Platelet aggregation function detection reagent, preparation method and detection method thereof
By preparing microencapsulated arachidonic acid reagent, the problem of easy oxidation after freeze-drying was solved, achieving high stability and high sensitivity of the reagent, which is suitable for detection by fully automated thromboembolic instruments.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- SUZHOU INST OF BIOMEDICAL ENG & TECH CHINESE ACADEMY OF SCI
- Filing Date
- 2023-06-26
- Publication Date
- 2026-06-23
AI Technical Summary
Existing platelet aggregation function test reagents are easily oxidized after lyophilization, resulting in reduced activity and failing to meet the testing requirements of fully automated thromboembolic analyzers. Furthermore, improper use of lyophilized reagents leads to waste.
Microencapsulated arachidonic acid reagents were prepared using arachidonic acid as the core material, cyclodextrin and gum arabic as the wall material, and emulsifiers, antioxidants, freeze-drying protectants, preservatives and buffer salts were added. The reagents were then processed by low-temperature freeze-drying.
It improves the stability and sensitivity of arachidonic acid, ensures the accuracy and precision of detection results, reduces waste, and is suitable for high-throughput detection in fully automated thromboembolic analyzers.
Smart Images
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Abstract
Description
Technical Field
[0001] This invention relates to the field of coagulation detection technology, and in particular to a platelet aggregation function detection reagent, its preparation method and detection method. Background Technology
[0002] Cardiovascular and cerebrovascular diseases have become the leading cause of death and disability in humans. The pathological basis of most cardiovascular and cerebrovascular diseases is abnormal platelet activation leading to arterial thrombosis. Aspirin, a common antiplatelet drug, is widely used and is a common medication for reducing the risk of cardiovascular and cerebrovascular diseases. However, due to individual differences and the complexity and diversity of diseases, some patients do not respond well to conventional doses of aspirin and still experience thrombotic events, a condition known as aspirin resistance. Currently, thromboelastography has become the main method for evaluating the efficacy of aspirin.
[0003] Thromboelastography is used to monitor the efficacy of antiplatelet drugs such as aspirin through platelet aggregation function assays. These assays include kaolin activator, fibrin activator (F), and platelet activators such as arachidonic acid (AA) and adenosine diphosphate (ADP). When AA and ADP are added, the platelet thromboxane A2 or ADP receptor channels are activated, leading to platelet aggregation. Since aspirin blocks the A2 receptor channel, adding AA activates platelets not inhibited by aspirin, thus enabling monitoring of aspirin efficacy. The main ingredient in the AA activator, arachidonic acid, is an unsaturated fatty acid that is easily oxidized under light and high temperatures, even at room temperature through free radical chain reactions. Without proper treatment, activation will fail, directly affecting the test results and failing to reflect the true drug effect.
[0004] Typically, AA reagents are used for platelet activation in lyophilized form. Some are lyophilized in vials and reconstituted with water before testing. Other manufacturers lyophilize them in test cups, which, while more convenient, results in higher packaging costs and requires significant storage space for high-throughput automated elastography equipment. Therefore, commercially available platelet activation reagents are primarily lyophilized in vials. These lyophilized reagents usually need to be used within 8 hours of reconstitution; otherwise, reduced activity or oxidation can lead to inaccurate test results. Reagents reconstituted beyond 8 hours must be discarded, resulting in substantial waste.
[0005] Therefore, there is an urgent need for arachidonic acid reagents with high sensitivity, good antioxidant properties, and good open-bottle stability to assess platelet function in patients after medication, thereby effectively detecting patients with platelet drug resistance, providing clinical reference, and also meeting the requirements of fully automated thromboelastography for detecting platelet aggregation function.
