Use of Systematic Approach for Development of RP-HPLC Method for Simultaneous Determination of Lopinavir and Ritonavir

AbstractA simple and robust reverse phase high-performance liquid chromatographic (RP-HPLC) method was developed and validated for the quantitative estimation of Lopinavir and Ritonavir in bulk and formulation. The systematic approach, one of the parts of QbD was use for the analytical method development. Chromatographic separation was conducted on Phenomenex Gemini C18 (250 mm×4.6 mm, 5 µ) column at 25°C using Sodium dihydrogen phosphate buffer: acetonitrile (pH adjusted to 5.3±0.1 with acetic acid) in the ratio of 45:55 v/v. The flow rate was 1 ml/min and UV detection was carried out at 262 nm. The retention times of Lopinavir and Ritonavir were 8.90 min and 5.36 min respectively. The method was found to be linear in the range of 5 30 µg/ml for lopinavir and 2 10 µg/ml for ritonavir. The developed method was validated for specificity, accuracy, precision, LOD, LOQ, robustness as per ICH guidelines.