Simultaneous Estimation of Raltegravir and Lamivudine in Bulk and Pharmaceutical Dosage Forms by RP-HPLC: Method Development and Validation
Authors: Baratam Manisha, KEV NAGOJI, G. DIVYA, DOLA MANJU, KORADA KRISHNA, mendi TARUN
Keywords: Raltegravir, Lamivudine, RP-HPLC, method validation, ICH Q2(R1).
Abstract:
Raltegravir (RAV), an HIV-1 integrase inhibitor, and Lamivudine (LAV), a nucleoside reverse transcriptase inhibitor, are widely used in combination therapy for the treatment of HIV/AIDS. Their simultaneous estimation in fixed-dose formulations is essential for quality control and regulatory compliance. The present study describes the development and validation of a simple, accurate, and robust reverse-phase high-performance liquid chromatographic (RP-HPLC) method for the concurrent quantification of RAV and LAV in bulk and tablet dosage forms.
Chromatographic separation was achieved on an Inertsil ODS 3V column (250 × 4.6 mm, 5 μm) using a mobile phase of phosphate buffer (pH 3.0) and acetonitrile (55:45, v/v) at a flow rate of 1.0 mL/min, with detection at 275 nm. The method provided sharp and well-resolved peaks for both analytes within 10 minutes. Linearity was observed in the concentration range of 150–450 µg/mL for RAV and 50–150 µg/mL for LAV, with correlation coefficients of 0.9998 and 0.9986, respectively. Recovery studies confirmed the accuracy of the method, yielding mean values of 100.68% for RAV and 102.23% for LAV. Precision data indicated %RSD values below 2.0, while assay results for tablet formulations were 102.50% (RAV) and 99.54% (LAV). The method demonstrated adequate sensitivity, with LOD/LOQ values of 28.11/85.20 µg/mL for RAV and 0.433/1.313 µg/mL for LAV, and remained robust under deliberate variations in flow rate and wavelength. The validated method fulfills ICH Q2(R1) requirements and can be reliably applied for routine quality control and stability analysis of RAV and LAV in combined pharmaceutical dosage forms.
