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METHOD DEVELOPMENT AND VALIDATION FOR THE SIMULTANEOUS ESTIMATION OF RITONAVIR AND LOPINAVIR IN PHARMACEUTICAL DOSAGE FORMS BY RP-HPLC
Gorja Ashok*, Jahnavi Bandla
ABSTRACT A simple, rapid, accurate and precise method was developed and validated for the simultaneous estimation of Ritonavir and Lopinavir in pharmaceutical dosage form. The method was based on RP-HPLC. Chromatographic separation was performed on Zorbax SB-C18 (150mm x 4.6mm, 3.5μm particle size) column using a mobile phase consisting of a mixture of KH2PO4 buffer (pH 2.5 with dilute orthophosphoric acid): Acetonitrile (50:50%v/v) in an isocratic mode. The following system conditions were maintained throughout development and validation i.e., flow rate 1.2mL/min, column was maintained at room temperature and the detected by a UV-wave length at 260nm. The Ritonavir and Lopinavir were well resolved on the stationary phase and the retention times were 6.9 and 8.0minutes respectively. The method was validated; Ritonavir and Lopinavir were shown to be linear over a range of 0.5-1.5mg/mL and 2.0-6.0mg/mL respectively. The limit of quantification was 0.16mg/mL for Ritonavir and 0.53mg/mL for Lopinavir and the limit of detection was 0.05mg/mL for Ritonavir and 0.18mg/mL for Lopinavir. The Precision, Accuracy, Specificity, Ruggedness and Robustness were determined to validate the method. Keywords: Ritonavir, Lopinavir, RP-HPLC, Method development, Validation and C18 Column. [Download Article] [Download Certifiate] |