Zuojin pill component content determination method based on UPLC-TQ-MS or MS method

Abstract

The invention relates to the technical field of Zuojin pill content detection, in particular to a Zuojin pill component content determination method based on a UPLC-TQ-MS or MS method. The method comprises the following steps: S1, selecting Zuojin pill samples needing component content determination, wherein the Zuojin pill samples are samples of Jiangxi Youchun Kangyuan Pharmaceutical Co., Jiangxi Yangdu Cinnamomum camphora Pharmaceutical Co., Jiangxi Yangdu Cinnamomum camphora Pharmaceutical Co., Beijing Nanzhongtang Pharmaceutical Co. And Shandong Guangyutang National Drug Co S2, preparing a mixed reference substance solution; s3, preparing a test solution; S4, determining chemical components in the mixed reference substance and the test solution by adopting a UPLC-TQ-MS or MS method, so as to determine the component content of the Zuojin pill; and S41, detecting by adopting an ultra-high performance liquid chromatograph. According to the method for determining the content of the components of the Zuojin pills based on the UPLC-TQ-MS or MS method, the UPLC-TQ-MS or MS method is established, and the content of nine components in the modified Zuojin pills of different manufacturers is determined at the same time; and comparing the difference of the flavored Zuojin pills in different batches in combination with clustering analysis and principal component analysis.

Description

Technical Field

[0001] The invention relates to the technical field of Zuojin Pill content detection, and in particular to a method for determining the content of Zuojin Pill components based on UPLC-TQ-MS or MS method. Background Art

[0002] Zuojin Pills are from Danxi's Heart Method. They are composed of two medicinal materials, coptis chinensis and evodia rutaecarpa. They have the effects of clearing away heat and detoxifying, drying dampness and relieving pain, and soothing the liver and stomach. Based on Zuojin Pills, Jiawei Zuojin Pills add 14 medicinal materials, including scutellaria baicalensis, bupleurum, costusroot, vinegar-curcuma cyperus, curcuma, white peony root, vinegar-curcuma chinensis, bran-fried aurantium, dried orange peel, vinegar-curcuma corydalis, angelica sinensis, and liquorice, to enhance and expand its efficacy. Clinically, Jiawei Zuojin Pills have good therapeutic effects in the treatment of liver-stomach disharmony, chronic superficial gastritis, gastroesophageal reflux disease, and gastric ulcers. Berberine and coptis chinensis, the main components of coptis chinensis, have anti-inflammatory, antioxidant, and cell apoptosis effects. Evodiamine and rutaecarpine in evodia rutaecarpa have the effects of regulating inflammation, anti-cancer, and anti-atherosclerosis. Therefore, it is necessary to detect the content of the main components of coptis chinensis and evodia rutaecarpa in Jiawei Zuojin Pills.

[0003] UPLC-TQ-MS or MS analysis technology is currently one of the means to quickly analyze the chemical components in Chinese patent medicines, Chinese medicinal materials and Chinese medicinal extracts. This technology has the characteristics of high sensitivity, high efficiency and high accuracy, so it has been widely used in quantitative research of Chinese medicine.

[0004] To this end, a method for determining the content of Zuojin Pills ingredients based on UPLC-TQ-MS or MS was designed to provide another technical solution to the above technical problems. Summary of the invention

[0005] Based on this, it is necessary to provide a method for determining the content of Zuojin Pill ingredients based on UPLC-TQ-MS or MS to solve the technical problems raised in the above background technology.

[0006] In order to solve the above technical problems, the present invention adopts the following technical solutions:

[0007] A method for determining the content of Zuojin Pills ingredients based on UPLC-TQ-MS or MS, the steps are as follows:

[0008] S1: Select Zuojin pill samples that need to be tested for component content;

[0009] S2: prepare mixed reference solution;

[0010] S3: Preparation of test solution

[0011] S4: Use UPLC-TQ-MS or MS method to determine the chemical components in the mixed reference substance and the test solution, so as to determine the component content of Zuojin Pills;

[0012] S41: Detect by ultra-high performance liquid chromatograph;

[0013] S42: Detect by ultra-high performance liquid chromatography-triple quadrupole mass spectrometry.

