to solve the problems existing in the prior art, broaden the dosage form of Shexiang Xintongning preparations, and help improve the quality of Shexiang Xintongning preparations.

[0007] The present invention realizes the above purpose by adopting the following technical solutions:

[0008] On the one hand, the present invention provides a preparation process of Shexiang Xintongning Pills, including the following steps:

[0009] (1) Mix the Corydalis Rhizoma slices, Ligustici Chuanxiong Rhizoma slices, and Ginseng Radix slices, add ethanol with a weight - volume concentration of 50% - 70% at 8 - 12 times the weight, and reflux - extract 1 - 2 times, with each reflux for 1 - 2 hours. Combine the extraction solutions, filter, and after recovering the ethanol from the filtrate, concentrate it under reduced pressure at 60 - 85°C to obtain a concentrated medicinal liquid with a concentration of 0.7 - 1.2 g of crude drug / mL at 60 ± 5°C;

[0010] (2) Purify the concentrated medicinal liquid through macroporous adsorption resin, then elute the purified liquid with ethanol having a volume concentration of 60% - 70%. Collect the eluate, evaporate it to dryness, and dry it to a constant weight to obtain a paste;

[0011] (3) Grind Moschus and Borneolum Syntheticum separately, grind the paste, mix the paste, Moschus, Borneolum Syntheticum, and Styrax Liquidambaris, and then add 3 - 5 times the weight of the matrix. Stir until melted and uniform under a water - bath heating at 80 - 90°C. The matrix is a mixture of PEG4000 and PEG6000 with a weight ratio of 3 - 4:1;

[0012] (4) Add the melted medicinal liquid to a dropping pill machine for dropping. The temperature of the coolant is 6 - 14°C, the dropping distance is 4 - 8 cm, the dropping speed is 25 - 35 drops / min. Collect the dropping pills and dry them to prepare the Moschus Xintongning dropping pills.

[0013] Preferably, in step (2), the macroporous adsorption resin is X - 5 type macroporous adsorption resin; in step (3), the coolant uses dimethyl silicone oil.

[0014] Preferably, in step (2), the specific purification method: Pack the macroporous adsorption resin according to a height - to - diameter ratio of 1:8, load the medicinal liquid by wet method at 1 - 2 bed volumes (BV), perform dynamic adsorption at a flow rate of 8 - 10 BV / h, and wash away the residual impurities on the surface of the macroporous adsorption resin with more than 2.5 BV of pure water.

[0015] Preferably, in step (2), the specific elution method: Elute with ethanol having a volume concentration of 60% - 70% at a flow rate of 8 - 10 BV / h with a volume of more than 8 BV.

[0016] Preferably, the weight parts of each raw material in the Moschus Xintongning dropping pills are: Corydalis Rhizoma 220 - 260 parts, Artificial Moschus 4 - 12 parts, Styrax Liquidambaris 4 - 12 parts, Ligustici Chuanxiong Rhizoma 100 - 140 parts, Ginseng Radix 100 - 140 parts, Borneolum Syntheticum 6 - 18 parts.

[0017] Furthermore, the present invention provides a Moschus Xintongning dropping pill prepared according to the above - mentioned preparation process of the Moschus Xintongning dropping pill.

[0018] On the other hand, the present invention provides a quality detection method for Shexiang Xintongning Pills, including a method for determining the content of ginsenoside Rg1 and / or a method for determining the contents of tetrahydropalmatine, cinnamic acid, and ferulic acid and / or a thin-layer identification method for Corydalis Rhizoma, Ligustici Chuanxiong Rhizoma, Ginseng Radix et Rhizoma, Styrax, and Borneolum Syntheticum.

[0019] Among them, the method for determining the content of ginsenoside Rg1 includes the following steps:

[0020] (1) Prepare control solutions with different concentrations:

[0021] Take the ginsenoside Rg1 reference substance, dissolve it in methanol to make a stock solution of ginsenoside Rg1, and then dilute it with methanol to prepare a series of control solutions of ginsenoside Rg1 with different mass concentrations;

[0022] (2) Prepare the test solution:

[0023] Take 0.5 g of the powder of Shexiang Xintongning Pills, place it in a stoppered conical flask, add 25 mL of methanol, ultrasonically treat for 30 min, let it cool to room temperature, filter, dilute to 25 mL with methanol, shake well, and filter through a 0.45 μm filter membrane to obtain the test solution;

[0024] (3) Prepare the negative control solution:

[0025] Add pure matrix to the pill dropping machine for dropping, prepare negative pills from the pure matrix according to the method in step (4) of claim 1, take 0.5 g, place it in a stoppered conical flask, add 25 mL of methanol, ultrasonically treat for 30 min, let it cool to room temperature, filter, dilute to 25 mL with methanol, shake well, and filter through a 0.45 μm filter membrane to obtain the negative control solution;

[0026] (4) Liquid chromatography conditions:

[0027] Chromatographic column: C18 chromatographic column; Mobile phase: 0.05% phosphoric acid water (A) - acetonitrile (B); Gradient elution program: 0 - 10 min: 80% A, 20% B; 10 - 25 min: 80% → 76% A, 20% → 24% B; 25 - 33 min: 76% A, 24% B; Flow rate: 1.0 mL / min; Column temperature: 30 °C; Detection wavelength: 203 nm; Injection volume: 10 μL;

[0028] (5) Determination:

[0029] Take the control solution, negative control solution, and test solution, inject them into the liquid chromatograph, and perform determination according to the said chromatographic conditions. Calculate the content of ginsenoside Rg1 based on the peak area.

[0030] Among them, the method for determining the contents of tetrahydropalmatine, cinnamic acid, and ferulic acid includes the following steps:

[0031] (1) Preparation of mixed reference substance solutions with different concentrations:

[0032] Take tetrahydropalmatine reference substance, cinnamic acid reference substance, and ferulic acid reference substance, add methanol to make up the volume, prepare the mother solution of the tetrahydropalmatine reference substance, and then dilute it with methanol to prepare a series of tetrahydropalmatine reference substance solutions, cinnamic acid reference substance solutions, and ferulic acid reference substance solutions with a series of mass concentrations;

[0033] (2) Preparation of test solution:

[0034] Take 0.5 g of Shexiang Xintongning Pills powder, place it in a stoppered conical flask, add 25 mL of methanol, ultrasonically treat it for 30 min, let it cool to room temperature, filter, add methanol to make up the volume to 25 mL, shake well, filter through a 0.45 μm filter membrane, and dilute it 4 times with methanol to obtain;

[0035] (3) Preparation of negative control solution:

[0036] Add pure matrix to the pill dropping machine for dropping, prepare negative pills from the pure matrix according to the method in step (4) of claim 1, take 0.5 g, place it in a stoppered conical flask, add 25 mL of methanol, ultrasonically treat it for 30 min, let it cool to room temperature, filter, add methanol to make up the volume to 25 mL, shake well, filter through a 0.45 μm filter membrane to obtain;

[0037] (4) Liquid chromatography conditions:

[0038] Chromatographic column: C18 chromatographic column; Mobile phase: 0.1% formic acid water (A) - acetonitrile (B); Gradient elution program: 0 - 10 min: 90% → 80% A, 10% → 20% B; 10 - 20 min: 80% → 70% A, 20% → 30% B; 20 - 23 min: 70% → 65% A, 30% → 35% B; 23 - 30 min: 65% → 48% A, 35% → 52% B; Flow rate 1.0 mL / min; Column temperature: 30 °C; Detection wavelength: 280 nm for tetrahydropalmatine and cinnamic acid, 321 nm for ferulic acid; Injection volume 10 μL;

[0039] (5) Determination:

[0040] Take the reference substance solution, negative control solution, and test solution, inject them into the liquid chromatograph, perform determination according to the said chromatographic conditions, and calculate the contents of tetrahydropalmatine, cinnamic acid, and ferulic acid based on the peak areas.

[0041] Among them, the method for determining the content of muskone includes the following steps:

[0042] (1) Preparation of reference substance solution:

[0043] Take an appropriate amount of muskone reference substance, dissolve it in ethyl acetate and make up to 25 mL to prepare a mother solution of the reference substance with a concentration of 406.4 μg / mL, and dilute it with ethyl acetate to prepare a series of muskone reference substance solutions with different mass concentrations;

[0044] (2) Preparation of the test solution:

[0045] Take 1 g of the powder of Shexiang Xintongning Pills, add 20 mL of ethyl acetate, extract ultrasonically for 30 min, filter, make up to 20 mL in a volumetric flask, and filter through a 0.22 μm filter membrane to obtain the solution;

[0046] (3) Preparation of the negative control solution:

[0047] Add the pure matrix to the pill dropping machine for dropping, and prepare negative pills from the pure matrix according to the method in step (4) of claim 1. Take 1 g, add 20 mL of ethyl acetate, extract ultrasonically for 30 min, filter, make up to 20 mL in a volumetric flask, and filter through a 0.22 μm filter membrane to obtain the solution;

[0048] (4) Gas chromatography conditions:

[0049] Chromatographic column: HP-5 capillary column; Temperature programming: Initial temperature 80 °C, hold for 1 min, increase the temperature at a rate of 20 °C / min to 160 °C, hold for 1 min, then increase the temperature at a rate of 20 °C / min to 260 °C and hold for 2 min; Detector: Flame ionization detector; Carrier gas: Nitrogen, carrier gas flow rate: 13.5 mL / min; Combustion-supporting air flow rate: 400 mL / min; Combustible gas hydrogen flow rate: 20 mL / min; Nitrogen tail gas blow flow rate: 25 mL / min; Injection volume: 1 μL; Split ratio: 10:1; Vaporization chamber temperature: 220 °C, detector temperature: 260 °C;

[0050] (5) Determination:

[0051] Take the reference substance solution, negative control solution and test solution, inject them into the gas chromatograph, and carry out the determination according to the said chromatographic conditions, and calculate the content of muskone according to the peak area.

[0052] Among them, the thin-layer identification method for Corydalis yanhusuo includes the following steps:

[0053] Preparation of test solution, control crude drug solution, negative control solution, and tetrahydropalmatine reference substance solution: Weigh 0.5 g of the powdered Shexiang Xintongning Pills after grinding, add 20 mL of petroleum ether with a boiling range of 60 - 90°C, then add 1.5 mL of concentrated ammonia water, perform ultrasonic extraction for 30 min, filter, evaporate the filtrate to dryness at 60°C, dissolve the residue with 1 mL of petroleum ether at 60 - 90°C to obtain the test solution; Weigh 0.5 g of the control crude drug of Corydalis yanhusuo and 0.5 g of the pill powder without Corydalis yanhusuo respectively, and prepare the control crude drug solution and negative control solution according to the same method as for the preparation of the test solution; Take tetrahydropalmatine reference substance, dissolve it with methanol to prepare a tetrahydropalmatine reference substance solution with a mass concentration of 0.2 mg / mL.

