Method for analyzing DSHP serum migration components and heart tissue distribution components based on myocardial infarction disease model

Abstract

The invention belongs to the technical field of medicine analysis, and particularly relates to a method for analyzing serum migration components and heart tissue distribution components of a Danshen-Shanzha herb pair (DSHP) based on a myocardial infarction disease model. The invention discloses a method for analyzing DSHP serum migration components and heart tissue distribution components in an MI model by adopting an ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry technology, according to the method, 22 prototype components of DSHP extracts are successfully identified in a rat body, and 10 of the prototype components are serum and heart tissue co-distribution components. The invention establishes a systematic analysis method for the medicinal components of DSHP in a pathological state, can realize comprehensive detection of DSHP serum migration components and heart tissue distribution components, and also can provide key technical support for clarification of medicinal material basis, reveal of action mechanism and establishment of quality control standard.

Description

Technical Field

[0001] The invention belongs to the technical field of drug analysis, and particularly relates to an analysis method for DSHP serum migration components and cardiac tissue distribution components based on a disease model. Background Art

[0002] Ischemic heart disease (IHD) is the leading cause of death worldwide. Its prevention and treatment face clinical challenges such as poor long-term prognosis and significant drug side effects. Ischemic heart disease falls under the category of "chest pain" in traditional Chinese medicine. Traditional Chinese medicine, based on the pathogenesis theory of "blood stasis leading to paralysis", has demonstrated unique advantages in the prevention and treatment of IHD. The Danshen-Shanzha Herb Pair (DSHP), composed of Danshen (Salviae Miltiorrhizae Radix et Rhizoma) and Hawthorn (Crataegi Fructus), is a classic combination for the treatment of "chest pain" in Traditional Chinese Medicine. Danshen has the effects of promoting blood circulation, removing blood stasis, unblocking meridians and relieving pain, while Hawthorn has the functions of promoting qi and dispersing stagnation, clearing turbidity and lowering lipids. The two can synergistically significantly improve the symptoms of blood stasis in the heart and reduce myocardial ischemic damage. However, the pharmacological material basis of DSHP has not been fully elucidated, especially the lack of systematic research on the transmission rules of its active components in the body, which has seriously restricted its precise clinical application and internationalization process.

[0003] Active ingredients in traditional Chinese medicine (TCM) formulas that can be absorbed into the bloodstream and targeted to cardiac tissue can directly influence the pathological progression of IHD. Currently, research on the in vivo delivery of DSHP pharmacodynamics is limited to two key limitations: first, chemical analysis of DSHP is often limited to a few components of a single herbal remedy, failing to capture the overall effects of the drug pair combination; second, studies of DSHP pharmacological compounds employ normal animal models, neglecting the specificity of drug pharmacokinetic analysis under conditions of myocardial ischemia. Therefore, establishing an analytical method for DSHP serum migration components and cardiac tissue distribution components based on an IHD disease model would not only elucidate its core pharmacological components and targeting mechanisms for IHD intervention but also provide a key scientific basis for optimizing related formulation processes and developing precision drug delivery systems, which is of great significance for promoting the modernization of traditional Chinese medicine. Summary of the Invention

[0004] Based on the above technical problems, the present invention innovatively takes the drug distribution characteristics under pathological conditions as the starting point, and systematically analyzes the distribution characteristics of serum migration components and cardiac tissue targeting components after oral administration of DSHP extract through chromatography-mass spectrometry technology, and establishes a dynamic analysis method for DSHP active components based on a myocardial infarction disease model.

[0005] In order to fully and unambiguously understand the technical solution of the present invention, it is necessary to supplement that, in this application, DSHP (Danshen-Shanzha Herb Pair) refers to a composition composed of Salviae Miltiorrhizae Radixet Rhizoma and Crataegi Fructus in a mass ratio of 1:3.

[0006] On the one hand, the present invention provides a method for analyzing DSHP serum migration components and cardiac tissue distribution components based on a disease model. The method uses ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry to measure ion currents, and obtains DSHP serum migration components and cardiac tissue distribution components by analyzing the ion currents.