[0014] As a preferred implementation manner of the method for determining the component content of Zuojin Pills based on UPLC-TQ-MS or MS method provided by the present invention, in step S1, the selection of Zuojin Pills samples is as follows:

[0015] S11: Select three batches of Zuojin Pills from Jiangxi Qingchun Kangyuan Pharmaceutical Co., Ltd. as samples;

[0016] S12: Select two batches of Zuojin Pills from Jiangxi Yaodu Zhangshu Pharmaceutical Co., Ltd. as samples;

[0017] S13: Select two batches of Zuojin Pills from Beijing Tongrentang Pharmaceutical Co., Ltd. as samples;

[0018] S14: Select three batches of Zuojin Pills from Shandong Guangyutang Guoyao Co., Ltd. as samples.

[0019] As a preferred implementation manner of the method for determining the component content of Zuojin Pills based on UPLC-TQ-MS or MS method provided by the present invention, in step S2, the preparation of the reference substance solution is as follows:

[0020] S21: Weigh an appropriate amount of the reference substance;

[0021] S22: Respectively add methanol to prepare single reference substance stock solutions with a mass concentration of about 1 mg / mL;

[0022] S23: Precisely measure an appropriate amount of the single reference substance stock solution in step S22, and add methanol to prepare a mixed reference substance solution stock solution of 2.5 μg / mL;

[0023] S24: Precisely measure an appropriate amount of the mixed reference substance solution stock solution in step S23, and add methanol to prepare a mixed reference substance solution of 500 ng / mL.

[0024] As a preferred implementation manner of the method for determining the component content of Zuojin Pills based on UPLC-TQ-MS or MS method provided by the present invention, the reference substances are berberine, jatrorrhizine, epiberberine, evodiamine, rutaecarpine, dehydroevodiamine, limonin, coptisine and palmatine, and the purity of each is >98.0%.

[0025] As a preferred embodiment of the method for determining the content of the components of Zuojin Pills based on UPLC-TQ-MS or MS provided by the present invention, in step S3, the preparation of the test solution is carried out as follows:

[0026] S31: Weigh an appropriate amount of the medicinal materials, grind them finely, weigh and take 1 g, and place it in a stoppered conical flask;

[0027] S32: Precisely add 50 mL of 50% methanol to the conical flask in step S31;

[0028] S33: Place the conical flask in step S32 into an ultrasonic device and perform ultrasonic treatment for 30 min;

[0029] S34: Let the conical flask after ultrasonic treatment obtained in step S33 cool down, make up the lost mass with 50% methanol, and shake well;

[0030] S35: Dilute the solution obtained in step S34 to 1 mg / mL for the extract, filter it through a 0.22 μm microporous filter membrane, and take the subsequent filtrate to obtain the corresponding test solution.

[0031] As a preferred embodiment of the method for determining the content of the components of Zuojin Pills based on UPLC-TQ-MS or MS provided by the present invention, in step S41, detection is carried out using an ultra-high performance liquid chromatograph as follows:

[0032] Use a Waters ACQUITY BEH C 18 chromatographic column, with methanol:acetonitrile in a ratio of 1:1 and 0.1% formic acid aqueous solution as the mobile phase; gradient elution: 0 - 3 min, 5 - 20% A; 3 - 9 min, 20 - 40% A; 9 - 15 min, 40 - 95% A; 15 - 16 min, 95 - 5% A; 16 - 18 min, 5% A.

[0033] As a preferred embodiment of the method for determining the content of the components of Zuojin Pills based on UPLC-TQ-MS or MS provided by the present invention, the chromatographic conditions for detection using an ultra-high performance liquid chromatograph with UPLC-TQ-MS or MS method are a flow rate of 0.2 mL / min, a column temperature of 40 °C, and an injection volume of 3 μL.

