CN113156144A - Arachidonic acid cup detection reagent with platelet aggregation function and preparation method thereof - Google Patents
Arachidonic acid cup detection reagent with platelet aggregation function and preparation method thereof Download PDFInfo
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/88—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving prostaglandins or their receptors
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/86—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving blood coagulating time or factors, or their receptors
Abstract
The invention belongs to the technical field of blood coagulation detection, and particularly relates to a blood platelet aggregation function arachidonic acid detection reagent and a preparation method thereof. The arachidonic acid detection reagent comprises arachidonic acid sodium and a freeze-drying protective agent, wherein the freeze-drying protective agent comprises a saccharide freeze-drying protective agent and a polymer freeze-drying protective agent; the invention fully considers the different reactivities of the diagnostic reagent to the antiplatelet drugs and has better sensitivity to aspirin, ibuprofen and the like which are commonly used clinically at present. The arachidonic acid cup detection reagent is used for the anticoagulant blood detection of the heparin sodium after taking the medicine, and has good detection result accuracy and high precision in batch and between batches.
Description
Technical Field
The invention belongs to the technical field of blood coagulation detection, and particularly relates to a blood platelet aggregation function arachidonic acid detection reagent and a preparation method thereof.
Background
The Thromboelastogram (TEG) is a blood coagulation function analyzer which carries out real-time, dynamic and continuous monitoring on the whole processes of platelet aggregation, blood coagulation, fibrinolysis and the like, and carries out qualitative or quantitative analysis on the blood coagulation condition of a patient.
The invention of the thromboelastogram instrument by Harter of German in 1948 is applied to guide the blood transfusion in clinical operation, and is one of the most important means for detecting the blood coagulation function at present. The principle of the thromboelastogram for detecting the blood coagulation function is that a sensor probe is immersed in a blood sample, a viscous force exists between the blood sample and the sensor probe, the viscous force changes along with changes of platelet aggregation, blood coagulation, fibrinolysis and the like, a suspension wire of a sensor generates torsional deformation under the action of the viscous force, the torsional deformation is detected by a detection unit and is converted into an electric signal to be fed back to a control system, the control system carries out operation after receiving the changed signal, and an operation result is displayed on a computer screen in the form of a curve graph and real-time data.
Thromboelastogram (TEG) uses anticoagulated whole blood to activate the coagulation and fibrinolysis system from the extrinsic coagulation pathway, thereby reflecting the whole process of human coagulation factors, platelets, fibrin and fibrinolysis more comprehensively and truly.
The main functions of platelets are coagulation, hemostasis, and repair of damaged blood vessels. Platelet surface coagulation function related receptor ADP receptor, alpha and beta-adrenoreceptor, thrombin receptor, collagen receptor (GP Ia/IIa, GP IV, GP-VI), TXA2/PGH2Receptor, PGI2/PGE1Receptor, 5-hydroxytryptamine receptor, heparin receptor, PFA (blood)Platelet activating factor) receptor, fibrinogen receptor, Ca2+Receptor fibronectin receptors, platelet factor 4 receptors, angiotensin II receptors, LDL receptors, HDL receptors, vWF receptors (GPIb/IX) and GPIIb/IIIa (fibrinogen receptors), and the like. The GP IIb/IIIa membrane receptor is an adhesive glycoprotein on the platelet surface, a most abundant integrin on the platelet surface, with a size of about 5X 104～8×104A GPIIb/IIIa membrane receptor. ADP, AA, thrombin and the like can activate platelets to change the conformation of the GP IIb/IIIa membrane receptors on the surfaces of the platelets, so that the affinity of the GP IIb/IIIa membrane receptors to soluble fibrinogen, vWF and the like is increased, and finally the platelets are aggregated on bridging fibrin and vWF to form platelet thrombus.
