Development and validation of an HPLC method for quantification of dolutegravir in human plasma

Abstract

Dolutegravir (DTG) has been the first‐line drug in many human immunodeficiency virus/acquired immunodeficiency syndrome (HIV/AIDS) guidelines for the treatment of naïve and experienced HIV‐infected individuals, which calls for cost‐effective and convenient methods for quantitative detection of DTG in human plasma for pharmacokinetic studies and patient adherence evaluation. Here, an HPLC–ultraviolet method in combination with liquid–liquid extraction with isocratic elution was developed for the first time. The analysis was performed on a CLC‐ODS column (6 mm internal diameter × 15 cm, 5 μm) using a mixture of acetonitrile and phosphate buffer (40:60, v/v) as the mobile phase at the flow rate of 1 mL/min. Using triamcinolone as the internal standard, 100 μL of plasma sample was extracted by methyl tert‐butyl ether, followed by evaporating under nitrogen stream, re‐dissolving with 100 μL mobile phase, and injection of 20–40 μL of supernatant into the chromatographic system. The linearity of DTG was good in the range of 0.05–10 μg/mL (r = 0.9995), and the inter‐ and intra‐day variabilities were 0.4%–4.3% (n = 10) and 1.2%–6.2% (n = 10) for the lower limit of quantification, low‐, medium‐, and high‐concentration quality control samples (0.05, 0.1, 0.8, and 8 μg/mL), respectively, while the methodological and extraction recoveries were 98.0%–103.0% (n = 20) and 65.2%–75.7% (n = 3), respectively. This method was successfully applied to analyze DTG plasma concentration in 84 Chinese patients with HIV.