[0006] Patent CN113156144A (Zhengzhou Puwan) discloses an arachidonic acid reagent for platelet aggregation function, comprising sodium arachidonic acid salt and sugars and polymeric lyophilization protectants. Patent CN109884326B (Shenzhen Youdi) discloses a platelet aggregation function detection kit, comprising fibrin activator, intrinsic coagulation activator, and platelet inducer, each reagent stored in a sealed, lyophilized sample cup. However, this kit does not mention the processing of arachidonic acid and does not contain antioxidants, only bovine plasma albumin and conventional buffer solutions. Patent CN104914254A (Beijing Lepu) discloses a platelet detection method, which involves preparing a mixed reagent of fibrin activator and platelet activator, adding an antioxidant, dispensing it into sample cups, and then lyophilizing the sample cups in vials. However, commercially available arachidonic acid microcapsules typically use spray drying, resulting in significant raw material waste and insufficient activity to activate platelets. Summary of the Invention
[0007] The technical problem to be solved by the present invention is to provide a platelet aggregation function detection reagent, its preparation method and detection method, in order to address the shortcomings of the prior art.
[0008] To solve the above-mentioned technical problems, the technical solution adopted by the present invention is as follows: In its first aspect, the present invention provides a platelet aggregation function detection reagent. This detection reagent is a microencapsulated arachidonic acid detection reagent prepared with arachidonic acid as the core material, cyclodextrin and gum arabic as the wall material, and with the addition of emulsifiers, antioxidants, lyophilization protectants, preservatives, buffer salts, and water as auxiliary components. The detection reagent comprises the following raw material components by mass percentage:
[0009] Arachidonic acid: 0.1-1%; β-cyclodextrin: 0.1-1%; gum arabic: 0.1-1%; emulsifier: 0.1-2%; antioxidant: 0.01-0.1%; freeze-drying protectant: 0.1-10%; preservative: 0.01-0.1%; the remainder is buffer salt and water.
[0010] Preferably, the emulsifier is any one or a mixture of casein, whey protein, monoglyceride fatty acid glycerides, and gelatin.
[0011] Preferably, the antioxidant is one or a mixture of vitamin C, vitamin E, sodium thiosulfate, tea polyphenols, and dithiothreitol.
[0012] Preferably, the freeze-drying protectant is one or more of PEG8000, Proneix, BSA, lactose, trehalose, and sucrose.
[0013] Preferably, the cyclodextrin is β-cyclodextrin, the preservative is Proclin, and the buffer salt is one of phosphate buffer, Tris buffer, and MES buffer, with a concentration of 5-50 mM.
[0014] The emulsifier is casein, the antioxidant is a mixture of sodium thiosulfate and dithiothreitol, the lyophilization protectant is a mixture of Proneix and lactose, and the buffer salt is phosphate buffer.
[0015] A second aspect of the present invention provides a method for preparing the platelet aggregation function detection reagent as described above, comprising the following steps:
[0016] S1. Take cyclodextrin and gum arabic, dissolve them thoroughly in buffer salt solution, cool to room temperature, and then add emulsifier, antioxidant, freeze-drying protectant and preservative in sequence, stirring until completely dissolved.
[0017] S2. Preparation of pre-emulsion: Arachidonic acid is added to the product obtained in step S1 under stirring to obtain a pre-emulsion;
[0018] S3. Emulsification: The pre-emulsion obtained in step S1 is fully emulsified using a homogenizer, then packaged and pre-frozen.
[0019] S4. Freeze-drying. After freeze-drying, vacuum stopper the sample to obtain the platelet aggregation function detection reagent. Preferably, the preparation method of the platelet aggregation function detection reagent includes the following steps:
[0020] S1. Take cyclodextrin and gum arabic, dissolve them thoroughly in a buffer salt solution at 60-70℃, cool to room temperature, and then add emulsifier, antioxidant, freeze-drying protectant and preservative in sequence. Stir magnetically until completely dissolved.
[0021] S2. Preparation of pre-emulsion: Arachidonic acid is added to the product obtained in step S1 under magnetic stirring in an environment below 16°C to obtain a pre-emulsion;
[0022] S3. Emulsification: In an environment below 16°C, the pre-emulsion obtained in step S1 is emulsified using a homogenizer at 8000-16000 r / min for 5-30 min, then dispensed and pre-frozen at -80°C for 2-6 h.
