Stability Indicating Method Development And Validation Of Tenofovir And Lamivudine By Uv Spectrophotometer And Rp-Hplc

Abstract

This study aimed to develop and validate stability-indicating methods for the analysis of Tenofovir and Lamivudine using UV spectrophotometry and RP-HPLC. Optimization of chromatographic parameters including selection of method, ionic strengths of mobile phase, mobile phase ratio, mobile phase pH, and flow rate was conducted. A binary combination of water and methanol was utilized for drug concentration determination. Wavelengths of 265 nm for Tenofovir and 271 nm for Lamivudine were chosen. Methanol and water at a ratio of 60:40% v/v were found suitable as the mobile phase. The retention times for Tenofovir and Lamivudine were 2.7 and 2.67 minutes, respectively. The developed method adhered to ICH standards and exhibited linear calibration curves for Tenofovir within the concentration range of 10 to 30 µg/mL. The LOD and LOQ were found to be 0.105 µg/mL and 0.32 µg/mL, respectively. The method demonstrated high precision with low RSDs for intraday and interday accuracy. Recovery experiments confirmed method reproducibility. Stability studies revealed degradation under oxidative conditions. The UV method proved suitable for routine analysis, while the RP-HPLC method offered simplicity, selectivity, precision, and accuracy, with better sensitivity. Overall, both methods are applicable for Tenofovir and Lamivudine analysis, with RPHPLC being more sensitive and accurate. The stability studies indicated significant degradation under oxidative conditions compared to other degradation types.it is concluded that the developed UV and RP-HPLC methods offer reliable means for the analysis of Tenofovir and Lamivudine in formulations, complying with ICH guidelines. The RP-HPLC method demonstrated superior sensitivity and accuracy, although both methods are applicable. Stability studies highlighted the susceptibility of the drugs to oxidative degradation.