[0034] As a preferred embodiment of the method for determining the content of the components of Zuojin Pills based on UPLC-TQ-MS or MS provided by the present invention, in step S42, detection is carried out using an ultra-high performance liquid chromatography-triple quadrupole mass spectrometer as follows:

[0035] Use an electrospray ionization source and perform positive ion detection;

[0036] The capillary voltage is 3.0 kV, the desolvation gas temperature is 300 °C, the cone gas flow rate is 150 L / h, the desolvation gas flow rate is 1000 h / L, and argon is used as the collision gas.

[0037] It can be seen without doubt that through the above technical solutions of this application, the technical problems to be solved by this application can surely be solved.

[0038] Meanwhile, through the above technical solutions, the present invention has at least the following beneficial effects:

[0039] 1. A method for determining the component content of Zuojin Pills based on UPLC-TQ-MS or MS method provided by the present invention determines the contents of 9 components in Jiawei Zuojin Pills from different manufacturers simultaneously by establishing the UPLC-TQ-MS or MS method; combining cluster analysis and principal component analysis to compare the differences of Jiawei Zuojin Pills in different batches, so as to lay a foundation for improving the research on the quality standard of Jiawei Zuojin Pills and further expanding its application fields.

[0040] 2. The method established by the present invention has a good linear relationship in a certain range (r 2 ≥0.999), the average sample addition recovery rate is 94.82 - 104.14%, the RSD is 1.53 - 3.55%, and the precision, repeatability and stability all meet the requirements. Description of the Drawings

[0041] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings required for the description of the embodiments will be briefly introduced below. Obviously, the drawings in the following description are some embodiments of the present invention. For those of ordinary skill in the art, other drawings can be obtained based on these drawings without creative efforts.

[0042] Figure 1 It is a schematic diagram of the mixed reference substance solution in the MRM chromatogram of the present invention;

[0043] Figure 2 It is a schematic diagram of the test solution of item S1 in the MRM chromatogram of the present invention;

[0044] Figure 3 It is a dendrogram of cluster analysis of 10 batches of samples of the present invention;

[0045] Figure 4 It is a PCA score chart of 10 batches of samples of the present invention;

[0046] Figure 5 It is a loading chart of 9 components of the present invention. Detailed Embodiments

[0047] In order to make the objectives, technical solutions and advantages of the present invention clearer and more understandable, the present invention will be further described in detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention.

[0048] In order to enable those skilled in the art of this technology to better understand the solution of the present invention, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings.

[0049] It should be noted that, without conflict, the embodiments in the present invention and the features and technical solutions in the embodiments can be combined with each other.

[0050] It should be noted that similar reference numerals and letters indicate similar items in the following drawings. Therefore, once an item is defined in one drawing, it does not need to be further defined and explained in subsequent drawings.

[0051] Refer to Figures 1 - 5 , a method for determining the component content of Zuojin Pills based on UPLC-TQ-MS or MS method.

[0052] 1. Materials

[0053] 1.1 Instruments

[0054] Waters ACQUITY UPLC / XEVO TQ-XS ultra-high performance liquid chromatography-triple quadrupole mass spectrometry (Waters Corporation); M204 one-ten-thousandth electronic analytical balance (Mettler Corporation), KQ-500DB numerically controlled ultrasonic cleaner (Kunshan Ultrasonic Instruments Co., Ltd.), Milli-Q type ultrapure water system.

[0055] 1.2 Reagents and Samples

[0056] Chromatographic pure methanol and acetonitrile (Merck), chromatographic pure formic acid (Fisher).