[0023] S4. The vacuum degree should be controlled at 0.01-1.0 mbar for lyophilization for 10-24 hours. After lyophilization, the platelet aggregation function detection reagent is obtained by vacuum plugging.
[0024] Preferably, the preparation method of the platelet aggregation function detection reagent includes the following steps:
[0025] S1. Take 0.2 wt% cyclodextrin and 0.5% gum arabic and add them to 20 mM phosphate buffer. Dissolve them completely at 70°C and cool to room temperature. Then add 1 wt% casein, 0.08 wt% dithiothreitol, 0.05 wt% sodium thiosulfate, 2 wt% Proneix, 4 wt% lactose and 0.04 wt% Proclin in sequence and stir magnetically until completely dissolved.
[0026] S2. Preparation of pre-emulsion: Add 0.5 wt% arachidonic acid to the product obtained in step S1 under magnetic stirring in an environment below 16°C to obtain a pre-emulsion;
[0027] S3. Emulsification: In an environment below 16°C, the pre-emulsion obtained in step S1 is emulsified for 12 min at 10000 r / min using a homogenizer, then dispensed and pre-frozen at -80°C for 4 h.
[0028] S4. The vacuum degree should be controlled at 0.2 mbar for 18 hours for freeze drying. After freeze drying, the platelet aggregation function test reagent is obtained by vacuum plugging.
[0029] A third aspect of the present invention provides a method for detecting platelet aggregation function, comprising the following steps:
[0030] 1) Redissolve the platelet aggregation function test reagent as described above in a specified volume of deionized water, let stand for 10-30 minutes, and obtain the test reagent solution;
[0031] 2) Add fibrin activator, the test reagent solution obtained in step 1), and a sample containing aspirin to the test cup respectively, and perform thromboelastography test. The obtained thromboelastography MA value is the arachidonic acid cup clot strength.
[0032] Preferably, the aspirin-containing sample is a blood sample from a person who has taken aspirin, or a sample obtained by mixing blood from a person who has not taken aspirin with 0.001-0.1 mg / mL of aspirin.
[0033] The beneficial effects of this invention are:
[0034] This invention uses arachidonic acid as the core material and cyclodextrin and gum arabic as the wall material to moderately encapsulate arachidonic acid. After thorough emulsification and homogenization with a suitable emulsifier, protective agents, antioxidants, and preservatives are added. Microencapsulated arachidonic acid, i.e., a platelet aggregation function detection reagent, is obtained by low-temperature freeze-drying. This method not only ensures that the activity of arachidonic acid remains unchanged before and after freeze-drying, but also improves the stability of arachidonic acid.
[0035] The freeze-drying process for preparing microencapsulated arachidonic acid provided by this invention can reduce changes in the physicochemical properties of arachidonic acid caused by changes in the external environment without altering its platelet-activating activity, thereby improving storage stability and further enhancing the sensitivity and open-bottle stability of arachidonic acid reagents. Detailed Implementation
[0036] The present invention will be further described in detail below with reference to embodiments, so that those skilled in the art can implement it based on the description.
[0037] It should be understood that terms such as “having,” “comprising,” and “including” as used herein do not exclude the presence or addition of one or more other elements or combinations thereof.
[0038] Unless otherwise specified, the experimental methods used in the following examples are conventional methods. Unless otherwise specified, the materials and reagents used in the following examples are commercially available. For examples where specific conditions are not specified, conventional conditions or conditions recommended by the manufacturer are followed. For reagents or instruments whose manufacturers are not specified, they are all commercially available products.
