An Alternative HPLC with Ultraviolet Detection for Determination of Piperaquine in Plasma.

A simple, sensitive, selective and reproducible method based on liquid chromatography was developed for the determination of piperaquine in human plasma samples. Sample preparation involved a single step liquid-liquid extraction by organic solvents (hexane: tert-butyl methylether at the ratio of 1:1, v:v). Piperaquine was separated from the internal standard mefloquine on a reversed-phase C18 column, with the mobile phase consisting of a mixture of acetonitrile and 0.1% trichloroacetic acid (in water) (15:85, v:v) running at a flow rate of 1.0 mL/min. Retention times of piperaquine and mefloquine were 9.92 and 14.20 min, respectively. Ultraviolet detection was set at the wavelength 354 nm. Good precision and accuracy were obtained for both within-day repeatability and day-to-day reproducibility. Limit of quantification for piperaquine was accepted as 10 ng/mL using 150-μL plasma sample. The mean recoveries for piperaquine and internal standard were between 88.8% and 91.7%. The validated analytical method was successfully applied in twenty blinded spiked human plasma samples.