A method for analyzing alkaloid components in traditional Chinese medicine

Abstract

The present invention belongs to the field of analytical chemistry, and relates to an analysis method for alkaloid components in traditional Chinese medicine samples, and specifically relates to an analysis method for alkaloid active ingredients in traditional Chinese medicine based on a zwitterionic reverse phase chromatographic stationary phase (SAC) modified with phenylsuccinic anhydride and diamine. When the SAC stationary phase is used for the separation of alkaloids, it can provide a separation selectivity different from that of existing chromatographic separation materials, and provide a new means for the multidimensional separation of complex alkaloid components.

Description

Technical Field

[0001] The invention belongs to the field of analytical chemistry, and in particular relates to a method for analyzing alkaloid components in traditional Chinese medicine by high performance liquid chromatography. Technical Background

[0002] Alkaloids are the main active ingredients of many traditional Chinese medicines. With the continuous understanding of their biological functions, their research has received more and more attention. In order to deeply study the relationship between their structure and function, chromatographic separation and analysis is very necessary. However, due to their complex composition and unique structure, there are problems such as poor separation, easy tailing of peaks, and poor method stability during the separation process, which brings certain challenges to separation and analysis.

[0003] The main chromatographic analysis methods for alkaloids are reversed phase chromatography, ion exchange chromatography and hydrophilic interaction chromatography. With the advancement of analytical technology, it is found that there are great limitations in chromatographic peak capacity and separation selectivity in single-dimensional chromatographic analysis, which can no longer meet the requirements of separation and analysis of complex systems of traditional Chinese medicine. Therefore, multidimensional liquid chromatography technology has emerged. In recent years, a series of two-dimensional systems have been constructed for the separation and analysis of alkaloid components by using the most commonly used reversed phase chromatography for alkaloid analysis and other modes of chromatography with different action mechanisms (mixed mode chromatography and ion exchange chromatography, etc.). For example, Long et al. used the reversed phase mixed mode / strong cation exchange chromatography (XCharge C18 / SCX) two-dimensional system for the analysis of scopolamine alkaloids, and the target obtained a good peak shape and separation; Zhang et al. constructed a reversed phase / reversed phase (XCharge C18 / XAqua C3) two-dimensional separation system with good orthogonality for the separation and analysis of alkaloid components in lycopodiella. However, these two-dimensional separation systems all use non-volatile buffer salts, and post-processing is more troublesome.

[0004] At present, the types of separation materials for alkaloids are limited. Providing separation selectivity different from existing materials and constructing a highly orthogonal two-dimensional separation system compatible with mass spectrometry is expected to provide a new means for the separation of complex alkaloid components. The patent of this invention adopts a new zwitterionic reversed-phase chromatographic stationary phase to establish a separation method for alkaloids in traditional Chinese medicine. The target compound has different separation selectivity with the reversed-phase column, and the peak shape is good. The mobile phase system is compatible with mass spectrometry, providing a new technical means for the study of the material basis of complex systems of traditional Chinese medicine. Summary of the invention

[0005] The purpose of the present invention is to provide a chromatographic analysis method for alkaloids in traditional Chinese medicine, which can obtain a separation selectivity different from that of a reverse phase column.

[0006] The above-mentioned purpose of the present invention is achieved by the following technical solutions:

[0007] The extract or components of traditional Chinese medicine containing alkaloid components are dissolved in a solvent and then separated by liquid chromatography, with SAC as the stationary phase of the chromatographic column, water and an organic solvent, or a buffered saline solution and an organic solvent as the mobile phase, and eluted using an isocratic or gradient method, and then entering the detector for detection.

[0008] The solvent used to dissolve the Chinese medicine extract or component is one or more of methanol, acetonitrile, ethanol, water, ammonium formate, and ammonium acetate; the concentration of the ammonium formate and ammonium acetate is 2-50mM; the ratio of one or more of methanol, acetonitrile, ethanol to the water phase or salt phase is 5-95%.

[0009] The SAC chromatographic column is a zwitterionic group with phenylsuccinic anhydride modified diamine at the end of the bonded phase, and its surface provides pH-based selectivity control. The schematic structure of the filler is as follows:

[0010]

[0011] SAC is a zwitterionic reverse phase chromatographic stationary phase material.

