Anti-platelet active component screening method based on platelet protein ultrafiltration affinity mass spectrometry
Through a high-throughput screening method based on platelet protein ultrafiltration affinity mass spectrometry, the problem of low screening efficiency of antiplatelet active ingredients in traditional Chinese medicine is solved, and rapid and efficient screening of active ingredients is achieved, which is suitable for high-throughput screening of complex samples.
Patent Information
- Application Number
- CN202411823393.6
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-12-11
- Publication Date
- 2025-03-11
- Estimated Expiration
- 2044-12-11
AI Technical Summary
The screening efficiency of antiplatelet active ingredients in traditional Chinese medicine in prior art is low, the operation is complicated and the accuracy is low.
Using a high-throughput screening method based on ultrafiltration affinity mass spectrometry of platelet proteins, the polymer of platelet protein and potential affinity components was obtained by co-incubating platelet proteins with target compounds, and the polymer of platelet proteins and potential affinity components was obtained by ultrafiltration tubes, and then potential anti-platelet active components were screened through LC-MS detection and single-dimensional statistical analysis.
It realizes rapid and efficient screening of antiplatelet active ingredients in traditional Chinese medicine, improves the screening efficiency of active ingredients, and is suitable for high-throughput screening of active ingredients in complex samples.
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Figure CN119595795A_ABST
Abstract
Description
Technical Field
[0001] The present invention belongs to the field of high-throughput screening, and specifically relates to a method for discovering antiplatelet active components based on platelet protein ultrafiltration affinity mass spectrometry technology Background Art
[0002] Musk Heart Pill originated from Suhexiang Pill recorded in Taiping Huimin Mixture Bureau Prescription. The whole prescription is a modern Chinese medicine compound composed of seven Chinese medicines. It has the effects of aromatic warming, invigorating qi and strengthening the heart. In recent years, large-scale evidence-based studies on Chinese patent medicines for coronary heart disease have shown that Musk Heart Pill is safe and effective in treating coronary heart disease, and can significantly reduce the incidence of adverse cardiovascular events caused by antiplatelet drugs. Previous studies have also found that Musk Heart Pill can inhibit the expression of platelet activation-related proteins, so Musk Heart Pill is likely to achieve the therapeutic effect of treating coronary heart disease by inhibiting platelet activation. However, it is not clear which specific components in Musk Heart Pill have antiplatelet activity.
[0003] Establishing a high-throughput screening method to find the antiplatelet active ingredients in Shexiang Baoxin Pills is of great significance for the development of new drugs for cardiovascular diseases. At present, the conventional methods for discovering active ingredients of traditional Chinese medicine mainly rely on separation and purification and pharmacological models to separate and detect the activity of monomers. However, these methods are cumbersome to operate, have a long research cycle and low accuracy, resulting in low efficiency in discovering antiplatelet active ingredients. Ultrafiltration affinity mass spectrometry technology can provide a method for accurately and quickly identifying potential affinity ligands by identifying the interaction between drugs and biomacromolecules. It can obtain its affinity ligands from natural products through protein targets to screen the corresponding active ingredients. Most of the current antiplatelet drugs treat cardiovascular diseases by targeting platelet function receptors, and these targets are mainly located in the platelet membrane and its interior. In the past, there have been studies on the extraction of platelet membrane proteins to prepare biochromatography for antiplatelet active ingredients screening. Therefore, perhaps we can also extract platelet proteins for affinity mass spectrometry to screen antiplatelet active ingredients in natural products. Summary of the invention
[0004] In view of the above-mentioned technical problems in the prior art, the present invention provides a method for screening active ingredients based on platelet protein ultrafiltration affinity mass spectrometry. The method for screening active ingredients based on platelet protein ultrafiltration affinity mass spectrometry is intended to solve the problem of low efficiency in screening antiplatelet active ingredients in traditional Chinese medicine in the prior art.
[0005] The present invention provides a method for screening antiplatelet active components based on platelet protein ultrafiltration affinity mass spectrometry, comprising the following steps:
[0006] S1. Prepare a test solution containing the target compound;
[0007] S2, extracting platelet protein solution from the blood sample;
[0008] S3, using S1 test solution and S2 platelet protein solution to incubate;
[0009] S4, using an ultrafiltration tube to elute the co-incubation solution in S3, thereby obtaining a polymer of platelet protein and the platelet protein affinity small molecule in the test melt;
[0010] S5, dissociating the small molecules in S4 from the platelet protein polymer to obtain a potential active component solution; S6, detecting the dissociated small molecules in S5 using LC-MS;
[0011] S7. The mass spectrometry data obtained in S6 are screened by unidimensional statistical analysis of the control group and the platelet protein affinity group data to screen small molecules in the test solution that have affinity with platelet proteins, that is, potential antiplatelet active components.
