UPLC-Q-TOF-MS / MS technology-based detection method for chemical components of Taoji Chengqi decoction pharmaceutical composition

Abstract

The invention discloses a UPLC-Q-TOF-MS / MS (Ultra Performance Liquid Chromatography-Quantitative Time of Flight-Mass Spectrometry-Mass Spectrometry) technology-based method for detecting chemical components of a Taoji Chengqi decoction pharmaceutical composition, and belongs to the technical field Comprising the following steps: firstly, mashing scalded peach kernels into coarse grains, adding water into the scalded peach kernel coarse grains, rheum officinale, cassia twigs and fried licorice roots, decocting, filtering, then adding mirabilite, heating until boiling, freeze-drying, crushing to obtain dry paste powder, adding methanol, performing ultrasonic treatment, and filtering to obtain a test solution; preparing a single decoction piece sample solution; a UPLC-Q-TOF-MS / MS technology is used for detection, the chromatographic condition is that 0.1% formic acid solution-acetonitrile is used as a mobile phase, gradient elution is carried out, the mass spectrum condition is carried out in a positive and negative ion mode, the dry gas temperature is 250-400 DEG C, and the dry gas flow velocity is 5-15 L / min. According to the method, the peach pit chengqi decoction is selected for mass spectrum analysis, chemical components in a sample can be systematically and rapidly detected, and a basis is provided for establishment of a quality control system.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of analysis and detection of chemical components in traditional Chinese medicine compositions, in particular, a detection method for chemical components of Taohuochengqi Decoction based on UPLC-Q-TOF-MS / MS technology. BACKGROUND

[0002] Taohuochengqi Decoction is derived from Treatise on Cold Damage Diseases, which is a blood-removing agent composed of five medicinal ingredients, specifically, Tao Ren, Dahuang, Guizhi, Mangxiao and Fried Gancao. In the prescription, Tao Ren can activate blood and resolve stasis, Dahuang can remove stasis and clear heat, Mangxiao can clear heat and soften hardness, Guizhi can dredge collaterals and blood vessels, and Gancao can protect the stomach and calm the center. The combination of these several drugs can remove stasis and clear heat, and is used for treating abdominal discomfort, self-urination, fever, irritability, blood stasis, amenorrhea, dysmenorrhea and other symptoms caused by stasis and heat.

[0003] The prior art CN113484429B discloses a method for establishing a Taohuochengqi Decoction substance reference, which includes: using thin layer chromatography to identify Tao Ren, Dahuang, Rougui and Gancao, using physicochemical identification method to identify Mangxiao; constructing a characteristic spectrum to identify the components in Taohuochengqi Decoction; using high performance liquid chromatography to determine the content of D-bitter apricot kernel glycoside, total anthraquinone, free anthraquinone, cinnamic acid, cinnamyl aldehyde and glycyrrhizic acid in Taohuochengqi Decoction, and calculating the combined anthraquinone content; wherein, the combined anthraquinone content = total anthraquinone content - free anthraquinone content.

[0004] The prior art CN110118846B discloses a method for establishing a Taohuochengqi Decoction substance reference, which first prepares a traditional Taohuochengqi Decoction based on traditional preparation process, determines the extract rate, establishes a common mode of fingerprint spectrum by high performance liquid chromatography, identifies the chromatographic peaks by reference substances, and on this basis, determines the content of bitter apricot kernel glycoside, rhein, emodin, chrysophanol, physcion, aloe emodin, cinnamyl aldehyde and glycyrrhizic acid, which can be used for quality investigation of Taohuochengqi Decoction.

[0005] However, the above-mentioned method of "medicinal ingredient selection" is inconsistent with the content of the classic famous prescription key information.

[0006] In addition, the existing Taohuochengqi Decoction pharmaceutical composition quality control method extracts chemical components of a certain part, and the types of components that can be analyzed are relatively few and one-sided, and cannot truly and comprehensively analyze the main components in Taohuochengqi Decoction preparation.