Phospholipids in cell membranes in phospholipase A2Catalyzing to release arachidonic acid AA (combined on the 2-carbon of phospholipid glycerol), and catalyzing the AA by cyclooxygenase COX (namely prostaglandin H synthetase PGHS) to form PGG2And PGH2And the two metabolize to various prostaglandin PGs under the action of the corresponding prostaglandin isomerase. PGs include PGE, PGF, PGD, PGI2(produced by vascular endothelial cells, inducing vasodilation and inhibiting platelet aggregation) and TXA2(produced by platelets, induced vasoconstriction, a strong platelet agonist) and the like. Aspirin can promote the conversion of cyclooxygenase COX of blood platelet to acetylphthalein and inhibit thromboxane TXA2Synthesis, platelet release reaction (such as release caused by epinephrine, collagen, thrombin, etc.), and endogenous ADP, 5-hydroxytryptamine, etc., thereby inhibiting platelet aggregation
Aspirin, as a classic anti-platelet drug, has a wide range of clinical applications and a high frequency, far surpassing other anticoagulant drugs. Currently, Thromboelastography (TEG) lacks sufficient sensitivity in detecting platelet aggregation function, i.e., it is not effective in detecting patient specimens taking aspirin drugs, or it is effective in detecting, but does not reflect the extent of platelet inhibition as compared to other methods.
Disclosure of Invention
The invention provides a platelet aggregation function arachidonic acid detection reagent and a preparation method thereof, aiming at the defects and problems that the conventional Thromboelastogram (TEG) lacks enough sensitivity when detecting the platelet aggregation function, cannot effectively detect a patient sample taking aspirin medicine, and cannot truly reflect the degree of platelet inhibition compared with other methods although some patients can effectively detect the platelet aggregation function.
The technical scheme adopted by the invention for solving the technical problems is as follows: a platelet aggregation function arachidonic acid detection reagent comprises arachidonic acid sodium and a freeze-drying protective agent.
The platelet aggregation function arachidonic acid detection reagent comprises a saccharide freeze-drying protective agent and a polymer freeze-drying protective agent.
In the arachidonic acid detection reagent with platelet aggregation function, the saccharide freeze-drying protective agent is one or more of trehalose, sucrose, dextran and glucose.
In the arachidonic acid detection reagent with platelet aggregation function, the saccharide protective agent is trehalose, and the addition amount of the trehalose is 1%.
In the arachidonic acid detection reagent with platelet aggregation function, the polymer freeze-drying protective agent is one or more of bovine serum albumin, polyvinylpyrrolidone and polyethylene glycol.
According to the arachidonic acid detection reagent with the platelet aggregation function, the polymer freeze-drying protective agent is bovine serum albumin, and the addition amount of the bovine serum albumin is 2%.
The arachidonic acid detection reagent with the platelet aggregation function also comprises antioxidation, and the antioxidant is one or more of dithiothreitol, tert-butyl hydroquinone, dibutyl hydroxy toluene and sodium thiosulfate.
The invention also provides a preparation method of the arachidonic acid detection reagent with the platelet aggregation function, which comprises the following steps:
(1) weighing a freeze-drying protective agent and an antioxidant according to the formula amount, and adding purified water or purified water to fully dissolve to obtain a freeze-drying protective agent solution;
(2) weighing the arachidonic acid sodium according to the formula amount, pouring the arachidonic acid sodium into the freeze-drying protective agent solution, and uniformly mixing to obtain the arachidonic acid sodium detection reagent mother solution;
(3) subpackaging the detection reagent for detecting the arachidonic acid sodium into penicillin bottles according to 20-60 microliters per bottle to obtain a finished product of the detection reagent for detecting the arachidonic acid sodium;
(4) and (3) carrying out vacuum freeze drying on the sodium arachidonic acid detection reagent to obtain a freeze-dried product of the sodium arachidonic acid reagent.
The invention has the beneficial effects that: the invention fully considers the different reactivities of the diagnostic reagent to the antiplatelet drugs and has better sensitivity to aspirin, ibuprofen and the like which are commonly used clinically at present. The arachidonic acid cup detection reagent is used for the anticoagulant blood detection of the heparin sodium after taking the medicine, and has good detection result accuracy and high precision in batch and between batches.
Detailed Description
The present invention will be further described with reference to the following examples.
The invention provides a platelet aggregation function arachidonic acid detection reagent, which comprises arachidonic acid sodium, a freeze-drying protective agent and an antioxidant, wherein the freeze-drying protective agent comprises a saccharide freeze-drying protective agent and a polymer freeze-drying protective agent, and the saccharide freeze-drying protective agent can be any one or more of trehalose, sucrose, glucan and glucose; the polymer freeze-drying protective agent can be any one or more of bovine serum albumin, polyvinylpyrrolidone and polyethylene glycol; the antioxidant can be any one or more of dithiothreitol, tert-butyl hydroquinone, dibutyl hydroxy toluene and sodium thiosulfate.