[0057] Reference substances berberine (batch number Yz110920), jatrorrhizine (batch number Yz032022), epiberberine (batch number Yz101724), evodiamine (batch number Yz0224221), rutaecarpine (batch number Yz0301221), dehydroevodiamine (batch number Yz030823), limonin (batch number Yz012922) were purchased from Nanjing Yuanzhi Biotechnology Co., Ltd., coptisine (batch number 16081403), palmatine (batch number 16041003) were purchased from Qingdao Jieshikang Biotechnology Co., Ltd. The purity of the above 9 reference substances is >98.0%.

[0058] 10 batches of Zuojin Pills with Added Flavors from different manufacturers and different batch numbers were drawn from 4 production enterprises, as shown in Table 1; among them, there were 3 batches of drugs (S1 - S3) from Jiangxi Qingchun Kangyuan Pharmaceutical Co., Ltd., 2 batches of drugs (S4 - S5) from Jiangxi Yaodu Zhangshu Pharmaceutical Co., Ltd., 2 batches of drugs (S6 - S7) from Beijing Tongrentang Pharmaceutical Co., Ltd., and 3 batches of drugs (S8 - S10) from Shandong Guangyutang Chinese Medicine Co., Ltd.

[0059] Table 1: Source of Samples

[0060]

[0061] 2. Methods and Results

[0062] 2.1 Solution Preparation

[0063] 2.1.1 Preparation of Reference Solution

[0064] Weigh an appropriate amount of each reference substance accurately, and separately prepare single reference substance stock solutions with a mass concentration of about 1 mg / mL in methanol; accurately measure an appropriate amount of the above single reference substance stock solutions, add methanol to prepare a mixed reference solution stock solution with a concentration of 2.5 μg / mL, accurately measure the mixed reference solution stock solution, and add methanol to prepare a mixed reference solution with a concentration of 500 ng / mL.

[0065] 2.1.2 Test Solution

[0066] Weigh an appropriate amount of the medicinal materials of each batch number, grind them finely, accurately weigh 1 g, place them in a stoppered conical flask, accurately add 50 mL of 50% methanol, weigh, ultrasonicate for 30 min (power 250 W, frequency 40 kHz), let it cool, make up the lost mass with 50% methanol, shake well, dilute the extraction solution to 1 mg / mL, filter through a 0.22 μm microporous membrane, and take the subsequent filtrate to obtain the test solutions of the medicinal materials of each batch number.

[0067] 2.2 UPLC - TQ - MS or MS Analysis Conditions

[0068] 2.2.1 Liquid Phase Conditions

[0069] Use Waters ACQUITY BEH C 18A chromatographic column (100 mm × 2.1 mm, 1.7 μm), mobile phase: acetonitrile: methanol 1:1 (A) - 0.1% formic acid aqueous solution (B); flow rate: 0.2 mL / min; column temperature: 40 °C; detection wavelength: 256 nm; injection volume: 3 μL; gradient elution: 0 - 3 min, 5 - 20% A; 3 - 9 min, 20 - 40% A; 9 - 15 min, 40 - 95% A; 15 - 16 min, 95 - 5% A; 16 - 18 min, 5% A [Zhang Jiaying, Yin Yi, Xu Qianqian, et al. Simultaneous determination of 23 components in Zuojin Pills by UPLC-MS / MS [J]. Chinese Journal of Pharmaceutical Analysis, 2023, 43(07): 1110 - 1119. DOI: 10.16155 / j.0254 - 1793.2023.07.03].

[0070] 2.2.2 Mass spectrometry conditions

[0071] Detected by ultra - performance liquid chromatography - triple quadrupole mass spectrometry, electrospray ionization source (ESI), positive ion detection, multiple reaction monitoring mode (MRM). Capillary voltage 3.0 kV, desolvation gas temperature 300 °C, cone gas flow rate 150 L / h, desolvation gas flow rate 1000 h / L. Argon (99.99%) was used as the collision gas, and the mass spectrometry parameters of each compound are shown in Table 2.

[0072] Table 2: Mass spectrometry parameters of 9 compounds

[0073]

[0074]

[0075] Among them, CV is the cone voltage, and CE is the collision energy.