[0054] According to the thin-layer chromatography test, pipette 10 μL each of the test solution, control crude drug solution, negative control solution, and tetrahydropalmatine reference substance solution, and spot them on a silica gel GF254 thin-layer plate respectively. Use a mixed solution of n-hexane - chloroform - methanol with a volume ratio of 10:6:1 as the developing agent, pre-saturate for 15 min, develop, air-dry, place in an iodine cylinder for color development, and observe under ultraviolet light at a wavelength of 365 nm after the iodine has evaporated; In the chromatogram of the test sample, fluorescent spots of the same color appear at the positions corresponding to the control crude drug of Corydalis yanhusuo and the tetrahydropalmatine reference substance, and there is no interference from the negative control.

[0055] Among them, the thin-layer identification method for Ligusticum chuanxiong Hort. includes the following steps:

[0056] Preparation of test solution, control crude drug solution, negative control solution: Weigh 0.5 g of the powdered Shexiang Xintongning Pills and place it in a conical flask, add 20 mL of ether, perform ultrasonic extraction for 10 min, filter, evaporate the filtrate to dryness at 30°C, dissolve the residue with 1 mL of ethanol to obtain the test solution; Weigh 0.5 g of the control crude drug of Ligusticum chuanxiong Hort. and 0.5 g of the pill powder without Ligusticum chuanxiong Hort. respectively, and prepare the control crude drug solution and negative control solution according to the same method as for the preparation of the test solution;

[0057] According to the thin-layer chromatography test, pipette 10 μL each of the test solution, control crude drug solution, and negative control solution, and spot them on the same silica gel GF254 thin-layer plate respectively. Use a mixed solution of n-hexane - ethyl acetate with a volume ratio of 4:1 as the developing agent, develop, air-dry, and observe under ultraviolet light at a wavelength of 365 nm; In the chromatogram of the test sample, fluorescent spots of the same color appear at the positions corresponding to the control crude drug, and there is no interference from the negative control.

[0058] Among them, the thin-layer identification method for Panax ginseng C. A. Mey. includes the following steps:

[0059] Preparation of test solution, negative control solution, ginsenoside Rg1 reference solution, and ginsenoside Re reference solution: Take 1 g of Shexiang Xintongning Pills, grind them, add 20 mL of ethanol, let stand for 3 hours, shake during the standing process, then perform ultrasonic extraction for 20 min, place in the refrigerator for 24 h, filter, evaporate the filtrate to nearly dry at 70 °C, dissolve the residue in 2 mL of methanol to obtain the test solution; take 1 g of the pill powder without ginseng and prepare the negative control solution in the same manner as the test solution; separately take ginsenoside Rg1 reference substance and ginsenoside Re reference substance, add methanol to prepare 0.5 mg / mL ginsenoside Rg1 reference solution and ginsenoside Re reference solution;

[0060] According to the thin-layer chromatography test, absorb 10 μL each of the test solution, negative control solution, ginsenoside Rg1 reference solution, and ginsenoside Re reference solution, and spot them on the same silica gel GF254 thin-layer plate respectively; use the lower layer solution of the mixed solution of chloroform-ethyl acetate-methanol-water with a volume ratio of 7.5:20:11:5 when placed at a temperature below 10 °C as the developing agent, develop, air dry, spray with a 10% sulfuric acid ethanol solution, air dry, and heat at 105 °C until the spots are clearly developed; in the test solution chromatogram, at the positions corresponding to ginsenoside Rg1 reference substance and ginsenoside Re reference substance, there are the same pink spots, and there is no interference from the negative control.

[0061] Among them, the thin-layer identification method for styrax includes the following steps:

[0062] Preparation of test solution, negative control solution, cinnamic acid reference solution: Take 1 g of the ground Shexiang Xintongning Pills powder, add 10 mL of ether and perform ultrasonic treatment for 2 min, take the supernatant as the test solution; take 1 g of the pill powder without styrax and prepare the negative control solution in the same manner as the test solution; take cinnamic acid reference substance and add ether to prepare 1 mg / mL cinnamic acid reference solution;

[0063] According to the thin-layer chromatography test, absorb the test solution, negative control solution, and cinnamic acid reference solution, and spot them on the same silica gel GF254 thin-layer plate respectively. Use the mixed solution of petroleum ether (30 - 60 °C)-n-hexane-ethyl formate-formic acid with a volume ratio of 10:30:15:1 as the developing agent, develop, air dry, spray with a 10% sulfuric acid ethanol solution, dry by blowing, heat at 105 °C until the spots are clearly developed, and examine under ultraviolet light at a wavelength of 254 nm; in the test solution chromatogram, at the position corresponding to cinnamic acid reference substance, there are fluorescent spots of the same color, and there is no interference from the negative control.

[0064] Among them, the thin-layer identification method for borneol includes the following steps:

[0065] Preparation of test solution, negative control solution, and borneol reference substance solution: Take 0.3 g of the powdered Shexiang Xintongning Pills after grinding, add 15 mL of ethyl acetate, ultrasonically treat for 2 min, filter, evaporate the filtrate to nearly dry at 60°C, and redissolve with 0.5 mL of ethyl acetate to obtain the test solution; take 0.3 g of the pill powder without borneol and prepare the negative control solution in the same manner as the test solution; take the borneol reference substance and prepare a 2.5 mg / mL borneol reference substance solution with ethyl acetate.

[0066] According to the thin-layer chromatography test, pipette 2 μL each of the test solution, negative control solution, and borneol reference substance solution, and spot them on the same silica gel GF254 thin-layer plate respectively. Use a mixed solution of toluene - acetone with a volume ratio of 9:1 as the developing agent, spray a 5% vanillin sulfuric acid solution, and heat at 105°C until the spots are clearly developed; in the chromatogram of the test sample, a same yellowish-green fluorescent spot appears at the same position as the borneol reference substance, and there is no interference from the negative control.

[0067] The beneficial effects of this application include but are not limited to:

[0068] The preparation process of the Shexiang Xintongning Pills provided by the present invention determines the extraction method of medicinal materials, extraction solvent, extraction time, dropping parameters of the pills, etc., and can better extract the active ingredients in the medicinal materials, improving the purity, dissolution rate, and extraction rate of the active ingredients, thereby improving the utilization rate of the medicinal materials and the absorption rate of the active ingredients in the medicinal materials; the prepared pills have a round and regular appearance, are moderately hard and soft, and are not prone to adhesion.

[0069] For the Shexiang Xintongning Pills provided by the present invention, the active ingredients in the medicine are highly dispersed in the pills, improving the dissolution performance of the active ingredients. The pills have a short disintegration time limit and a fast dissolution rate of the active ingredients, thereby improving the bioavailability of the medicine in the body and enabling the medicine to exert a high-efficiency effect. The Shexiang Xintongning Pills can be taken orally or sublingually. Sublingual administration can directly absorb the medicine into the blood through the oral mucosa and take effect quickly.

[0070] The quality detection method of the Shexiang Xintongning Pills provided by the present invention uses an optimized solution preparation method and instrument detection method to quantitatively and qualitatively detect the main active ingredients contained in the medicine. The content determination methods of ginsenoside Rg1, tetrahydropalmatine, cinnamic acid, and ferulic acid established by the present invention have high precision, strong reproducibility, strong specificity, high stability, accurate results, and can ensure the quality stability of the components in the pills. The thin-layer identification methods of Corydalis Rhizoma, Ligustici Chuanxiong Rhizoma, Ginseng Radix, Styrax, and Borneol established by the present invention have strong specificity, clear chromatographic spots, and no interference from the negative control. Description of the Drawings

[0071] The accompanying drawings described herein are used to provide a further understanding of the present application and form a part of the present application. The schematic embodiments of the present application and their descriptions are used to explain the present application and do not constitute an improper limitation of the present application. In the drawings:

[0072] Figure 1 is the appearance photo of Shexiang Xintongning Pills;

[0073] Figure 2 is the influence of the concentration of the sample solution on the adsorption capacity;

[0074] Figure 3 is the influence of the adsorption flow rate of macroporous adsorption resin on the adsorption capacity;

[0075] Figure 4 is the adsorption leakage curve;

[0076] Figure 5 is the influence of the ethanol concentration on the concentration of the residual liquid;

[0077] Figure 6 is the HPLC chart of ginseng (A is the negative control, B is the reference substance of ginsenoside Rg1, C is the test sample; peak 1 is ginsenoside Rg1);

[0078] Figure 7 is the HPLC chart of Corydalis yanhusuo, Styrax, and ferulic acid (A is the negative control, B is the mixed reference substance, C is the test sample; peak 1 is tetrahydropalmatine, 2 is cinnamic acid);

[0079] Figure 8 is the TLC identification results of Corydalis yanhusuo (A), Ligusticum chuanxiong (B), ginseng (C), borneol (D), and Styrax (E). In A: 1 is the reference substance of tetrahydropalmatine, 2 is the test sample of the pills, 3 is the reference medicinal material of Corydalis yanhusuo, 4 is the negative control without Corydalis yanhusuo; in B: 1 is the test sample of the pills, 2 is the reference medicinal material of Ligusticum chuanxiong, 3 is the negative control without Ligusticum chuanxiong; in C: 1 is the reference substance of ginsenoside Rg1, 2 is the reference substance of ginsenoside Re, 3 is the test sample of the pills, 4 is the negative control without ginseng; in D: 1 is the reference substance of cinnamic acid, 2 is the test sample of the pills, 3 is the negative control without Styrax; in E: 1 is the reference substance of borneol, 2 is the test sample of the pills, 3 is the negative control without borneol. Detailed Description of the Invention

[0080] In the following content, the preparation process and quality inspection method of Shexiang Xintongning Pills provided by the present invention application will be further described in detail. However, it should be noted that the following specific embodiments only give specific operation examples of the present invention in an exemplary manner, but the protection scope of the present invention is not limited thereto.

[0081] I. Instruments and Materials

[0082] 1.1 Instruments:

[0083] Agilent 1260 high performance liquid chromatograph; ZB-1E intelligent disintegration tester (Tianjin Tianda Tianfa Technology Co., Ltd.); JOANLAB BHS-4 numerically controlled constant temperature water bath (Ningbo Yinzhou Qunan Experimental Instrument Co., Ltd.); SQP-SECURA125-1CN electronic balance (Sartorius Scientific Instruments (Beijing) Co., Ltd.).

