Bioanalytical method development and validation of a novel antiseizure agent Cenobamate using LC-MS/MS

Abstract

To quantify Cenobamate in human plasma, a precise and sensitive electron ionisation procedure in tandem mass spectrometry has been developed. The study was successfully confirmed by using Cenobamate -D4 as the internal norm and tert-butyl methyl ether in the testing of several positive ionic reactions. It has been used in liquid-liquid extraction to prepare samples as an extraction solvent. Cenobamate and Cenobamate – D4 (internal standard) were separated on an Eclipse C18 column (150 mm, 4.6 mm, 5 µm) by isocratic elution. This process was performed using a mobile phase of 0.1% formic acid in 20:80% v/v ratio of acetonitrile with a flow rate of 0.6 ml/min. The observed mass transitions for Cenobamate and D4 were m/z (amu) 268.23, 198.10 and 272.11 198.10 for each. The lower quantitative limit was 10 ng/ml based on 500 ng/ml plasma, with no chromatogram disturbance observed. A linear curve was observed with a correlation coefficient (r2) of 0.999 from 10 to 500 ng/ml. The intraday and interday precision variations were 15% and the accuracy values were 15% of the relative error values (RE). Extraction recovery rates are in the tolerance ranges of less than 15%. The new LC-MS/MS method complies with all regulatory requirements and demonstrates satisfactory accuracy and accuracy and is sensitive enough to detect Cenobamate in humans.