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Abstract
A sensitive, specific, and simple high performance liquid chromatography assay with fluorometric detection for the determination of the antiretroviral agent emtricitabine in human plasma is described. Using 500 µL of plasma and Oasis MAX columns, the solid phase extraction (SPE) method results in a clean baseline and high extraction efficiencies (107%). An Atlantis dC18 analytical column is used along with a 15 min linear gradient elution of phosphate buffer and methanol to provide sharp peaks for emtricitabine at excitation and emission wavelengths of 244 and 356 nm, respectively. The method was validated over the range 0.01 to 5.0 mg/L and is accurate (average accuracy of five different concentrations ranged from 100% to 107%) and precise (inter‐ and intra‐assay precision ranged from 0 to 3.7% and 1.8 to 5.3%, respectively). Participation in an external quality control program resulted in deviations for three different levels of less than 2.4% from the nominal concentrations. This method is suitable for use in clinical trials and therapeutic drug monitoring of HIV‐infected patients.