[0007] The conditions of the ultra-high performance liquid chromatography include:

[0008] Chromatographic column: Waters ACQUITY HSS T3 column;

[0009] Column temperature: 20-40°C;

[0010] Flow rate: 0.2-0.3 mL / min;

[0011] Injection volume: 2 μL;

[0012] Detection wavelength: 190~410nm;

[0013] Mobile phase: Phase A is acetonitrile, phase B is 0.1% formic acid water;

[0014] The elution gradient conditions are: 0-2 minutes, 95% phase B; 2-5 minutes, 95%-90% phase B; 5-18 minutes, 90%-65% phase B; 18-25 minutes, 65%-30% phase B; 25-30 minutes, 30%-10% phase B; 30-34 minutes, 10%-0 phase B; 34.1-40 minutes, 95% phase B.

[0015] The quadrupole time-of-flight mass spectrometer adopts electrospray ionization technology and performs detection in positive and negative ion modes respectively.

[0016] Furthermore, in the analysis method of DSHP serum migration components and cardiac tissue distribution components based on a disease model, the disease model is an animal cardiogenic disease model, and the animal cardiogenic disease model is an animal myocardial infarction (MI) model.

[0017] Furthermore, the disease model-based analysis method for DSHP serum migration components and cardiac tissue distribution components includes administering DSHP to a myocardial infarction model and collecting blood samples and cardiac tissue samples from animals after a predetermined period of time as a test group.

[0018] Furthermore, in the analysis method of DSHP serum migration components and cardiac tissue distribution components based on the disease model, the samples detected by the ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry method are blood samples and cardiac tissue samples of the experimental group and the control group.

[0019] Furthermore, in the analysis method of DSHP serum migration components and cardiac tissue distribution components based on the disease model, the blood sample preparation method is: collecting animal plasma, adding hydrochloric acid to adjust the plasma pH value, extracting the plasma with ethyl acetate, blowing the extract with nitrogen to dryness, removing the organic solvent, and re-dissolving it for use.

[0020] Furthermore, in the analysis method of DSHP serum migration components and cardiac tissue distribution components based on the disease model, the preparation method of the cardiac sample is: the cardiac tissue is minced and ultrasonically homogenized, hydrochloric acid and acetonitrile are added and vortexed in sequence, and then the homogenate is extracted with ethyl acetate, the extract is blown dry with nitrogen, and then dissolved and set aside for use.

[0021] Furthermore, in the analysis method of DSHP serum migration components and cardiac tissue distribution components based on the disease model, the test samples were vortexed before testing, centrifuged at 12000 rpm for 10 minutes, the supernatant was filtered with a 0.22 μm filter membrane, and 2 μL of the filtrate was injected into ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry analysis.

[0022] On the other hand, the present invention also provides the application of the analysis method of DSHP serum migration components and cardiac tissue distribution components based on the disease model in analyzing DSHP serum migration components in a myocardial infarction model.

[0023] In another aspect, the present invention also provides the application of the analysis method of DSHP serum migration components and cardiac tissue distribution components based on the disease model in analyzing the cardiac tissue distribution components of DSHP in a myocardial infarction model.

[0024] Finally, the present invention also provides an analysis method of DSHP serum migration components and cardiac tissue distribution components based on disease models for use in analyzing DSHP prototype components in myocardial infarction model rats or in analyzing the absorption and distribution of DSHP prototype components in normal rats and myocardial infarction model rats.