[0076] 2.3. Methodological investigation

[0077] 2.3.1 Specificity test

[0078] Respectively absorb 3 μL of the mixed reference substance solution at the lowest concentration under item "2.1.1" and the test solution of Zuojin Pills in item S1, and inject for analysis according to the conditions under item "2.2.1". The results are shown in Figure 1 and Figure 2 , it can be seen that under the multiple reaction monitoring mode, the peak shapes of each peak to be measured are good, and the resolution from adjacent chromatographic peaks > 1.5, indicating that this method has strong specificity.

[0079] 2.3.3 Linear relationship investigation

[0080] Take the mixed reference substance solution under item "2.1.1", dilute it step by step, inject the sample for determination under the conditions of item "2.2.1", perform linear regression with the corresponding reference substance peak area (Y) as the ordinate and the mass concentration (X) as the abscissa. The results in Table 3 show that the injection concentrations of the 9 components have good linear relationships within their respective linear ranges.

[0081] Table 3: Investigation on the linear relationships of each component

[0082]

[0083] 2.3.2 Precision determination

[0084] Take the mixed reference substance solution with the lowest concentration under item "2.1.1", continuously inject the sample for determination 6 times under the conditions of item "2.2.1", and calculate the RSD (n = 6) of the peak areas of each component, all of which are < 3.07% (see Table 4), indicating good instrument precision.

[0085] 2.3.4 Repeatability test

[0086] Take the modified Zuojin Pills of batch number S1, prepare 6 portions of test solution in parallel according to the method under item "2.1.1", inject the sample for determination under the conditions of item "2.2.1", and calculate the RSD (n = 6) of the contents of each component, all of which are < 3.73%, indicating good method repeatability (see Table 4).

[0087] 2.3.5 Stability test

[0088] Take the test solution of item S1, place it at room temperature for 0, 2, 4, 8, 12 h, then inject the sample for determination under the conditions of item "2.2.1", and calculate the RSD (n = 6) of the peak areas of each component, all of which are < 3.09% (see Table 4), indicating good stability of the test solution when placed at room temperature for 12 h.

[0089] 2.3.6 Recovery test

[0090] Precisely weigh 6 portions of the modified Zuojin Pills of batch number S1, prepare 6 portions of test solution according to the method under item "2.1.2", precisely add the mixed reference substance solution equivalent to 100% of the content of each component to be measured in the sample, inject the sample for determination under the conditions of item "2.2.1", and calculate the recovery rates of each component. The results are shown in Table 4.

[0091] Table 4: Results of precision, stability, reproducibility and recovery tests for 9 compounds

[0092]

[0093]

[0094] 2.3.7 Determination of sample content

[0095] The test solutions of Jiawei Zuojin Pills for batches S1 - S10 were prepared according to the methods described in items "2.2.1" and "2.2.2" respectively. Three parallel preparations were made for each batch, and the samples were injected for determination under the conditions described in item "2.1". The contents of each component were calculated according to the regression equation, and the results are shown in Table 5.

[0096] Table 5: Contents of 9 components in 10 batches of samples (mg / g)

[0097]

[0098] 2.4 Chemometric Analysis

[0099] 2.4.1 Cluster Analysis

[0100] Hierarchical Cluster Analysis is a commonly used method in cluster analysis. It can gradually merge or decompose data according to the degree of similarity, forming a tree - shaped clustering structure, that is, a clustering dendrogram. Using SPSS 26.0 software, the between - groups linkage method was used to analyze the contents of 9 components in 10 batches of samples. The results are shown in Figure 3 ... The 10 batches of samples were clustered into 4 categories according to each production enterprise. The first category is S1 - 3, Jiangxi Qingchun Kangyuan Pharmaceutical Co., Ltd.; the second category is S4 - 5, Jiangxi Yaodu Zhangshu Pharmaceutical Co., Ltd.; the third category is S6 - 7, Beijing Tongrentang Pharmaceutical Co., Ltd.; the fourth category is S8 and S10, Shandong Guangyutang Guoyao Co., Ltd. The clustering effect is obvious. Among them, the S9 sample from Shandong Guangyutang is a separate category with relatively large differences. Analyzing in combination with the contents of 9 components, it may be related to the relatively low contents of its various components.