[0007] The information disclosed in this BACKGROUND section is only intended to increase an understanding of the general context in which the present application can be practiced. It is not admitted that any of the information provided herein represents prior art against the present application, nor does it constitute an admission as to the correctness or furness of the published documents. SUMMARY

[0008] To address the technical problem that existing technologies and methods cannot accurately reflect the components of Taohe Chengqi Decoction in classic prescriptions and the analysis is not comprehensive enough, this invention provides a method for detecting chemical components in the Taohe Chengqi Decoction drug composition based on UPLC-Q-TOF-MS / MS (ultra-high performance liquid chromatography coupled with time-of-flight mass spectrometry). This method can rapidly, accurately, and comprehensively perform qualitative analysis of the chemical components in Taohe Chengqi Decoction dry extract powder.

[0009] To achieve the above objectives, this invention provides a method for detecting the chemical components of a Taohe Chengqi Decoction pharmaceutical composition based on UPLC-Q-TOF-MS / MS technology, comprising the following steps:

[0010] (1) Solution preparation:

[0011] First, crush the roasted peach kernels into coarse particles. Then, decoct the coarse roasted peach kernels, rhubarb, cinnamon twigs, and stir-fried licorice root in water, filter the decoction, add Glauber's salt, heat to boiling, freeze-dry, and pulverize to obtain a dry extract powder. Add methanol to the dry extract powder, sonicate, filter, and take the filtrate to obtain the test solution.

[0012] Preparation of single-herb decoction sample solution: Crush the roasted peach kernel into coarse particles, then add water to the coarse roasted peach kernel, rhubarb, cinnamon twig, and stir-fried licorice root, decoct, cool, take the supernatant and filter, filter, evaporate to dryness, add methanol to dissolve the residue, filter, take the filtrate to obtain the single-herb decoction sample solution.

[0013] (2) The solution obtained in step (1) was detected using ultra-high performance liquid chromatography-tandem quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS / MS).

[0014] Chromatographic conditions: 0.1% formic acid solution was used as mobile phase A, and acetonitrile was used as mobile phase B; the elution method was gradient elution, and the elution conditions for the gradient elution were as follows:

[0015]

[0016]

[0017] Mass spectrometry conditions: Performed in positive and negative ion modes, with a drying gas temperature of 250-400℃, a drying gas flow rate of 5-15 L / min, a nebulizing gas pressure of 25-45 psi, a sheath gas temperature of 300-400℃, and a sheath gas flow rate of 7-15 L / min; the capillary voltage in positive ion mode is 3500-5000 V, and the voltage in negative ion mode is 2500-4500 V.

[0018] Ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS / MS) technology, by employing quadrupole time-of-flight mass spectrometry, can provide higher precision and accuracy in mass analysis, while also possessing high sensitivity and high resolution, thereby improving the detection and analysis capabilities of components in the complex system of Taohe Chengqi Decoction.

[0019] The test solution, individual herbal decoction sample solutions, and reference solution of the Taohe Chengqi Decoction were analyzed by ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS / MS) to determine the types of components and their corresponding Chinese herbal medicine sources in the Taohe Chengqi Decoction. The chromatographic and mass spectrometric results of the test solution were compared with those of the individual herbal decoction sample solutions and the reference solution, and also compared with literature data to determine the types of components and their corresponding Chinese herbal medicine sources in the Taohe Chengqi Decoction. Alternatively, the chromatographic and mass spectrometric results of the test solution were compared with the information recorded in Table 1 of this invention to determine the types of components and their corresponding Chinese herbal medicine sources in the Taohe Chengqi Decoction.

[0020] In some preferred embodiments, in the preparation of the test solution and the single-herb decoction sample solution, the particle size of the coarse particles is 1-10 mm; preferably, the particle size of the coarse particles is 1-8 mm; most preferably, the particle size of the coarse particles is 2-6 mm.

[0021] In some preferred embodiments, in the preparation of the test solution, the above-mentioned decoction method is as follows: first, bring to a boil over high heat, and then simmer over low heat; preferably, the simmering time is 70-110 minutes; most preferably, the simmering time is 90 minutes.

[0022] In some preferred embodiments, the decoction time for adding water in the preparation of the single herb sample solution is 20-50 minutes; most preferably, the decoction time is 30 minutes.