[0025] Compared with the prior art, the technical solution provided by the present invention has at least the following beneficial effects or advantages:

[0026] The present invention uses ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry to establish an analytical method for DSHP serum migration components and cardiac tissue distribution components based on a myocardial infarction model. Using this method, 22 prototype components were identified in rats. Among them, 10 prototype components, including lithospermic acid and its isomers, rosmarinic acid, salvianolic acid A, quercetin, dimethyl lithospermate, lithospermic acid, salvianolic acid B, dihydrotanshinone I, cryptotanshinone, and tanshinone IIA, were present simultaneously in plasma and cardiac tissue. The myocardial infarction model promoted the absorption of rutin, neochlorogenic acid, lithospermic acid, salvianolic acid B, dihydrotanshinone I, cryptotanshinone, and tanshinone IIA into the blood, and promoted the distribution of salvianolic acid A, quercetin, and cryptotanshinone in cardiac tissue. The chemical component detection and analysis method based on ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry established in the present invention can comprehensively and systematically detect the in vivo and in vitro chemical components of DSHP, providing a reference for its subsequent research on the material basis of efficacy, mechanism of action and quality control. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 Figure 1 is a DSHP mass spectrum BPI diagram. A is a DSHP mass spectrum in DSHP negative ion mode; B is a DSHP mass spectrum in DSHP positive ion mode.

[0028] Figure 2 Figures 1 and 2 show TIC plots of plasma samples from MI model rats in positive and negative ion modes. A shows the TIC plot of MI blank plasma in positive ion mode; B shows the TIC plot of MI blank plasma in negative ion mode; C shows the TIC plot of MI drug-containing plasma in positive ion mode; and D shows the TIC plot of MI drug-containing plasma in negative ion mode.

[0029] Figure 3 Figures 1 and 2 show TIC images of MI model rat heart samples in positive and negative ion modes. A is the TIC image of an MI-free heart in positive ion mode; B is the TIC image of an MI-free heart in negative ion mode; C is the TIC image of an MI-treated heart in positive ion mode; and D is the TIC image of an MI-treated heart in negative ion mode.

[0030] Figure 4 The distribution diagram and EIC mass spectra of rutin, neochlorogenic acid, and lithospermic acid prototype components in healthy rats and MI model rats (BPI-). A is rutin; B is neochlorogenic acid; and C is lithospermic acid.

[0031] Figure 5 The distribution diagram and EIC mass spectrum (BPI-) of the prototype component of salvianolic acid A in healthy rats and MI model rats are shown. A is the distribution diagram; B is the EIC mass spectrum. DETAILED DESCRIPTION

[0032] The technical solutions of the present invention are described below with reference to the following embodiments; however, the present invention is not limited to the following embodiments.

[0033] In order to enable those skilled in the art to better understand and implement the technical solution of the present invention, the present invention is further described below with reference to specific embodiments and drawings, but the embodiments are not intended to limit the present invention.

[0034] The experimental methods and detection methods described in the following examples are conventional methods unless otherwise specified; the reagents and materials are commercially available unless otherwise specified.

[0035] The experimental animals involved in the present invention are rats, specifically SPF-grade male SD rats, weighing 160-180 g, purchased from the Experimental Animal Center of the Air Force Medical University. After the SD rats were purchased, they were adaptively raised for 3 days, given standard feed and drinking water, and maintained at room temperature of 23±2°C and relative humidity of 50% to 60%. They were fasted but not watered for 12 hours before the experiment. The experimental operations on all animals complied with the requirements of the Animal Ethics Committee of the Air Force Medical University (Ethics Approval Number: IACUC-20241174).

[0036] In order to fully and unambiguously understand the technical solution of the present invention, it should be supplemented that the MI model described in the present invention is a myocardial infarction model.

[0037] Example 1

[0038] This example is to construct a rat myocardial infarction (MI) model.

[0039] The myocardial infarction (MI) model in rats was induced by ligation of the left anterior descending coronary artery. The specific operation was as follows: After SD rats were anesthetized by 5% isoflurane inhalation, they were fixed in a supine position and connected to an electrocardiogram monitoring system. A 5mm longitudinal incision was made in the 4th intercostal space of the left chest wall, and a thoracotomy was placed to expose the heart. Ophthalmic non-invasive sutures were used to ligate 3mm from the root of the coronary artery. Local myocardial pallor, electrocardiogram ST segment arch elevation and QRS wave group widening were used as the success criteria of the model. After the heart was reset, the chest cavity was sutured layer by layer, and the monitoring equipment was removed after the animal recovered spontaneous breathing. At the same time, a sham operation group was set up, and the sham operation group only underwent threading without ligation. Penicillin (80×10 4 U / d) prevent infection.