Example 1: the total amount of the arachidonic acid detection reagent in this example is 40 microliters, and the amounts of the raw materials by volume ratio are respectively 0.5% of arachidonic acid sodium salt, 1.0% of trehalose, 2.0% of bovine serum albumin, 0.03% of dithiothreitol, 0.04% of dibutylhydroxytoluene, and the balance of purified water.
The preparation method comprises the following steps:
(1) weighing trehalose, bovine serum albumin, dithiothreitol and dibutyl hydroxy toluene according to the formula amount, and adding purified water or purified water to fully dissolve to obtain a freeze-drying protective agent solution;
(2) weighing sodium arachidonic acid according to the formula ratio, adding the sodium arachidonic acid into the prepared freeze-drying protective agent solution, and uniformly mixing to obtain a mother solution of the sodium arachidonic acid detection reagent;
(3) subpackaging the detection reagent for detecting the arachidonic acid sodium into penicillin bottles according to 40 microliter per bottle to obtain a finished product of the detection reagent for detecting the arachidonic acid sodium;
(4) and (3) carrying out vacuum freeze drying on the sodium arachidonic acid detection reagent to obtain a freeze-dried product of the sodium arachidonic acid reagent.
Example 2: the total amount of the arachidonic acid detection reagent in this example is 60 microliters, and the amounts of the raw materials by volume ratio are respectively 1.0% of arachidonic acid sodium salt, 2.0% of trehalose, 3.0% of bovine serum albumin, 0.06% of dithiothreitol, 0.06% of dibutylhydroxytoluene, and the balance of purified water.
The preparation method comprises the following steps:
(1) weighing trehalose, bovine serum albumin, dithiothreitol and dibutyl hydroxy toluene according to the formula amount, and adding purified water or purified water to fully dissolve to obtain a freeze-drying protective agent solution;
(2) weighing sodium arachidonic acid according to the formula ratio, adding the sodium arachidonic acid into the prepared freeze-drying protective agent solution, and uniformly mixing to obtain a mother solution of the sodium arachidonic acid detection reagent;
(3) subpackaging 60 microliters of the sodium arachidonic acid detection reagent into penicillin bottles to obtain a finished product of the sodium arachidonic acid detection reagent;
(4) and (3) carrying out vacuum freeze drying on the sodium arachidonic acid detection reagent to obtain a freeze-dried product of the sodium arachidonic acid reagent.
Example 3: the total amount of the arachidonic acid detection reagent in this example was 40 microliters, and the amounts of the raw materials were 0.5% arachidonic acid sodium, 2.5% trehalose, 2.0% bovine serum albumin, 0.09% dithiothreitol, 0.02% dibutylhydroxytoluene, and the balance purified water, respectively, by mass volume concentration.
The preparation method comprises the following steps:
(1) weighing trehalose, bovine serum albumin, dithiothreitol and dibutyl hydroxy toluene according to the formula amount, and adding purified water or purified water to fully dissolve to obtain a freeze-drying protective agent solution;
(2) weighing arachidonic acid sodium according to the formula ratio, adding the arachidonic acid sodium into the prepared freeze-drying protective agent solution, and uniformly mixing to obtain arachidonic acid detection reagent mother liquor;
(3) subpackaging the arachidonic acid sodium detection reagent into penicillin bottles according to 40 microliter per bottle to obtain finished arachidonic acid detection reagent;
(4) and (3) carrying out vacuum freeze drying on the arachidonic acid detection reagent to obtain an arachidonic acid reagent freeze-dried product.
Test example: the test example provides a reagent for detecting arachidonic acid with a whiteboard aggregation function, wherein the addition amount of the arachidonic acid in the reagent is 0.1-3.0%. In this example, arachidonic acid sodium is measured at seven levels of 0.1%, 0.5%, 1.0%, 1.5%, 2.0%, 2.5%, and 3.0%. The experimental groups are shown in table 1 below.
TABLE 1 arachidonic acid cup main raw material addition optimization grouping
| Experiment number | Detecting an object |
| 1 | Arachidonic acid sodium salt (0.1%) |
| 2 | Arachidonic acid sodium salt (0.5%) |
| 3 | Arachidonic acid sodium salt (1.0%) |
| 4 | Arachidonic acid sodium salt (1.5%) |
| 5 | Arachidonic acid sodium salt (2.0%) |
| 6 | Arachidonic acid sodium salt (2.5%) |
| 7 | Arachidonic acid sodium salt (3.0%) |
TABLE 2 Effect of different amounts of arachidonic acid cup on the respective indices of the thromboelastogram
Note:
as can be seen from the data in table 2, the relative deviation of the measurement results of R, K, Angle and MA with the reference reagent in the No. 2 experimental group was the smallest and satisfied that the relative deviation was not more than 10%. Therefore, the dosage of the arachidonic acid sodium in the reagent is preferably 0.5 percent.