[0039] This invention provides a platelet aggregation function detection reagent. The reagent is a microencapsulated arachidonic acid detection reagent prepared with arachidonic acid as the core material, cyclodextrin and gum arabic as wall materials, and with the addition of emulsifiers, antioxidants, lyophilization protectants, preservatives, buffer salts, and water as auxiliary components. The reagent comprises the following raw material components by mass percentage:
[0040] Arachidonic acid: 0.1-1%; β-cyclodextrin: 0.1-1%; gum arabic: 0.1-1%; emulsifier: 0.1-2%; antioxidant: 0.01-0.1%; freeze-drying protectant: 0.1-10%; preservative: 0.01-0.1%; the remainder is buffer salt and water.
[0041] In a preferred embodiment, the emulsifier is any one or a mixture of casein, whey protein, monoglycerides of fatty acids, and gelatin. The antioxidant is any one or a mixture of vitamin C, vitamin E, sodium thiosulfate, tea polyphenols, and dithiothreitol. The freeze-drying protectant is any one or a mixture of PEG8000, Proneix, BSA, lactose, trehalose, and sucrose.
[0042] In a further preferred embodiment, the cyclodextrin is β-cyclodextrin, the preservative is Proclin, the buffer salt is one of phosphate buffer, Tris buffer, and MES buffer, and the concentration of the buffer salt is 5-50 mM; the emulsifier is casein, the antioxidant is a mixture of sodium thiosulfate and dithiothreitol, the lyophilization protectant is a mixture of Proneix and lactose, and the buffer salt is phosphate buffer.
[0043] This invention also provides a method for preparing the above-mentioned platelet aggregation function detection reagent, comprising the following steps:
[0044] S1. Take cyclodextrin and gum arabic, dissolve them thoroughly in a buffer salt solution at 60-70℃, cool to room temperature, and then add emulsifier, antioxidant, freeze-drying protectant and preservative in sequence. Stir magnetically until completely dissolved.
[0045] S2. Preparation of pre-emulsion: Arachidonic acid is added to the product obtained in step S1 under magnetic stirring in an environment below 16°C to obtain a pre-emulsion;
[0046] S3. Emulsification: In an environment below 16°C, the pre-emulsion obtained in step S1 is emulsified using a homogenizer at 8000-16000 r / min for 5-30 min, then dispensed and pre-frozen at -80°C for 2-6 h.
[0047] S4. The vacuum degree should be controlled at 0.01-1.0 mbar for lyophilization for 10-24 hours. After lyophilization, the platelet aggregation function test reagent is obtained by vacuum plugging.
[0048] This invention also provides a method for detecting platelet aggregation function, comprising the following steps:
[0049] 1) Redissolve the platelet aggregation function test reagent as described in any one of claims 1-5 in a specified volume of deionized water, let stand for 10-30 minutes, and obtain the test reagent solution;
[0050] 2) Add fibrin activator, the test reagent solution obtained in step 1), and a sample containing aspirin to the test cup respectively, and perform thromboelastography test. The obtained thromboelastography MA value is the arachidonic acid cup clot strength.
[0051] Among them, the aspirin-containing samples are blood samples from people who have taken aspirin, or blood samples from people who have not taken aspirin mixed with 0.001-0.1 mg / mL of aspirin.
[0052] The above is the overall concept of the present invention. Detailed embodiments and comparative examples are provided below.
[0053] Example 1
[0054] A platelet aggregation function detection reagent, the preparation method of which includes the following steps:
[0055] S1. Take 0.2 wt% cyclodextrin and 0.5% gum arabic and add them to 20 mM phosphate buffer. Dissolve them completely at 70°C and cool to room temperature. Then add 1 wt% casein, 0.08 wt% dithiothreitol, 0.05 wt% sodium thiosulfate, 2 wt% Proneix, 4 wt% lactose and 0.04 wt% Proclin in sequence and stir magnetically until completely dissolved.
