Estimation of Gliclazide in Human Plasma by LCMS/MS

Gliclazide and Glipizide (IS) were separated on a Reverse phase chromatography using the mobile phase mixture of ammonium acetate and methanol at a flow rate of 1.0 ml/min after extraction from plasma by liquid – liquid extraction method. Both the analyte and the IS were detected in API 4000 Mass spectrometer in the positive atmospheric pressure Ionization (API) mode with multiple reactions monitoring (MRM). The MRM transitions were monitored by following m/z for parent ion 324.1 and daughter ion 127.2 (Gliclazide), and m/z 446.1 and daughter 321.1 (Glipizide, IS). A linear calibration plot of Gliclazide was achieved in the concentration ranges of 7.776 ng/ml to 6000 ng/ml. Mean recovery was 75.67%.This method was fully validated for specificity, precision, accuracy, reproducibility and other criteria as per regulations.