Method for analyzing safflower preparation by liquid chromatography-mass spectrometry and application thereof

Abstract

The present application provides a kind of safflower preparation liquid chromatography-mass spectrometry analysis method and application, belong to the technical field of drug detection, the liquid chromatography-mass spectrometry analysis method is using ultra-high performance liquid chromatography tandem mass spectrometry technology with acetonitrile-carbonic acid aqueous solution as mobile phase, by gradient elution, the chemical composition in safflower preparation is characterized;The characterization result obtained in safflower preparation is used for the whole process quality evaluation or control of safflower preparation research / development / production / clinical application.The present application is characterized by adjusting chromatographic condition and mass spectrometry condition, using ultra-high performance liquid chromatography tandem mass spectrometry technology to safflower preparation, active ingredient in safflower preparation can be effectively separated and detected, and 42 kinds of chemical compositions are successfully analyzed.

Description

Technical Field

[0001] This invention relates to the field of drug detection technology, and in particular to a liquid chromatography-mass spectrometry (LC-MS) analysis method and its application for safflower preparations. Background Technology

[0002] The chemical composition of safflower, a traditional Chinese medicine, is complex. Since the early 20th century, scholars have been studying its chemical components. Safflower preparations, made from safflower using modern pharmaceutical processes, possess the effects of promoting blood circulation, regulating menstruation, dispersing blood stasis, and relieving pain. Clinically, they are widely used to treat various diseases such as cerebrovascular diseases, angina pectoris, arrhythmia, pulmonary heart disease, and hyperlipidemia. Due to the complexity and diversity of active ingredients in safflower preparations, and the mutual interference between these components, the pharmacodynamic material basis remains unclear. Therefore, analyzing the chemical components in safflower preparations is crucial for studying their pharmacodynamic material basis. However, because the active ingredients in safflower preparations interact and vary significantly in content, some active ingredients are difficult to isolate and detect, thus affecting the assessment of the pharmacological activity of safflower preparations. In practical research, it is difficult to effectively analyze the chemical components in safflower preparations and conduct fundamental research on their pharmacodynamic material basis. Summary of the Invention

[0003] To address the above problems, this invention provides a liquid chromatography-mass spectrometry (LC-MS) analysis method for safflower preparations and its application.

[0004] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0005] A liquid chromatography-mass spectrometry (LC-MS) analysis method for safflower preparations, wherein the LC-MS analysis method uses ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS / MS) to characterize the chemical components in safflower preparations;

[0006] The ultra-high performance liquid chromatography-tandem mass spectrometry technique uses acetonitrile as mobile phase A and formic acid aqueous solution with a concentration of 0.1-0.12 wt% as mobile phase B.

[0007] The elution method is gradient elution; the elution conditions for the gradient elution are:

[0008] 0–5 min, 5% → 15% mobile phase A, 95% → 85% mobile phase B;

[0009] 5–15 min, 15% → 20% mobile phase A, 85% → 80% mobile phase B;

[0010] 15–30 min, 20% → 30% mobile phase A, 80% → 70% mobile phase B;

[0011] 30–50 min, 30% → 70% mobile phase A, 70% → 30% mobile phase B;

[0012] 50–50.5 min, 70% → 5% mobile phase A, 30% → 95% mobile phase B;

[0013] 50.5–60 min, 5% mobile phase A, 95% mobile phase B.

[0014] Furthermore, the liquid chromatography-mass spectrometry (LC-MS) analysis method also includes:

[0015] Hydroxysafflower yellow A, N1,N5,N10-triscoumaylspermethyleneamine, adenosine, guanosine, isoleucine, tyrosine, caffeic acid, p-coumaric acid, baicalin, isoquercitrin, sucrose, quercetin, rutin, kaempferol 3-O-rutinoside, uridine, tryptophan, phenylalanine, ferulic acid, p-hydroxybenzoic acid, luteolin, D-anhydrous glucose, kaempferol, and chlorogenic acid were used as reference standards. The same chromatographic and mass spectrometric conditions were used to characterize the components in the safflower preparation to determine the corresponding component types.

[0016] Furthermore, the detection wavelength of the ultra-high performance liquid chromatography-tandem mass spectrometry technique is 190–410 nm, the column temperature is 38–42 °C, and the flow rate is 0.18–0.22 mL / min.