[0056] S2. Preparation of pre-emulsion: 0.5 wt% arachidonic acid is slowly added to the product obtained in step S1 under magnetic stirring in an environment below 16°C to obtain a pre-emulsion;
[0057] S3. Emulsification: In an environment below 16°C, the pre-emulsion obtained in step S1 is emulsified for 12 minutes at 10000 r / min using a homogenizer, and then dispensed into vials (the dispensing amount depends on the specific specifications and models), and placed at -80°C for pre-freezing for 4 hours.
[0058] S4. Transfer the pre-frozen sample from S4 to a lyophilization chamber and run the drying program: the vacuum degree should be controlled at 0.2 mbar and the running time is 18 h. After the lyophilization is completed, vacuum stopper the sample to obtain the platelet aggregation function detection reagent.
[0059] Comparative Example 1
[0060] To verify the stability of microencapsulated arachidonic acid after homogenization with added wall material and emulsifier, this comparative example was set up as lyophilized arachidonic acid reagent with only antioxidant, protective agent, and preservative added. The specific steps are as follows:
[0061] Add 0.08wt% dithiothreitol, 0.05wt% sodium thiosulfate, 2wt% Proneix, 4wt% lactose, and 0.04wt% Proclin sequentially to S1 and 20mM PBS buffer, and stir until completely dissolved.
[0062] S2. Place the solution obtained in S1 in an ice bath, control the temperature of the S1 solution to be less than 16℃, and slowly add 0.5wt% arachidonic acid while magnetically stirring, and mix thoroughly.
[0063] S3. Dispense the contents of S2 into vials (the dispensing amount depends on the specific specifications and models), and pre-freeze them in an ultra-low temperature freezer at -80℃ for 4 hours;
[0064] S4. Transfer the pre-frozen sample from S3 to a freeze dryer and run the drying program: control the vacuum at 0.2 mbar and run for 18 hours. After freeze drying, press the stopper under vacuum to obtain the platelet aggregation function detection reagent.
[0065] Example 2: Accuracy Test of Test Reagents Before and After Lyophilization
[0066] For the test reagents obtained in Example 1 and Comparative Example 1 before and after lyophilization (the product of step S2 before lyophilization and the product of S4 after lyophilization), the following method was used for testing, and the accuracy of the test reagents before and after lyophilization was evaluated based on the obtained arachidonic acid cup blood clot strength; wherein, AA reagent (purchased from American Blood Technologies, brand name and company) from a commercially available platelet aggregation function test kit was used as a comparison reagent for comparison.
[0067] 1) Redissolve each sample in the specified volume of deionized water and let stand for 10 minutes;
[0068] 2) Add fibrin activator, the test reagent solution obtained in step 1), and a sample containing aspirin to the test cup respectively, and perform thromboelastography test. The obtained thromboelastography MA value (maximum amplitude of thrombus formation) is the arachidonic acid cup clot strength.
[0069] The test results are shown in Table 1 below.
[0070] Table 1. Accuracy test results before and after freeze-drying
[0071]
[0072]
[0073] The results above show that, whether before or after freeze-drying, the relative deviations of the test results of Example 1 and Comparative Example 1 from those of the standard reagent are controlled within 10%, proving that Example 1 and Comparative Example 1 can be used for subsequent detection, and that the freeze-drying process after microencapsulation does not affect the activity of arachidonic acid.
[0074] Example 3 Precision Test
[0075] 1) Reconstitute the lyophilized sample with the specified volume of deionized water and let it stand for 10 minutes.
[0076] 2) Add fibrin activator, the test reagent solution obtained in step 1), and a sample containing aspirin (a mixture of healthy human blood and a certain concentration of aspirin) to the test cup, respectively, and perform thromboelastography test. The obtained thromboelastography MA value (maximum amplitude of thrombus formation) is the arachidonic acid cup clot strength. Repeat the test 10 times; the test results are shown in Table 2 below.
[0077] Table 2 Precision Test Results
[0078]
[0079]
[0080] Precision test results show that the CVs of both Example 1 and Comparative Example 1 are within 10%, with Example 1 exhibiting better precision. While Comparative Example 1's precision is within 10%, it is already within the critical range. The reason for this is that emulsification resulted in a more uniform distribution of arachidonic acid, leading to higher precision in the test results.
