Development and Validation of UPLC-MS/MS Bioanalytical Method for Simultaneous Quantification of the Antiretroviral Drugs Dolutegravir, Lamivudine, and Tenofovir in Human Plasma

Abstract

The current proposed validated bio-analytical UPLC-MS/MS method for assessing components like dolutegravir, lamivudine, and tenofovir in human plasma using bictigravir as an internal standard with a mobile phase of formic acid (0.1%), 2 Mm ammonium formate in water and acetonitrile (30:70, v/v), and optimized flow rate of 0.5 mL/min, the separation was performed on an Agilent XDB C18 column (250 mm X 4.6, 5μ). The three components were analyzed by using LC-MS/MS in positive ion mode owing to the presence of primary groups. For the technique validation for the concentration range of 20 to 1000ng/mL for dolutegravir, lamivudine, and tenofovir in human plasma, bitigravir was employed as the internal control. The three-drug moieties average recoveries from spiking plasma samples were discovered to be reproducible. Based on the data displayed above, it was determined that the procedure was rapid and dependable, with a minimum total run duration of 3.0 minutes. The stability studies were assessed as a consequence of the current approach’s successful validation in accordance with the food and drug administration (FDA), european medicines agency (EMA), and international council for harmonisation (ICH) requirements.