[0040] Example 2

[0041] This example is a method for analyzing DSHP serum migration components and cardiac tissue distribution components.

[0042] 1. Animal drug administration, biological sample collection and processing.

[0043] 1.1 Dosage regimen for healthy rats and collection of biological samples

[0044] Twenty-four healthy male Sprague-Dawley (SD) rats were randomly divided into four groups (n=6): a normal control group, a normal control group plus Danshen (DS) (2.7 g / kg), a normal control group plus Crataegus pinnatifida (SZ) (8.1 g / kg), and a normal control group plus a 1:3 Danshen-Crataegus pinnatifida (DSHP) (10.8 g / kg). The dose in each group was approximately twice the human clinical dose, with an oral gavage volume of 20 mL / kg for seven consecutive days. The normal control group received an equal volume of normal saline. On the final day of dosing, the time of day was recorded, and orbital blood was collected at 0.5, 1, 2, and 4 hours, followed by centrifugation at 3500 rpm for 10 minutes to obtain plasma, which was then frozen at -80°C until use.

[0045] 1.2 Dosage regimen for MI model rats and collection of biological samples

[0046] Thirty-nine healthy male Sprague-Dawley rats were randomly divided into four groups: MI (control group), MI plus Danshen (DS) (2.7 g / kg), MI plus Crataegus pinnatifida (SZ) (8.1 g / kg), and MI plus Danshen-Crataegus pinnatifida (DSHP) (10.8 g / kg). The MI plus DSHP (1:3) group consisted of 15 rats, while the remaining groups consisted of 6 rats each. Drug administration began on the day of modeling and continued for seven consecutive days. The MI group received an equal volume of normal saline (20 mL / kg) via gavage. On the final day of drug administration, the time of administration was recorded. Orbital blood was collected 0.5, 1, 2, and 4 hours after administration. Plasma was collected by centrifugation at 3500 rpm for 10 minutes and frozen at -80°C until use. Heart tissue was collected from three animals at each time point in the MI plus DSHP (1:3) group. After washing with normal saline, blood and tissue contents were removed, and the tissue was then blotted with filter paper to remove water. The tissue was then stored at -80°C until use.

[0047] 1.3 Plasma sample processing method

[0048] The plasma collected in Examples 1.1 and 1.2 was processed, specifically as follows: 400 μL of plasma sample and 200 μL of 1 mol / L hydrochloric acid were added to a centrifuge tube, vortexed for 30 s, and then 1.5 mL of ethyl acetate was added after standing, mixed with a fast mixer for 3 min, centrifuged at 4000 r / min for 15 min, the supernatant was aspirated, 1.5 mL of ethyl acetate was added to the lower precipitate, vortexed for 3 min, and centrifuged at 4000 r / min for 15 min. The supernatants of the two extractions were combined and dried with a nitrogen blow dryer in a 40°C water bath. The residue was re-dissolved with 200 μL of the initial mobile phase, mixed with a fast mixer for 5 min, centrifuged at 12000 r / min, 4°C for 15 min, and the supernatant was aspirated and filtered with a 0.22 μm filter membrane for use.

[0049] 1.4 Pretreatment of cardiac tissue samples

[0050] Accurately weigh 100 mg of heart tissue and add physiological saline at a W / V ratio of 1:3. The tissue was minced and then ground. The resulting ground homogenate was sonicated at 4000 r / min and centrifuged at 4°C for 10 min. 200 μL of the supernatant was accurately aspirated, and 20 μL of hydrochloric acid solution (2 mol / L) and 400 μL of acetonitrile were added in sequence. The mixture was vortexed for 5 min, and 800 μL of ethyl acetate was added. The mixture was vortexed for 5 min, mixed thoroughly, and extracted. The mixture was centrifuged at 4000 r / min and 4°C for 10 min. The supernatant was collected (extracted twice). The extracts were combined and dried with nitrogen at 35°C. The residue was reconstituted with 200 μL of acetonitrile-0.15% formic acid aqueous solution (18:82), vortexed for 5 min, and centrifuged at 12000 r / min and 4°C for 15 min. The supernatant was collected for later use.