[0012] S8, platelet aggregation experiment was used to verify the potential antiplatelet active components in S7 in vitro.
[0013] Further, step S1 is specifically as follows:
[0014] Use an appropriate extraction method to extract the components in the target sample as comprehensively as possible, evaporate the solvent, and re-dissolve with buffer (25mM Tris-HCl (pH 8.0), 150mM NaCl, 1mM dithiothreitol and 10mM MgCl2) to obtain the test solution for use.
[0015] Further, step S2 is specifically as follows:
[0016] 1) Whole blood extraction: SD rats were anesthetized with sodium pentobarbital at a mass percentage concentration of 2%, and blood was collected from the abdominal aorta of the SD rats. The whole blood was placed in a centrifuge tube containing 5% ethylenediaminetetraacetic acid at a mass percentage concentration;
[0017] 2) Platelet-rich plasma extraction: a blood sample anticoagulated with 5% ethylenediaminetetraacetic acid by mass is centrifuged at room temperature, and the upper layer after centrifugation is platelet-rich plasma, and the lower layer is precipitated white blood cells and red blood cells;
[0018] 3) Platelet extraction: transfer platelet-rich plasma into a new centrifuge tube, centrifuge, discard the upper plasma, and precipitate pure platelets;
[0019] 4) Platelet protein extraction: Pure platelets are resuspended in a buffer solution, and an appropriate amount of 1% by mass concentration of phenylmethylsulfonyl fluoride protease inhibitor is added. After ultrasonic rupture, a platelet protein suspension is obtained for use.
[0020] Further, step S3 is specifically as follows:
[0021] The platelet protein suspension was mixed with the test solution in a volume percentage of 1:1 and incubated at 37°C.
[0022] Further, step S4 is specifically as follows:
[0023] 1) Preparation of ultrafiltration tube: Use a 10kDa ultrafiltration centrifuge tube, add ultrapure water before use, and allow the water to completely pass through the membrane. Precool in an ice bath or refrigerator for 10 minutes;
[0024] 2) Add the incubation solution to the ultrafiltration tube: operate on ice, pour out the water, and add the above S3 incubation solution to the 10kDa ultrafiltration tube;
[0025] 3) Centrifugation and washing: centrifuge the 10 kDa ultrafiltration centrifuge tube with the incubation solution at 4°C, stop the centrifugation, wash the mixture with 50 mM ammonium acetate solution and centrifuge again;
[0026] Further, step S5 is specifically as follows:
[0027] The S4 mixture was transferred to a new EP tube, and the compounds bound to platelet proteins were extracted using methanol. The tube was then centrifuged at 4°C, the protein precipitate was discarded, and the tube was dried and concentrated using a nitrogen stream, and then re-dissolved for testing.
[0028] Further, step S6 is specifically as follows:
[0029] The test solution is obtained by high-resolution mass spectrometry detection S5.
[0030] Further, step S7 is specifically as follows:
[0031] After unidimensional statistical analysis using Progenesis QI analysis software, the differential ions between the control group and the affinity group were screened to obtain potential antiplatelet active components:
[0032] Further, step S8 is specifically as follows:
[0033] In order to verify the antiplatelet activity of the potential components screened, antiplatelet aggregation experiments were performed on them.