The arachidonic acid sodium salt is extremely unstable and is easy to be oxidized and deteriorated in the air. It is contemplated to add antioxidant protectors to their formulations.
The sugar freeze-drying protective agent added into the arachidonic acid cup detection reagent is one or more of trehalose, sucrose, glucan and glucose, and the pre-experimental result shows that the freeze-drying effect is best when the trehalose accounting for 0.5-2.5% of the mass volume of the reagent is added, and the freeze-drying protective agent has no obvious influence on all indexes of the arachidonic acid cup detection reagent. Trehalose without reducing group is added, so that the sodium arachidonic acid can be protected from deterioration and inactivation caused by low temperature and dehydration in the whole freeze-drying process.
The polymer freeze-drying protective agent added into the arachidonic acid cup detection reagent is one or more of bovine serum albumin, polyvinylpyrrolidone and polyethylene glycol, and an experimental result shows that the freeze-drying effect is best when the bovine serum albumin with the mass volume of 0.1-4.0% of the reagent is added, and the freeze-drying protective agent has no obvious influence on all indexes of the arachidonic acid cup detection reagent. The bovine serum albumin is added to protect the arachidonic acid sodium from being influenced by low temperature and dehydration on the activity of the arachidonic acid sodium in freeze drying.
The antioxidant added into the arachidonic acid cup detection reagent is one or more of dithiothreitol, tert-butyl hydroquinone, dibutyl hydroxy toluene and sodium thiosulfate, and the experimental result shows that the freeze-drying effect is best when 0.02-0.1% of dithiothreitol and 0.02-0.1% of dibutyl hydroxy toluene in mass volume of the reagent are added, and the indexes of the arachidonic acid cup detection reagent are not obviously influenced. Dithiothreitol and dibutyl hydroxy toluene are preferably used as a combined antioxidant system, so that the activity loss caused by the oxidation of arachidonic acid sodium can be effectively prevented.
Bovine serum albumin, trehalose, dithiothreitol and dibutyl hydroxy toluene are used as freeze-drying protective agents, and the effect of activity maintenance of the arachidonic acid sodium is the most ideal. After preliminary experiments, the dosage of each freeze-drying protective agent is temporarily determined as follows: trehalose was selected at three levels of 1.0%, 1.5% and 2.0%; bovine serum albumin was selected at three levels, 1.0%, 2.0% and 3.0%; dithiothreitol is selected at three levels of 0.03%, 0.06% and 0.09%; three levels of 0.02%, 0.04% and 0.06% were selected for dibutylhydroxytoluene.
By using L9(43) Orthogonal test design is carried out on an orthogonal table, trehalose is a factor 1 and occupies the first column of the orthogonal table, 1.0% is horizontal (i), 1.5% is horizontal (ii), and 2.0% is horizontal (i 1); bovine serum albumin is a factor 2, occupies the second column of the orthogonal table, and 1.0% is horizontal (i), 2.0% is horizontal (0), and 3.0% is horizontal (iii); dithiothreitol is a factor 3, occupies the third column of the orthogonal table, and 0.03 percent is horizontal (i), 0.06 percent is horizontal (ii), and 0.09 percent is horizontal (iii); dibutyl hydroxy toluene is a factor 4, occupies the fourth column of the orthogonal table, 0.02 percent is horizontal (i), 0.04 percent is horizontal (ii), and 0.06 percent is horizontal (iii); the levels of each test, the combination of factors are shown in Table 3 belowShown in the figure.
TABLE 3 arachidonic acid cup test reagent formulation optimization test design protocol
Taking an imported reagent as a reference reagent, each sample is respectively carried out in parallel three times by using the reagent and the reference reagent, and the detection results are shown in the following table 4.
TABLE 4 arachidonic acid cup test reagent optimization test results
Note:
as can be seen from the data in Table 4, R, K, Angle and MA in the test group No. 6 have the smallest relative deviation from the measurement result of the reference reagent, so that the amount of trehalose in the reagent of the present invention is preferably 1.0%, the amount of bovine serum albumin is preferably 2.0%, the amount of dithiothreitol is preferably 0.03%, and the amount of dibutylhydroxytoluene is preferably 0.04%.