[0017] Furthermore, the ultra-high performance liquid chromatography-tandem mass spectrometry technique uses a Waters ACQITY UPLC BEH C18 column.

[0018] Furthermore, the mass spectrometry conditions for the ultra-high performance liquid chromatography-tandem mass spectrometry technique are as follows: an electrospray ionization source is used, with positive and negative ion modes, a nebulizer gas flow rate of 780–820 L / h, a desolvation gas temperature of 580–620 °C, an ion source temperature of 115–125 °C, and a capillary voltage of 3.0 kV.

[0019] Furthermore, in the mass spectrometry conditions of the ultra-high performance liquid chromatography-tandem mass spectrometry technique, the cone voltage is 38–42V and the compensation voltage is 78–82V.

[0020] Furthermore, the mass spectrometry conditions for the ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS / MS) technique are as follows: detection is performed in MSE scanning mode; the transmission collision energy is 6 eV during low-energy scans and 30–60 eV during high-energy scans; and the nebulizer pressure is 6.4–6.6 × 10⁻⁶ eV. 5 Pa, air curtain volumetric flow rate is 48-52 L / h, and scanning range is 50-1500 m / z.

[0021] Furthermore, the liquid chromatography-mass spectrometry (LC-MS) analysis method specifically includes the following steps:

[0022] Prepare the test solution from safflower preparations;

[0023] Prepare a mixed reference solution I by taking hydroxysafflower yellow A, N1,N5,N10-triscoumaylspermidine, adenosine, guanosine, isoleucine, tyrosine, caffeic acid, p-coumaric acid, baicalin, isoquercitrin, sucrose and quercetin.

[0024] Prepare a mixed reference solution II by taking rutin, kaempferol 3-O-rutin glycoside, uridine, tryptophan, phenylalanine, ferulic acid, p-hydroxybenzoic acid, luteolin, D-anhydrous glucose, kaempferol and chlorogenic acid;

[0025] The test solution, mixed reference solution I and mixed reference solution II were respectively subjected to ultra-high performance liquid chromatography-tandem mass spectrometry. The entire detection process was carried out in the wavelength range of 190-410 nm without specifying a wavelength. The chemical components contained in the safflower preparation were determined by analysis.

[0026] The analysis involved comparing the chromatographic results and mass spectrometry results of the positive and negative electrodes obtained from the detection of the test sample solution with the chromatographic results and mass spectrometry results of the positive and negative electrodes obtained from the detection of mixed reference solution I and mixed reference solution II. The chemical components contained in the safflower preparation were identified by precise molecular weight, fragment ions, and relevant literature reports.

[0027] The analysis can also be performed by comparing the chromatographic results of the test sample solution and the mass spectrometry results of the positive and negative electrodes with the information recorded in Table 1 of this invention to confirm the types of chemical components contained in the safflower preparation.

[0028] The chromatographic results are information such as the peak numbers and corresponding retention times of all chromatographic peaks in the 190–410 nm range obtained by ultra-high performance liquid chromatography-tandem mass spectrometry without a specified wavelength.

[0029] Mass spectrometry results are mass spectra of the positive and negative electrodes obtained by ultra-high performance liquid chromatography-tandem mass spectrometry, or information from the mass spectra of the positive and negative electrodes; the information in the mass spectra of the positive and negative electrodes includes the measured molecular weight, theoretical molecular weight, error, adducts, etc.

[0030] Furthermore, the solvent used in the test solution is an aqueous methanol solution with a concentration of 58-62 vol%.

[0031] The characterization results of chemical components in safflower preparations obtained by the above liquid chromatography-mass spectrometry analysis method are applied in the quality evaluation or control of the entire process of research / development / production / clinical application of safflower preparations. The characterization results of the chemical components show that 42 chemical components were isolated and detected in safflower preparations.

[0032] The characterization results of the chemical components are the types of chemical components contained in safflower preparations.

[0033] The beneficial effects of the liquid chromatography-mass spectrometry (LC-MS) analysis method for safflower preparations of the present invention and its application are as follows:

[0034] This invention utilizes ultra-high performance liquid chromatography-tandem mass spectrometry to detect and analyze safflower preparations by adjusting chromatographic and mass spectrometric conditions. It can effectively separate and detect the active ingredients in safflower preparations and successfully analyzes 42 chemical components.