[0084] 1.2 Materials:

[0085] Tetrahydropalmatine (batch number Y21S7Y17091), ferulic acid (batch number L03A9D57744), ginsenoside Rg1 (batch number G30N10Y104330) were all purchased from Shanghai Yuanye Bio-Technology Co., Ltd. (purity ≥ 98%);

[0086] Ginsenoside Re (batch number 52286-59-6), cinnamic acid (batch number 140-10-3), muskone (batch number B18N10D103404), borneol (batch number 23356-29-8) were all purchased from Shanghai Yuanye Bio-Technology Co., Ltd. (purity ≥ 98%);

[0087] Rhizoma Chuanxiong reference crude drug (batch number 120918-201813), Rhizoma Corydalis reference crude drug (batch number 120928-201609), Radix Ginseng reference crude drug (batch number 120917-201712) were all purchased from National Institutes for Food and Drug Control;

[0088] Purified Rhizoma Corydalis (batch number 20060011), purified Rhizoma Chuanxiong (batch number 20040011), purified Radix Ginseng (batch number 18040016) were respectively purchased from Anhui Weibai Traditional Chinese Medicine Co., Ltd., Inner Mongolia Mengyuantang Pharmaceutical Technology Co., Ltd., Anhui Helin Traditional Chinese Medicine Decoction Pieces Technology Co., Ltd.; artificial musk (China National Traditional Chinese Medicine Co., Ltd.); borneol (Fujian Qingsong Co., Ltd.); styrax (Hebei Yiren Pharmaceutical Co., Ltd.).

[0089] Shexiang Xintongning dropping pills, batches 20220429, 20220430, 20220501; dropping pills without Rhizoma Corydalis, dropping pills without Rhizoma Chuanxiong, dropping pills without Radix Ginseng, dropping pills without styrax, dropping pills without borneol were respectively prepared by the same preparation process as Shexiang Xintongning dropping pills after removing Rhizoma Corydalis, Rhizoma Chuanxiong, Radix Ginseng, styrax, borneol from the raw materials.

[0090] Silica gel GF254 thin layer plates were purchased from Qingdao Ocean Chemical Co., Ltd.; analytical grade methanol, chromatographic grade formic acid, phosphoric acid and acetonitrile were all purchased from Sinopharm Chemical Reagent Co., Ltd.; Wahaha pure water was purchased from Hangzhou Wahaha Group Co., Ltd.

[0091] Phosphoric acid, chromatographic grade acetonitrile (Sinopharm Chemical Reagent Co., Ltd.); triethylamine (Tianjin Fuyu Fine Chemical Co., Ltd.); alcohol (Jilin Xintianlong Industry Co., Ltd.); AB-8, X-5, D101, HPD100, HPD400 macroporous adsorption resins (Shanghai Yuanye Bio-Technology Co., Ltd.).

[0092] PEG 4000, PEG 6000 (Shanghai Macklin Biochemical Co., Ltd.); dimethyl silicone oil (Tianjin Damao Chemical Reagent Factory); liquid paraffin (Tianjin Fuyu Fine Chemical Co., Ltd.).

[0093] II. Preparation process of Shexiang Xintongning Pills

[0094] Example 1:

[0095] The Shexiang Xintongning Pills provided in this example are made from the following raw materials by weight: 239 g of Corydalis Rhizoma slices, 8 g of artificial musk, 8 g of Styrax, 119 g of Ligusticum Chuanxiong Rhizoma slices, 119 g of Ginseng Radix slices, and 12 g of Borneolum Syntheticum.

[0096] The preparation process of the Shexiang Xintongning Pills provided in this example is as follows:

[0097] (1) Mix the Corydalis Rhizoma slices, Ligusticum Chuanxiong Rhizoma slices, and Ginseng Radix slices, and reflux and extract them twice with 50% ethanol. Each time, add 10 times the amount of 50% ethanol, reflux for 1.5 hours each time, combine the extraction solutions, filter, and concentrate the filtrate by recovering ethanol to a concentrated medicinal liquid with a concentration of 1 g of crude drug / mL;

[0098] (2) Purify the concentrated medicinal liquid through X-5 macroporous adsorption resin (belonging to styrene-type non-polar copolymer). The purification parameters are as follows: Pack the macroporous adsorption resin according to a height-diameter ratio of 1:8, load the medicinal liquid wetly at about 2 BV, perform dynamic adsorption at a flow rate of 10 BV / h, wash the residual impurities on the surface of the macroporous adsorption resin with 3 BV of pure water, and then elute with about 9 BV of 70% ethanol at a flow rate of 10 BV / h. Collect the eluate, evaporate it to dryness, and dry it to constant weight to obtain a paste. The paste yield is about 2.17%;

[0099] (3) Grind the paste finely, and according to the conversion of "the daily dosage of the original process is converted into crude drug amounts of 1.434 g of Corydalis Rhizoma, 48 mg of musk, 48 mg of Styrax, 0.714 g of Ligusticum Chuanxiong Rhizoma, 0.714 g of Ginseng Radix, and 72 mg of Borneolum Syntheticum", add musk, Borneolum Syntheticum, and Styrax respectively. According to drug:matrix = 1:4, add the matrix (PEG4000:PEG 6000 = 3:1), and stir and melt it in a water bath at 85°C;

[0100] (4) Add the molten medicinal liquid to a pill dropping machine for dropping. The temperature of the coolant is 6 - 14°C, the dropping distance is 6 cm, and the dropping speed is 30 drops / min for dropping. Collect the pills and dry them to prepare the Shexiang Xintongning Pills.

[0101] The daily raw drug dosage of Corydalis yanhusuo, Ligusticum chuanxiong, and Panax ginseng is 1.434 + 0.714 + 0.714 = 2.862 g. For the preparation process provided by the present invention, the yield after purification according to the process results is approximately 2.17% of the raw drug dosage. The purified mass of the above three medicinal materials contained in the Shexiang Xintongning dropping pills prepared is 2.862 × 2.17% = 62 mg, 48 mg of Moschus, 48 mg of Styrax, and 72 mg of Borneolum Syntheticum.

[0102] The Shexiang Xintongning dropping pills prepared by the present invention have a pill weight of 35 mg. Take 11 pills once, three times a day, and the daily weight of the taken dropping pills is 1150 mg.

[0103] 2.1 Characters of the Shexiang Xintongning dropping pills prepared by the present invention

[0104] Through observation and tasting, the characters of the Shexiang Xintongning dropping pills are all dark brown dropping pills, with a slight aroma and a slightly bitter taste, and the appearance is as shown in Figure 1 .

[0105] 2.2 Dissolution time limit

[0106] Check the dissolution time limit of the Shexiang Xintongning dropping pills according to the general rules in Part IV of the Chinese Pharmacopoeia (2020 Edition), as shown in Table 1.

[0107] Table 1 Dissolution time limit of the Shexiang Xintongning dropping pills

[0108]

[0109]

[0110] It can be seen that the average dissolution time limit of the Shexiang Xintongning dropping pills prepared by the present invention is 3.73 min, which is less than 30 min, meeting the requirements of the pharmacopoeia. The dissolution time limit of the Shexiang Xintongning tablets is 10 - 20 min.

[0111] 2.3 Verification of the preparation process of the dropping pills:

[0112] 2.3.1 Verification of the extraction process of Corydalis yanhusuo, Ligusticum chuanxiong, and Panax ginseng

[0113] 2.3.1.1 Content determination method of the active ingredients in the extracts of Corydalis yanhusuo, Ligusticum chuanxiong, and Panax ginseng:

[0114] (1) Preparation of the reference substance solution: Weigh appropriate amounts of tetrahydropalmatine, ferulic acid, and ginsenoside Rg1 reference substances separately, dissolve them in methanol and make the volume constant to prepare a tetrahydropalmatine reference substance solution with a concentration of 0.686 mg / mL, a ferulic acid reference substance solution with a concentration of 1.056 mg / mL, and a ginsenoside Rg1 reference substance solution with a concentration of 0.689 mg / mL;

[0115] (2) Preparation of the test solution: Weigh the processed products of Corydalis yanhusuo, Ligusticum chuanxiong, and Ginseng according to the prescription amount of the dripping pills. Add 6 times the amount of 70% ethanol, reflux and extract once for 1 hour. Filter and make up the volume to 100 mL in a volumetric flask. Accurately measure 20 mL and transfer it to an evaporating dish, evaporate to dryness in a water bath, dissolve it ultrasonically with 70% methanol, centrifuge, take the supernatant and transfer it to a 5 mL volumetric flask, make up the volume with 70% methanol, and filter through a 0.45 μm microporous filter membrane to obtain the solution.

[0116] Another accurately measure 20 mL of the extract and transfer it to an evaporating dish. Evaporate to dryness in a water bath and then transfer it to a drying oven to dry to a constant weight. Transfer it to a desiccator and wait for the temperature to cool down before weighing to calculate the extract yield.

[0117] (3) Chromatographic conditions: Chromatographic column: Thermo Hypersil GOLD C18 column (250 mm × 4.6 mm, 5 μm); The mobile phase for determining tetrahydropalmatine is acetonitrile (A) - 0.02% triethylamine (B), with gradient elution (0 - 10 min, 30% - 55% A; 10 - 25 min, 55% - 57% A); Flow rate: 1.0 mL / min; Detection wavelength: 280 nm; Column temperature: 30 °C; Injection volume: 10 μL; For the determination of ferulic acid and ginsenoside Rg1, a dual-wavelength detection method is used. The mobile phase is acetonitrile (A) - 0.2% phosphoric acid (B), with gradient elution (0 - 10 min, 18% - 20% A; 10 - 40 min, 20% - 23% A); Flow rate: 1.0 mL / min; Detection wavelengths: 321 nm (ferulic acid), 203 nm (ginsenoside Rg1); Column temperature: 35 °C; Injection volume: 10 μL.

[0118] (4) Determine the test solution according to the above chromatographic conditions, calculate the contents of the active ingredients in the extracts of Corydalis yanhusuo, Ligusticum chuanxiong, and Ginseng, and further calculate the transfer rate.

[0119] 2.3.1.2 Orthogonal experiment

[0120] Combined with the properties of the components and the actual production situation, select 4 factors that have a greater impact on the extraction results, namely ethanol concentration (A), extraction times (B), extraction time (C), and solid-liquid ratio (D), as the investigation factors. Select the L9(3 4 ) factor level table for the experiment. The factor level tables of each factor are shown in Table 2. Arrange 9 extraction processes according to the orthogonal design table, as shown in Table 3.

[0121] Table 2 Orthogonal design factor level table

[0122]

[0123]

[0124] Table 3 Orthogonal experiment arrangement and results

[0125]

[0126] Taking the comprehensive score of the transfer rates of the active ingredients tetrahydropalmatine, ferulic acid, and ginsenoside Rg1 in the prescription as the index, and according to the compatibility concept of monarch, minister, assistant, and envoy in the prescription and the relative importance of each index in the prescription, the weight coefficients are assigned. The weight coefficients of each index are 50, 30, and 20 points in sequence. Comprehensive score = tetrahydropalmatine transfer rate / maximum transfer rate × 50 + ferulic acid transfer rate / maximum transfer rate × 30 + ginsenoside Rg1 transfer rate / maximum transfer rate × 20.

