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Objective: The objective of the study was to develop and validate a sensitive, precise, and accurate stability indicating reverse-phase (RP) high-performance liquid chromatography method for the quantification of saquinavir, ritonavir, and amprenavir simultaneously. Methods: The determination of saquinavir, ritonavir, and amprenavir in their mixtures was done using a mobile phase consisted of 0.1M phosphate buffer (pH 3.5) and methanol (70:30, v/v). The method is based on the simultaneous separation of studied drugs in a RP Inertsil ODS C18 (4.6 mm×100 mm, 5 μm) column at ambient temperature. Detection and quantitation were achieved with photodiode array detector set at 260 nm. Results: Saquinavir, ritonavir, and amprenavir showed linearity over a concentration range of 40–200 μg/ml (R2-0.9994), 20–100 μg/ml (R2-0.9992), and 30–150 μg/ml (R2-0.9990), respectively. The limit of quantification was 0.64 μg /ml, 0.57 μg /ml, and 0.53 μg /ml for saquinavir, ritonavir, and amprenavir, respectively. The accuracies for the three drugs were in the range of 99.40–100.53% (saquinavir), 99.45-100.47% (ritonavir), and 100.03–100.53% (amprenavir). The percentage relative standard deviations for the studied drugs were 0.785–0.848% (saquinavir), 0.338–0.499% (ritonavir), and 0.336–0.775% (amprenavir). No peaks were observed at the retention time of saquinavir, ritonavir, and amprenavir in placebo blank, mobile phase blank and stress degraded samples which suggested that the proposed was selective and specific. Conclusion: The method was found to be suitable for the regular analysis of saquinavir, ritonavir, and amprenavir simultaneously in the presence of their stress degradation products. Objective: The objective of the study was to develop and validate a sensitive, precise, and accurate stability indicating reverse-phase (RP) high-performance liquid chromatography method for the quantification of saquinavir, ritonavir, and amprenavir simultaneously. Methods: The determination of saquinavir, ritonavir, and amprenavir in their mixtures was done using a mobile phase consisted of 0.1M phosphate buffer (pH 3.5) and methanol (70:30, v/v). The method is based on the simultaneous separation of studied drugs in a RP Inertsil ODS C18 (4.6 mm×100 mm, 5 μm) column at ambient temperature. Detection and quantitation were achieved with photodiode array detector set at 260 nm. Results: Saquinavir, ritonavir, and amprenavir showed linearity over a concentration range of 40–200 μg/ml (R 2 -0.9994), 20–100 μg/ml (R 2 -0.9992), and 30–150 μg/ml (R 2 -0.9990), respectively. The limit of quantification was 0.64 μg /ml, 0.57 μg /ml, and 0.53 μg /ml for saquinavir, ritonavir, and amprenavir, respectively. The accuracies for the three drugs were in the range of 99.40–100.53% (saquinavir), 99.45-100.47% (ritonavir), and 100.03–100.53% (amprenavir). The percentage relative standard deviations for the studied drugs were 0.785–0.848% (saquinavir), 0.338–0.499% (ritonavir), and 0.336–0.775% (amprenavir). No peaks were observed at the retention time of saquinavir, ritonavir, and amprenavir in placebo blank, mobile phase blank and stress degraded samples which suggested that the proposed was selective and specific. Conclusion: The method was found to be suitable for the regular analysis of saquinavir, ritonavir, and amprenavir simultaneously in the presence of their stress degradation products.

asian journal of pharmaceutical and clinical research

DEVELOPMENT AND VALIDATION OF STABILITY INDICATING REVERSE-PHASE HIGHPERFORMANCE LIQUID CHROMATOGRAPHY METHOD FOR THE SIMULTANEOUS QUANTIFICATION OF SAQUINAVIR, RITONAVIR, AND AMPRENAVIR