Zeitschrift für Chromatographie und Trenntechniken
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High-Performance Liquid Chromatography with Diode Array Detection for the Simultaneous Quantification of Piperaquine and Mefloquine in Human Plasma in Vietnamese Uncomplicated Malaria Patients

Pham Van Toi*, Truong Le Phuc Nhi, Nguyen Hoang Chau, Nguyen Thanh Tong, Guy. E. Thwaites, Tran Tinh Hien

A bioanalytical method using high-performance liquid chromatography with diode array detection was developed and validated for quantification of piperaquine and mefloquine in human plasma. After extraction from 200 μL of plasma by using solid phase extraction on MCX-96 well plate; piperaquine, mefloquine and quinine (internal standard) were separated using a mobile phase consisting of trichloroacetic acid 1 mM and trimethylamine 0.025%, pH 3.4-Acetonitrile applying gradient elution on a Poroshell 120, C8 end capped 100 × 4.6 mm, 2.7 μm column at a flow rate of 1.0 mL/min at 35°C. The peaks were monitored with a diode array detection set at a wavelength of 343 nm for piperaquine, quinine and 283 nm for mefloquine. The limit of quantification was 10 ng/mL, 50 ng/ mL for piperaquine and mefloquine, respectively. The method was linear over the range of 10 to 2,000 ng/ml for piperaquine and 50 to 10,000 ng/ml for mefloquine in plasma. The intra, inter assay precision were less than 8% overall quality control ranges for these analytes. The accuracies varied between 99.85% and 104.6% at all quality control levels for piperaquine and mefloquine. The validated method was applied to measure the concentrations of piperaquine, mefloquine in samples collected from 12 uncomplicated malaria patients in Binh Phuoc province, Vietnam.