[0127] Table 4 Results of variance analysis

[0128]

[0129] Note: F 0.05 (2,2) = 19.000.

[0130] From the results of intuitive analysis (Table 3) and variance analysis (Table 4), the degree of influence of factors is B > C > A > B, that is, the number of extraction times > extraction time > ethanol concentration > solid-liquid ratio. The optimal extraction process is A2B2C3D2, that is, extracting with 50% ethanol 2 times, 1.5 h each time, and the solid-liquid ratio is 1:10.

[0131] 2.3.1.3 Extraction process verification experiment

[0132] Three verification tests were carried out with the optimized process. As shown in Table 5, the calculated transfer rates of tetrahydropalmatine, ferulic acid, and ginsenoside Rg1 were 96.15%, 88.50%, and 97.68% respectively, and the RSDs were 0.38%, 2.40%, and 0.98% respectively, all within 3%, indicating that the optimized process is stable and feasible.

[0133] Table 5 Results of verification experiment (n = 3)

[0134]

[0135] 2.3.2 Investigation of the purification process of the extract

[0136] 2.3.2.1 Content determination methods of tetrahydropalmatine, ferulic acid, and ginsenoside Rg1 in the extract:

[0137] (1) Preparation of reference substance solutions: Weigh appropriate amounts of reference substances of tetrahydropalmatine, ferulic acid, and ginsenoside Rg1 accurately, dissolve them in methanol and make up the volume to a constant volume to prepare tetrahydropalmatine reference substance solution, ferulic acid reference substance solution, and ginsenoside Rg1 reference substance solution with concentrations of 0.235, 0.201, and 0.433 mg / mL respectively;

[0138] (2) Preparation of the test solution: Weigh Corydalis yanhusuo, Ligusticum chuanxiong, and Ginseng slices according to the prescription amount of the dripping pills. Extract them twice with 50% ethanol for 1.5 hours each time. The material-liquid ratio is 1:10, and heat under reflux for extraction. After concentration under reduced pressure (60 °C), centrifuge (12,000 rpm, 20 min). Take an appropriate amount of the extract, purify it through a macroporous adsorption resin column, and filter the residual liquid behind the column through a 0.45 μm microporous filter membrane to obtain the solution.

[0139] (3) Chromatographic conditions: Chromatographic column: Thermo Hypersil GOLD C18 column (250 mm × 4.6 mm, 5 μm); The mobile phase for determining tetrahydropalmatine is acetonitrile (A) - 0.02% triethylamine (B), with gradient elution (0 - 10 min, 30% - 55% A; 10 - 25 min, 55% - 57% A); Flow rate: 1.0 mL / min; Detection wavelength: 280 nm; Column temperature: 30 °C; Injection volume: 10 μL; For determining ferulic acid and ginsenoside Rg1, the dual-wavelength detection method is used. The mobile phase is acetonitrile (A) - 0.2% phosphoric acid (B), with gradient elution (0 - 10 min, 18% - 20% A; 10 - 40 min, 20% - 23% A); Flow rate: 1.0 mL / min; Detection wavelengths: 321 nm (ferulic acid), 203 nm (ginsenoside Rg1); Column temperature: 35 °C; Injection volume: 10 μL.

[0140] (4) Measure the test solution under the above-mentioned chromatographic conditions, and calculate the contents of the active ingredients in the extracts of Corydalis yanhusuo, Ligusticum chuanxiong, and Ginseng.

[0141] 2.3.2.2 Investigation of macroporous adsorption resins

[0142] Pre-treat five macroporous adsorption resins, namely AB-8, X-5, D101, HPD100, and HPD400. Take 1.0 g of each and place them in a stoppered conical flask with a ground glass stopper. Add 20 mL of the medicinal liquid with a concentration of 0.75 g of crude drug / mL, and shake on a constant temperature shaker for 24 h (25 °C, 100 r / min) for static adsorption. After adsorption, filter, measure the concentrations of tetrahydropalmatine, ferulic acid, and ginsenoside Rg1 in the filtrate, and calculate the static adsorption rate of each macroporous adsorption resin; Then, desorb the macroporous adsorption resin with 20 mL of 70% (V / V) ethanol, place it on a constant temperature shaker for 24 h (25 °C, 100 r / min), measure the concentrations of tetrahydropalmatine, ferulic acid, and ginsenoside Rg1 in the desorbing solution, and calculate the static desorption rate of each macroporous adsorption resin. The results are shown in Table 6 below.

[0143] Table 6 Screening results of macroporous adsorption resins (n = 3)

[0144]

[0145] Note: Adsorption rate % = (C0V0 - C1V1) / C0V0, desorption rate % = C2V2 / (C0V0 - C1V1). C0 is the concentration before adsorption, C1 is the concentration after adsorption, V0 is the volume of the solution before adsorption, V1 is the volume of the solution after adsorption. C2 is the concentration of the desorbing solution, and V2 is the volume of the desorbing solution.

[0146] Comprehensive score = (Adsorption rate of Corydalis yanhusuo + Desorption rate) × 0.5 + (Adsorption rate of Ferulic acid + Desorption rate) × 0.3 + (Adsorption rate of Ginsenoside Rg1 + Desorption rate) × 0.2.

[0147] Based on the comprehensive comparison of the experimental results of adsorption and desorption, it can be seen that the X-5 macroporous adsorption resin has good adsorption and desorption rates for the three components and the highest comprehensive score.

[0148] 2.3.2.3 Investigation of the sample loading concentration

[0149] Weigh 7.5 g of pretreated X-5 macroporous adsorption resin, pack it into a column by wet method according to the diameter-height ratio of 1:8 (column size 1.2 cm × 25 cm), with a flow rate of 10 BV / h and a sample loading volume of 8 BV, and investigate the influence of different sample loading solution concentrations on the adsorption capacity; the concentrations are sequentially selected as 0.5 g crude drug / mL, 1.0 g crude drug / mL, and 1.5 g crude drug / mL. The results are shown in Figure 2 , and it can be seen that the specific adsorption capacity of the X-5 macroporous adsorption resin first increases and then decreases with the increase of the sample loading concentration. When the concentration is 1.0 g / mL, the specific adsorption capacities of the three components reach the highest values, and when the concentration is further increased, the specific adsorption capacities of all three decrease.

[0150] 2.3.2.4 Investigation of the adsorption flow rate

[0151] Weigh 7.5 g of pretreated macroporous adsorption resin, pack it into a column by wet method according to the diameter-height ratio of 1:8 (column size 1.2 cm × 25 cm), load the sample with the optimal sample loading solution concentration of 1.0 g crude drug / mL, and the sample loading volume of 8 BV, and investigate the influence of different adsorption flow rates on the adsorption capacity. The flow rates are sequentially selected as 5 BV / h, 10 BV / h, and 20 BV / h. The results are shown in Figure 3 , and it can be seen that with the increase of the flow rate, the leakage of the three components becomes more serious, but if the flow rate is too slow, it will cause the disadvantages of a long adsorption cycle and high overall process cost.

[0152] 2.3.2.5 Investigation of the sample loading volume

[0153] Weigh 7.5 g of pretreated macroporous adsorption resin and pack it into a column by wet method according to the diameter-height ratio of 1:8 (column size 1.2 cm × 25 cm), load the sample with the optimal sample loading solution concentration of 1.0 g crude drug / mL, and pass the column at an adsorption flow rate of 10 BV / h. Collect the effluent every 1 BV and measure the concentrations of tetrahydropalmatine, ferulic acid, and ginsenoside Rg1 in it. The results are shown in Figure 4, it can be seen that as the sample loading amount increases, the concentration of the residual liquid becomes higher and higher, indicating that the leakage becomes more and more serious. From the perspectives of process production and pharmacoeconomics, the sample loading amount provided by the present invention can avoid multiple process cycles and waste of medicinal materials.

[0154] 2.3.2.6 Selection of elution solvent

[0155] Weigh 7.5 g of pretreated X-5 macroporous adsorption resin, wet-pack the column according to the diameter-height ratio of 1:8 (column size 1.2 cm × 25 cm), with a flow rate of 10 BV / h and a sample loading amount of 4 BV. After the macroporous adsorption resin is completely adsorbed, wash the column with 3 BV of pure water, and then elute successively with 1 BV each of 10%, 30%, 50%, 70%, and 90% (V / V) ethanol at an elution flow rate of 10 BV / h. Collect the elution residual liquid and measure the concentrations of tetrahydropalmatine, ferulic acid, and ginsenoside Rg1 therein. The results are shown in Figure 5 , it can be seen that as the ethanol concentration of the eluent increases, the concentrations of all three components show a trend of first increasing and then decreasing. The residual liquid concentrations of tetrahydropalmatine and ferulic acid are the highest when the ethanol concentration is 70%, that is, the desorption rate is the highest. The desorption rate of ginsenoside Rg1 is the highest at 50% ethanol concentration and slightly decreases at 70% ethanol.

[0156] 2.3.2.7 Verification of purification process

[0157] Use pretreated X-5 macroporous adsorption resin, weigh 7.5 g, wet-pack the column according to the diameter-height ratio of 1:8 (column size 1.2 cm × 25 cm), with a flow rate of 10 BV / h and a sample loading amount of 2 BV. After the macroporous adsorption resin is completely adsorbed, wash the column with 3 BV of pure water, and then elute with 9 BV of 70% (V / V) ethanol at an elution flow rate of 10 BV / h. Collect the eluate and measure the contents of tetrahydropalmatine, ferulic acid, and ginsenoside Rg1 therein, and calculate the elution rate and transfer rate. In addition, accurately measure 50 mL from a 100 mL volumetric flask, evaporate it to dryness in a water bath, and dry it to a constant weight, and calculate the paste rate. The results are shown in Table 7.

[0158] Table 7 Results of verification test (n = 3)

[0159]

[0160] The results show that under the purification process of the present invention, the elution rates of tetrahydropalmatine, ferulic acid, and ginsenoside Rg1 all reach more than 90%, and the transfer rates of all three components reach more than 67%.

[0161] 2.3.3 Study on the dropping pill forming process

[0162] Weigh Corydalis Rhizome, Chuanxiong Rhizome, and Ginseng according to the proportion of the dripping pill prescription. Add 50% ethanol at a material-liquid ratio of 1:10, and extract by heating under reflux twice, each time for 1.5 h. Concentrate the extract under reduced pressure to recover ethanol (60 °C) to obtain an aqueous solution (1 g of crude drug / mL). Purify it through X-5 macroporous adsorption resin, evaporate to dryness in a water bath, and dry to constant weight to obtain the extracts of Corydalis Rhizome, Chuanxiong Rhizome, and Ginseng.