[0101] 2.4.2 Principal Component Analysis

[0102] Principal Component Analysis was performed on the measurement results using SIMCA 14.1 software. The unsupervised pattern recognition method was used to observe the automatic aggregation of samples. The model automatically selected 4 principal components, and the cumulative contribution rate was 99.7%. This indicates that the 4 principal components in the model can better represent most of the information content of the 9 analysis variables in Jiawei Zuojin Pills, and the prediction degree of the model is good. After principal component analysis, the PCA score plot of the samples and the load contribution plot of the drug components were obtained, as shown in Figure 4 and Figure 5 ... The farther the distance from the origin, the more important it is for classification. Among them, jatrorrhizine, palmatine, evodiamine, and rutaecarpine are the farthest from the origin, indicating that they may be the main components causing the quality differences of different batches of Jiawei Zuojin Pills.

[0103] 3. Results

[0104] The content determination results were evaluated by cluster analysis and PCA. The results showed that there were differences in the amounts of some important compounds in Zuojin Pills with added flavor from different manufacturers and different batches. Compounds such as jatrorrhizine, palmatine, evodiamine, and rutaecarpine were the key components for differentiating various batches of Zuojin Pills with added flavor.

[0105] The contents of 9 components in Zuojin Pills with added flavor from different manufacturers and different batches were determined by UPLC-TQ-MS or MS method, and the contents of 9 components in different batches of Zuojin Pills with added flavor were obtained. There were obvious differences in the contents of each batch, which might be attributed to the uneven quality of the cut herbs used in the samples of different batches, and also related to the preparation processes of different manufacturers. Therefore, in order to ensure the stable content of drug components, the quality of each component in each process of drug production should be strictly controlled, and the content of the main components should be dynamically monitored to ensure the stable quality of Zuojin Pills with added flavor and guarantee the effectiveness and safety of clinical medication.

[0106] 4. Conclusion

[0107] An UPLC-TQ-MS or MS method for determining 9 components in Zuojin Pills with added flavor was established, which had high accuracy and high separation efficiency. The established method was applied to determine Zuojin Pills with added flavor from different manufacturers and different batches. Cluster analysis and PCA were used to evaluate the quantitative determination results. The results showed that there were differences in the amounts of some components in Zuojin Pills with added flavor from different manufacturers and different batches. Therefore, this method can be used for the quality control of related preparations of Zuojin Pills with added flavor, and can also provide reference for further research on the pharmacodynamic material basis.

[0108] The preferred embodiments of the present invention disclosed above are only used to help illustrate the present invention. The preferred embodiments do not describe all the details in detail, nor limit the present invention to the specific embodiments described. Obviously, many modifications and variations can be made according to the content of this specification. These embodiments are selected and specifically described in this specification to better explain the principles and practical applications of the present invention, so that those skilled in the art can understand and utilize the present invention well. The present invention is only limited by the claims and their full scope and equivalents.

Claims

1. A method for determining the component content of Zuojin Pills based on UPLC-TQ-MS or MS method, characterized in that, The steps are as follows: S1: Select the Zuojin Pills samples for which the component content needs to be determined; S2: Prepare the mixed reference substance solution; S3: Prepare the test solution S4: Use UPLC-TQ-MS or MS method to determine the chemical components in the mixed reference substance and the test solution, so as to determine the component content of Zuojin Pills; S41: Detect by ultra-high performance liquid chromatograph; S42: Detect by ultra-high performance liquid chromatography-triple quadrupole mass spectrometer.