[0035] This invention employs ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS / MS) for detection and analysis, which significantly improves detection efficiency. A single UHPLC-MS / MS analysis can successfully identify 42 chemical components in safflower preparations. The setting of detection indicators is more conducive to the assessment of the pharmacological activity and safety of safflower preparations.

[0036] This invention employs ultra-high performance liquid chromatography-tandem mass spectrometry for detection and analysis, which has extremely high sensitivity and excellent qualitative capabilities;

[0037] The 42 chemical components in the safflower preparation analyzed in this invention have subtle structural differences, and the resulting mass spectrometry feature fragments have significant differences, which can accurately identify the active ingredients.

[0038] This invention employs ultra-high performance liquid chromatography-tandem mass spectrometry for detection and analysis. It only requires the construction of standard chromatographic results and standard mass spectrum library on the instrument. In subsequent analysis, no reference standard is needed. Only the sample needs to be tested. The sample test results are compared with the standard chromatographic results and standard mass spectrum library to confirm whether the sample contains the target active ingredient.

[0039] The liquid chromatography-mass spectrometry (LC-MS) analysis method for safflower preparations of the present invention is applicable to the detection of active ingredients in various safflower preparations (preparations made solely from safflower as the raw material), thereby determining the pharmacological activity of safflower preparations;

[0040] The liquid chromatography-mass spectrometry (LC-MS) analysis method for safflower preparations of the present invention is simple, accurate, stable, reproducible, and effectively separates active ingredients, and can be used as an important method for examining the pharmacological activity of safflower preparations. Attached Figure Description

[0041] Figure 1 These are BPI mass spectra obtained in negative ion mode in Example 1 of the present invention, wherein (A) is the BPI mass spectrum obtained by the test solution (batch number 140307C1) in negative ion mode, (B) is the BPI mass spectrum obtained by the blank solvent in negative ion mode, (C) is the BPI mass spectrum obtained by mixed reference solution I in negative ion mode, and (D) is the BPI mass spectrum obtained by mixed reference solution II in negative ion mode.

[0042] Figure 2These are BPI mass spectra obtained in positive ion mode in Example 1 of the present invention, wherein (A) is the BPI mass spectrum obtained in positive ion mode for the test solution (batch number 140307C1), (B) is the BPI mass spectrum obtained in positive ion mode for the blank solvent, (C) is the BPI mass spectrum obtained in positive ion mode for mixed reference solution I, and (D) is the BPI mass spectrum obtained in positive ion mode for mixed reference solution II. Detailed Implementation

[0043] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Many specific details are set forth in the following description to provide a thorough understanding of the present invention. However, the present invention may also be implemented in other ways different from those described herein. Those skilled in the art can make similar extensions without departing from the spirit of the present invention. Therefore, the present invention is not limited to the specific embodiments disclosed below.

[0044] Example 1: A liquid chromatography-mass spectrometry (LC-MS) analysis method for safflower preparations and its application

[0045] In this embodiment, safflower injection was used as the safflower preparation for detection and analysis by ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-QTOF-MS, i.e., liquid chromatography-mass spectrometry). Waters Acquity I-CLASS was used in this embodiment. TM The detection and analysis were performed using an ultra-high performance liquid chromatography (UPLC) system in tandem with a Waters Xevo G2-XS quadrupole time-of-flight mass spectrometer. The specific analytical method is as follows:

[0046] 1) Take 1 mL of safflower injection (batch number 140307C1) and place it in a 5 mL volumetric flask. Dilute to volume with 60 vol% methanol aqueous solution and filter through a 0.22 μm microporous membrane to obtain the test solution.

[0047] 2) Take appropriate amounts of hydroxysaffron yellow A, N1,N5,N10-triscoumoylspermethyleneamine, adenosine, guanosine, isoleucine, tyrosine, caffeic acid, p-coumaric acid, baicalin, isoquercitrin, sucrose, and quercetin, dissolve them in 50 vol% methanol aqueous solution, filter through a 0.22 μm microporous membrane to prepare mixed reference solution I; wherein, mixed reference solution I contains hydroxysaffron yellow A 2.06 μg / mL, N1,N5,N10-triscoumaylsperidin 4.25 μg / mL, adenosine 0.88 μg / mL, guanosine 0.96 μg / mL, isoleucine 1.02 μg / mL, tyrosine 1.13 μg / mL, caffeic acid 2.12 μg / mL, p-coumaric acid 0.86 μg / mL, baicalin 3.54 μg / mL, isoquercitrin 0.58 μg / mL, sucrose 0.09 μg / mL, and quercetin 0.98 μg / mL;