[0163] Weigh the drugs (extracts of Corydalis Rhizome, Chuanxiong Rhizome, and Ginseng, artificial musk, styrax, and borneol) and the matrix according to a certain proportion, melt and mix them at a certain temperature. Keep a certain dropping distance, and at a certain dropping speed, use a rubber head dropper (outer diameter of the dropper tip is 3 mm) to drop the material liquid into a condenser filled with a coolant. The liquid drops are condensed and formed to obtain Shexiang Xintongning dripping pills.

[0164] 2.3.3.1 Appearance quality indicators and scoring criteria of dripping pills

[0165] Table 8 Appearance quality inspection indicators and scoring criteria

[0166]

[0167] 2.3.3.2 Investigation of matrix types

[0168] Use liquid paraffin as the coolant, with a cooling temperature of 6 °C, a liquid medicine temperature of 80 °C, a dropping distance of 6 cm, and a dropping speed of 30 drops / min. Prepare blank dripping pills with pure matrix without drugs, screen the matrix ratio according to different ratios, and investigate the influence of different matrix ratios on the formation of dripping pills, as shown in Table 9.

[0169] Table 9 Investigation results of matrix types

[0170]

[0171] It was found in the experiment that the larger the proportion of PEG6000, the harder the dripping pills, but the roundness becomes worse and they are prone to adhesion.

[0172] 2.3.3.3 Selection of coolant types

[0173] With a cooling temperature of 6 °C, a liquid medicine temperature of 80 °C, a dropping distance of 6 cm, and a dropping speed of 30 drops / min, without drugs, blank dripping pills. Use the dropping speed and formation situation of the dripping pills as indicators to investigate the coolant. It was found in the experiment that when liquid paraffin is used as the coolant, the dripping pills are well formed and the dropping speed is fast; when dimethyl silicone oil is used as the coolant, the dripping pills are well formed but the dropping speed is slow.

[0174] 2.3.3.4 Investigation of dropping distance range

[0175] The drug-to-matrix ratio is 1:3, the matrix ratio of PEG4000:6000 = 3:1, the liquid medicine temperature is 80°C, the coolant is liquid paraffin, the cooling temperature is 6°C, the dropping speed is 30 drops / min, and the dropping distances of 2 cm, 4 cm, 6 cm, 8 cm, and 10 cm are investigated for their effects on the formation of drop pills, as shown in Table 10.

[0176] Table 10 Investigation results of dropping distance range

[0177]

[0178]

[0179] The results show that too small a dropping distance will lead to poor roundness and adhesion during the sedimentation process, while too large a dropping distance will also result in poor formation and adhesion. Moreover, in actual operation, if the dropping distance is too high, it is difficult to control steadily by hand, and the drop pills are likely to drip onto the inner wall.

[0180] 2.3.3.5 Investigation of dropping speed range

[0181] The drug-to-matrix ratio is 1:3, the matrix ratio of PEG4000:6000 = 3:1, the liquid medicine temperature is 80°C, the coolant is liquid paraffin, the cooling temperature is 6°C, the dropping distance is 6 cm, and the dropping speeds of 10 drops / min, 20 drops / min, 30 drops / min, 40 drops / min, 50 drops / min, and 60 drops / min are investigated for their effects on the formation of drop pills, as shown in Table 11.

[0182] Table 11 Investigation results of dropping speed range

[0183]

[0184] The results show that too fast a dropping speed is likely to cause adhesion and the drop pills are relatively flat, while too slow a dropping speed will result in too low production efficiency.

[0185] 2.3.3.6 Investigation of liquid medicine temperature range

[0186] The drug-to-matrix ratio is 1:3, the matrix ratio of PEG4000:6000 = 3:1, the coolant is liquid paraffin, the cooling temperature is 6°C, the dropping distance is 6 cm, the dropping speed is 30 drops / min, and the liquid medicine temperatures of 60°C, 70°C, 80°C, and 90°C are investigated for their effects on the formation of drop pills, as shown in Table 12.

[0187] Table 12 Investigation results of liquid material temperature range

[0188]

[0189] The results show that when the liquid material temperature is low, the roundness of the drop pills is poor and adhesion is likely to occur; however, when the liquid material temperature is high, the hardness is poor and tailing phenomenon is likely to occur.

[0190] 2.3.3.7 Orthogonal Test

[0191] Taking the ratio of drug to matrix, the temperature of the drug solution, and the dropping distance as the investigation factors, and the appearance quality and the dissolution time limit as the investigation indexes, the comprehensive scoring method was used for scoring to select the best molding process. Among them, the scoring method for appearance quality was the same as that under "2.3"; the scoring regulations for the dissolution time limit were as follows: less than 4 minutes was 20 points, 4 - 5 minutes was 16 points, 5 - 6 minutes was 12 points, 6 - 7 minutes was 8 points, and 8 - 9 minutes was 4 points. The comprehensive score = appearance quality score + dissolution time limit score. The dissolution time limit was in accordance with the provisions of "Dissolution Time Limit" for dripping pills under the general rules of Part IV of the Pharmacopoeia of the People's Republic of China (2020 Edition). A hanging basket with a sieve aperture of 2.0 mm was selected. Six test samples were taken and placed in the glass tubes of the hanging basket respectively, without baffles, and the disintegration tester was started for inspection. The dripping pills should be completely dissolved within 30 minutes.

[0192] The L9(3 4 ) factor level table was selected. Taking the ratio of drug to matrix (A), the temperature of the drug solution (B), and the dropping distance (C) as the investigation factors, and the factor level tables are shown in Table 13. According to the orthogonal design table, 9 molding processes were arranged, as shown in Table 14.

[0193] Table 13 Factor Level Table

[0194]

[0195] Table 14 Orthogonal Test Arrangement and Results

[0196]

[0197]

[0198] Table 15 ANOVA Results

[0199]

[0200] Note: F 0.05 (2, 2) = 19.000.

[0201] Range analysis showed that the ratio of drug to matrix had the greatest influence on the experimental results and was significant. Followed by the dropping distance and the temperature of the drug solution, which was consistent with the results of the ANOVA. From the results of the intuitive analysis (Table 14) and the ANOVA (Table 15), the degree of factor influence was A > C > B. The best molding process was A3B3C2, that is, the ratio of drug to matrix was 1:4, the temperature of the drug solution was 85 °C, and the dropping distance was 6 cm.

[0202] 2.3.3.8 Verification Experiment

[0203] Repeat the experiment 3 times according to the above optimization conditions, conduct an appearance inspection on the test samples, and determine the disintegration time limit. The measurement results are shown in Table 16.

[0204] Table 16 Verification experiment results (n = 3)

[0205]

[0206] III. Quality detection method of Shexiang Xintongning Pills

[0207] 3.1 Determination of the content of ginsenoside Rg1

[0208] 3.1.1 Determination method:

[0209] (1) Preparation of ginsenoside Rg1 reference substance solution

[0210] Take an appropriate amount of ginsenoside Rg1 reference substance, dissolve it in methanol and make up to 25 mL to prepare a stock solution with a mass concentration of 83.6 μg / mL, and dilute it with methanol to prepare ginsenoside Rg1 reference substance solutions with mass concentrations of 8.36, 16.72, 25.08, 33.44, 41.8, 50.16, 58.52, 66.88, 83.6 μg / mL respectively;

[0211] (2) Preparation of test solution

[0212] Weigh 0.5 g of Shexiang Xintongning Pills powder, place it in a stoppered conical flask, add 25 mL of methanol, sonicate for 30 min, cool to room temperature, filter, make up to 25 mL with methanol, shake well, and filter through a 0.45 μm filter membrane to obtain the test solution;

[0213] (3) Preparation of negative control solution

[0214] Take 0.5 g of negative pills made of pure matrix and prepare a negative control solution in the same way as the test solution;

[0215] (4) Liquid chromatography conditions

[0216] Chromatographic column: Thermo Hypersil GOLD C18 column (4.6×250 mm, 5 μm); Mobile phase: 0.05% phosphoric acid water (A) - acetonitrile (B), gradient elution program is shown in Table 17; Flow rate: 1.0 mL / min, Column temperature: 30 °C, Detection wavelength: 203 nm, Injection volume: 10 μL;

[0217] Table 17 Program elution table

[0218]

[0219] (5) Content determination

[0220] Take 3 batches of Shexiang Xintongning Pills (20220429, 20220430, 20220501), prepare the test solution, determine according to the chromatographic conditions, record the peak area, and calculate the content of ginsenoside Rg1. See Table 18.

[0221] Table 18 Content determination of three batches of pills

[0222]

[0223] 3.1.2 Methodology investigation:

[0224] (1) Specificity

[0225] Respectively take the reference substance solution of ginsenoside Rg1, the test solution, and the negative control solution, inject samples according to the chromatographic conditions, and record the HPLC chromatogram. See Figure 6 . The results show that under these chromatographic conditions, the chromatographic peak of ginsenoside Rg1 in the test sample has a good shape, the resolution is greater than 1.5, and the number of theoretical plates is greater than 60000. There is no interference from the negative sample.

[0226] (2) Linear range

[0227] Respectively accurately pipette 10 μL of a series of concentration reference substance solutions of ginsenoside Rg1 for injection, determine according to the chromatographic conditions, record the peak areas of the corresponding chromatographic peaks. Taking the mass concentration as the abscissa (X) and the peak area as the ordinate (Y), perform linear regression to obtain the regression equation Y = 2.3922X - 3.6926 (R 2 = 0.9992). The results show that ginsenoside Rg1 has a good linear relationship in the range of 8.36 - 83.6 μg / mL.

[0228] (3) Precision

[0229] Accurately pipette 10 μL of the reference substance solution of ginsenoside Rg1, determine according to the chromatographic conditions, inject samples continuously for 6 times, and record the peak areas of each chromatographic peak. See Table 19.

[0230] Table 19 Precision results (n = 6)

[0231]

[0232] It can be seen that the RSD of the peak area of ginsenoside Rg1 is 0.55%, indicating good precision of the instrument.

[0233] (4) Stability

[0234] Accurately pipette 10 μL of the test solution, and inject samples for detection at 0, 2, 4, 8, 12, and 24 h after preparation respectively, record the peak areas of the chromatographic peaks. See Table 20.

[0235] Table 20 Stability results

[0236]

[0237] It can be seen that the RSD of the peak area of ginsenoside Rg1 is 3.43%, indicating that the test solution has good stability within 24 hours.

[0238] (5) Repeatability

[0239] Six test solutions were prepared and determined under the chromatographic conditions. The peak areas were recorded and the content of ginsenoside Rg1 was calculated, as shown in Table 21.

[0240] Table 21 Repeatability results (n = 6)

[0241]

[0242] It can be seen that the RSD of the content of ginsenoside Rg1 is 3.10%, indicating that the method has good repeatability.