Application example:
the arachidonic acid detection reagent of the invention can be used for whole blood detection and anticoagulated whole blood detection in the technology of Thromboelastogram (TEG).
The Thromboelastogram (TEG) test feature quantities are explained below:
blood coagulation time R: the blood coagulation factors such as thrombin are sufficiently activated, fibrin clot formation starts, and the time is required until the amplitude reaches 2mm as measured by a thromboelastography. An extended R-value, indicative of a deficiency of certain coagulation factors; the decrease in R value indicates a high blood coagulation status.
Clot formation time K: the time from the end of the R value time to the time at which the thromboelastogram traces the amplitude to 20mm reflects the interaction of fibrin and platelets at the beginning of clotting formation. The length of the K value reflects the fibrinogen level.
Hemagglutination rate Angle: the thromboelastography instrument traces the included angle between the tangent line of the maximum radian point of the amplitude curve and the horizontal line, and reflects the forming speed of the blood clot. When the blood is in a severe low-coagulation state, the amplitude of the tracing meter does not reach 20mm, and the K value cannot be determined.
Blood clot strength MA: the maximum profilometer amplitude for the thromboelastogram, which is the binding of fibrin to the platelet GPIIb/IIIa receptor, represents the maximum strength of the fibrin/platelet clot.
Inhibition ratio%: less than 50, indicating that the medicine can not play a better anticoagulation effect and has a certain degree of medicine resistance.
Inhibition ratio%: more than or equal to 50, prompting the drug to take effect and obviously inhibit.
The arachidonic acid cup detection reagent directly detects anticoagulated whole blood treated by heparin sodium, and the collected anticoagulated whole blood is placed at room temperature for not more than 24 hours, preferably within 2 hours.
Wherein: arachidonic acid sodium salt: SIGMA;
bovine serum albumin manufacturer: BioRuler;
trehalose manufacturers: shanghai-sourced leaf Biotechnology, Inc.;
dithiothreitol manufacturer: beijing Solaibao science and technology, Inc.;
dibutyl hydroxy toluene manufacturer: shanghai Michell chemical technology, Inc.
The method for using the platelet aggregation function detection kit for whole blood detection comprises the following steps:
1) opening application software, inputting a patient name on a patient information interface, and selecting a test type, wherein the test type comprises a common cup 'CK-Cirtated kaolin', a batroxobin cup 'A-Activated', and an arachidonic acid cup 'AA-Activated + AA';
2) loading a common cup, a batroxobin cup and an arachidonic acid cup on a channel of the thromboelastography instrument; note and test type corresponds!
3) Taking out the batroxobin cup detection reagent, the arachidonic acid cup detection reagent and the purified water, respectively transferring 40 mu l of the purified water, adding the reagents of the batroxobin cup detection reagent and the arachidonic acid cup detection reagent into bottles, covering the bottles, and fully shaking up;
4) transferring 1ml of the anticoagulated whole blood sample of sodium citrate into a reagent 1 bottle by using an lml pipettor, turning the reagent 1 bottle upside down for 5 times, fully mixing uniformly, and standing for 4 minutes to activate blood;
5) and (3) detecting a common cup: transferring 20 mul of reagent 2 to the bottom of a common cup, transferring 340 mul of blood activated in the step 4) to the common cup, pinching a cup holder, pushing the common cup to the top end, shifting a test rod to a test position, selecting a CK-circled kaolin channel, clicking a software interface start or a keyboard F10, and starting testing;
6) batroxobin cup detection: transferring 10 mu l of liquid from the redissolving reagent A in the step 2) into a batroxobin cup, adding 360 mu l of heparinized blood sample, pumping three times in the cup, uniformly mixing, pushing the batroxobin cup to the top end, shifting a test rod to a test position, selecting an A-Activated channel, clicking a software interface 'start' or a keyboard F10, and starting testing;
7) and (3) detecting arachidonic acid in a cup: transferring 10 mu l of liquid from the redissolving reagent A in the step 2), sucking 10 mu l of liquid from the redissolving reagent C, adding the liquid into an arachidonic acid cup, adding 360 mu l of heparinized blood sample, sucking for three times in the cup, uniformly mixing, pushing the arachidonic acid cup to the top end, dialing a test rod to a test position, selecting an AA-Activated + AA channel, clicking a software interface 'start' or a keyboard F10, and starting testing;
8) the clotting process was finished for about 30 minutes;
9) after the test is finished, clicking stop on a software interface, unloading the test cup, and disposing according to the laboratory management regulation;
10) inhibition rate: and clicking 'composite' on a software interface, selecting a test result graph of three channels of a common cup, a batroxobin cup and an arachidonic acid cup, and clicking 'finish' to check the inhibition rate information of the arachidonic acid pathway.