[0048] Take appropriate amounts of rutin, kaempferol 3-O-rutinoside, uridine, tryptophan, phenylalanine, ferulic acid, p-hydroxybenzoic acid, luteolin, D-anhydrous glucose, kaempferol, and chlorogenic acid, dissolve them in 50 vol% methanol aqueous solution, and filter through a 0.22 μm microporous membrane to prepare mixed reference solution II; wherein mixed reference solution II contains 4.88 μg / mL of rutin, 4.68 μg / mL of kaempferol 3-O-rutinoside, 0.25 μg / mL of uridine, 1.36 μg / mL of tryptophan, 0.98 μg / mL of phenylalanine, 1.64 μg / mL of ferulic acid, 0.76 μg / mL of p-hydroxybenzoic acid, 4.19 μg / mL of luteolin, approximately 0.13 μg / mL of D-anhydrous glucose, 2.34 μg / mL of kaempferol, and 2.76 μg / mL of chlorogenic acid.

[0049] 60 vol% methanol-water was used as the blank solvent;

[0050] It should be noted that, since this invention is a qualitative analysis, the concentrations of the components contained in mixed reference solution I and mixed reference solution II will not have a significant impact on the analytical results. Therefore, the concentrations in this embodiment are not intended to limit the concentrations of the components contained in mixed reference solution I and mixed reference solution II of this invention.

[0051] 3) Take the test solution, blank solvent, mixed reference solution I and mixed reference solution II respectively for liquid chromatography-mass spectrometry analysis (UPLC-QTOF-MS analysis, which is ultra-high performance liquid chromatography tandem mass spectrometry detection and analysis);

[0052] The chromatographic conditions are as follows:

[0053] Chromatographic column: Waters ACQITY UPLC BEH C18 column (2.1×100mm, 1.7μm);

[0054] Detection wavelength: 190–410 nm; no specific wavelength is specified during the entire detection process.

[0055] Column temperature: 40℃;

[0056] Flow rate: 0.2 mL / min;

[0057] Injection volume: The injection volume of the test sample is 5 μL, and the injection volumes of mixed reference solution I and mixed reference solution II are 2 μL respectively;

[0058] Mobile phase A is acetonitrile, and mobile phase B is a 0.1 wt% aqueous solution of formic acid;

[0059] The elution method is gradient elution, and the specific elution procedure is as follows:

[0060] 0–5 min, 5% → 15% mobile phase A, 95% → 85% mobile phase B;

[0061] 5–15 min, 15% → 20% mobile phase A, 85% → 80% mobile phase B;

[0062] 15–30 min, 20% → 30% mobile phase A, 80% → 70% mobile phase B;

[0063] 30–50 min, 30% → 70% mobile phase A, 70% → 30% mobile phase B;

[0064] 50–50.5 min, 70% → 5% mobile phase A, 30% → 95% mobile phase B;

[0065] 50.5–60 min, 5% mobile phase A, 95% mobile phase B.

[0066] Mass spectrometry conditions are:

[0067] Electrospray ionization (ESI) source, positive and negative ion modes, desolvation gas flow rate of 800 L / h, desolvation gas temperature of 600℃, ion source temperature of 120℃, capillary voltage of 3.0 kV, sampling cone voltage of 40 V, and source offset voltage of 80 V. E During scanning mode detection, the transmitted collision energy is 6 eV in low-energy scanning and 30–60 eV in high-energy scanning, with a sprayer pressure of 6.5 × 10⁻⁶. 5 Pa, cone gas flows of 50 L / h, scan range m / z 50–1500. Leucine-enkephalin (m / z 554.2615, 556.2771) was used as an external standard (Lock Spray). TM Real-time quality correction was performed, and the volumetric flow rate was set to 5 μL / min.