[0034] The present invention uses a high-throughput screening method for antiplatelet active ingredients based on platelet protein ultrafiltration affinity mass spectrometry to screen antiplatelet active ingredients in traditional Chinese medicine. Antiplatelet drugs usually interfere with the aggregation or activation process of platelet surface proteins or specific proteins in platelets by binding to them, thereby achieving the purpose of inhibiting platelet function and reducing thrombosis. Platelet aggregation is a key process in thrombosis, so finding small molecule drugs that can effectively interfere with platelet activity is an important direction for developing antiplatelet drugs. Ultrafiltration affinity separation technology uses platelet protein as an affinity phase to separate antiplatelet active ingredients in traditional Chinese medicine ingredients by selectively adsorbing compounds that are affinity with platelet protein. This process achieves efficient screening of active ingredients through the binding characteristics of platelet protein with candidate compounds. Combined with mass spectrometry technology, traditional Chinese medicine ingredients that bind to platelet protein can be accurately identified. Mass spectrometry can provide molecular mass information and structural characteristics of each compound, helping researchers identify potential antiplatelet active ingredients. At the same time, mass spectrometry technology has high sensitivity and high resolution, and can process complex libraries of traditional Chinese medicine components and quickly screen out molecules with potential activity. Therefore, the advantages of the high-throughput screening method for antiplatelet active components based on platelet protein ultrafiltration affinity mass spectrometry enable it to effectively identify potential antiplatelet components in traditional Chinese medicine, providing new ideas and tools for the discovery of antiplatelet drugs.
[0035] The present invention has the following advantages over the prior art:
[0036] 1. The present invention can quickly screen and obtain potential antiplatelet active ingredients, thereby improving the screening efficiency of active ingredients.
[0037] 2. The present invention is based on the ultrafiltration affinity mass spectrometry technology of platelet proteins and is suitable for high-throughput screening of active ingredients in complex samples.
[0038] For better understanding and implementation, the present invention is described in detail below with reference to the accompanying drawings. BRIEF DESCRIPTION OF THE DRAWINGS
[0039] Figure 1 Workflow diagram of the high-throughput screening method for antiplatelet active components based on platelet protein ultrafiltration affinity mass spectrometry.
[0040] Figure 2 Method validation of antiplatelet positive drug affinity mass spectrometry for platelet proteins.
[0041] Figure 3 Screening of components with affinity to platelet proteins in Musk Heart Pills.
[0042] Figure 4 Platelet aggregation assay validation of representative active ingredients. DETAILED DESCRIPTION
[0043] Embodiment 1:
[0044] A method for screening antiplatelet active components based on platelet protein ultrafiltration affinity mass spectrometry.
[0045] See also Figure 1 , which is a workflow diagram of the high-throughput screening method for anti-platelet active components based on platelet protein ultrafiltration affinity mass spectrometry. First, the test solution of the target sample is extracted; then the platelet protein solution is extracted from the blood sample; then the test solution is mixed with the platelet protein solution and co-incubated; then the polymer of platelet protein and potential affinity components in the test solution is obtained by elution through an ultrafiltration tube; then the platelet protein is dissociated by an organic solvent to obtain a potential active component solution; the obtained control solution and the potential active component solution are further subjected to mass spectrometry detection; then a single-dimensional statistical analysis is performed to screen and identify the active components. Finally, the present invention also selects several representative components among the potential active components for platelet aggregation experimental verification.
[0046] This example conducts high-throughput screening of potential active ingredients in Shexiang Baoxin Pills, specifically comprising the following steps:
[0047] (1) Preparation of buffer (25 mM Tris-HCl, 150 mM NaCl, 1 mM dithiothreitol, and 10 mM MgCl2): dilute 1 mL of 1 M Tris-HCl 40-fold with ultrapure water to 40 mL, add 348 mg NaCl, 6.0 mg dithiothreitol, and 83.4 mg MgCl2 6H2O;
[0048] Preparation of 150 mM ammonium acetate solution: Take 40 mL of ultrapure water and add 461.1 mg of ammonium acetate.
[0049] (2) Preparation of Musk Heart Pill Solution: Take 4 Musk Heart Pills (25 mg each) and add 1 mL of methanol for ultrasonic extraction for 0.5 h. Centrifuge at 13,000 rpm for 10 min at 4°C. Take 100 μL of the supernatant and dissolve it in 10 mL of buffer (25 mM Tris-HCl, 150 mM NaCl, 1 mM dithiothreitol and 10 mM MgCl2).
[0050] (3) Preparation of positive drug solution (positive drug solution is a solution of two antiplatelet drug standards commonly used on the market, which are compounds known to have affinity with platelet proteins. The positive drug solution provided here is to verify the feasibility of the method with compounds with known affinity. The control group solution is a buffer solution without platelet proteins added).
[0051] 0.02 mM positive drug stock solutions of ticagrelor and tirofiban were prepared with 100% methanol respectively, 100 μL of each of the positive drug stock solutions were taken and mixed, and then diluted with 10 mL of buffer (containing 25 mM Tris-HCl, 150 mM NaCl, 1 mM dithiothreitol and 10 mM MgCl2) to obtain positive drug solution preparation solutions.
