Reversed-phase liquid chromatographic method with spectrophotometric detection for the determination of antiretroviral drugs Original Research Article

In the present paper, a new chromatographic method for the determination of acquired immune deficiency syndrome (AIDS) drugs in plasma samples is proposed. The method consists of solid-phase extraction for sample pretreatment and further chromatographic analysis. Drugs have been separated on a C18 column using an elution gradient based on an increase in the acetonitrile percentage. Analytes have been detected spectrophotometrically at 240, 250, 260 and 280 nm. Chromatographic conditions have been thoroughly optimized using experimental design and multicriteria functions. Analytical parameters of the method have been established for both synthetic and plasma samples. Limits of detection are around 5 ng mL−1 for reverse transcriptase inhibitors (nucleoside and non-nucleoside) and 20 ng mL−1 for protease inhibitors. The method has been validated through a spiking/recovery procedure at three concentration levels. Results obtained are highly satisfactory, with recovery values around 100% for all drugs. The method has been applied to the determination of various drug mixtures of medical interest in plasma samples.