[0051] 2. Biological sample testing

[0052] The above biological samples were detected by ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry.

[0053] 2.1 Solution preparation

[0054] Accurately weigh 10 reference samples of D-fructose and 5-hydroxymethylfurfural into a 25 mL volumetric flask and dilute to the mark with 50% methanol to create mixed reference solution 1. Add 18 reference samples of L-malic acid and syringic acid to a 25 mL volumetric flask and dilute to the mark with 50% methanol to create mixed reference solution 2. Add 12 reference samples of danshensu and 9'-salvianolic acid B monomethyl ester to a 25 mL volumetric flask and dilute to the mark with 50% methanol to create mixed reference solution 3. Add 150 mg of the aforementioned Danshen:crataegi (1:3) extract to a 50 mL volumetric flask and sonicate in 80% methanol for 30 minutes before diluting to the mark to create the test solution. Vortex all reference and test solutions, centrifuge at 12,000 rpm for 10 minutes, filter the supernatant with a 0.22 μm filter, and analyze 2 μL of the filtrate by ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry.

[0055] 2.2 Chromatographic conditions

[0056] The mixed reference and test solutions were detected in positive and negative ion modes according to the following chromatographic conditions and quality parameters.

[0057] Chromatographic column: Waters ACQUITY HSS T3 column (2.1×100 mm, 1.8 μm);

[0058] Column temperature: 40°C;

[0059] Flow rate: 0.3 mL / min;

[0060] Injection volume: 2 μL;

[0061] Detection wavelength: 190~410nm;

[0062] Mobile phase: acetonitrile as phase A, 0.1% formic acid water as phase B;

[0063] The elution gradient is shown in Table 1.

[0064] Table 1 Elution gradient

[0065] Time (min) Acetonitrile (%) 0.1 Formic acid water (%) 0 5 95 2 5 95 5 10 90 18 35 65 25 70 30 30 90 10 34 100 0 34.1 5 95 40 5 95

[0066] 2.3 Mass spectrometry conditions

[0067] Electrospray ionization (ESI) was used for detection in both positive and negative ion modes. The experimental parameters were set as follows: nebulizer gas flow rate 600 L / h, desolvation gas temperature 500°C, ion source temperature 120°C, capillary voltage 3.0 kV. The cone voltage and offset voltage were set to 40 V and 80 V, respectively. E In scanning mode, the collision energy of low energy scanning was 6 eV, and the collision energy gradient of high energy scanning was set to 30-60 eV. The spray pressure was maintained at 6.5×10 5 Pa, the curtain gas flow rate was 50 L / h, and the mass scan range was set to m / z 50 to 1500. Leucine-enkephalin (m / z 554.2615 [M–H] – ) and (m / z 556.2771[M+H] + ) as external standard (Lock Spray TM ) for real-time mass calibration, and the volume flow rate was set to 5 μL / min.

[0068] 3. Data Analysis

[0069] By consulting CNKI, SciFinder, Web of Science, ChemicalBook, PubChem and HMDB and other platforms or databases, the relevant information on the chemical components of Danshen and Crataegus pinnatifida was collected and sorted out to construct the DSHP chemical component database. TM The Chinese medicine small molecule screening software comprehensively conducts qualitative analysis of the chemical components in DSHP through a self-built medicinal material database and reference material comparison system.

[0070] 3.1 Analysis of Chemical Components of DSHP Entering Blood and Its Distribution in Heart Tissue

[0071] Ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry was used to detect the test solution and the mixed reference solution, and the base peak chromatograms of the best compatible DSHP extract in positive and negative ion modes were obtained. The base peak chromatograms are shown in Figure 2. Figure 1 A total of 96 compounds were identified (36 of which were consistent with the reference), of which 36 were derived from Salvia miltiorrhiza, 31 from Crataegus pinnatifida, and 29 were shared by both herbs. The identification results of each component are shown in Table 2.