[0012] Preparation process of SAC column stationary phase:

[0013] (1) Modification of polyamine substituents on the surface of silica gel: Modification of polyamino groups by a one-step silanization reaction of polyaminosilane, or first bonding halogen groups or epoxy groups on the surface of silica gel by a silanization reaction of halogen silane / epoxy silane, and then modifying polyamino groups by a two-step method of reacting them with polyamine compounds. The bonding method of the silanization reaction can be a single-point or multi-point bonding method, or a horizontal copolymerization method.

[0014] Or a. One-step reaction modification of polyamino: Add silica gel to a hydrochloric acid or nitric acid solution with a concentration of 1% to 38%, heat and reflux with stirring for 1 to 48 hours, filter, wash with water until neutral, and dry at 100 to 160°C to constant weight. Place in a reactor containing an organic solvent under a nitrogen atmosphere, stir evenly, add polyaminosilane reagent, stir and reflux at 80 to 120°C for 1 to 48 hours, filter under reduced pressure and wash with toluene, dichloromethane, water, tetrahydrofuran, and methanol, and dry at 60 to 150°C to constant weight to obtain polyamino silica gel.

[0015] Or b. Two-step reaction modification of polyamino groups: Add silica gel to a hydrochloric acid or nitric acid solution with a concentration of 1% to 38%, heat and reflux with stirring for 1 to 48 hours, filter, wash with water until neutral, and dry at 100 to 160°C to constant weight. Place in a reactor containing an organic solvent under a nitrogen atmosphere, stir evenly, add halogen silane or epoxy silane reagent, stir and reflux at 80 to 120°C for 1 to 48 hours, filter under reduced pressure and wash with anhydrous toluene, dichloromethane, water, tetrahydrofuran, and methanol, and dry at 60 to 150°C to constant weight to obtain a halogen group or epoxy group modified carrier. Continue to add the above one-step modified silica gel to an organic solvent, add polyamine compounds, stir and reflux at 50 to 120°C for 1 to 48 hours, filter under reduced pressure and wash with toluene, dichloromethane, water, tetrahydrofuran, and methanol, and dry at 60 to 150°C to constant weight to obtain polyamino modified silica gel.

[0016] The halogen silane is one or more of chloropropyltrichlorosilane, chlorohexyltrimethylsilane and bromopropyltrichlorosilane;

[0017] The epoxysilane reagent is one or more of 3-(2,3-epoxypropyloxy)propyltrimethoxysilane, 3-(2,3-epoxypropyloxy)heptyltrimethoxysilane, trimethoxy[2-(7-oxabicyclo[4.1.0]hept-3-yl)ethyl]silane, and 2-(3,4-epoxycyclohexyl)methyldiethoxysilane;

[0018] The polyaminosilane reagent is one or more of N-(β-aminoethyl-γ-aminopropyl)trimethoxysilane, N-(2-aminopropyl)-aminopropyltrimethoxysilane, N-(2-N-benzylaminoethyl)-3-aminopropyltrimethoxysilane, and N-(2-N-benzylaminovinyl)-3-aminopropyltriethoxysilane;

[0019] The organic solvent is one or more of toluene, tetrahydrofuran, ethylbenzene, N,N-dimethylformamide, and dimethyl sulfoxide;

[0020] (2) Amidation reaction: Add polyamino silica gel to an organic solvent, then add a certain amount of phenylsuccinic anhydride, a base catalyst and an acid binding agent, and stir at 20-120°C for 1-72 hours. Filter under reduced pressure and wash with anhydrous toluene, dichloromethane, water, tetrahydrofuran and methanol, and dry at 60-150°C until constant weight to obtain a zwitterionic reverse phase chromatography stationary phase (SAC).

[0021] The organic solvent is one or more of toluene, tetrahydrofuran, ethylbenzene, N,N-dimethylformamide, and dimethyl sulfoxide;

[0022] The base catalyst is one or more of 4-dimethylaminopyridine, 1,8-diazacyclo[5,4,0]undecene-7, 1,5-diazabicyclo[4.3.0]non-5-ene, and diisopropylethylamine;

[0023] The acid-binding agent is one or more of pyridine, triethylamine and imidazole.

[0024] The mobile phase additive comprises one or more of ammonium formate, ammonium acetate, formic acid, acetic acid and ammonia water, the concentration of the additive is 2-50mM, and the pH value of the mobile phase is 2-8; the organic phase in the mobile phase is one or more of acetonitrile, methanol and acetone, and the proportion of the organic phase in the mobile phase is 5%-95%.

