Novel UPLC-TQ-MS (Ultra Performance Liquid Chromatography-Thermal Quadrature Mass Spectrometry) or MS (Mass Spectrometry) method-based method

Abstract

The invention relates to the technical field of Zuojin pill content detection, in particular to a novel UPLC-TQ-MS or MS method-based Zuojin pill component content determination method. The method comprises the following steps: S1, selecting Zuojin pill samples of which the component content needs to be measured, wherein the Zuojin pill samples are respectively samples of Hubei coptis chinensis rural limited responsibility company, Hubei Nowinsheng Pharmaceutical Co., Ltd. And Wenzhou Hicrane Pharmaceutical Co., Ltd. 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; s41, detecting by adopting an ultra-performance liquid chromatograph; and S42, carrying out detection by using an ultra-high performance liquid chromatography-triple quadrupole mass spectrometer. According to the novel 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 present invention relates to the technical field of the content detection of Zuojin Pills, and particularly relates to a novel method for determining the component content of Zuojin Pills based on UPLC-TQ-MS or MS method. Background Art

[0002] Zuojin Pills originated from "Danxi's Heart Method" by Zhu Danxi, a medical expert in the Yuan Dynasty. It is composed of two medicinal materials, Coptis chinensis and Evodia rutaecarpa, and has the effects of clearing heat and detoxifying, drying dampness and relieving pain, soothing the liver and regulating the stomach. Research shows that the main components of Coptis chinensis are berberine, jatrorrhizine, and epiberberine, which have anti-inflammatory, hypoglycemic, and anti-tumor effects; Evodia rutaecarpa contains evodiamine, rutaecarpine, and dehydroevodiamine, which have anti-inflammatory, antibacterial, and cardiovascular protection effects.

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

[0004] Therefore, a novel method for determining the component content of Zuojin Pills based on UPLC-TQ-MS or MS method is designed to provide a technical solution for the above technical problems. Summary of the Invention

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

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

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

[0008] S1: Select the Zuojin Pills sample to be determined for component content;

[0009] S2: Prepare the mixed reference substance solution;

[0010] S3: Prepare the 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 with an ultra-high performance liquid chromatograph;

[0013] S42: Detect with an ultra-high performance liquid chromatography-triple quadrupole mass spectrometer.

[0014] As a preferred embodiment of the novel method for determining the component content of Zuojin Pills based on UPLC-TQ-MS or MS 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 Hubei Huanglian Township Co., Ltd. as samples;

[0016] S12: Select one batch of Zuojin Pills from Hubei Nuodesheng Pharmaceutical Co., Ltd. as a sample;

[0017] S13: Select three batches of Zuojin Pills from Wenzhou Haihe Pharmaceutical Co., Ltd. as samples.

[0018] As a preferred embodiment of the novel method for determining the component content of Zuojin Pills based on UPLC-TQ-MS or MS provided by the present invention, in step S2, the preparation of the reference substance solution is as follows:

[0019] S21: Weigh an appropriate amount of the reference substance;

[0020] S22: Respectively add methanol to prepare single reference substance stock solutions with a mass concentration of about 1 mg / mL;

[0021] 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 with a concentration of 2.5 μg / mL;

[0022] 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 with a concentration of 500 ng / mL.

[0023] As a preferred embodiment of the novel method for determining the component content of Zuojin Pills based on UPLC-TQ-MS or MS 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%.

[0024] As a preferred embodiment of the novel method for determining the component content 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 as follows:

[0025] S31: Weigh an appropriate amount of the medicinal material, grind it finely, weigh and take 1 g, and place it in a stoppered conical flask;

[0026] S32: Precisely add 50 mL of 50% methanol to the conical flask in step S31;

[0027] S33: Place the conical flask in step S32 into an ultrasonic device and ultrasonicate for 30 min;

[0028] S34: Let the ultrasonically treated conical flask obtained in step S33 cool down, make up the lost mass with 50% methanol, and shake well;

[0029] S35: Dilute the solution obtained in step S34 to 1 mg / mL with the extractant, filter through a 0.22 μm microporous membrane, and take the subsequent filtrate to obtain the corresponding test solution.

[0030] As a preferred embodiment of the method for determining the component content 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, and the steps are as follows:

[0031] Use a Waters ACQUITY BEH C 18 chromatographic column, with a mobile phase of methanol:acetonitrile in a ratio of 1:1 and 0.1% formic acid aqueous solution; 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.

