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J Vet Cardiol. 2017 Aug;19(4):384-395. doi: 10.1016/j.jvc.2017.03.004. Epub 2017 Jun 9.

Validation of a method for quantitation of the clopidogrel active metabolite, clopidogrel, clopidogrel carboxylic acid, and 2-oxo-clopidogrel in feline plasma.

Author information

1
Program in Individualized Medicine (PrIMe), Department of Veterinary Clinical Sciences, College of Veterinary Medicine, PO Box 647060, Washington State University, WA 99164-7060, USA.
2
Program in Individualized Medicine (PrIMe), Department of Veterinary Clinical Sciences, College of Veterinary Medicine, PO Box 647060, Washington State University, WA 99164-7060, USA. Electronic address: pamelamlee@vetmed.wsu.edu.

Abstract

INTRODUCTION:

The clopidogrel active metabolite (CAM) is unstable and challenging to quantitate. The objective was to validate a new method for stabilization and quantitation of CAM, clopidogrel, and the inactive metabolites clopidogrel carboxylic acid and 2-oxo-clopiodgrel in feline plasma.

ANIMALS:

Two healthy cats administered clopidogrel to demonstrate assay in vivo utility.

MATERIALS AND METHODS:

Stabilization of CAM was achieved by adding 2-bromo-3'methoxyacetophenone to blood tubes to form a derivatized CAM (CAM-D). Method validation included evaluation of calibration curve linearity, accuracy, and precision; within and between assay precision and accuracy; and compound stability using spiked blank feline plasma. Analytes were measured by high performance liquid chromatography with tandem mass spectrometry. In vivo utility was demonstrated by a pharmacokinetic study of cats given a single oral dose of 18.75mg clopidogrel.

RESULTS:

The 2-oxo-clopidogrel metabolite was unstable. Clopidogrel, CAM-D, and clopidogrel carboxylic acid appear stable for 1 week at room temperature and 9 months at -80°C. Standard curves showed linearity for CAM-D, clopidogrel, and clopidogrel carboxylic acid (r > 0.99). Between assay accuracy and precision was ≤2.6% and ≤7.1% for CAM-D and ≤17.9% and ≤11.3% for clopidogrel and clopidogrel carboxylic acid. Within assay precision for all three compounds was ≤7%. All three compounds were detected in plasma from healthy cats receiving clopidogrel.

DISCUSSION:

This methodology is accurate and precise for simultaneous quantitation of CAM-D, clopidogrel, and clopidogrel carboxylic acid in feline plasma but not 2-oxo-clopidogrel.

CONCLUSIONS:

Validation of this assay is the first step to more fully understanding the use of clopidogrel in cats.

KEYWORDS:

Antiplatelet; Antithrombotic; Cat; Plavix; Thromboprophylaxis

PMID:
28602635
PMCID:
PMC5557681
DOI:
10.1016/j.jvc.2017.03.004
[Indexed for MEDLINE]
Free PMC Article

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