[0025] The detector is UV, CAD, ELSD or MS.

[0026] The Chinese medicine extracts or components include Corydalis yanhusuo, Sophora flavescens, Ephedra sinica, Radix Sophorae flavescentis and the like.

[0027] When the SAC stationary phase is used for the separation of alkaloids, it can provide a separation selectivity different from that of existing chromatographic separation materials, providing a new means for the multidimensional separation of complex alkaloid components.

[0028] The present invention has the following advantages:

[0029] (1) Good separation selectivity. The chromatographic method established in the present invention has a separation selectivity different from that of reverse phase chromatographic materials when separating alkaloid components in traditional Chinese medicine, laying a foundation for the separation, analysis and purification preparation of complex compounds in traditional Chinese medicine.

[0030] (2) The analytical method is simple, stable and easy to operate. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1 This is a diagram showing the separation of the alkaloid samples described in Example 2 of the present invention on different reverse phase chromatography columns.

[0032] Figure 2 The SAC column described in Example 3 of the present invention is used to separate alkaloid samples under different pH conditions.

[0033] Figure 3 The separation spectra of the Corydalis yanhusuo extract described in Example 4 of the present invention on different chromatographic columns are shown.

[0034] Figure 4 The separation spectra of the Corydalis yanhusuo component samples described in Example 5 of the present invention on different chromatographic columns. DETAILED DESCRIPTION

[0035] The technical solution of the present invention is further described below through specific implementation methods.

[0036] Example 1

[0037] Preparation process of SAC column stationary phase:

[0038] (1) 10 g silica gel (3.5 μm particle size, The pore size was added into 150 mL of 10% hydrochloric acid solution, heated under reflux and stirred for 12 hours, filtered, washed with water until neutral, and dried at 160°C to constant weight. In a nitrogen atmosphere, it was placed in a reactor containing 80 mL of toluene, stirred evenly, and 1 mL of N-(β-aminoethyl-γ-aminopropyl)methyldimethoxysilane was added, stirred and refluxed at 120°C for 24 hours, filtered under reduced pressure, and washed with toluene, dichloromethane, water, tetrahydrofuran, and methanol in turn, and dried at 150°C to constant weight to obtain polyamino silica gel.

[0039] (2) Amidation reaction: 10 g of polyamino silica gel was added to 50 mL of N,N-dimethylformamide, followed by 1 g of phenylsuccinic anhydride, 0.1 g of 4-dimethylaminopyridine and 0.1 g of pyridine, and the mixture was stirred at 120°C for 24 hours. The mixture was filtered under reduced pressure and washed with anhydrous toluene, dichloromethane, water, tetrahydrofuran and methanol in sequence, and dried at 150°C until constant weight was obtained to obtain a zwitterionic reverse phase chromatography stationary phase (SAC), with a bonding density of 3.07 μmol / m 2 .

[0040] The product was used as the stationary phase of the SAC chromatography column in the following examples.

[0041] Example 2

[0042] Evaluation of the orthogonality of different reversed-phase columns for alkaloid reference substances

[0043] Appropriate amounts of alkaloid reference substances colchicine, camptothecin, capsaicin, tropaeotaxel, tetrahydropalmatine, jatrorrhizine and quinine were weighed respectively, prepared into 1 mg / mL solutions, and equal volumes were mixed to prepare alkaloid mixed standard samples, which were separated by different reverse phase columns for selectivity evaluation between different chromatographic columns.

[0044] Chromatographic analysis conditions are as follows: chromatographic columns SAC (diameter * length = 4.6 * 150mm, silica gel particle size 3.5μm), PFP (diameter * length = 4.6 * 100mm, silica gel particle size 3.5μm, Agilent) (control group 1) and Symmetry C18 (diameter * length 2.1 * 150mm, silica gel particle size 3.5μm, Waters) (control group 2); mobile phase A-acetonitrile; B-100mM ammonium formate; C-water; flow rate is 1.0 (SAC and control group 1) or 0.2mL / min (control group 2); gradient elution is used, and the elution conditions (volume ratio) are 0-30min, 20-65% A, B is 10% volume ratio unchanged, and the rest C;

[0045] Chromatographic column XCharge C18 (diameter*length=3.0*100mm, silica gel particle size 3.5μm, Waters) (control group 3), mobile phase A-0.1% formic acid acetonitrile (volume concentration); B-0.1% formic acid water (volume concentration; flow rate is 0.4mL / min; gradient elution is adopted, and the elution conditions (volume ratio, the same below) are 0-20min, 5-40%A; 20-30min, 40-75%A;

[0046] The UV detector was used for detection at 254 nm.

