The objective of this study was to validate a simple and precise Ultra Performance liquid chromatographic method with Tandem Mass Spectrometry-
(AB SCIEX) method for the determination of Asenapine and N-Desmethyl Asenapine (metabolite) in human plasma using Asenapine Maleate 13C D3 as Internal Standard (IS). The precision and accuracy data have to fulfill the requirements for quantification of the analytes in biological matrices to generate
data for bioequivalence, bioavailability, pharmacokinetic or toxicology investigations. A Hypersil GOLD C18, 5μ column having 4.6 x 50 mm internal
diameter in binary gradient mode with flow rate was 0.6 mL/min of mobile phase containing acetonitrile and ammonium formate (90:10) were used. The experiments were performed by loading in UPLC with a triple quadruple mass spectrometer, operating in the multiple reaction monitoring (MRM) modes. The
method was validated over the concentration range of 0.1080 – 35.314 ng/mL(ANALYTE) and 0.1060 – 34.47 ng/mL (METABOLITE), by using 500
μL plasma samples.The mean recovery of Asenapine (81%) and N-Desmethyl
Asenapine (80%) from spiked plasma samples was consistent and reproducible.
The method was validated for linearity, accuracy, precision, specificity, and robustness. The intra- and inter-day precision and accuracy values were found
to be within the assay variability limits as per the FDA guidelines. The developed assay method was applied to a clinical pharmacokinetic study in human volunteers