[0032] As a preferred embodiment of the method for determining the component content 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 are a flow rate of 0.2 mL / min, a column temperature of 40 °C, and an injection volume of 3 μL.

[0033] As a preferred embodiment of the method for determining the component content 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, and the steps are as follows:

[0034] Use an electrospray ionization source and positive ion detection;

[0035] 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.

[0036] It can be undoubtedly seen that through the above technical solutions of the present application, the technical problems to be solved by the present application can surely be solved.

[0037] Meanwhile, through the above technical solutions, the present invention has at least the following beneficial effects:

[0038] A novel method for determining the component contents of Zuojin Pills based on UPLC-TQ-MS or MS provided by the present invention determines the contents of 9 components in Jiawei Zuojin Pills from different manufacturers 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 provide a reference for improving the quality standard research of Jiawei Zuojin Pills and lay a foundation for further expanding its application fields.

[0039] The method established in the present invention has a good linear relationship within a certain range (r 2 ≥0.999), the average sample addition recovery rate is 94.82-104.14%, and the RSD is 1.53-3.55%. The precision, repeatability and stability all meet the requirements. Description of the Drawings

[0040] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the following will briefly introduce the drawings required for the description of the embodiments. Obviously, the following drawings are some embodiments of the present invention. For those of ordinary skill in the art, other drawings can be obtained according to these drawings without creative work.

[0041] Figure 1 It is a schematic diagram of the mixed reference substance solution in the MRM chromatogram of the present invention;

[0042] Figure 2 For the present invention Figure 1 Continued table, which is a schematic diagram of the test solution of item S1 in the MRM chromatogram;

[0043] Figure 3 It is a dendrogram of cluster analysis of 7 batches of samples of the present invention;

[0044] Figure 4 It is a PCA score chart of 7 batches of samples of the present invention;

[0045] Figure 5 It is a loading chart of 9 components of the present invention. Detailed Embodiments

[0046] In order to make the purpose, technical solutions and advantages of the present invention clearer, the following will further describe the present invention in detail with reference to the drawings and embodiments. It should be understood that the specific embodiments described here are only used to explain the present invention and are not used to limit the present invention.

[0047] In order to enable those in the technical field to better understand the solution of the present invention, the following will clearly and completely describe the technical solutions in the embodiments of the present invention with reference to the drawings.

[0048] 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.

[0049] 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.

[0050] Refer to Figures 1 - 5 , a new method for determining the component content of Zuojin Pills based on UPLC-TQ-MS or MS method.

[0051] 1. Materials

[0052] 1.1 Instruments

[0053] Waters ACQUITY UPLC / XEVO TQ-XS ultra-high performance liquid chromatography-triple quadrupole mass spectrometry (Waters); M204 one ten-thousandth electronic analytical balance (Mettler), KQ-500DB numerically controlled ultrasonic cleaner (Kunshan Ultrasonic Instruments Co., Ltd.), Milli-Q ultrapure water system.

[0054] 1.2 Reagents and Samples

[0055] Chromatographic pure methanol and acetonitrile (Merck), chromatographic pure formic acid (Fisher).

[0056] 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%.

[0057] 7 batches of Zuojin Pills were drawn from business and production units across the country, involving 3 production enterprises, as shown in Table 1; among them, there are 3 batches of samples from Hubei Huanglianzhixiang Co., Ltd. (S1-S3), 1 batch of samples from Hubei Nuodesheng Pharmaceutical Co., Ltd. (S4), and 3 batches of samples from Wenzhou Haihe Pharmaceutical Co., Ltd. (S5-S7).

[0058] Table 1: Sample Sources

[0059]

[0060] 2. Methods and Results

[0061] 2.1 Solution Preparation

[0062] 2.1.1 Preparation of Reference Substance Solution

[0063] Accurately weigh appropriate amounts of various reference substances, and separately add methanol to prepare single reference substance stock solutions with a mass concentration of approximately 1 mg / mL; accurately measure appropriate amounts of the above single reference substance stock solutions, add methanol to prepare a mixed reference substance solution stock solution with a concentration of 2.5 μg / mL, accurately measure the mixed reference substance solution stock solution, and add methanol to prepare a mixed reference substance solution with a concentration of 500 ng / mL.

[0064] 2.1.2 Test Solution

[0065] Weigh appropriate amounts of 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 the mass, 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 extract to 1 mg / mL, filter through a 0.22 μm microporous membrane, and take the subsequent filtrate to obtain the test solutions of medicinal materials of each batch number.