[0047] Figure 1 Comparison of alkaline samples analyzed by different chromatographic columns (1-colchicine, 2-camptothecin, 3-capsaicin, 4-tetrahydropalmatine, 5-homotaxitrine, 6-jatrorrhizine, 7-quinine)

[0048] The zwitterionic stationary phase (SAC) in the present invention has a large difference in selectivity from other reversed-phase columns and a good peak shape, and can be used as a new separation material for analyzing alkaloid samples in traditional Chinese medicine.

[0049] Example 3

[0050] Analysis of alkaloid samples by SAC column under different pH conditions

[0051] Appropriate amounts of alkaloid reference substances colchicine, camptothecin, capsaicin, scutellariae, leonurine, jatrorrhizine, berberine and quinine were weighed and prepared into 2 mg / mL solutions respectively, and equal volumes were mixed to prepare alkaloid mixed standard samples, which were separated by SAC column under different pH buffer salt systems.

[0052] The chromatographic analysis conditions are as follows: chromatographic column SAC (diameter*length=4.6*150mm, silica gel particle size 3.5μm); mobile phase A-acetonitrile; B-100mM ammonium formate, pH 6.2 or 7.5; C-water; flow rate is 1.0mL / min; gradient elution is adopted, and the elution conditions are 0-40min, 10-40% A, B is 10% constant in volume ratio, and the rest is C; detection is carried out at 254nm by UV detector.

[0053] Figure 2 The chromatograms of alkaloid samples separated by SAC column under different pH conditions. Comparison of alkaline samples analyzed under different pH conditions (1-colchicine, 2-camptothecin, 3-capsaicin, 4-homotaxitol, 5-leonurine, 6-jatrorrhizine, 7-berberine, 8-quinine)

[0054] SAC columns are zwitterionic fillers containing amino and carboxyl groups, and their surfaces can provide pH-based selectivity control, thereby achieving changes in separation and selectivity between samples. Figure 2 As shown, the retention of some compounds was significantly enhanced by adjusting the pH value.

[0055] Example 4

[0056] Analysis of the total extract of Corydalis yanhusuo alkaloids

[0057] Weigh an appropriate amount of methanol water extract sample of Corydalis yanhusuo medicinal material, prepare a methanol solution with a concentration of 10 mg / mL, and separate it using SAC column (diameter * length = 4.6 * 150mm, silica gel particle size 3.5μm) and Waters Symmetry C18 (diameter * length = 2.1 * 150mm, silica gel particle size 3.5μm) columns. Chromatographic conditions: A-10% acetonitrile water; B-80% acetonitrile water, A and B phases both contain 10mM ammonium acetate, gradient elution, elution conditions: 0-40min, 0-35% B, 40-60min, 35-40% B, 60-65min, 40-90B. Column temperature 35℃, injection volume 10μL, MS detection.

[0058] Figure 3 The separation spectra of Corydalis yanhusuo extract on different chromatographic columns.

[0059] There is a great difference in selectivity between SAC column and C18 column, which shows its better analytical advantage in multidimensional analysis.

[0060] Example 5

[0061] Analysis of alkaloid samples in the components of Corydalis yanhusuo

[0062] Weigh an appropriate amount of the component samples of the water extract of Corydalis yanhusuo after segmentation separation by cation exchange column, prepare a methanol solution with a concentration of 8 mg / mL, and separate them using SAC column (diameter * length = 4.6 * 150mm, silica gel particle size 3.5μm) and Waters Symmetry C18 column (diameter * length = 2.1 * 150mm, silica gel particle size 3.5μm). The separation chromatographic conditions are as follows: mobile phase A-acetonitrile, B-water, C-100mM ammonium acetate, acetonitrile as a strong elution solvent, gradient elution, elution conditions: 0-30min, 85%-50% A, C keeps the volume ratio of 10% unchanged, and the rest is B. The column temperature is 35℃, and the injection volume is 10μL. The UV detector is detected at 254nm.

[0063] Figure 4 The separation spectra of Corydalis yanhusuo component samples on different chromatographic columns also reflect the difference in selectivity between SAC column and C18 column.