[0243] (6) Spiked recovery experiment

[0244] Approximately 0.5 g of Shexiang Xintongning Pills powder was accurately weighed, and the ginsenoside Rg1 reference substance equal to the sample content was added to prepare the test solution. The test solution was determined under the chromatographic conditions, and the spiked recovery rate of ginsenoside Rg1 was calculated, as shown in Table 22.

[0245] Table 22 Spiked recovery results of ginsenoside Rg1

[0246]

[0247] It can be seen that the spiked recovery rate of ginsenoside Rg1 is between 98.47% and 103.59%, and the RSD is 2.19%, indicating that the spiked recovery rate of this method is good.

[0248] 3.2 Determination of the contents of tetrahydropalmatine, cinnamic acid and ferulic acid

[0249] 3.2.1 Determination method:

[0250] (1) Preparation of the mixed reference substance solution

[0251] An appropriate amount of tetrahydropalmatine and cinnamic acid reference substances were accurately weighed and dissolved in methanol to a volume of 50 mL to prepare a stock solution containing 29.6 μg / mL of tetrahydropalmatine, 22.8 μg / mL of cinnamic acid and 28 μg / mL of ferulic acid. A series of mixed reference substance solutions with different mass concentrations were prepared by diluting with methanol;

[0252] (2) Preparation of the test solution

[0253] Weigh 0.5 g of Shexiang Xintongning Pills powder, place it in a stoppered conical flask, add 25 mL of methanol, ultrasonicate for 30 min, let it cool to room temperature, filter, make up the volume to 25 mL with methanol, shake well, filter through a 0.45 μm filter membrane, and dilute 4-fold to prepare the test solution.

[0254] (3) Preparation of negative control solution

[0255] Take 0.5 g of negative pills made of pure matrix to prepare the negative control solution.

[0256] (4) HPLC conditions

[0257] Chromatographic column: Thermo Hypersil GOLD C18 column (4.6×250 mm, 5 μm), mobile phase: 0.1% formic acid in water (A) - acetonitrile (B), gradient elution is shown in Table 23, flow rate 1.0 mL·min -1 , column temperature: 30 °C, detection wavelengths 280 nm (tetrahydropalmatine, cinnamic acid), 321 nm (ferulic acid), injection volume 10 μL.

[0258] Table 23 Gradient elution program

[0259]

[0260] (5) Content determination

[0261] Take the mixed reference solution, test solution, and negative control solution, carry out the determination according to the chromatographic conditions, record the peak areas, and calculate the contents of tetrahydropalmatine, cinnamic acid, and ferulic acid.

[0262] 3.2.2 Results of content determination of tetrahydropalmatine, cinnamic acid, and ferulic acid

[0263] Take 3 batches of Shexiang Xintongning Pills (20220429, 20220430, 20220501), add 20 mL of ethyl acetate, ultrasonically extract for 30 min, filter, make up the volume to 20 mL in a volumetric flask, filter through a 0.22 μm filter membrane to prepare the test solution; carry out the determination according to the chromatographic conditions, record the peak areas of each component, and calculate the contents of tetrahydropalmatine, cinnamic acid, and ferulic acid, as shown in Table 24.

[0264] Table 24 Determination of the contents of active ingredients in three batches of pill samples

[0265]

[0266]

[0267] Taking 80% of the average content as the limit for the determination of the content of the active ingredients in Shexiang Xintongning Pills, each gram of the pills should contain no less than 1.62 mg of Corydalis Rhizoma calculated as tetrahydropalmatine, no less than 1.35 mg of Styrax calculated as cinnamic acid, and no less than 0.52 mg of Chuanxiong Rhizoma calculated as ferulic acid. The Shexiang Xintongning Pills prepared by the present invention meet the requirements.

[0268] 3.2.3 Methodology investigation:

[0269] (1) Specificity

[0270] Respectively take the mixed reference substance solution, the test solution, and the negative control solution, inject samples according to the chromatographic conditions, and record the HPLC chromatogram as shown in Figure 7 . The results show that under these chromatographic conditions, the chromatographic peaks of tetrahydropalmatine, cinnamic acid, and ferulic acid in the test solution have good shapes, the resolution is greater than 1.5, the number of theoretical plates is greater than 70000, and the negative sample has no interference.

[0271] (2) Linear range

[0272] Precisely pipette 10 μL of a series of concentration mixed reference substance solutions respectively for injection, perform the determination according to the chromatographic conditions, record the corresponding chromatographic peak areas, take the mass concentration as the abscissa (X) and the peak area as the ordinate (Y), and perform linear regression to obtain the regression equation: Y 延胡索 = 8.525X + 2.1405 (R 2 = 0.9993), indicating that tetrahydropalmatine has a good linear relationship in the range of 2.96 - 29.6 μg / mL; Y 肉桂酸 = 122.77X + 19.22 (R 2 = 0.9994), the results show that cinnamic acid has a good linear relationship in the range of 2.28 - 22.8 μg / mL; Y 阿魏酸 = 68.08X - 13.26 (R 2 = 0.9998), the results show that cinnamic acid has a good linear relationship in the range of 2 - 28 μg / mL.

[0273] (3) Precision

[0274] Precisely pipette 10 μL of the mixed reference substance solution, perform the determination according to the chromatographic conditions, inject samples continuously for 6 times, and record the chromatographic peak areas of each, as shown in Table 25.

[0275] Table 25 Precision results (n = 6)

[0276]

[0277]

[0278] It can be seen that the RSDs of the peak areas of tetrahydropalmatine, cinnamic acid, and ferulic acid are 0.24%, 0.27%, and 0.94% respectively, indicating that the instrument precision is good.

[0279] (4) Stability

[0280] Precisely pipette 10 μL of the test solution and inject it for detection at 0, 2, 4, 8, 12, and 24 h after preparation, and record the peak areas of the chromatographic peaks, as shown in Table 26.

[0281] Table 26 Stability results

[0282]

[0283] It can be seen that the RSDs of the peak areas of tetrahydropalmatine, cinnamic acid, and ferulic acid are 1.77%, 2.15%, and 1.38% respectively, indicating that the test solution has good stability within 24 h.

[0284] (5) Repeatability

[0285] Prepare 6 portions of the test solution, determine according to the chromatographic conditions, record the peak areas, and calculate the contents, as shown in Table 27.

[0286] Table 27 Repeatability results (n = 6)

[0287]

[0288] It can be seen that the RSDs of the contents of tetrahydropalmatine and cinnamic acid are 2.22%, 3.11%, and 2.16% respectively, indicating that the method has good repeatability.

[0289] (6) Spiked recovery experiment

[0290] Precisely weigh 0.5 g of 6 portions of Shexiang Xintongning Pills powder, add the standard substance equal to the sample content, prepare the test solution, determine according to the chromatographic conditions, and calculate the spiked recoveries of tetrahydropalmatine, cinnamic acid, and ferulic acid, as shown in Tables 28 - 30.

[0291] Table 28 Spiked recovery results of tetrahydropalmatine (n = 6)

[0292]

[0293] Table 29 Spiked recovery results of cinnamic acid (n = 6)

[0294]

[0295] Table 30 Spiked recovery results of ferulic acid (n = 6)

[0296]

[0297] The recoveries of tetrahydropalmatine and cinnamic acid were in the ranges of 97.57% - 103.89%, 95.51% - 99.11%, and 96.84% - 100.82% respectively, and the RSDs were 2.93%, 1.52%, and 1.49% respectively. The results showed that the accuracy of this method met the requirements.

[0298] 3.3 Determination of muscone content

[0299] 3.3.1 Determination method:

[0300] (1) Preparation of reference solution:

[0301] Take an appropriate amount of muscone reference substance, dissolve it in ethyl acetate and make the volume up to 25 mL to prepare a mother solution of reference substance with a concentration of 406.4 μg / mL, and dilute it with ethyl acetate to prepare a series of reference solutions of muscone with different mass concentrations;

[0302] (2) Preparation of test solution:

[0303] Take 1 g of Shexiang Xintongning Pills powder, add 20 mL of ethyl acetate, extract it by ultrasonic for 30 min, filter, make the volume up to 20 mL in a volumetric flask, and filter through a 0.22 μm filter membrane to obtain the test solution;

[0304] (3) Preparation of negative control solution:

[0305] Add pure matrix into the pill dropping machine and drop pills according to the same dropping method as Shexiang Xintongning Pills to prepare negative pills. Take 1 g of negative pills and prepare a negative control solution according to the same method as the test solution;

[0306] (4) Gas chromatography conditions:

[0307] The chromatographic column is HP-5 capillary column (5% Phenyl Methyl Siloxane, 30 m × 320 μm × 0.25 μm); the detector is flame ionization detector; the carrier gas is nitrogen, and the flow rate of the carrier gas is 13.5 mL / min; the air flow rate is 400 mL / min; the hydrogen flow rate is 20 mL / min; the tail gas blowing flow rate of nitrogen is 25 mL / min; the injection volume is 1 μL; the split ratio is 10:1; the programmed temperature is shown in Table 31, and the total running time is 18 min; the vaporization chamber temperature is 220 °C, and the detector temperature is 260 °C.

[0308] Table 31 Programmed temperature table

[0309]

[0310] (5) Determination:

[0311] Take 3 batches of Shexiang Xintongning Pills (20220429, 20220430, 20220501), prepare the test solution, conduct the determination according to the described chromatographic conditions, record the peak area, calculate the content of muskone, as shown in Table 32. Take 80% of the average content as the limit for the content determination of Shexiang Xintongning Pills. Each gram of the pills contains artificial musk calculated as muskone, not less than 0.86 mg. The Shexiang Xintongning Pills prepared by the present invention meet the requirements.

[0312] Table 32 Determination of Muskone Content in Three Batches of Pills

[0313]

[0314] 3.4 Thin Layer Identification Method

[0315] 3.4.1 Identification of Corydalis Rhizoma:

[0316] Take 0.5 g of the powdered Shexiang Xintongning Pills after grinding, add 20 mL of petroleum ether with a boiling range of 60 - 90 °C, then add 1.5 mL of concentrated ammonia water, extract ultrasonically for 30 min, filter, evaporate the filtrate to dryness at 60 °C, add 1 mL of petroleum ether (60 - 90 °C) to dissolve the residue, and use it as the test solution;

[0317] Weigh 0.5 g of the reference medicinal material of Corydalis Rhizoma, and prepare the reference medicinal material solution according to the same steps. Take 0.5 g of the powdered pills without Corydalis Rhizoma, and prepare the negative control solution according to the same steps;

[0318] In addition, take tetrahydropalmatine reference substance, and prepare a reference substance solution with a mass concentration of 0.2 mg / mL with methanol;

[0319] According to the test of thin layer chromatography (General Rule 0502), use a capillary to absorb 10 μL of each of the above four solutions, and spot them on a silica gel GF254 thin layer plate from left to right in sequence. Prepare the developing agent solution according to the volume ratio of n - hexane - chloroform - methanol of 10:6:1, and shake well. Pre - saturate for 15 min, develop, air - dry, place in an iodine cylinder for color development, and view under ultraviolet light at a wavelength of 365 nm after the iodine has volatilized. The test solution of Shexiang Xintongning Pills shows fluorescent spots of the same color at the corresponding positions as the reference medicinal material of Corydalis Rhizoma and the tetrahydropalmatine reference substance, and the negative control has no interference, as Figure 8 shown in Figure A.

