Method development and validation of few anti-retroviral drugs in bulk and pharmaceutical dosage form by using UV visible spectroscopy

Abstract

A comprehensive study was undertaken to develop and validate multiple analytical methods for the simultaneous and individual estimation of antiviral drugs Dolutegravir, Lamivudine, Emtricitabine, and Tenofovir Alafenamide in bulk and pharmaceutical dosage forms. The methods included UV-Visible spectrophotometry (Methods 1–4), reversed-phase high-performance liquid chromatography (RP-HPLC; Methods 5–6), and liquid chromatography-tandem mass spectrometry (LC-MS/MS; Methods 7–8). Spectrophotometric methods, including Q-absorbance ratio and colorimetric techniques, demonstrated simplicity, precision, and cost-effectiveness. Absorbance maxima and is absorptive points were strategically utilized for drug quantification with acceptable limits of detection and quantification, making them suitable for routine quality control. RP-HPLC methods, employing O-Phosphoric acid and acetic acid buffer systems, achieved rapid separation with excellent linearity, accuracy, and robustness. LC-MS/MS techniques provided the highest sensitivity and selectivity, with ng/ml detection limits, minimal sample preparation, and high throughput analysis capabilities. Each method was validated as per ICH guidelines, ensuring precision, linearity, accuracy, and ruggedness. The developed methodologies offer scalable, reliable tools for routine analysis, quality assurance, and regulatory compliance in pharmaceutical environments, with method selection tailored to resource availability and sensitivity requirements.