Simultaneous Determination of Raltegravir, Dolutegravir, Elvitegravir, and Bictegravir in Human Plasma Using High-performance Liquid Chromatography-tandem Mass Spectrometry

In this study, a highly sensitive method to simultaneously quantify the integrase strand transfer inhibitors (INSTIs) raltegravir, dolutegravir, elvitegravir, and bictegravir, which are recommended drugs in the HIV treatment guidelines, was established using liquid chromatography–tandem mass spectrometry (LC-MS/MS). Raltegravir-d3 was used as the internal standard substance. The plasma samples were deproteinized with methanol and analyzed by LC-MS/MS. Chromatographic separation was performed using the gradient method with a mobile phase A (20 mmol/l ammonium formate water) and mobile phase B (20 mmol/l ammonium formate methanol). In addition, an InertSustain C18 column (3 μm, 100 × 2.1 mm), a ow rate of 0.45 ml/min, and a measurement time of 10 minutes were used. The calibration curve showed linearity (r> 0.9904) within the range of 0.51,250 ng/ml, and the limit of quanti cation was 0.5 ng/ml for all drugs. The mean intraand inter-day accuracy was 99.6%± 7.2% and 101.0%± 5.0%, respectively, and the coef cient of variation (CV) was ≤ 18.5% and ≤ 10.3%, respectively. This method enables the highly sensitive simultaneous analysis of INSTIs and is useful for con rming the ef cacy and safety of drugs in clinical practice.