[0081] Example 4 Stability Test Results
[0082] Open-bottle stability: For the arachidonic acid reagent obtained in Example 1 and Comparative Example 1, after being fully dissolved, it was placed at 2-8℃ and tested at 2 (KP2h), 8 (KP8h), 24 (KP24h), 48 (KP48h), and 72h (KP72h), respectively. The results were compared with the test results at 0h after opening (KP0h).
[0083] Accelerated stability testing: The arachidonic acid reagents obtained in Example 1 and Comparative Example 1 were stored in water baths at 2-8°C and 37°C for 7 days, respectively. Three samples from healthy individuals with aspirin were tested, and TEG arachidonic acid reagent (from the same source as in Example 2) was used as a standard reagent for comparative testing.
[0084] The test results are shown in Tables 3 and 4 (all results in the tables are MA values obtained from the tests):
[0085] Table 3 Results of bottle-opening stability test
[0086]
[0087]
[0088] The results of the opening tests show that at 2h, 4h, and 8h after opening, the test results of Example 1 and Comparative Example 1 are basically consistent with the test results at 0h, with no significant changes. Comparative Example 1 began to turn yellow and its activity began to decrease after 24h after opening, indicating that the decrease in arachidonic acid activity was due to partial oxidation. In contrast, the activity of the three samples tested in Example 1 was basically consistent with the results of the newly opened test within 72h after opening, which fully demonstrates that the antioxidant performance and stability of the microencapsulated arachidonic acid in the examples increased after opening.
[0089] Table 4 Accelerated Stability Test Results
[0090]
[0091] The comparison reagent TEG was prepared and used immediately, with its test data serving as a benchmark for comparison.
[0092] As shown in Table 4, the accelerated stability test results of the example after 7 days of accelerated testing at 37℃ are basically consistent with those of the 2-8℃ and TEG reagent tests. However, the activity of the comparative example decreased significantly after 7 days of accelerated testing at 37℃. This indicates that the arachidonic acid in Example 1, after microencapsulation, exhibits better heat resistance and higher stability.
[0093] Although the embodiments of the present invention have been disclosed above, they are not limited to the applications listed in the specification and embodiments. They can be applied to various fields suitable for the present invention. For those skilled in the art, other modifications can be easily made. Therefore, without departing from the general concept defined by the claims and their equivalents, the present invention is not limited to the specific details.
Claims
1. A reagent for detecting platelet aggregation function, characterized in that, This test reagent is a microencapsulated arachidonic acid test reagent prepared with arachidonic acid as the core material, cyclodextrin and gum arabic as the wall material, and with the addition of emulsifiers, antioxidants, lyophilization protectants, preservatives, buffer salts and water as auxiliary components. The test reagent comprises the following raw material components by mass percentage: Arachidonic acid: 0.1-1%; β-cyclodextrin: 0.1-1%; Gum arabic: 0.1-1%; Emulsifier: 0.1-2%; Antioxidant: 0.01-0.1%; Lyophilization protectant: 0.1-10%; Preservatives: 0.01-0.1%; the remainder is buffer salts and water.
2. The platelet aggregation function detection reagent according to claim 1, characterized in that, The emulsifier is any one or a mixture of casein, whey protein, monoglyceride fatty acid glycerides, and gelatin.
3. The platelet aggregation function detection reagent according to claim 2, characterized in that, The antioxidant is one or more of vitamin C, vitamin E, sodium thiosulfate, tea polyphenols, and dithiothreitol, or a mixture thereof.
4. The platelet aggregation function detection reagent according to claim 3, characterized in that, The freeze-drying protectant is any one or a mixture of PEG8000, Proneix, BSA, lactose, trehalose, and sucrose.
