Novel liquid chromatography-tandem mass spectrometry method for simultaneous detection of three anti-HIV drugs tenofovir, lamivudine, and dolutegravir in plasma.

Currently, the world health organization (WHO) recommends a fixed-dose combination therapy called TLD, composed of tenofovir, lamivudine, and dolutegravir (TLD) for human immunodeficiency virus (HIV) treatment. About 23 of 39 million people living with HIV/AIDS (PLWHA) are on a daily TLD pill. In this study, we successfully developed a simple, efficient, and sensitive method to extract and enable the simultaneous quantification of tenofovir, lamivudine, and dolutegravir in rats, non-human primates (NHP), and human plasma samples. The 3 HIV drugs in small volume, 100 µL plasma were extracted after protein precipitation, and all the analytes were quantified using a novel liquid chromatography-tandem mass spectrometry. The assay provided analysis of all three analytes in rat, NHP, and human plasma. This method was used to evaluate time-plasma drug concentrations in two NHPs (Macaca nemestrina). The reported bioanalytical method demonstrated high sensitivity, reproducibility, and robustness for quantifying the 3 HIV drugs with efficiency. Lower limit of quantification (LLOQ) for tenofovir (TFV), lamivudine (3TC) and dolutegravir (DTG) were between 0.3 - 2.6 ng/mL equivalent to 30 - 260 pg in 100 µL of rats, NHPs, or human plasma samples. This assay may be adapted for evaluating drug concentrations across multiple species to support preclinical, clinical, and translational studies for developing long-acting products.