[0023] In some preferred embodiments, the above chromatographic conditions use a Waters ACQUITY UPLC HSS T3 column (2.1×100mm, 1.8μm); detection wavelength 250-260nm, column temperature 32-38℃, flow rate 0.3-0.6ml / min; preferably, the detection wavelength is 254nm, column temperature 35℃, flow rate 0.4ml / min; and injection volume is 1μl.

[0024] In some preferred embodiments, the above-mentioned mass spectrometry conditions are achieved by using a combination of primary mass spectrometry analysis and secondary mass spectrometry scanning to obtain ion fragment information data of the chemical components in the test sample solution.

[0025] In some preferred embodiments, the first-stage mass spectrometer uses MS mode with a mass scan range of 100-1200 m / z; the second-stage mass spectrometer uses Auto-MS / MS mode; and the collision voltage is 5 eV-35 eV, preferably 10 eV, 15 eV, 20 eV, or 30 eV.

[0026] In some preferred embodiments, the aforementioned ion fragment information data is acquired by Agilent Mass Hunter Qualitative Analysis software, and data processing is performed using Agilent software Qualitative Navigator (B.08.00) and Qualitative Workflows (B.08.00).

[0027] In some preferred embodiments, the mass spectrometry conditions are as follows: drying gas temperature: 350°C, drying gas flow rate: 10 L / min, nebulizing gas pressure: 35 psi, sheath gas temperature: 350°C, sheath gas flow rate: 12 L / min; capillary voltage in positive ion mode: 4500 V, and voltage in negative ion mode: 3500 V.

[0028] In some preferred embodiments, a total of 35 chemical components were characterized in the above-mentioned Taohe Chengqi Decoction pharmaceutical composition, including: 10 flavonoids, 16 anthraquinones, 1 alkaloid, 5 saponins, 2 phenylpropanoids, and 1 organic acid.

[0029] The inventors identified the above 35 chemical components as drugs derived from four medicinal herbs: peach kernel, rhubarb, cinnamon twig, and licorice.

[0030] The 35 compounds mentioned above are specifically gallic acid, amygdalin, catechin, veselin-2, epicatechin gallate, glycyrrhizin, apigenin glycyrrhizin, aloe-emodin glucoside, 2-cinnamoyl glucose, rhein-8-O-β-D-glucoside, monazine, glycyrrhizin or its isomers, cinnamic acid, vitexin, 6-methyl-rhein-8-O-β-D-glucopyranoside, and torachrysone-8-O-β-D-glucopyranoside. oside, rhein-1-O-β-D-glucoside, rhein-8-β-D-glucoside, rhein-1-O-glucoside, rhein-8-β-D-glucoside, rhein-1-O-(6-carboxyacetyl)-β-D-glucopyranoside, hydroxyemodin, 6-methylemodin, aloe-emodin, ural glycyrrhizin F, rhein, glycyrrhizin A3, glycyrrhizin alcohol or its isomers, glycyrrhizin, glycyrrhizin G2, glycyrrhizic acid, glycyrrhizic acid isomers, rhein, glycyrrhizin, rhein.

[0031] Among them, the flavonoids are catechin (peak 3), vezidin 2 (peak 4), epicatechin gallate (peak 5), glycyrrhizin (peak 6), apigenin glycyrrhizin (peak 7), glycyrrhizin or its isomer (peak 12), vitexin (peak 14), glycyrrhizin flavonol or its isomer (peak 28), glycyrrhizin (peak 29), and glycyrrhizin (peak 34).