[0066] 2.2 UPLC-TQ-MS or MS Analysis Conditions

[0067] 2.2.1 Liquid Phase Conditions

[0068] Use a Waters ACQUITY BEH C 18 (100 mm × 2.1 mm, 1.7 μm) chromatographic column, 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 [张佳颖,尹艺,徐倩倩,等.UPLC-MS / MS方法同时测 Determination of the Contents of 23 Components in Dingzuojin Pills [J]. Chinese Journal of Pharmaceutical Analysis, 2023, 43(07): 1110 - 1119. DOI: 10.16155 / j.0254 - 1793.2023.07.03].

[0069] 2.2.2 Mass Spectrometry Conditions

[0070] It was detected by ultra-high performance liquid chromatography-triple quadrupole mass spectrometry. Electrospray ionization source (ESI), positive ion detection, multiple reaction monitoring mode (MRM). The capillary voltage was 3.0 kV, the desolvation gas temperature was 300 °C, the cone gas flow rate was 150 L / h, and the desolvation gas flow rate was 1000 h / L. Argon (99.99%) was used as the collision gas. The mass spectrometry parameters of each compound are shown in Table 2.

[0071] Table 2: Mass spectrometry parameters of 9 compounds

[0072]

[0073]

[0074] Among them, CV is the cone voltage, and CE is the collision energy.

[0075] 2.3. Methodological investigation

[0076] 2.3.1 Specificity test

[0077] 3 μL of the mixed reference solution at the lowest concentration under item "2.1.1" and the test solution of Zuojin Pills in item S1 were respectively aspirated and injected for analysis under the conditions in 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 is > 1.5, indicating that this method has strong specificity.

[0078] 2.3.3 Investigation of linear relationship

[0079] The mixed reference solution under item "2.1.1" was taken and diluted step by step, and injected for determination under the conditions in item "2.2.1". Linear regression was performed with the corresponding reference 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.

[0080] Table 3: Investigation of linear relationships of each component

[0081]

[0082]

[0083] 2.3.2 Precision determination

[0084] The mixed reference solution at the lowest concentration under item "2.1.1" was taken and continuously injected for determination 6 times under the conditions in item "2.2.1". The RSD (n = 6) of the peak areas of each component was calculated to be < 3.07% (see Table 4), indicating good precision of the instrument.

[0085] 2.3.4 Repeatability test

[0086] Take Zuojin Pills of item S1, prepare 6 portions of test solutions in parallel according to the method in item "2.1.1", inject samples for determination under the conditions in item "2.2.1", and the RSD (n = 6) of the contents of each component is < 3.73%, indicating that the method has good repeatability (see Table 4).

[0087] 2.3.5 Stability test

[0088] Take the test solution of item S1, inject samples for determination under the conditions in item "2.2.1" after standing at room temperature for 0, 2, 4, 8, and 12 h. The RSD (n = 6) of the peak areas of each component is < 3.09% (see Table 4), indicating that the test solution has good stability when standing at room temperature for 12 h.

[0089] 2.3.6 Recovery test

[0090] Precisely weigh 6 portions of Zuojin Pills of item S1, prepare 6 portions of test solutions according to the method in item "2.1.2", precisely add mixed reference substance solutions equivalent to 100% of the contents of each component to be measured in the sample respectively, inject samples for determination under the conditions in item "2.2.1", and calculate the recovery 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] 2.3.7 Determination of sample content

[0094] Prepare test solutions of Zuojin Pills of S1 - S7 respectively according to the methods in items "2.2.1" and "2.2.2", prepare 3 portions in parallel for each batch, inject samples for determination under the conditions in item "2.1", and calculate the content of each component according to the regression equation. The results are shown in Table 5.

[0095] Table 5: Contents of 9 compounds in 7 batches of samples (mg / g)

[0096]

[0097] 2.4 Chemometric analysis

[0098] 2.4.1 Cluster analysis

[0099] Hierarchical cluster analysis (HCA) is an unsupervised pattern analysis method, which can more intuitively show the differences in chemical components among sample batches, using the average values of the contents of each component as the original data. Using SPSS 26.0 software, the group average linkage method is used for the contents of 9 components in 7 batches of samples. The results are shown in Figure 3。The 7 batches of samples were grouped into 3 categories according to each manufacturing enterprise. The results showed that there were certain differences in the product quality among different manufacturing enterprises, and there were also differences in the product quality of different batches from the same enterprise.