[0052] (4) Platelet protein extraction: Take an appropriate amount of SD male rats and anesthetize them with sodium pentobarbital at a mass percentage concentration of 2%, and then collect blood from the abdominal aorta of SD male rats. Place the whole blood in a centrifuge tube containing 5% ethylenediaminetetraacetic acid at a mass percentage concentration. Centrifuge at 1200rpm for 10 minutes, take the upper platelet-rich plasma after centrifugation, and centrifuge at 3500rpm for 5 minutes. Discard the upper plasma and take the precipitate as platelets. Resuspend the platelets in a buffer (containing 25mM Tris-HCl, 150mMNaCl, 1mM dithiothreitol and 10mM MgCl2), add an appropriate amount of 1% phenylmethylsulfonyl fluoride protease inhibitor, and mix after ultrasonic rupture for 30 minutes to obtain a platelet protein solution.
[0053] (5) Co-incubation of the test solution with platelet protein: Take 100 μL of platelet protein solution and 100 μL of positive drug solution and incubate them at 37°C for 30 min for method validation of the positive drug.
[0054] Six portions of 100 μL platelet protein solution were taken and incubated with 100 μL Musk Heart Pill solution at 37°C for 30 min as affinity group samples. At the same time, six portions of 100 μL control buffer solution (containing 25 mM Tris-HCl, 150 mM NaCl, 1 mM dithiothreitol and 10 mM MgCl2) were taken and incubated with 100 μL Musk Heart Pill solution at 37°C for 30 min as control group samples.
[0055] (6) Ultrafiltration tube elution: Add the above incubation solution to a 0.5 mL 10 kDa ultrafiltration tube. Centrifuge the ultrafiltration centrifuge tube and the incubation solution at 13,000 rpm at 4°C for 10 min. Wash the mixture five times with 200 μL of 150 mM ammonium acetate solution and centrifuge to 100 μL.
[0056] (7) Dissociation of active ingredients: The above concentrates were transferred to new EP tubes respectively, and the active ingredients bound to platelet proteins were extracted using 300 μL of methanol. The tubes were then centrifuged at 13,000 rpm at 4°C for 10 min, and the supernatant was collected for testing.
[0057] (8) Chromatographic conditions: The chromatographic separation of the samples was performed using a Waters Acquity UPLC system. The chromatographic column was a Waters Acquity UPLC HSS T3 (2.1 mm × 100 mm, 1.8 μm) column, the column temperature was 40°C, the mobile phase consisted of water (A) containing 0.1% formic acid by volume and acetonitrile (B), the flow rate was 0.3 mL / min, and the gradient elution program was as follows: 0-1 min, 5%-5% B; 1-13 min, 5%-100% B; 13-15 min, 100%-100% B; 15-15.1 min, 100%-5% B; 15.1-20 min, 5%-5% B; the injection volume was 2 μL.
[0058] (9) Mass spectrometry conditions: Mass spectrometry detection was performed on a SYNAPT G2-Si HDMS system. Data were collected in the fastDDA mode in both positive and negative ion modes. The mass spectrometry conditions were set as follows: drying gas flow rate, 800 L / h; drying gas temperature, 400 °C; ion source temperature, 120 °C; capillary voltage, 2.5 kV (positive ion mode), 2.0 kV (negative ion mode); cone voltage, 40 V, source offset, 80 V; cone gas flow rate, 50 L / h; mass range, 50-1500 Da; MS scan rate, 0.2 s. The parameters of the fast DDA mode were set as follows: low-mass collision energy was 10-40 V, and high-mass collision energy was 40-120 V. A 200 pg / μL leucine enkephalin solution was used for real-time data calibration. Data acquisition was obtained by MassLynx V4.1.
[0059] (10) Method validation: In order to verify the reliability of the platelet protein affinity mass spectrometry method, two antiplatelet positive drugs were used for affinity incubation with platelet proteins, and the affinity of the positive drugs with platelet proteins was verified by comparing the abundance difference between the positive drugs and the control group.
