A Novel UPLC–MS/MS Method for Determination of γ-Amino Butyric acid Analogue in Human Plasma: Application to Pharmacokinetic Study.

ABSTRACT

Pregabalin, a structural analogue of γ-amino butyric acid, is an anticonvulsant used for neuropathic pain. To facilitate its pharmacokinetic study in human subjects, ultra-performance liquid chromatography–mass spectrom etric method employing positive electrospray ionization was developed for the determination of Pregabalin concentration in human plasma. Pregabalin together with the internal standard was extracted from 125μl of human plasma by solid phase extraction. The chromatography was performed using an amide column and multiple reaction monitoring detection mode was used for the quantification of Pregabalin in plasma.The validation of the method including sensitivity, linearity, reproducibility and stability was examined.The lower limit of quantification (LLOQ) of the developed method for Pregabalin was 0.1 μg/mL and the linear calibration curve was acquired with r > 0.99 between 0.1 and 20 μg/ml. The intra-day and inter-day variation of the current assay was evaluated with the coefficient of variations within 10.73% at LLOQ and 9.8% for other quality control samples, whereas the mean accuracy ranged from 96.8% to 107.6%. The present method provides a robust, rapid and sensitive analytical tool for Pregabalin in human plasma and has been successfully applied to a clinical pharmacokinetic study where Pregabalin was orally administered in healthy subjects.