[0064] Example 6

[0065] Analysis of alkaloids in Sophora flavescens

[0066] Weigh an appropriate amount of the water extract of Sophora flavescens and prepare a 90% methanol solution with a concentration of 20 mg / mL. Separation was performed using SAC and BEH C18 columns (Waters) (diameter * length = 4.6 * 150 mm, silica gel particle size 3.5 μm). The separation chromatographic conditions were as follows: mobile phase A-acetonitrile, B-water, C-50 mM ammonium formate, pH 8.0, acetonitrile as a strong elution solvent, gradient elution, elution conditions: 0-30 min, 85%-20% A, C kept the volume ratio of 20% unchanged, and the rest B. The column temperature was 30 ° C, the injection volume was 50 μL, and CAD detection showed that SAC and BEH C18 columns showed different peak orders, and the two stationary phases were complementary.

[0067] Example 7

[0068] Analysis of alkaloids in the roots of Sophora flavescens

[0069] Weigh an appropriate amount of ethanol extract of the root of northern bean, prepare a methanol solution with a concentration of 20 mg / mL, and separate it using a SAC column and an XCharge C18 (Acchrom) column (diameter*length=4.6*150 mm, silica gel particle size 3.5 μm). The separation chromatographic conditions are as follows: mobile phase A-acetonitrile, B-water, C-100 mM ammonium acetate, acetonitrile as a strong elution solvent, gradient elution, elution conditions: 0-30 min, 85%-50% A, C keeps the volume ratio of 10% unchanged, and the rest is B. The column temperature is 35 ° C, the injection volume is 100 μL, and ELSD detection is used. The order of chromatographic peaks in the extract is different on the two chromatographic columns, and the SAC column separates more chromatographic peaks.

[0070] The applicant declares that the present invention illustrates the detailed method of the present invention through the above embodiments, but the present invention is not limited to the above detailed method, that is, it does not mean that the present invention must rely on the above detailed method to be implemented. Those skilled in the art should understand that any improvement of the present invention, equivalent replacement of the raw materials of the product of the present invention, addition of auxiliary components, selection of specific methods, etc., all fall within the protection scope and disclosure scope of the present invention.