[0072]

[0073]

[0074]

[0075]

[0076]

[0077]

[0078] 3.2 Analysis and identification of the prototype components of DSHP in MI model rats

[0079] The same liquid chromatography and mass spectrometry conditions were used for the characterization of in vivo and in vitro chemical components. The spectra of the plasma samples of the MI + Danshen-Hawthorn drug pair (Danshen:Hawthorn mass ratio = 1:3) treatment group (10.8 g / kg), i.e., the model drug-containing plasma samples, and the plasma samples of the model blank group were compared. Combined with the DSHP in vitro chemical component mass spectrometry analysis data, a total of 22 prototype components absorbed into the blood were found and identified in the plasma of MI model rats based on the chromatographic and mass spectrometric information. The TIC diagrams of the model blank plasma and drug-containing plasma samples are shown in Figure 2. Figure 2 The information of serum migration components is shown in Table 3.

[0080] Table 3 Blood migration component information table

[0081]

[0082]

[0083] *Identified * by * comparison * with * reference * standards. * B: enters the blood; * H: Enters the heart (heart tissue distribution)

[0084] The results showed that 10 prototype components were found and identified in the hearts of MI model rats. The TIC images of the model blank hearts and the heart samples of the drug-treated groups were as follows: Figure 3 The distribution information of cardiac tissue components is shown in Table 2. A total of 10 prototype components, including tanshinone IIA, lithospermic acid and its isomers, rosmarinic acid, salvianolic acid A, quercetin, dimethyl lithospermate, lithospermic acid, salvianolic acid B, dihydrotanshinone I, and cryptotanshinone, are present in both serum and cardiac tissue, indicating that these components can be rapidly distributed to cardiac tissue through the blood circulation and exert a cardioprotective effect.

[0085] 3.3 Absorption and distribution of different prototype components of DSHP in normal and MI rats

[0086] There are significant differences in the absorption and metabolism of drugs in animals under normal and pathological conditions. At present, conventional serum drug chemistry studies focus more on normal animals. This example explores the distribution characteristics of drugs under pathological conditions. By comparing the distribution diagrams and extracted ion current (EIC) mass spectra of each prototype component of DSHP in normal and MI rats, it was found that the response values ​​of rutin, neochlorogenic acid, lithospermic acid, tanshinone B, dihydrotanshinone I, cryptotanshinone and tanshinone IIA in the model drug-administered plasma were significantly higher. Among them, the top three rutin, neochlorogenic acid and lithospermic acid are shown in the following diagrams: Figure 4 As shown, the absorption of DSHP in the body increased after modeling, suggesting that these components are the key active components that exert myocardial protective effects.

[0087] It is worth noting that salvianolic acid A was not detected in normal heart tissue, but was detected in model heart tissue, suggesting that modeling can promote its transfer to heart tissue. In addition, it was found that modeling promoted the transfer of quercetin and cryptotanshinone to heart tissue, among which the detection results of salvianolic acid A were as follows: Figure 5 shown.

[0088] In summary, this example identified 22 prototype components of DSHP extract in rats, of which 10 prototype components, including lithospermic acid and its isomers, rosmarinic acid, salvianolic acid A, quercetin, dimethyl lithospermate, lithospermic acid, salvianolic acid B, dihydrotanshinone I, cryptotanshinone, and tanshinone IIA, were simultaneously present in plasma and heart tissue. The model can promote the absorption of rutin, neochlorogenic acid, lithospermic acid, salvianolic acid B, dihydrotanshinone I, cryptotanshinone, and tanshinone IIA into the blood; and promote the distribution of salvianolic acid A, quercetin, and cryptotanshinone in heart tissue. The analytical method based on ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry established in the present invention can comprehensively and systematically detect the chemical components of DSHP in vivo and in vitro, providing a reference for its subsequent pharmacological material basis, mechanism of action, and quality control research.

