A Rapid and Sensitive RP-UPLC Method for Simultaneous Determination of Zidovudine, Lamivudine and Nevirapine in Tablet Dosage form

The objective of the current study was to develop a validated ultra performance liquid chromatographic (UPLC) method for the quantitation of the novel antiretroviral agents zidovudine, lamivudine and nevirapine in tablet dosage form. The method utilizes sample preparation step followed by separation on a waters C¹⁸ column consisting high strength silica (ACQUITY UPLC HSS-T3), 100 × 2.1 mm, 1.8 mm particle size, in gradient mode using 0.1%, v/v trifluoroacetic acid in water and acetonitrile as the mobile phase. The analytical column was thermostated at 50°C and flow rate was set at 0.4 mL/min. The eluted peaks were detected by photo diode array (PDA) detector at a wavelength of 266 nm. The method was validated with respect to linearity, accuracy, precision and robustness. The responses were linear over the ranges of 100-500, 50-250 and 100-180 mg/mL for zidovudine, lamivudine and nevirapine, respectively. The limit of detection and limit of quantification were found 0.279, 0.068, 0.360 and 0.846, 0.205, 1.091 mg/mL for zidovudine, lamivudine and nevirapine, respectively. The utility of the procedure was verified by its application to formulations and found that the developed method is fast, accurate, precise, selective and reproducible.