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Simultaneously accurate and quantitative analysis of human plasma vicagrelor and its main metabolite detection method and its kit

A technology for quantitative detection of metabolites, applied in the analysis of materials, measurement devices, material separation, etc., can solve problems that have not been reported, and achieve good accuracy and repeatability

Active Publication Date: 2020-05-19
普灵生物(南京)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the optimized method for stabilizing vicagrelor in whole blood or plasma and the method for simultaneous detection of the above three compounds in human plasma have not been reported yet

Method used

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  • Simultaneously accurate and quantitative analysis of human plasma vicagrelor and its main metabolite detection method and its kit
  • Simultaneously accurate and quantitative analysis of human plasma vicagrelor and its main metabolite detection method and its kit
  • Simultaneously accurate and quantitative analysis of human plasma vicagrelor and its main metabolite detection method and its kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] Embodiment 1 The kit for simultaneously detecting the original drug of vicagrelor in human plasma and its main metabolites includes the following components:

[0067] Reference solution A: vicagrel: 8 μg / mL

[0068] Reference solution B: 2-oxo-clopidogrel: 8μg / mL

[0069] Reference solution C: MP-H4 (MPB derivative of H4; commercially available): 30 μg / mL

[0070] Internal standard (IS) solution: namely clopidogrel (clopidogrel): 300ng / mL

[0071] Stabilizer A: citric acid (CA) solution: 5M

[0072] Stabilizer B: sodium fluoride (NaF) solution: 1M

[0073] Stabilizer C: MPB (2-bromo-3'-methoxyacetophenone) solution: 500mM

[0074] Calibrator dilution (i.e., stabilized human blank plasma): stabilized human blank plasma, prepared by adding Stabilizer A, Stabilizer B, and Stabilizer C to K 2 EDTA anticoagulated, fresh human blank whole blood, mixed thoroughly, and centrifuged at 4000rpm for 10min to separate the plasma, which is the stabilized human blank plasma. Whe...

Embodiment 2

[0077] Example 2 Kit usage method

[0078] LC-MS / MS method for determination, the specific operation is as follows.

[0079] 1. Whole blood collection and plasma preparation

[0080] 1) Inject stabilizer A (1:100; v / v), stabilizer B (1:100; v / v), and stabilizer C (1:50; v / v) into K in sequence 2 Put EDTA anticoagulant in the vacuum blood collection tube, be careful not to break its vacuum. The volume ratio of stabilizer A to collected whole blood is 1:100; the volume ratio of stabilizer B to collected whole blood is 1:100; the volume ratio of stabilizer C to collected whole blood is 1:50;

[0081] 2) Collect the whole blood to the mark line of the blood collection tube, mix it upside down immediately, centrifuge at 4000rpm at 4°C for 10min, separate the plasma and store it at -80°C for testing.

[0082] 2. Preparation of internal standard working solution and mobile phase

[0083] 1) Preparation of internal standard working solution: pipette the required amount of internal...

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Abstract

The invention discloses a detection method and kit for accurately and quantitatively analyzing Vicagrel in human plasma and the main metabolites thereof. The kit for accurately and quantitatively analyzing Vicagrel in human plasma and the main metabolites thereof is characterized in that the kit comprises citric acid, sodium fluoride and 2-bromine-3'-methoxyacetophenone, or comprises citric acid,sodium fluoride, 2-bromine-3'-methoxyacetophenone and a K2EDTA anticoagulant vacuum blood collection tube. The invention provides optimization schemes for maintaining the stability of the Vicagrel andthe main metabolites thereof in human whole blood or plasma, and provides a detection method capable of accurately and quantitatively analyzing Vicagrel in human plasma and the main metabolites thereof simultaneously. The lowest limit of quantitation, specificity, recovery rate, matrix effect, precision, accuracy, stability and residual effect of the detection method meet the standard requirements of biological sample detection, which indicates that the method has good accuracy and repeatability.

Description

technical field [0001] The invention belongs to the field of biological detection, and relates to a detection method and a kit for simultaneously accurately and quantitatively analyzing the original drug of vicargrel and its main metabolites in human plasma. Background technique [0002] Clopidogrel (clopidogrel) is an antiplatelet drug widely used in acute coronary syndrome (acute coronary syndrome, ACS) or percutaneous coronary intervention (percutaneous coronary intervention, PCI) after stent implantation to prevent coronary thrombotic events . However, a large number of clinical studies have found that after taking conventional doses of clopidogrel, some patients still cannot obtain the expected antiplatelet effect, manifested as low or no response to clopidogrel, and severe thrombosis, ischemia, etc. This phenomenon is called "clopidogrel resistance (clopidogrel resistance, CR)". [0003] Vicagrel, a clopidogrel acetate analogue, is a new antiplatelet drug designed to...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/06G01N30/88
CPCG01N30/06G01N30/88
Inventor 谢红光邰婷周缓贾雨萌吉金子米琼宇
Owner 普灵生物(南京)有限公司