[0089] As described above, the basic principles, main features and advantages of the present invention are well described. The above embodiments and descriptions are merely descriptions of preferred embodiments of the present invention, and the present invention is not limited to the above embodiments. Various changes and improvements made to the technical solutions of the present invention by those skilled in the art without departing from the spirit and scope of the present invention should fall within the scope of protection determined by the present invention.

Claims

1. A method for analyzing DSHP serum migration components and cardiac tissue distribution components based on a disease model, characterized in that: The method uses ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry to measure ion currents, and obtains DSHP serum migration components and heart tissue distribution components by analyzing the ion currents. The conditions of the ultra-high performance liquid chromatography include: Chromatographic column: Waters ACQUITY HSS T3 column; Column temperature: 20-40°C; Flow rate: 0.2-0.3 mL / min; Injection volume: 2 μL; Detection wavelength: 190~410nm; Mobile phase: Phase A is acetonitrile, phase B is 0.1% formic acid water; The elution gradient conditions are: 0-2 minutes, 95% phase B; 2-5 minutes, 95%-90% phase B; 5-18 minutes, 90%-65% phase B; 18-25 minutes, 65%-30% phase B; 25-30 minutes, 30%-10% phase B; 30-34 minutes, 10%-0 phase B; 34.1-40 minutes, 95% phase B. The quadrupole time-of-flight mass spectrometer adopts electrospray ionization technology and performs detection in positive and negative ion modes respectively.

2. The method for analyzing DSHP serum migration components and cardiac tissue distribution components based on the disease model according to claim 1, characterized in that: The disease model is an animal cardiogenic disease model, and the animal cardiogenic disease model is an animal myocardial infarction model.

3. The method for analyzing DSHP serum migration components and cardiac tissue distribution components based on the disease model according to claim 2, characterized in that: The method includes administering DSHP to the myocardial infarction model, and collecting blood samples and heart tissue samples from the animals after a predetermined period of time to serve as the experimental group.

4. The method for analyzing DSHP serum migration components and cardiac tissue distribution components based on the disease model according to claim 3, characterized in that: The samples detected by ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry were blood samples and heart tissue samples from the experimental group and the control group.

5. The method for analyzing DSHP serum migration components and cardiac tissue distribution components based on the disease model according to claim 4, characterized in that: The blood sample preparation method comprises: collecting animal plasma, adding hydrochloric acid to adjust the pH value of the plasma, extracting the plasma with ethyl acetate, drying the extract with nitrogen, removing the organic solvent, and re-dissolving the plasma for use.

6. The method for analyzing DSHP serum migration components and cardiac tissue distribution components based on the disease model according to claim 4, characterized in that: The preparation method of the heart tissue sample is as follows: the heart tissue is minced and then ultrasonically homogenized, hydrochloric acid and acetonitrile are added in sequence and vortexed, and then the homogenate is extracted with ethyl acetate, the extract is blown dry with nitrogen, and then redissolved for use.

7. The method for analyzing DSHP serum migration components and cardiac tissue distribution components based on a disease model according to claim 4, characterized in that: The test samples were vortexed and mixed before testing, and after high-speed centrifugation at 12000 rpm for 10 min, the supernatant was filtered with a 0.22 μm filter membrane, and 2 μL of the filtrate was injected into ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry for analysis.

8. Use of the method for analyzing DSHP serum migration components and cardiac tissue distribution components based on a disease model according to any one of claims 1 to 7 in analyzing DSHP serum migration components in a myocardial infarction model.

9. Use of the method for analyzing the serum migration component and cardiac tissue distribution component of DSHP based on a disease model according to any one of claims 1 to 7 in analyzing the cardiac tissue distribution component of DSHP in a myocardial infarction model.

10. Use of the method for analyzing the serum migration component and cardiac tissue distribution component of DSHP based on a disease model according to any one of claims 1 to 7 in analyzing the prototype component of DSHP in rats with myocardial infarction model or in analyzing the absorption and distribution of the prototype component of DSHP in normal rats and rats with myocardial infarction model.

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