[0068] UPLC-QTOF-MS analysis was performed on the test solution, blank solvent, mixed reference solution I, and mixed reference solution II. Chromatographic results and mass spectra of the positive and negative electrodes were acquired, respectively. This yielded the chromatographic results and mass spectra of the test solution, blank solvent, mixed reference solution I, and mixed reference solution II. The mass spectra of the positive and negative electrodes are attached. Figures 1-2 .

[0069] The chromatographic results are information such as the peak numbers and corresponding retention times of all chromatographic peaks in the 190–410 nm range obtained by ultra-high performance liquid chromatography-tandem mass spectrometry without a specified wavelength.

[0070] Mass spectrometry results are mass spectra of the positive and negative electrodes obtained by ultra-high performance liquid chromatography-tandem mass spectrometry, or information from the mass spectra of the positive and negative electrodes; the information in the mass spectra of the positive and negative electrodes includes the measured molecular weight, theoretical molecular weight, error, adducts, etc.

[0071] The chromatographic results and mass spectrometry results of the test sample and the positive and negative electrodes were compared with the chromatographic results and mass spectrometry results of the blank solvent, the mixed reference solution I, and the mixed reference solution II. The chemical components in the safflower injection were identified by precise molecular weight, fragment ions, and relevant literature reports. The results showed that 42 compounds were identified in the safflower injection. Among them, 10 compounds were identified by comparison with reference standards: D-anhydrous glucose (compound 1), sucrose (compound 2), uridine (compound 5), hydroxysafflower yellow A (compound 6), p-coumaric acid (compound 7), rutin (compound 16), isoquercitrin (compound 17), luteolin (compound 19), kaempferol 3-O-rutin (compound 22), and N1,N5,N10-tris-p-coumaryl spermidine (compound 42). The remaining compounds were identified by precise molecular weight, fragment ions, and relevant literature reports. The specific results are shown in Table 1.

[0072] Table 1. Identification results of chemical components of safflower injection.

[0073]

[0074]

[0075] *Indicates identification based on comparison with a reference standard.

[0076] As shown in Table 1, this invention identified 42 chemical components from safflower injection. In subsequent testing of the safflower preparations, the above method was used to detect the safflower preparations. The obtained chromatographic and mass spectrometric results of the safflower preparations were directly compared and analyzed with the retention time, measured molecular weight, and theoretical molecular weight information in Table 2 (without the need for preparation and detection of blank solvents, mixed reference solution I, and mixed reference solution II). This allowed for the determination of the types of chemical components in the safflower preparations and, consequently, the quality of the safflower preparations.

[0077] When the safflower preparation is a solid safflower preparation (i.e., a solid preparation made solely from safflower as a raw material), the solid safflower preparation needs to be pulverized and sieved, and an appropriate amount (5.65 mg) should be added to a volumetric flask (20 mL). A methanol aqueous solution with a concentration of 58-62 vol% should be used as the solvent. The solution should be dissolved at 80 °C, diluted to a fixed volume, filtered, and the test solution obtained.

[0078] The characterization results of the chemical components obtained in safflower preparations are used for quality evaluation or control throughout the entire process of research, development, production, and clinical application of safflower preparations.

[0079] Examples 2-3: Liquid chromatography-mass spectrometry analysis method for safflower preparations

[0080] Examples 2 and 3 are liquid chromatography-mass spectrometry (LC-MS) analytical methods for safflower preparations. Their steps are basically the same as in Example 1, differing only in some parameters, as detailed in Table 2.

[0081] Table 2. Summary of parameters in Examples 2-3

[0082]

[0083] The contents of Examples 2 and 3 are the same as those of Example 1, and the results of the liquid chromatography-mass spectrometry analysis are also the same as those of Example 1, so they will not be repeated here.

[0084] Obviously, the described embodiments are only some, not all, of the embodiments of the present invention. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without inventive effort are within the scope of protection of the present invention.

