CHEMOMETRIC OPTIMIZATION FOR HPLC ANALYSIS OF SIMULTANEOUS DETERMINATION OF TENOFOVIR AND EMTRICITABINE IN HUMAN PLASMA

Abstract

This research paper dealt with chemometric optimization for HPLC analysis of simultaneous determination of the nucleotide tenofovir (TFV) and nucleoside emtricitabine (FTC) reverse transcriptase inhibitors in human plasma. Central composite design and Derringer’s desirability function were used for optimization. The ranges of independent variables used for the optimization were MeOH concentration (A) (50- 60%), phosphate buffer molarity (B) (pH 4.0) (18-22) and flow rate (C) (0.8-.2 mL/min). The influence of this independent variable on the output responses were the capacity factor for 1st peak i.e. Tenofovir (k1), the resolution between two pairs of lamivudine (IS), emtricitabine (Rs2, 3) and the retention time of last peak i.e. emtricitabine (tR3). Using this strategy, mathematical model were defined and response surface were derived for the separation. Optimum condition chosen for assay were methanol concentration 60% v/v, phosphate buffer molarity (pH 4.0)18.35 mM and flow rate 0.8 mL/min. Plasma samples were prepared by liquid liquid extraction of the analyte and lamivudine used as internal standard. Peak area ratio of the analyte and internal standard was used for the quantification of the plasma samples. The method was fully validated for its sensitivity, selectivity, accuracy, precision, recovery and stability. The method was found to be simple and hence it could be applied in bioavailability studies.