[0060] (11) Unidimensional statistical analysis: The control group and affinity group data sets were peak aligned and extracted, and a characteristic ion quantification table containing all data grouping information, retention time, m / z, and peak area was exported. The characteristic ion quantification table was corrected and normalized by StatTarget software and then imported into the MetaboAnalyst platform to screen potential platelet affinity-active ions with the criteria of p < 0.05 and FC > 2, and then identified.
[0061] (12) Platelet aggregation test verification: Platelet aggregation test verification of two representative active ingredients (cinnamic acid-CA and taurocholic acid-TCA) was performed by platelet aggregation instrument turbidimetry. First, when preparing TB solution, 504.6 mg NaHCO3, 4.032 g NaCl, 108 mg KCl, 203 mg MgCl2·6H2O, 32.76 mg NaH2PO4·H2O, and 1192 mg 4-hydroxyethylpiperazineethanesulfonic acid were dissolved in 500 mL ultrapure water, and the pH value was adjusted to 7.4 after preparation, and stored at 4°C. Then, blood from the abdominal aorta of SD rats was collected. The collected blood was added with physiological saline in a volume ratio of 1:1 and adenosine triphosphate diphosphodiesterase in a volume ratio of 400:1, and centrifuged at 870 rpm for 20 minutes to obtain platelet-rich plasma. Subsequently, ethylenediaminetetraacetic acid was added and centrifuged at 1900 rpm for 10 minutes to obtain platelet-poor plasma. After discarding the supernatant, the platelets settle at the bottom of the aggregation tube. To prepare the washed platelet solution, TB solution was mixed with glucose in a 1:1 volume ratio. Subsequently, platelet aggregation was detected by platelet aggregometer turbidimetry. In the aggregometer preheated to 37°C, 300 μL of washed platelet solution was added, followed by 1.5 μL of active ingredient solution. After incubation for 5 minutes, the aggregometer was adjusted to the baseline, the agonist ADP was added, and the changes in the aggregation curve were recorded.
[0062] Figure 2 This example verifies the platelet protein affinity mass spectrometry method for platelet-positive drugs: the two positive drugs, ticagrelor and tirofiban, both have significant affinity with platelet proteins, thus confirming the feasibility of high-throughput screening of antiplatelet active ingredients in traditional Chinese medicine based on platelet protein affinity mass spectrometry.
[0063] Figure 3 This example is the screening of components with affinity to platelet proteins in Musk Heart Pills: the control group and affinity group data sets were subjected to unidimensional statistical analysis, and the components with affinity to platelets in Musk Heart Pills were screened and identified using the criteria of p<0.05, FC>2, and ultimately 17 potential active ingredients were screened out from Musk Heart Pills.
[0064]
[0065]
[0066] Figure 4 The platelet aggregation experiment for the representative active ingredients of this example was verified: two representative ingredients, cinnamic acid (CA) and taurocholic acid (TCA), were selected from 17 screened potential antiplatelet active ingredients for platelet aggregation experiments. The results showed that within a certain concentration range, cinnamic acid and taurocholic acid could inhibit platelet aggregation to a certain extent.
[0067] In summary, this embodiment adopts a high-throughput screening method for antiplatelet active ingredients based on platelet protein ultrafiltration affinity mass spectrometry to screen the antiplatelet active ingredients in Musk Heart Pills. Antiplatelet drugs often achieve antiplatelet effects by binding to platelet proteins. Therefore, by screening the antiplatelet active ingredients in Musk Heart Pills that have affinity with platelet proteins, potential antiplatelet active ingredients can be quickly and high-throughput screened from the Musk Heart Pills composed of complex components, and finally the platelet aggregation experiment can be used to verify the activity of the screened active ingredients.
[0068] The above-mentioned embodiment only expresses one implementation mode of the present invention, and its description is relatively specific and detailed, but it cannot be understood as limiting the scope of the invention patent. It should be pointed out that for ordinary technicians in this field, several modifications and improvements can be made without departing from the concept of the present invention, which all belong to the protection scope of the present invention.