Claims

1. A method for analyzing alkaloid components in traditional Chinese medicine, wherein the extract or component of the traditional Chinese medicine containing alkaloid components is dissolved in a solvent and then separated by a liquid chromatography method, wherein the liquid chromatography uses SAC as a chromatographic column stationary phase, and the mobile phase is water and an organic solvent, or the mobile phase is a buffered saline solution and an organic solvent for elution, and then enters a detector for detection; The stationary phase of the SAC chromatographic column is based on silica gel, and the bonding phase terminal of the silica gel surface is a zwitterionic group of phenylsuccinic anhydride modified diamine. The schematic structural formula is as follows: ； SAC is a reverse phase chromatography stationary phase material with zwitterionic properties; Preparation process of SAC column stationary phase: (1) Modification of polyamine substituents on the surface of silica gel: Modification of polyamine groups by one-step reaction of polyaminosilane or two-step reaction. The first step is to bond halogen groups or epoxy groups on the surface of silica gel by silylation reaction of halogen silane / epoxy silane, and the second step is to modify the polyamino groups by two-step method by reacting them with polyamine compounds. The bonding method of silanization reaction can be single-point or multi-point bonding method, or horizontal copolymerization method. a. One-step reaction modification of polyamino groups: Add silica gel to a 1% to 38% hydrochloric acid or nitric acid solution, heat and reflux with stirring for 1 to 48 hours, filter, wash with water until neutral, and heat at 100 to 160 o C to constant weight; placed in a reactor containing an organic solvent under a nitrogen atmosphere, stirred evenly, added a polyaminosilane reagent, 80-120 o C, stirred and refluxed for 1 to 48 hours, filtered under reduced pressure and washed with toluene, dichloromethane, water, tetrahydrofuran and methanol, and heated at 60 to 150 o Polyamino silica gel can be obtained by drying to constant weight under C conditions; b. Two-step reaction modification of polyamino groups: Add silica gel to a 1% to 38% hydrochloric acid or nitric acid solution, heat and reflux with stirring for 1 to 48 hours, filter, wash with water until neutral, and heat at 100 to 160 o C to constant weight; placed in a reactor containing an organic solvent under a nitrogen atmosphere, stirred evenly, added halogen silane or epoxy silane reagent, 80-120 o C, stirred and refluxed for 1 to 48 hours, filtered under reduced pressure and washed with anhydrous toluene, dichloromethane, water, tetrahydrofuran and methanol, and heated at 60 to 150 o C drying conditions until constant weight is obtained to obtain a halogen group or epoxy group modified carrier; the above-mentioned one-step modified silica gel is continued to be added to an organic solvent, and a polyamine compound is added at 50-120 o C, stirred and refluxed for 1 to 48 hours, filtered under reduced pressure and washed with toluene, dichloromethane, water, tetrahydrofuran and methanol, and heated at 60 to 150 o C drying conditions to constant weight to obtain polyamino-modified silica gel; The halogen silane is one or more of chloropropyltrichlorosilane, chlorohexyltrimethylsilane and bromopropyltrichlorosilane; The epoxysilane reagent is one or more of 3-(2,3-epoxypropyloxy)propyltrimethoxysilane, 3-(2,3-epoxypropyloxy)heptyltrimethoxysilane, trimethoxy[2-(7-oxabicyclo[4.1.0]hept-3-yl)ethyl]silane, and 2-(3,4-epoxycyclohexyl)methyldiethoxysilane; The polyaminosilane reagent is one or more of N-(β-aminoethyl-γ-aminopropyl)trimethoxysilane, N-(2-aminopropyl)-aminopropyltrimethoxysilane, N-(2-N-benzylaminoethyl)-3-aminopropyltrimethoxysilane, and N-(2-N-benzylaminovinyl)-3-aminopropyltriethoxysilane; The organic solvent is one or more of toluene, tetrahydrofuran, ethylbenzene, N,N-dimethylformamide, and dimethyl sulfoxide; (2) Amidation reaction: Add polyamino silica gel to an organic solvent, then add phenyl succinic anhydride, a base catalyst and an acid binding agent, 20-120 o C, stirring for 1 to 72 hours; filtering under reduced pressure and washing with anhydrous toluene, dichloromethane, water, tetrahydrofuran, and methanol; o C drying condition until constant weight is obtained to obtain the zwitterionic reverse phase chromatography stationary phase SAC; The organic solvent is one or more of toluene, tetrahydrofuran, ethylbenzene, N,N-dimethylformamide, and dimethyl sulfoxide; The base catalyst is one or more of 4-dimethylaminopyridine, 1,8-diazacyclo[5,4,0]undecene-7, 1,5-diazabicyclo[4.3.0]non-5-ene, and diisopropylethylamine; The acid-binding agent is one or more of pyridine, triethylamine and imidazole; The detector is ultraviolet, electrospray CAD, evaporative light scattering ELSD or mass spectrometry MS.

2. The analysis method according to claim 1, Features: The particle size of silica gel is 2-10 μm, and the bonding amount of phenylsuccinic anhydride on the silica gel surface is 2-8 μmol / m 2 .

3. The analysis method according to claim 1, Features: The solvent used to dissolve the Chinese herbal medicine extract or component is an aqueous solution, to which one or more of methanol, acetonitrile, ethanol, ammonium formate, and ammonium acetate may be added; The concentration of one or both of the aqueous solutions of ammonium formate or ammonium acetate is 2-50 mM; The volume ratio of the organic solvent in the aqueous solution of one or two of methanol, acetonitrile and ethanol is 5-95%.

4. The analysis method according to claim 1, Features: The mobile phase additive includes one or more aqueous solutions of ammonium formate, ammonium acetate, formic acid, acetic acid, and ammonia water, the concentration of the additive is 2-50 mM, and the pH value of the mobile phase is 2-8; the organic phase in the mobile phase is one or more of acetonitrile, methanol, and acetone, the volume proportion of the organic phase in the mobile phase is 5%-95%, and the rest is an aqueous solution.

5. The analysis method according to claim 1, Features: The Chinese medicine extract or component comprises one or more extracts or components of Corydalis yanhusuo, Sophora flavescens, Ephedra sinica and Radix Sophorae flavescentis.

6. The analysis method according to claim 1, Features: The Chinese medicine extract is an extract of one or more of the Chinese medicine in methanol, ethanol, and water; one or more of formic acid and acetic acid are added to methanol, ethanol, and water, and the added volume ratio is 0-10%.

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