A simple, fast, and precise reverse phase, High Performance Liquid Chromatography (HPLC) method was developed for the separation and quantification of Lamivudine, Zidovudine and Nevirapine in bulk drug and pharmaceutical dosage form. The quantification was carried out using C18 (250 × 4.6mm, 5μm) column and mobile phase comprising of methanol and phosphate buffer (pH 3) in proportion of 45:55 (v/v). Flow rate was maintained at rate of 0.8 ml/min and the effluent was monitored at 280 nm. The retention time of Lamivudine Zidovudine and Nevirapine were 3.12, 5.01 and 7.08 min respectively. Method was validated in terms of linearity, precision, accuracy, specificity, limit of detection and limit of quantitation. Linearity was 5.0-100.47 μg/ml for lamivudine, 10.0-170.0 μg/ml for zidovudine and 5.0-60.0 μg/ml for nevirapine respectively. Percentage recoveries of the drugs from the tablet formulation were found to be 99.7.%, 99.6% and 99.3% for Lamivudine, Zidovudine and Nevirapine respectively. Hence, proposed method was found to be suitable for simultaneous determination of Lamivudine Zidovudine and Nevirapine in pharmaceutical dosage form and bulk drug when compared to the reported method in terms of sensitivity, precision and accuracy.
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