Claims

1. A liquid chromatography-mass spectrometry (LC-MS) method for analyzing safflower preparations, characterized in that, The liquid chromatography-mass spectrometry (LC-MS) analysis method uses a single-pass ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS / MS) technique to characterize the chemical components in safflower preparations; the characterization results show that 42 chemical components were isolated and detected in the safflower preparations. The ultra-high performance liquid chromatography-tandem mass spectrometry technique uses acetonitrile as mobile phase A and formic acid aqueous solution with a concentration of 0.1~0.12wt% as mobile phase B. The elution method is gradient elution; the elution conditions for the gradient elution are: 0~5min, 5%→15% mobile phase A, 95%→85% mobile phase B; 5~15min, 15%→20% mobile phase A, 85%→80% mobile phase B; 15~30 min, 20%→30% mobile phase A, 80%→70% mobile phase B; 30~50 min, 30%→70% mobile phase A, 70%→30% mobile phase B; 50~50.5 min, 70%→5% mobile phase A, 30%→95% mobile phase B; 50.5~60 min, 5% mobile phase A, 95% mobile phase B; The chromatographic column was a Waters ACQITY UPLC BEH C18 column, 2.1×100mm, 1.7μm; The mass spectrometry conditions for the ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS / MS) technique are as follows: an electrospray ionization source is used, with positive and negative ion modes, a nebulizer gas flow rate of 780-820 L / h, a desolvation gas temperature of 580-620 °C, an ion source temperature of 115-125 °C, and a capillary voltage of 3.0 kV. Hydroxysafflower yellow A, N1,N5,N10-triscoumaylsperidin, adenosine, guanosine, isoleucine, tyrosine, caffeic acid, p-coumaric acid, baicalin, isoquercitrin, sucrose, quercetin, rutin, kaempferol 3-O-rutinoside, uridine, tryptophan, phenylalanine, ferulic acid, p-hydroxybenzoic acid, luteolin, D-anhydrous glucose, kaempferol, and chlorogenic acid were used as reference standards. The same chromatographic and mass spectrometric conditions were used to characterize the chemical components in the safflower preparation to determine the corresponding component types.

2. The liquid chromatography-mass spectrometry (LC-MS) analysis method for safflower preparations according to claim 1, characterized in that, The ultra-high performance liquid chromatography-tandem mass spectrometry technique has a detection wavelength of 190~410nm, a column temperature of 38~42℃, and a flow rate of 0.18~0.22mL / min.

3. The liquid chromatography-mass spectrometry (LC-MS) analysis method for safflower preparations according to claim 1, characterized in that, In the mass spectrometry conditions of the ultra-high performance liquid chromatography-tandem mass spectrometry technique, the cone voltage is 38~42V and the compensation voltage is 78~82V.

4. The liquid chromatography-mass spectrometry (LC-MS) analysis method for safflower preparations according to claim 1, characterized in that, The mass spectrometry conditions for the ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS / MS) technique are as follows: MSE scanning mode detection; transmission collision energy of 6 eV for low-energy scans; transmission collision energy of 30–60 eV for high-energy scans; and nebulizer pressure of 6.4–6.6 × 10⁻⁶ eV. 5 Pa, air curtain volumetric flow rate is 48~52 L / h, and scanning range is 50~1500 m / z.

5. The liquid chromatography-mass spectrometry (LC-MS) analysis method for safflower preparations according to claim 1, characterized in that, The liquid chromatography-mass spectrometry (LC-MS) analysis method specifically includes the following steps: Prepare the test solution from safflower preparations; Prepare a mixed reference solution I by taking hydroxysafflower yellow A, N1,N5,N10-triscoumaylspermidine, adenosine, guanosine, isoleucine, tyrosine, caffeic acid, p-coumaric acid, baicalin, isoquercitrin, sucrose and quercetin. Prepare a mixed reference solution II by taking rutin, kaempferol 3-O-rutin glycoside, uridine, tryptophan, phenylalanine, ferulic acid, p-hydroxybenzoic acid, luteolin, D-anhydrous glucose, kaempferol and chlorogenic acid; The test solution, mixed reference solution I, and mixed reference solution II were respectively subjected to ultra-high performance liquid chromatography-tandem mass spectrometry detection and analysis to determine the types of chemical components contained in the safflower preparation.

6. The liquid chromatography-mass spectrometry (LC-MS) analysis method for safflower preparations according to claim 5, characterized in that, The solvent used for the test sample solution is a methanol aqueous solution with a concentration of 58-62 vol%.

7. The application of the characterization results of chemical components in safflower preparations obtained by the liquid chromatography-mass spectrometry analysis method according to any one of claims 1-6 in the quality evaluation or control of the entire process of research / development / production / clinical application of safflower preparations, characterized in that, The characterization results of the chemical components showed that 42 chemical components were isolated and detected in the safflower preparation.

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