Analytical method validation for tenofovir alafenamide and known impurities

Abstract

This study comprehensively validates an analytical method for assessing Tenofovir alafenamide and its related impurities using UPLC. The results demonstrate that the method is highly specific, accurate, precise, linear, and robust. Specificity analysis confirms that the method exhibits high selectivity, with no interfering peaks observed at the retention time of Tenofovir alafenamide or its specified impurities when compared to blank diluents. The peaks for Tenofovir alafenamide and its known impurities are well resolved from each other and from potential blank peaks, as confirmed by peak purity analysis. Linearity is established for various impurities in relation to the limit of detection (LOD) and limit of quantification (LOQ). Accuracy testing reveals that most impurities exhibit recoveries within acceptable ranges at different concentration levels, with low %RSD values indicating method accuracy. The method's range is justified for concentrations spanning from the LOQ to 150% of the test concentration, supported by precision, linearity, and accuracy characteristics within this range. Robustness is confirmed through parameter variations, ensuring consistent results under different conditions. Stability studies show that the standard preparation remains stable for up to 13 hours when stored at 8°C, with immediate injection recommended for system suitability and test preparations. The mobile phase remains stable for 79 hours at room temperature. In conclusion, this validated UPLC method is suitable for routine analysis of Tenofovir alafenamide Fumarate and its related substances. It meets all necessary criteria for reliable analysis and exhibits high selectivity and robustness.