[0100] 2.4.2 Principal Component Analysis

[0101] Principal Component Analysis (PCA) was performed on the measurement results using SIMCA 14.1 software. The unsupervised pattern recognition method was adopted to observe the automatic aggregation of samples. The model automatically selected 3 principal components by fitting, and the cumulative contribution rate was 99.3%. This indicates that the 3 principal components in the model can well represent most of the information content of the 9 analytical variables in Zuojin Pills, and the prediction degree of the model is good. After PCA, 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 respectively.

[0102] 3. Results

[0103] The contents of 9 components in Zuojin Pills from different manufacturers and different batches were determined by UPLC-TQ-MS or MS method, and the contents of 9 components in Zuojin Pills of different batches were obtained. Through cluster analysis, the products S1-3 of Hubei Huanglianzhixiang Co., Ltd. were grouped into one category as A, the product S4 of Hubei Nuodesheng Pharmaceutical Co., Ltd. was grouped into one category as B, and the products S5-7 of Wenzhou Haihe Pharmaceutical Co., Ltd. were grouped into one category as C. The clustering effect was obvious, indicating that there were significant differences in the component contents of Zuojin Pills produced by different enterprises. To further study the relationship between different manufacturers and components, the PCA score plot of 7 batches of samples and the load plot of 9 components were obtained by principal component analysis. From the results of the PCA score plot, it can be seen that there are differences in the product quality of the same manufacturer but different batches. Through the analysis of the component load plot, berberine, epiberberine, and palmatine may be the main components leading to the quality differences of Zuojin Pills of different batches. According to Part I of the Pharmacopoeia of the People's Republic of China (2020 Edition), the preparation process of Zuojin Pills is to take Coptis chinensis Franch. and Evodia rutaecarpa (Juss.) Benth. herbs in a ratio of 6:1, crush, sieve, mix evenly, and make pills with water. The content determination mainly measures the content of berberine hydrochloride in the samples, and the contents of berberine, epiberberine, and palmatine are not specified. Based on the experimental results, the quality control of Zuojin Pills should consider the content of berberine in Coptis chinensis Franch., and at the same time determine the contents of berberine, epiberberine, and palmatine.

[0104] 4. Conclusions

[0105] The present invention establishes a UPLC-TQ-MS or MS method for determining nine components in Zuojin Pills, which has high accuracy and high separation efficiency. The established method is used to determine Zuojin Pills from different manufacturers and different batches. Cluster analysis and PCA are used to evaluate the quantitative determination results. The results show that there are differences in the amounts of some components in Zuojin Pills from different manufacturers and different batches. Therefore, this method can be used for the quality control of related preparations of Zuojin Pills and can also provide a reference for further research on the pharmacodynamic material basis thereof.

[0106] 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 do they 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 well understand and utilize the present invention. 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 using an ultra-high performance liquid chromatograph; S42: Detect using an 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 the Zuojin Pills samples, the steps are as follows: S11: Select three batches of Zuojin Pills from Hubei Huanglianzhixiang Co., Ltd. as samples; S12: Select one batch of Zuojin Pills from Hubei Nuodesheng Pharmaceutical Co., Ltd. as samples; S13: Select three batches of Zuojin Pills from Wenzhou Haihe Pharmaceutical Co., Ltd. as samples.

3. A method for determining the content of the components of Zuojin Pills based on UPLC-TQ-MS or MS method according to claim 1, characterized in that, In step S2, for the preparation of the reference substance solution, the steps are as follows: S21: Weigh an appropriate amount of the reference substance; S22: Respectively add methanol to prepare 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. The method for determining the component content of Zuojin Pills based on UPLC-TQ-MS or MS method according to claim 3, wherein, The reference substances are berberine, jatrorrhizine, epiberberine, evodiamine, rutaecarpine, dehydroevodiamine, limonin, coptisine, and palmatine, and the purity of each is >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, characterized in that, In step S3, for the preparation of the test solution, the steps are as follows: S31: Weigh an appropriate amount of the medicinal material, grind it finely, weigh and take 1 g, and place it 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 content of the components of Zuojin Pills based on UPLC-TQ-MS or MS method according to claim 1, characterized in that, In step S41, for the detection using an 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 content of the components of Zuojin Pills based on UPLC-TQ-MS or MS method according to claim 6, characterized in that, The chromatographic conditions for detection using an ultra-high performance liquid chromatograph by 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 using an 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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