[0320] 3.4.2 Identification of Chuanxiong Rhizoma:

[0321] Take 0.5 g of the powdered Shexiang Xintongning Pills after grinding and place them in a conical flask, add 20 mL of ether, extract ultrasonically for 10 min, filter, evaporate the filtrate to dryness at 30 °C, and dissolve the residue with 1 mL of ethanol to use it as the test solution.

[0322] Separately take 0.5 g of the prepared Ligusticum wallichii reference medicinal material, and prepare a reference medicinal material solution by the same method;

[0323] Then take 0.5 g of the dripping pill powder without Ligusticum wallichii, and still prepare a negative control solution according to the same method. According to the thin-layer chromatography method (General Rule 0502) test, use a capillary tube to absorb 10 μL of each of the above three solutions, and spot them on the same silica gel GF254 thin-layer plate from left to right in sequence; Prepare the developing solvent according to the volume ratio of n-hexane-ethyl acetate of 4:1, and observe under ultraviolet light at a wavelength of 365 nm; The test solution of Shexiang Xintongning dripping pills and the Ligusticum wallichii reference medicinal material show the same blue fluorescent spots at the corresponding positions, and the negative control has no interference, as Figure 8 shown in Figure B.

[0324] 3.4.3 Identification of ginseng:

[0325] Take 1 g of Shexiang Xintongning dripping pills, grind them, add 20 mL of ethanol, let it stand for 3 hours, shake it constantly during the standing process, then perform ultrasonic extraction for 20 min and put it in the refrigerator for refrigeration for 24 h, filter, evaporate the filtrate to nearly dryness at 70 °C, dissolve the residue in 2 mL of methanol, and use it as the test solution;

[0326] Take 1 g of the dripping pill powder without ginseng, and prepare a negative control solution according to the same method;

[0327] Take reference substances of ginsenoside Rg1 and ginsenoside Re, and separately prepare reference substance solutions of 0.5 mg / mL with methanol;

[0328] According to the thin-layer chromatography method (General Rule 0502) test, use a capillary tube to absorb 10 μL of each of the above four solutions, and spot them on the same silica gel GF254 thin-layer plate from left to right in sequence; Use the lower layer solution of chloroform-ethyl acetate-methanol-water (7.5:20:11:5) placed below 10 °C as the developing solvent for development, take it out, dry it, spray an appropriate amount of 10% sulfuric acid ethanol solution, dry it and heat it at 105 °C until the spots are clearly developed; The test solution of Shexiang Xintongning dripping pills and the ginseng reference substance show the same pink spots at the corresponding positions, and the negative control has no interference, as Figure 8 shown in Figure C.

[0329] 3.4.4 Identification of styrax:

[0330] Take 1 g of the ground Shexiang Xintongning dripping pill powder, add 10 mL of ether and perform ultrasonic treatment for 2 min, and take the supernatant as the test solution;

[0331] Take 1 g of the dripping pill powder without styrax, and prepare a negative control solution according to the same method;

[0332] Separately take reference substance cinnamic acid, and prepare a reference substance solution of 1 mg / mL with ether;

[0333] Test according to the thin layer chromatography method (General Principles 0502). Use a capillary tube to aspirate appropriate amounts of the above-mentioned test solution, negative control solution, and reference substance solution, and spot them successively on the same silica gel GF254 thin layer plate. Use petroleum ether (30 - 60°C) - n-hexane - ethyl formate - formic acid (10:30:15:1) as the developing solvent for development. After drying, spray an appropriate amount of 10% sulfuric acid ethanol solution, dry it with a blower, and then place it in an oven at 105°C to heat until the spots are clearly developed. Examine it under an ultraviolet lamp at a wavelength of 254 nm; the test sample of Shexiang Xintongning Pills and the chromatogram of cinnamic acid reference substance show fluorescent spots of the same color at the corresponding positions, and the negative control has no interference. As Figure 8 shown in D.

[0334] 3.4.5 Identification of Borneol:

[0335] Take 0.3 g of the powdered Shexiang Xintongning Pills after grinding, add 15 mL of ethyl acetate, ultrasonically treat for 2 minutes, filter, evaporate the filtrate to nearly dry at 60°C, and redissolve it with 0.5 mL of ethyl acetate to obtain the test solution;

[0336] Take 0.3 g of the powdered pills without borneol and prepare a negative control solution in the same manner as above;

[0337] Take an appropriate amount of dextrorotatory borneol reference substance and dissolve it in ethyl acetate to prepare a reference substance solution with a concentration of 2.5 mg / mL;

[0338] Test according to the thin layer chromatography method (General Principles 0502). Use a capillary tube to aspirate 2 μL of each of the above three solutions and spot them successively on the same silica gel GF254 thin layer plate. Prepare a developing solvent solution according to the volume ratio of toluene - acetone of 9:1. Spray an appropriate amount of 5% vanillin sulfuric acid solution and then place it in an oven at 105°C to heat until the spots are clearly developed; the test sample of Shexiang Xintongning Pills and the borneol reference substance show the same yellowish green fluorescent spots at the same positions, and the negative control has no interference. As Figure 8 shown in E.

[0339] The above specific implementation manners cannot be used as a limitation on the protection scope of the present invention. For those skilled in the art of this technology, any alternative improvement or transformation made to the implementation manner of the present invention falls within the protection scope of the present invention.

[0340] Where the present invention is not described in detail, it is all common knowledge for those skilled in the art of this technology.

Claims

1. A quality inspection method for Shexiang Xintongning Pills, characterized in that, Including the method for determining the content of ginsenoside Rg1 and / or the method for determining the content of tetrahydropalmatine, cinnamic acid, and ferulic acid and / or the thin-layer identification method for Corydalis yanhusuo, Ligusticum chuanxiong, Panax ginseng, Styrax, and Borneol; The method for determining the content of ginsenoside Rg1 includes the following steps: (1) Prepare control solutions with different concentrations: Take the ginsenoside Rg1 reference substance, dissolve it in methanol to make a stock solution of ginsenoside Rg1, and then dilute it with methanol to prepare a series of control solutions of ginsenoside Rg1 with different mass concentrations; (2) Prepare the test solution: Take 0.5 g of Shexiang Xintongning Pills powder, place it in a stoppered conical flask, add 25 mL of methanol, ultrasonically treat for 30 min, cool to room temperature, filter, dilute to 25 mL with methanol, shake well, and filter through a 0.45 μm filter membrane to obtain the test solution; (3) Prepare the negative control solution: Add pure matrix to a pill dropping machine for dropping, prepare negative pills from the pure matrix according to the method in step (4) of claim 1, take 0.5 g, place it in a stoppered conical flask, add 25 mL of methanol, ultrasonically treat for 30 min, cool to room temperature, filter, dilute to 25 mL with methanol, shake well, and filter through a 0.45 μm filter membrane to obtain the negative control solution; (4) Liquid chromatography conditions: Chromatographic column: C18 chromatographic column; Mobile phase: 0.05% phosphoric acid water (A) - acetonitrile (B); Gradient elution program: 0 - 10 min: 80% A, 20% B; 10 - 25 min: 80% → 76% A, 20% → 24% B; 25 - 33 min: 76% A, 24% B; Flow rate: 1.0 mL / min; Column temperature: 30 °C; Detection wavelength: 203 nm; Injection volume: 10 μL; (5) Determination: Take the control solution, negative control solution, and test solution, inject them into the liquid chromatograph, and perform determination according to the above chromatographic conditions. Calculate the content of ginsenoside Rg1 based on the peak area; The preparation process of the Shexiang Xintongning Pills includes the following steps: (1) Mix the slices of Corydalis yanhusuo, Ligusticum chuanxiong, and Panax ginseng, add 8 - 12 times the weight of ethanol with a volume concentration of 50% - 70%, reflux extract 1 - 2 times, each time for 1 - 2 h, combine the extraction solutions, filter, recover ethanol from the filtrate, and concentrate it under reduced pressure at 60 - 85 °C to a concentrated medicinal liquid with a concentration of 0.7 - 1.2 g of crude drug / mL at 60 ± 5 °C; (2) Purify the concentrated medicinal liquid through macroporous adsorption resin, then elute the purified liquid with ethanol with a volume concentration of 60% - 70%, collect the eluate, evaporate it to dryness, and dry it to a constant weight to obtain a paste; (3) Grind Moschus and Borneol separately, grind the paste, mix the paste, Moschus, Borneol, and Styrax, and add 3 - 5 times the weight of the matrix. Stir at 80 - 90 °C in a water bath until it melts evenly. The matrix is a mixture of PEG4000 and PEG6000 with a weight ratio of 3 - 4:1; (4) Add the molten medicinal liquid to a pill dropping machine for dropping, with the coolant temperature at 6 - 14 °C, the dropping distance at 4 - 8 cm, the dropping speed at 25 - 35 drops / min, collect the pills, and dry them to prepare the Shexiang Xintongning Pills.

2. The quality inspection method of Shexiang Xintongning Pills according to claim 1, characterized in that, The method for determining the contents of tetrahydropalmatine, cinnamic acid, and ferulic acid comprises the following steps: (1) Preparation of mixed reference substance solutions with different concentrations: Take reference substances of tetrahydropalmatine, cinnamic acid, and ferulic acid, make up the volume with methanol to prepare the mother reference substance solution of tetrahydropalmatine, and then dilute with methanol to prepare a series of reference substance solutions of tetrahydropalmatine, cinnamic acid, and ferulic acid with different mass concentrations; (2) Preparation of test sample solution: Take 0.5 g of the powder of Shexiang Xintongning Pills, place it in a stoppered conical flask, add 25 mL of methanol, ultrasonically treat for 30 min, let it cool to room temperature, filter, make up the volume with methanol to 25 mL, shake well, filter through a 0.45 μm filter membrane, and dilute 4 times with methanol to obtain; (3) Preparation of negative control solution: Add pure matrix to a pill dropping machine for dropping, prepare negative pills from the pure matrix according to the method in step (4) of claim 1, take 0.5 g, place it in a stoppered conical flask, add 25 mL of methanol, ultrasonically treat for 30 min, let it cool to room temperature, filter, make up the volume with methanol to 25 mL, shake well, filter through a 0.45 μm filter membrane to obtain; (4) Liquid chromatography conditions: Chromatographic column: C18 chromatographic column; Mobile phase: 0.1% formic acid water (A) - acetonitrile (B); Gradient elution program: 0 - 10 min: 90% → 80% A, 10% → 20% B; 10 - 20 min: 80% → 70% A, 20% → 30% B; 20 - 23 min: 70% → 65% A, 30% → 35% B; 23 - 30 min: 65% → 48% A, 35% → 52% B; Flow rate 1.0 mL / min; Column temperature: 30 °C; Detection wavelength: 280 nm for tetrahydropalmatine and cinnamic acid, 321 nm for ferulic acid; Injection volume 10 μL; (5) Determination: Take the reference substance solution, negative control solution, and test sample solution, inject them into a liquid chromatograph, carry out determination according to the said chromatographic conditions, and calculate the contents of tetrahydropalmatine, cinnamic acid, and ferulic acid based on the peak areas.

