Stereoselective High-Performance Liquid Chromatography Tandem Mass Spectrometry Method for Determination of R- and S-Acenocoumarol in Human Plasma and Its Application in a Pharmacokinetics Study

A stereoselective, sensitive and rapid liquid chromatography-tandem mass spectrometry (LCMSMS) method for the determination of R-acenocoumarol and S-acenocoumarol in human plasma was developed and validated at IPRC bioanalytical labs. The procedure involved solid phase extraction of both enantiomers and their corresponding internal standard. The chromatographic separation was accomplished employing a chiral column and proper mobile phase. Detection was carried out using Waters Micromass® Quattro Premier mass spectrometer in multiple reaction monitoring (MRM) mode using turbo ion spray with negative ionization. The method was validated over a linear range of 0.40 40.00 ng/ml for R-acenocoumarol and 0.20 20.00 ng/ml for the S-acenocoumarol. Method validation covered different parameters such as linearity, accuracy, precision and stability. The method was successfully applied for the determination of R and Sacenocoumarol in plasma samples of 28 healthy subjects who participated in a pharmacokinetics study.