[0032] Anthraquinone compounds include aloe-emodin glucoside (peak 8), rhein-8-O-β-D-glucopyranoside (peak 10), rhein-1-O-β-D-glucopyranoside (peak 11), 6-methyl-rhein-8-O-β-D-glucopyranoside (peak 15), Torachrysone-8-O-β-D-glucopyranoside (peak 16), and rhein-1-O-β-D-glucopyranoside (peak 17). Peaks: emodin-8-β-D-glucoside (peak 18), emodin-1-O-glucoside (peak 19), emodin-8-β-D-glucoside (peak 20), emodin-1-O-(6-carboxyacetyl)-β-D-glucopyranoside (peak 21), hydroxyemodin (peak 22), 6-methylemodin (peak 23), aloe-emodin (peak 24), emodin (peak 26), emodin (peak 33), emodin (peak 35);

[0033] The alkaloid compound is amygdalin (peak 2);

[0034] The saponin compounds are glycyrrhizin F (peak 25), glycyrrhizin A3 (peak 27), glycyrrhizin G2 (peak 30), glycyrrhizic acid (peak 31), and glycyrrhizic acid isomer (peak 32).

[0035] The phenylpropanoid compounds are 2-cinnamoyl glucose (peak 9) and cinnamic acid (peak 13);

[0036] The organic acid compound is gallic acid (peak 1).

[0037] Compared with the prior art, the present invention has the following beneficial effects:

[0038] (1) This invention uses the same medicinal slices as Taohe Chengqi Decoction in the ancient classic prescription "Treatise on Febrile Diseases" for sample preparation. The dry powder of Taohe Chengqi Decoction is selected for mass spectrometry analysis. UPLC-Q-TOP-MS / MS technology is used to analyze the overall chemical composition of the Taohe Chengqi Decoction drug composition, rather than the chemical composition of a certain extraction part. The established qualitative analysis method can systematically and rapidly detect the chemical composition in the sample. An analytical method that conforms to the original chemical composition of Taohe Chengqi Decoction is established, which solves the problems of inaccurate dosage and processing method of medicinal slices and only examines the chemical composition of a certain part in the existing technology.

[0039] (2) This invention, by adjusting chromatographic and mass spectrometric conditions, employs ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS / MS) to detect and analyze the Taohe Chengqi Decoction preparation. Simultaneously, by comparing and analyzing the LC-MS results of the Taohe Chengqi Decoction drug composition test solution and the solutions of individual medicinal herbs, the medicinal source attribution of 35 components and the qualitative analysis of the main chromatographic peaks were clarified. These compounds, in positive mode, can generate [M+H]. + [M+Na] + [M+K] + and [M+NH4] + Isadductions, in negative mode, can generate [MH]. - [M+COOH] - and [M+Cl] - Ions; knowing the medicinal source of each component is helpful in judging the pharmacological activity and safety of Taohe Chengqi Decoction preparations, and can make an overall quality evaluation of Taohe Chengqi Decoction preparations.

[0040] (3) The detection method of the present invention can quickly and effectively determine 35 components in the Taohe Chengqi Decoction composition at the same time. It can quickly, efficiently and accurately detect the main chemical components in the composition, identify the components of the complex system of traditional Chinese medicine composition, and can comprehensively reflect the medicinal flavor in the drug formula, realize the comprehensive control of its material group, provide a reliable reference for the quality control in the production process of Taohe Chengqi Decoction preparation, and provide a basis for the establishment of subsequent quality control system. Attached Figure Description

[0041] Figure 1 These are the UPLC-UV chromatograms and UPLC-TOF-MS total ion chromatograms (positive and negative modes) of the Taohe Chengqi Decoction composition. Detailed Implementation

[0042] The specific embodiments of the present invention will now be described in detail with reference to the accompanying drawings, but it should be understood that the scope of protection of the present invention is not limited to the specific embodiments.

[0043] The following specific examples illustrate the implementation of the present invention. Those skilled in the art can easily understand other advantages and effects of the present invention from the content disclosed in this specification. The present invention can also be implemented or applied through other different specific embodiments, and various details in this specification can also be modified or changed based on different viewpoints and applications without departing from the spirit of the present invention.