11) And after the detection is finished, closing the instrument power switch.
The test results are shown in tables 5 to 9.
TABLE 5 high and Low inhibitionDetection result of enterprise reference
TABLE 6 results of anticoagulated whole blood assays in 3 random healthy persons
TABLE 7 clot Strength MA reference interval
As can be seen from tables 5, 6 and 7, the detection results (relative deviation) of the enterprise reference products with high and low inhibition rates are within the range of +/-10%; the detection results of 3 random healthy human anticoagulated whole blood samples are all in the specified range, and the detection results of the kit are consistent with the actual conditions of patients.
The results of the measurement of the in-batch precision and the inter-batch precision of the adenosine diphosphate detection reagent of the present invention are shown in tables 8 and 9.
TABLE 8 in-batch precision
As can be seen from Table 8, the variation coefficients of the precision detection results of the common cup, the batroxobin cup and the arachidonic acid cup on the fresh sodium citrate and heparin sodium anticoagulated whole blood in batch are all less than 10%, and the precision in batch is high.
TABLE 9 inter-batch precision
As is clear from the results in Table 8, the relative deviations in the measurement results of the inter-lot precision of the respective characteristic amounts were less than 10%, and the inter-lot precision was high.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents and improvements made within the spirit and scope of the present invention are intended to be covered thereby.
Claims (8)
1. A platelet aggregation function arachidonic acid detection reagent is characterized in that: comprises arachidonic acid sodium and a freeze-drying protective agent.
2. A platelet aggregation function arachidonic acid detecting reagent in accordance with claim 1, wherein: the freeze-drying protective agent comprises a saccharide freeze-drying protective agent and a polymer freeze-drying protective agent.
3. A platelet aggregation function arachidonic acid detecting reagent in accordance with claim 2, wherein: the saccharide freeze-drying protective agent is one or more of trehalose, sucrose, glucan and glucose.
4. A platelet aggregation function arachidonic acid detecting reagent in accordance with claim 3, wherein: the saccharide protective agent is trehalose, and the addition amount of trehalose is 1%.
5. A platelet aggregation function arachidonic acid detecting reagent in accordance with claim 3, wherein: the polymer freeze-drying protective agent is one or more of bovine serum albumin, polyvinylpyrrolidone and polyethylene glycol.
6. A platelet aggregation function arachidonic acid detecting reagent in accordance with claim 5, wherein: the polymer freeze-drying protective agent is bovine serum albumin, and the addition amount of the bovine serum albumin is 2%.
7. A platelet aggregation function arachidonic acid detecting reagent in accordance with claim 1, wherein: the antioxidant is one or more of dithiothreitol, tert-butyl hydroquinone, dibutyl hydroxy toluene and sodium thiosulfate.
8. A method for producing a platelet aggregation function arachidonic acid detecting reagent according to any one of claims 1 to 7, wherein: the method comprises the following steps:
(1) weighing a freeze-drying protective agent and an antioxidant according to the formula amount, and adding purified water or purified water to fully dissolve to obtain a freeze-drying protective agent solution;
(2) weighing the arachidonic acid sodium according to the formula amount, pouring the arachidonic acid sodium into the freeze-drying protective agent solution, and uniformly mixing to obtain the arachidonic acid sodium detection reagent mother solution;
(3) subpackaging the detection reagent for detecting the arachidonic acid sodium into penicillin bottles according to 20-60 microliters per bottle to obtain a finished product of the detection reagent for detecting the arachidonic acid sodium;
(4) and (3) carrying out vacuum freeze drying on the sodium arachidonic acid detection reagent to obtain a freeze-dried product of the sodium arachidonic acid reagent.
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| 耿诗涵等: "注射用丹参多酚酸与阿司匹林联合应用对抗凝效果和出血风险的影响", 《中国药学杂志》 * |
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