2. The method for determining the component content of Zuojin Pills based on UPLC-TQ-MS or MS method according to claim 1, characterized in that, In step S1, for the selection of Zuojin Pills samples, the steps are as follows: S11: Select three batches of Zuojin Pills from Jiangxi Qingchun Kangyuan Pharmaceutical Co., Ltd. as samples; S12: Select two batches of Zuojin Pills from Jiangxi Yaodu Zhangshu Pharmaceutical Co., Ltd. as samples; S13: Select two batches of Zuojin Pills from Beijing Tongrentang Pharmaceutical Co., Ltd. as samples; S14: Select three batches of Zuojin Pills from Shandong Guangyutang Guoyao Co., Ltd. as samples.

3. The method for determining the component content of Zuojin Pills based on UPLC-TQ-MS or MS method according to claim 1, wherein In step S2, for the preparation of the reference substance solution, the steps are as follows: S21: Weigh an appropriate amount of reference substance; S22: Respectively add methanol to make single reference substance stock solutions with a mass concentration of about 1 mg / mL; S23: Precisely measure an appropriate amount of the single reference substance stock solution in step S22, and add methanol to prepare a 2.5 μg / mL mixed reference substance solution stock solution; S24: Precisely measure an appropriate amount of the mixed reference substance solution stock solution in step S23, and add methanol to prepare a 500 ng / mL mixed reference substance solution.

4. A method for determining the component content of Zuojin Pills based on UPLC-TQ-MS or MS method according to claim 3, characterized in that, The reference substances are berberine, jatrorrhizine, epiberberine, evodiamine, rutaecarpine, dehydroevodiamine, limonin, coptisine and palmatine, and their purities are all >98.0%.

5. The method for determining the component content of Zuojin Pills based on UPLC-TQ-MS or MS method according to claim 1, wherein, In step S3, for the preparation of the test solution, the steps are as follows: S31: Weigh an appropriate amount of medicinal materials, grind them finely, weigh 1 g, and place them in a stoppered conical flask; S32: Precisely add 50 mL of 50% methanol to the conical flask in step S31; S33: Place the conical flask in step S32 into an ultrasonic device and ultrasonicate for 30 min; S34: Let the conical flask after ultrasonication in step S33 cool down, make up the lost mass with 50% methanol, and shake well; S35: Dilute the solution in step S34 to 1 mg / mL for the extract, filter it through a 0.22 μm microporous filter membrane, and take the subsequent filtrate to obtain the corresponding test solution.

6. The method for determining the component content of Zuojin Pills based on UPLC-TQ-MS or MS method according to claim 1, wherein In step S41, for the detection by ultra-high performance liquid chromatograph, the steps are as follows: Using a Waters ACQUITY BEH C 18 chromatographic column, with a mobile phase of methanol:acetonitrile in a ratio of 1:1 and an aqueous solution of 0.1% formic acid; gradient elution: 0 - 3 min, 5 - 20% A; 3 - 9 min, 20 - 40% A; 9 - 15 min, 40 - 95% A; 15 - 16 min, 95 - 5% A; 16 - 18 min, 5% A.

7. A method for determining the component content of Zuojin Pills based on UPLC-TQ-MS or MS method according to claim 6, characterized in that, The chromatographic conditions for detection by ultra-high performance liquid chromatograph using UPLC-TQ-MS or MS method are a flow rate of 0.2 mL / min, a column temperature of 40 °C, and an injection volume of 3 μL.

8. A method for determining the component content of Zuojin Pills based on UPLC-TQ-MS or MS method according to claim 1, characterized in that, In step S42, for the detection by ultra-high performance liquid chromatography-triple quadrupole mass spectrometer, the steps are as follows: Use an electrospray ionization source for positive ion detection; The capillary voltage is 3.0 kV, the desolvation gas temperature is 300 °C, the cone gas flow rate is 150 L / h, the desolvation gas flow rate is 1000 h / L, and argon is used as the collision gas.

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