[0044] The reagents used in the following examples and their sources are as follows:

[0045] Example 1: Preparation of the test solution of the Taohe Chengqi Decoction drug composition

[0046] The preparation steps include: first, crushing the roasted peach kernels into coarse particles of 2-6 mm; then, weighing 13.50 g of roasted peach kernel coarse particles, 55.20 g of rhubarb, 27.60 g of cinnamon twig, and 27.60 g of stir-fried licorice root into a clay pot, adding 1400 ml of water, bringing to a boil over high heat, then simmering over low heat until about 500 ml remains, filtering, placing the filtrate into a clay pot, adding 27.60 g of Glauber's salt, heating to a boil, freeze-drying, and pulverizing to obtain a dry extract powder; taking about 1.0 g of the dry extract powder, accurately weighing it, placing it in a stoppered conical flask, accurately adding 25 mL of methanol, sealing tightly, weighing, ultrasonicating (power 500 W, frequency 40 kHz) for 30 minutes, removing, cooling, weighing again, replenishing the lost weight with methanol, shaking well, filtering, and taking the filtrate to obtain the test solution of the Taohua Chengqi Decoction drug composition.

[0047] Example 2: Preparation of single-herb medicinal slice sample solution

[0048] The preparation method is as follows: First, crush the roasted peach kernel slices into coarse particles of 2-6 mm; then accurately weigh 1.0 g each of roasted peach kernel slices, rhubarb slices, and cinnamon twig slices, as well as 0.5 g of stir-fried licorice slices, place them in round-bottom flasks, add 50 mL of water, decoct for 30 minutes, cool, filter, evaporate to dryness, dissolve the residue in methanol, transfer to a 20 mL volumetric flask, dilute with methanol to the mark, shake well, filter, and take the filtrate to obtain the sample solution of each single herb slice.

[0049] Example 3: Detection and Identification of Chemical Components in the Peach Kernel Decoction Composition

[0050] (1) Chromatographic conditions

[0051] A Waters ACQUITY UPLC HSS T3 column (2.1 × 100 mm, 1.8 μm) equipped with an online filter was used. The mobile phase was 0.1% formic acid solution (A) and acetonitrile (B). The detection wavelength was 254 nm, the column temperature was 35 °C, the flow rate was 0.4 mL / min, and the injection volume was 1 μL. Gradient elution was used, and the elution conditions were as follows:

[0052]

[0053] (2) Mass spectrometry conditions

[0054] The tests were performed in both positive and negative ion modes, with a drying gas temperature of 350℃, a drying gas flow rate of 10 L / min, a nebulizing gas pressure of 35 psi, a sheath gas temperature of 350℃, a sheath gas flow rate of 12 L / min, and capillary voltages of 4500 V (positive mode) and 3500 V (negative mode). The primary mass spectrometry (MS) mode was used, with a mass scan range of 100–1200 m / z. The secondary mass spectrometry (Auto-MS / MS) mode was used, with collision voltages of 10 eV, 15 eV, 20 eV, and 30 eV.

[0055] (3) Identification

[0056] The solutions prepared in the examples were analyzed under the chromatographic and mass spectrometric detection conditions of this example to obtain the total ion current chromatogram (TIC) of the test solution of the Taohe Chengqi Decoction drug composition in positive and negative ion modes, as shown in the figure. Figure 1 .

[0057] The data were acquired using Agilent Mass Hunter software and processed using Agilent QualitativeNavigator (B.08.00) and Qualitative Workflows (B.08.00). A comprehensive chemical composition database was established with reference to relevant literature. The molecular formulas of the compounds were deduced by comparing the precise relative molecular masses of the main chromatographic peaks, and the peaks were identified and assigned based on mass spectrometry fragment information. A total of 35 chemical components of the Taohe Chengqi Decoction were identified, as shown in Table 1.

[0058] Table 1. Results of Primary and Secondary Mass Spectrometry Analysis of Taohe Chengqi Decoction as a Material Reference.

[0059]

[0060]

[0061]

[0062]

[0063] The traditional Chinese medicine composition of Taohe Chengqi Decoction was analyzed by primary mass spectrometry and secondary mass spectrometry to obtain ion fragment information of the compounds. Then, based on the fragmentation patterns of reference standards and literature data, the obtained compounds were identified and inferred. A total of 35 compounds were identified and inferred, and their identification and classification into corresponding single herbs were confirmed by comparison with reference standards.