5. The platelet aggregation function detection reagent according to claim 4, characterized in that, The cyclodextrin is β-cyclodextrin, the preservative is Proclin, and the buffer salt is one of phosphate buffer, Tris buffer, and MES buffer, with a concentration of 5-50 mM. The emulsifier is casein, the antioxidant is a mixture of sodium thiosulfate and dithiothreitol, the lyophilization protectant is a mixture of Proneix and lactose, and the buffer salt is phosphate buffer.
6. A method for preparing a platelet aggregation function detection reagent as described in any one of claims 1-5, characterized in that, Includes the following steps: S1. Take cyclodextrin and gum arabic, dissolve them thoroughly in buffer salt solution, cool to room temperature, and then add emulsifier, antioxidant, freeze-drying protectant and preservative in sequence, stirring until completely dissolved. S2. Preparation of pre-emulsion: Arachidonic acid is added to the product obtained in step S1 under stirring to obtain a pre-emulsion; S3. Emulsification: The pre-emulsion obtained in step S1 is fully emulsified using a homogenizer, then packaged and pre-frozen. S4. Freeze-drying. After freeze-drying, vacuum stopper is used to obtain the platelet aggregation function detection reagent.
7. The method for preparing the platelet aggregation function detection reagent according to claim 6, characterized in that, Includes the following steps: S1. Take cyclodextrin and gum arabic, dissolve them thoroughly in a buffer salt solution at 60-70℃, cool to room temperature, and then add emulsifier, antioxidant, freeze-drying protectant and preservative in sequence. Stir magnetically until completely dissolved. S2. Preparation of pre-emulsion: Arachidonic acid is added to the product obtained in step S1 under magnetic stirring in an environment below 16°C to obtain a pre-emulsion. S3. Emulsification: In an environment below 16°C, the pre-emulsion obtained in step S1 is emulsified using a homogenizer at 8000-16000 r / min for 5-30 min, then dispensed and pre-frozen at -80°C for 2-6 h. S4. The vacuum degree should be controlled at 0.01-1.0 mbar for lyophilization for 10-24 hours. After lyophilization, the platelet aggregation function detection reagent is obtained by vacuum plugging.
8. The method for preparing the platelet aggregation function detection reagent according to claim 7, characterized in that, Includes the following steps: S1. Take 0.2 wt% cyclodextrin and 0.5% gum arabic and add them to 20 mM phosphate buffer. Dissolve them completely at 70°C and cool to room temperature. Then add 1 wt% casein, 0.08 wt% dithiothreitol, 0.05 wt% sodium thiosulfate, 2 wt% Proneix, 4 wt% lactose and 0.04 wt% Proclin in sequence and stir magnetically until completely dissolved. S2. Pre-emulsion preparation: 0.5 wt% arachidonic acid is added to the product obtained in step S1 under magnetic stirring in an environment below 16°C to obtain a pre-emulsion; S3. Emulsification: In an environment below 16°C, the pre-emulsion obtained in step S1 is emulsified using a homogenizer at 10000r / min for 12min, then dispensed and pre-frozen at -80°C for 4h. S4. The vacuum degree should be controlled at 0.2 mbar for 18 hours for freeze drying. After freeze drying, the platelet aggregation function test reagent is obtained by vacuum plugging.
9. A method for detecting platelet aggregation function, characterized in that, Includes the following steps: 1) Redissolve the platelet aggregation function detection reagent as described in any one of claims 1-5 in a specified volume of deionized water, let stand for 10-30 minutes, and obtain the detection reagent solution; 2) Add fibrin activator, the test reagent solution obtained in step 1), and a sample containing aspirin to the test cup respectively, and perform thromboelastography test. The obtained thromboelastography MA value is the arachidonic acid cup clot strength.
10. The method for detecting platelet aggregation function according to claim 9, characterized in that, in, The aspirin-containing sample is a blood sample from a person who has taken aspirin, or a sample obtained by mixing blood from a person who has not taken aspirin with 0.001-0.1 mg / mL of aspirin.