3. The quality inspection method of Shexiang Xintongning Pills according to claim 1, characterized in that, The method for determining the content of muscone comprises the following steps: (1) Preparation of reference substance solution: Take an appropriate amount of muscone reference substance, make up the volume with ethyl acetate to 25 mL to prepare a mother reference substance solution of 406.4 μg / mL, and dilute with ethyl acetate to prepare a series of reference substance solutions of muscone with different mass concentrations; (2) Preparation of test sample solution: Take 1 g of the powder of Shexiang Xintongning Pills, add 20 mL of ethyl acetate, ultrasonically extract for 30 min, filter, make up the volume to a 20 mL volumetric flask, and filter through a 0.22 μm filter membrane to obtain; (3) Preparation of negative control solution: Add pure matrix to a pill dropping machine for dropping, prepare negative pills from the pure matrix according to the method in step (4) of claim 1, take 1 g, add 20 mL of ethyl acetate, ultrasonically extract for 30 min, filter, make up the volume to a 20 mL volumetric flask, and filter through a 0.22 μm filter membrane to obtain; (4) Gas chromatography conditions: Chromatographic column: HP-5 capillary column; Temperature programming: initial temperature 80°C, hold for 1 min, increase temperature at 20°C / min to 160°C, hold for 1 min, then increase temperature at 20°C / min to 260°C, hold for 2 min; Detector: flame ionization detector; Carrier gas: nitrogen, carrier gas flow rate 13.5 mL / min; Combustion-supporting air flow rate 400 mL / min; Combustible gas hydrogen flow rate 20 mL / min; Nitrogen make-up gas flow rate 25 mL / min; Injection volume 1 μL; Split ratio 10:1; Vaporization chamber temperature 220°C, detector temperature 260°C; (5) Determination: Take the reference substance solution, negative control solution, and test sample solution, inject into the gas chromatograph, and determine according to the said chromatographic conditions, and calculate the content of muskone based on the peak area.

4. The quality inspection method of Shexiang Xintongning Pills according to claim 2, characterized in that, The thin-layer identification method for Corydalis yanhusuo includes the following steps: Prepare the test sample solution, reference crude drug solution, negative control solution, and tetrahydropalmatine reference substance solution: Take 0.5 g of the powdered Shexiang Xintongning Pills, add 20 mL of petroleum ether with a boiling range of 60 - 90°C, then add 1.5 mL of concentrated ammonia water, ultrasonically extract for 30 min, filter, evaporate the filtrate to dryness at 60°C, add 1 mL of petroleum ether at 60 - 90°C to dissolve the residue, as the test sample solution; Take 0.5 g of the reference crude drug of Corydalis yanhusuo and 0.5 g of the pill powder without Corydalis yanhusuo respectively, and prepare the reference crude drug solution and negative control solution in the same method as preparing the test sample solution; Take tetrahydropalmatine reference substance, dissolve it with methanol to prepare a tetrahydropalmatine reference substance solution with a mass concentration of 0.2 mg / mL. According to the thin-layer chromatography test, suck 10 μL of the test sample solution, reference crude drug solution, negative control solution, and tetrahydropalmatine reference substance solution respectively, spot them on a silica gel GF254 thin-layer plate, use a mixed solution of n-hexane - chloroform - methanol with a volume ratio of 10:6:1 as the developing agent, pre-saturate for 15 min, dry after development, place in an iodine cylinder for color development, and examine under ultraviolet light at a wavelength of 365 nm after the iodine has volatilized; In the test sample chromatogram, fluorescent spots of the same color appear at the positions corresponding to the reference crude drug of Corydalis yanhusuo and the tetrahydropalmatine reference substance, and there is no interference from the negative control. The thin-layer identification method for Ligusticum chuanxiong Hort. includes the following steps: Prepare the test sample solution, reference crude drug solution, negative control solution: Take 0.5 g of the powdered Shexiang Xintongning Pills and place it in a conical flask, add 20 mL of ether, ultrasonically extract for 10 min, filter, evaporate the filtrate to dryness at 30°C, dissolve the residue with 1 mL of ethanol, as the test sample solution; Take 0.5 g of the reference crude drug of Ligusticum chuanxiong Hort. and 0.5 g of the pill powder without Ligusticum chuanxiong Hort. respectively, and prepare the reference crude drug solution and negative control solution in the same method as preparing the test sample solution. For the test by thin-layer chromatography, 10 μL of the test solution, the solution of the control crude drug, and the negative control solution were each taken and spotted on the same silica gel GF254 thin-layer plate. A mixed solution of n-hexane-ethyl acetate with a volume ratio of 4:1 was used as the developing solvent. After development, it was dried in air and observed under ultraviolet light at a wavelength of 365 nm. In the chromatogram of the test solution, the same fluorescent spots were shown at the corresponding positions as those in the chromatogram of the control crude drug, and there was no interference from the negative control.

5. The quality inspection method of Shexiang Xintongning Pills according to claim 2, characterized in that, The thin-layer identification method for ginseng includes the following steps: Preparation of the test solution, negative control solution, reference substance solution of ginsenoside Rg1, and reference substance solution of ginsenoside Re: Take 1 g of Shexiang Xintongning Pills, crush them, add 20 mL of ethanol, let stand for 3 hours, shake during the standing process, then perform ultrasonic extraction for 20 min, place in the refrigerator for refrigeration for 24 h, filter, evaporate the filtrate to nearly dry at 70 °C, dissolve the residue in 2 mL of methanol to obtain the test solution; take 1 g of the pill powder without ginseng, and prepare the negative control solution in the same way as the test solution; take the reference substance of ginsenoside Rg1 and the reference substance of ginsenoside Re respectively, and add methanol to prepare a reference substance solution of ginsenoside Rg1 with a concentration of 0.5 mg / mL and a reference substance solution of ginsenoside Re; For the test by thin-layer chromatography, 10 μL of the test solution, the negative control solution, the reference substance solution of ginsenoside Rg1, and the reference substance solution of ginsenoside Re were each taken and spotted on the same silica gel GF254 thin-layer plate. A lower layer solution of a mixed solution of chloroform-ethyl acetate-methanol-water with a volume ratio of 7.5:20:11:5 when placed at a temperature below 10 °C was used as the developing solvent. After development, it was dried in air, sprayed with a sulfuric acid ethanol solution with a mass concentration of 10%, dried in air, and heated at 105 °C until the spots developed clearly. In the chromatogram of the test solution, the same pink spots were shown at the corresponding positions as those in the chromatograms of the reference substances of ginsenoside Rg1 and ginsenoside Re, and there was no interference from the negative control; The thin-layer identification method for styrax includes the following steps: Preparation of the test solution, negative control solution, and reference substance solution of cinnamic acid: Take 1 g of the powdered Shexiang Xintongning Pills after grinding, add 10 mL of ether, perform ultrasonic treatment for 2 min, and take the supernatant as the test solution; take 1 g of the pill powder without styrax, and prepare the negative control solution in the same way as the test solution; take the reference substance of cinnamic acid, and add ether to prepare a reference substance solution of cinnamic acid with a concentration of 1 mg / mL; For the test by thin-layer chromatography, 10 μL of the test solution, the negative control solution, and the reference substance solution of cinnamic acid were each taken and spotted on the same silica gel GF254 thin-layer plate. A mixed solution of petroleum ether (30-60 °C)-n-hexane-ethyl formate-formic acid with a volume ratio of 10:30:15:1 was used as the developing solvent. After development, it was dried in air, sprayed with a sulfuric acid ethanol solution with a mass concentration of 10%, dried by blowing, heated at 105 °C until the spots developed clearly, and inspected under ultraviolet light at a wavelength of 254 nm. In the chromatogram of the test solution, the same fluorescent spots of the same color were shown at the corresponding position as that of the reference substance of cinnamic acid, and there was no interference from the negative control; The thin-layer identification method for borneol includes the following steps: Preparation of test solution, negative control solution, and borneol reference substance solution: Take 0.3 g of the powdered Shexiang Xintongning Pills after grinding, add 15 mL of ethyl acetate, ultrasonically treat for 2 min, filter, evaporate the filtrate to near dryness at 60 °C, and redissolve with 0.5 mL of ethyl acetate to obtain the test solution; take 0.3 g of the pill powder without borneol and prepare the negative control solution in the same manner as the test solution; take the borneol reference substance and prepare a 2.5 mg / mL borneol reference substance solution with ethyl acetate; According to the thin-layer chromatography test, respectively pipette 2 μL of the test solution, negative control solution, and borneol reference substance solution, spot them on the same silica gel GF254 thin-layer plate, use a mixed solution of toluene - acetone with a volume ratio of 9:1 as the developing agent, spray a 5% vanillin sulfuric acid solution, and heat at 105 °C until the spots are clearly developed; in the chromatogram of the test sample, a same yellowish-green fluorescent spot appears at the same position as the borneol reference substance, and there is no interference from the negative control.

6. The quality inspection method of Shexiang Xintongning Pills according to claim 1, characterized in that, In the preparation process step (2), the macroporous adsorption resin is X-5 type macroporous adsorption resin; In the preparation process step (3), the coolant is dimethyl silicone oil.

7. The quality inspection method of Shexiang Xintongning Pills according to claim 1, characterized in that, In the preparation process step (2), the specific purification method: Pack the macroporous adsorption resin according to a height-diameter ratio of 1:8, load the medicinal solution by the wet method at 1 - 2 BV, perform dynamic adsorption at a flow rate of 8 - 10 BV / h, and wash away the residual impurities on the surface of the macroporous adsorption resin with more than 2.5 BV of pure water; The specific elution method: Elute with more than 8 BV of ethanol with a volume concentration of 60 - 70% at a flow rate of 8 - 10 BV / h.

8. The quality inspection method of Shexiang Xintongning Pills according to claim 1, characterized in that, The weight parts of each raw material in the Shexiang Xintongning Pills are as follows: Corydalis yanhusuo 220 - 260 parts, artificial musk 4 - 12 parts, styrax 4 - 12 parts, chuanxiong rhizome 100 - 140 parts, ginseng 100 - 140 parts, borneol 6 - 18 parts.

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