[0064] The compound derived from roasted peach kernels is amygdalin;

[0065] The compounds derived from roasted licorice are vezin-2, glycyrrhizin, apigenin, styracin, glycyrrhizin F, glycyrrhizin A3, glycyrrhizin flavonol or its isomer, glycyrrhizin, glycyrrhizin G2, glycyrrhizic acid, glycyrrhizic acid isomer, glycyrrhizin, glycyrrhizin or its isomer.

[0066] Compounds derived from rhubarb include gallic acid, catechin, epicatechin gallate, aloe-emodin glucoside, rhein-8-O-β-D-glucoside, vitexin, 6-methyl-rhein-8-O-β-D-glucopyranoside, Torachrysone-8-O-β-D-glucopyranoside, rhein-1-O-β-D-glucoside, rhein-8-β-D-glucoside, rhein-1-O-glucoside, rhein-8-β-D-glucoside, rhein-1-O-(6-carboxyacetyl)-β-D-glucopyranoside, hydroxyemodin, 6-methylemodin, aloe-emodin, rhein, rhein, and rhein.

[0067] Compounds derived from cinnamon twigs include 2-cinnamonyl glucose and cinnamic acid.

[0068] Examples 4 and 5: Detection methods for chemical components in Taohe Chengqi Decoction pharmaceutical compositions based on UPLC-Q-TOF-MS / MS technology

[0069] Examples 4 and 5 are methods for detecting chemical components in the Taohe Chengqi Decoction pharmaceutical composition based on UPLC-Q-TOF-MS / MS technology. The only differences are in a few parameters in the solvent preparation step, chromatographic conditions, and mass spectrometry conditions; otherwise, they are the same as in Examples 1-3. See Table 2 for specific methods.

[0070] Table 2 shows some parameters of the solvent preparation steps, chromatographic conditions, and mass spectrometry conditions in Examples 4 and 5.

[0071]

[0072]

[0073] The analytical results obtained in Examples 4 and 5 are basically the same as those in Example 4.

[0074] The foregoing description of specific exemplary embodiments of the invention is for illustrative and explanatory purposes. These descriptions are not intended to limit the invention to the precise forms disclosed, and it will be apparent that many changes and variations can be made in accordance with the foregoing teachings. The exemplary embodiments were chosen and described in order to explain the specific principles of the invention and its practical application, thereby enabling those skilled in the art to implement and utilize various different exemplary embodiments of the invention, as well as various different choices and variations. The scope of the invention is intended to be defined by the claims and their equivalents.

Claims

1. A method for detecting chemical components of a Taohechengqi decoction pharmaceutical composition based on UPLC-Q-TOF-MS / MS technology, characterized in that, The detection method comprises the following steps: (1) Preparation of solution: First, the Semen Pruni Persicae is crushed into coarse particles, and then the Semen Pruni Persicae coarse particles, Rhubarb, Cassia twig, and roasted licorice are decocted with water, filtered, and then sodium sulfate is added, heated to boiling, freeze-dried, and pulverized to obtain dry powder; methanol is added to the dry powder, ultrasonically treated, filtered, and the filtrate is taken to obtain the test solution; Preparation of single herb sample solution: the Semen Pruni Persicae is crushed into coarse particles, and then the Semen Pruni Persicae coarse particles, Rhubarb, Cassia twig, and roasted licorice are decocted with water, cooled, and the supernatant is filtered, dried, and the residue is dissolved in methanol, filtered, and the filtrate is taken to obtain the single herb sample solution; (2) The solution obtained in step (1) is detected by using ultra-high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS / MS) technology: The chromatographic conditions are as follows: 0.1% formic acid solution is used as mobile phase A, and acetonitrile is used as mobile phase B; the elution mode is gradient elution, and the elution conditions of the gradient elution are as follows: The mass spectrometry conditions are as follows: in positive and negative ion modes, the drying gas temperature is 250-400℃, the drying gas flow rate is 5-15L / min, the atomization gas pressure is 25-45psi, the sheath gas temperature is 300-400℃, and the sheath gas flow rate is 7-15L / min; the capillary voltage in the positive ion mode is 3500-5000V, and the voltage in the negative ion mode is 2500-4500V.