Claims
1. A method for screening antiplatelet active ingredients based on platelet protein ultrafiltration affinity mass spectrometry, characterized in that: The following steps are involved: S1. Prepare a test solution containing the target compound; S2, extracting platelet protein solution from the blood sample; S3, using S1 test solution and S2 platelet protein solution to incubate; S4, using an ultrafiltration tube to elute the co-incubation solution in S3, thereby obtaining a polymer of platelet protein and the platelet protein affinity small molecule in the test melt; S5, dissociating the small molecules in S4 from the platelet protein polymer to obtain a potential active ingredient solution; S6, using LC-MS to detect the small molecules after dissociation in S5; S7. The mass spectrometry data obtained in S6 are screened by unidimensional statistical analysis of the control group and the platelet protein affinity group data, thereby screening small molecules in the test solution that have affinity with platelet proteins, that is, potential antiplatelet active components. S8, platelet aggregation experiment was used to verify the potential antiplatelet active components in S7 in vitro.
2. The method for screening antiplatelet active components based on platelet protein ultrafiltration affinity mass spectrometry according to claim 1, characterized in that: Step S1 is specifically as follows: The extracted target sample is re-dissolved in a buffer solution containing 25 mM Tris-HCl, 150 mM NaCl, 1 mM dithiothreitol and 10 mM MgCl2, the components in the target sample are extracted, and the solvent is evaporated to obtain a test solution for use.
3. The method for screening antiplatelet active components based on platelet protein ultrafiltration affinity mass spectrometry according to claim 1, characterized in that: Step S2 is specifically as follows: 1) Whole blood extraction: SD rats were anesthetized with sodium pentobarbital at a mass percentage concentration of 2%, and blood was collected from the abdominal aorta of the SD rats. The whole blood was placed in a centrifuge tube containing 5% ethylenediaminetetraacetic acid at a mass percentage concentration; 2) Platelet-rich plasma extraction: a blood sample anticoagulated with 5% ethylenediaminetetraacetic acid by mass is centrifuged at room temperature, and the upper layer after centrifugation is platelet-rich plasma, and the lower layer is precipitated white blood cells and red blood cells; 3) Platelet extraction: transfer platelet-rich plasma into a new centrifuge tube, centrifuge, discard the upper plasma, and precipitate pure platelets; 4) Platelet protein extraction: The pure platelets are resuspended in a buffer solution, and a 1% by weight concentration of phenylmethylsulfonyl fluoride protease inhibitor is added. After ultrasonic rupture, a platelet protein suspension is obtained for use.
4. The method for screening antiplatelet active components based on platelet protein ultrafiltration affinity mass spectrometry according to claim 1, characterized in that: Step S3 is as follows: The platelet protein suspension was mixed with the test solution in a volume percentage of 1:1 and incubated at 37°C.
5. The method for screening antiplatelet active components based on platelet protein ultrafiltration affinity mass spectrometry according to claim 1, characterized in that: Step S4 is specifically as follows: 1) Preparation of ultrafiltration tube: Use a 10kDa ultrafiltration tube, add ultrapure water before use, and allow the water to completely pass through the membrane. Precool in an ice bath or refrigerator for 10 minutes; 2) Add the incubation solution to the ultrafiltration tube: operate on ice, pour out the water, and add the above S3 incubation solution to the 10kDa ultrafiltration tube; 3) Centrifugation and washing: centrifuge the 10 kDa ultrafiltration centrifuge tube and the incubation solution at 4°C, stop the centrifugation, wash the mixture with 50 mM ammonium acetate solution, and then centrifuge.
6. The method for screening antiplatelet active components based on platelet protein ultrafiltration affinity mass spectrometry according to claim 1, characterized in that: Step S5 is specifically as follows: The S4 mixture was transferred to a new EP tube, and the compounds bound to platelet proteins were extracted using methanol. The tube was then centrifuged at 4°C, the protein precipitate was discarded, and the tube was dried and concentrated using a nitrogen stream, and then re-dissolved for testing.
7. The method for screening antiplatelet active components based on platelet protein ultrafiltration affinity mass spectrometry according to claim 1, characterized in that: Step S6 is specifically as follows: The solution to be tested is obtained by high-resolution mass spectrometry.
8. The method for screening antiplatelet active components based on platelet protein ultrafiltration affinity mass spectrometry according to claim 1, characterized in that: Step S7 is specifically as follows: After unidimensional statistical analysis using Progenesis QI analysis software, the differential ions between the control group and the platelet protein affinity group were screened to obtain potential antiplatelet active components.
9. The method for screening antiplatelet active components based on platelet protein ultrafiltration affinity mass spectrometry according to claim 1, characterized in that: Step S8 is specifically as follows: In order to verify the antiplatelet activity of the potential components screened, antiplatelet aggregation experiments were performed on them.