Determination of clopidogrel in human plasma by liquid chromatography/tandem mass spectrometry: application to a clinical pharmacokinetic study

Abstract

A rapid and sensitive LC/MS/MS assay was developed and validated for the determination of clopidogrel in human plasma. Clopidogrel was extracted by single liquid-liquid extraction with pentane, and chromatographic separations were achieved on a C(18) column. The method was validated to demonstrate the specificity, linearity, recovery, lower limit of quantification (LLOQ), stability, accuracy and precision. The multiple reaction monitoring was based on m/z transition of 322.2 --> 211.9 for clopidogrel and 264.1 --> 125.1 for ticlopidine (internal standard). The total analytical run time was relatively short (3 min), and the LLOQ was 10 pg/mL using 0.5 mL of human plasma. The assay was linear over a concentration range from 10 to 10,000 pg/mL (r > 0.999). The intra- and inter-day accuracies were 101.3-108.8 and 98.4-103.5%, respectively, and the intra- and inter-day assay precisions were 1.9-5.5 and 4.4-8.1%, respectively. The developed assay method was applied to a pharmacokinetic study in human volunteers after oral administration of clopidogrel at a dose of 150 mg.

Sensitivity enhancement and matrix effect evaluation during summation of multiple transition pairs—case studies of clopidogrel and ramiprilat

“…m/z 322.2 to m/z 212.0 for clopidogrel and m/z 389.2 to m/z 206.1 for ramiprilat) contributed more signifi cantly for the summed response factor. It was not surprising that these single transition pairs have been considered for the quantifi cation of the two compounds (Shin and Yoo, 2007;Nirogi et al, 2006;Lu et al, 2006;Zhu et al, 2002). However, the other lesser contributing MRM transition pair (i.e.…”

“…Several methods are available for the determination of clopidogrel (Shin and Yoo, 2007;Nirogi et al, 2006;Lainesse et al, 2004) and ramiprilat (Zhu et al, 2002) using tandem mass spectrometry. Both compounds produce stable and intense multiple MRM transition pairs as evidenced by the positive ion mass spectral data and therefore are amenable for the proposed summation approach of multiple MRM transition pairs to improve sensitivity.…”

Clopidogrel: review of bioanalytical methods, pharmacokinetics/pharmacodynamics, and update on recent trends in drug–drug interaction studies

“…However, the advancement in the LC-MS/MS technology and availability of plethora of options for optimization and enhanced detection has paved the way for the better understanding of the pharmacokinetics of clopidogrel as well as its active metabolite, and also applicability to important drugdrug interaction studies. Therefore, although the measurement of the active moiety has been extremely challenging, a number of analytical methods have been published in recent years that have the ability to measure the parent drug, clopidogrel (Taubert et al, 2004;Von Beckerath et al, 2005;Nirogi et al, 2006a;Shin and Yoo, 2007;Robinson et al, 2007); while some of them have the ability to even measure the active moiety at extremely low concentrations (Taubert et al, 2004;Von Beckerath et al, 2005). Table 1 lists the various methods that has been published for the quantification of clopidogrel and metabolites (active/inactive) in a period of over 10 years-it also provides detailed information on the separation conditions, validation routine with relevant parameters investigated and applicable conclusions.…”

Development and validation of high-throughput liquid chromatography–tandem mass spectrometric method for simultaneous quantification of Clopidogrel and its metabolite in human plasma

“…Analytical methods so far reported for quantification of Clopidogrel, employing tandem mass spectrometric detection [8][9][10] and for quantification of carboxylic acid metabolite, employing HPLC coupled ultraviolet detection [11,12] in human plasma and in human serum [13], mass spectrometric detection [14] and tandem mass spectrometric detection [15]. No method was reported for simultaneous quantification of Clopidogrel and its metabolite.…”