2. The detection method according to claim 1, characterized in that, In the preparation of the test solution and the single herb sample solution, the particle size of the coarse particles is 1-10mm; preferably, the particle size of the coarse particles is 1-8mm; most preferably, the particle size of the coarse particles is 2-6mm.

3. The method of claim 1, wherein, In the preparation of the test solution, the decoction mode is as follows: first, use a strong fire to boil, and then use a weak fire to decoct; preferably, the time of the weak fire decoction is 70-110min; most preferably, the time of the weak fire decoction is 90min. In the preparation of the single herb sample solution, the decoction time is 20-50min; most preferably, the decoction time is 30min.

4. The method of claim 1, wherein In the chromatographic conditions, a Waters ACQUITY UPLC HSS T3 column (2.1x100mm, 1.8μm) is used; the detection wavelength is 250-260nm, the column temperature is 32-38℃, the flow rate is 0.3-0.6ml / min, preferably, the detection wavelength is 254nm, the column temperature is 35℃, and the flow rate is 0.4ml / min; the injection volume is 1μl.

5. The method of claim 1, wherein In the mass spectrometry conditions, the ion fragment information data of the chemical components in the test solution are obtained by using a combination of primary mass spectrometry analysis and secondary mass spectrometry scanning.

6. The detection method according to claim 5, characterized in that, The primary mass spectrometry selects the MS mode, and the mass scan range is 100-1200m / z; the secondary mass spectrometry selects the Auto-MS / MS mode; the collision voltage is 5eV-35eV, preferably 10eV, 15eV, 20eV, or 30eV.

7. The detection method according to claim 5, characterized in that, The ion fragment information data is collected by Agilent Mass Hunter Qualitative Analysis software, and data processing adopts Agilent software Qualitative Navigator (B.08.00) and Qualitative Workflows (B.08.00).

8. The method of claim 1, wherein, In the mass spectrometry condition, the drying gas temperature is 350℃, the drying gas flow rate is 10L / min, the atomization gas pressure is 35psi, the sheath gas temperature is 350℃, and the sheath gas flow rate is 12L / min; the capillary voltage in the positive ion mode is 4500V, and the voltage in the negative ion mode is 3500V.

9. The assay of any one of claims 1-8, wherein, 35 chemical components are characterized in the Taohewanchigutang pharmaceutical composition, including 10 flavonoids, 16 anthraquinones, 1 alkaloid, 5 saponins, 2 phenylpropanoids and 1 organic acid.

10. The detection method according to claim 9, characterized in that, The flavonoids are catechin (peak 3), vitexin 2 (peak 4), epicatechin gallate (peak 5), glycyrrhizin (peak 6), apiosyl glycyrrhizin (peak 7), glycyrrhizin or its isomer (peak 12), vitexin (peak 14), glycyrrhizin flavonol or its isomer (peak 28), glycyrrhizin renotone (peak 29) and glycyrrhizin (peak 34); The anthraquinones are aloe-emodin 8-O-glucopyranoside (peak 8), rhein-8-O-β-D-glucopyranoside (peak 10), ononin (peak 11), 6-methyl-rhein-8-O-β-D-glucopyranoside (peak 15), Torachrysone-8-O-β-D-glucopyranoside (peak 16), chrysophanol-1-O-β-D-glucopyranoside (peak 17), chrysophanol-8-β-D-glucopyranoside (peak 18), emodin-1-O-glucopyranoside (peak 19), emodin-8-β-D-glucopyranoside (peak 20), emodin-1-O-(6-carboxyacetyl)-β-D-glucopyranoside (peak 21), hydroxyemodin (peak 22), 6-methylrhein (peak 23), aloe-emodin (peak 24), rhein (peak 26), emodin (peak 33) and chrysophanol (peak 35); The alkaloid is amygdalin (peak 2); The saponins are uralsaponin F (peak 25), glycyrrhizin A3 (peak 27), glycyrrhizin G2 (peak 30), glycyrrhizic acid (peak 31) and glycyrrhizic acid isomer (peak 32); The phenylpropanoids are 2-cinnamoylglucose (peak 9) and cinnamic acid (peak 13); The organic acid is gallic acid (peak 1).

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