Abstract

DEVELOPMENT AND VALIDATION OF A LIQUID CHROMATOGRAPHY-MASS SPECTROMETRY METHOD FOR QUANTIFICATION OF CARBAMAZEPINE IN HUMAN PLASMA

Shravan Goski*, Dr. P. Venkatesh, Upendra Chowdary and N. Ajay Sai

ABSTRACT

This study presents the development and validation of a sensitive and accurate liquid chromatography-mass spectrometry (LC-MS/MS) method for the quantification of carbamazepine in human plasma samples. Chromatographic separation was achieved on a Zorbax SB C18, 4.6x75mm, 3.5μm column using a mobile phase consisting of acetonitrile and water with 0.1% formic acid. Liquid-liquid extraction (LLE) was optimized for efficient extraction of Carbamazepine from plasma matrices, ensuring minimal matrix interference and high recovery. Method validation was performed according to International Council for Harmonisation (ICH) guidelines, demonstrating satisfactory results for specificity, linearity, accuracy, precision, sensitivity, dilution integrity, and stability. Detection and
quantification were performed using mass spectrometryin positive ionization mode, utilizing multiple reaction monitoring (MRM) for enhanced specificity and sensitivity. The method exhibited excellent linearity over a wide concentration range of 10.0-5000.0 ng/mL, with a correlation coefficient (r) 0.99. The lower limit of quantification (LLOQ) was determined to be suitable for clinical analysis. Validation of the method was conducted following international guidelines, including assessments of specificity, accuracy, precision, and stability. The validated method was successfully applied to the analysis of human plasma samples from patients undergoing carbamazepine therapy, demonstrating its reliability and utility for therapeutic drug monitoring and pharmacokinetic studies. This LC-MS/MS method offers a robust and efficient approach for the quantitative determination of carbamazepine in human plasma, providing valuable support for clinical decision-making and pharmacological research in neurological and psychiatric disorders.

Keywords: Carabamazepine, LC-MS/MS, Human Plasma(K3EDTA).