High throughput assay for the determination of piperaquine in plasma.

Journal article

A high throughput assay for the determination of the antimalarial piperaquine in plasma has been developed and validated. The assay utilises 96-wellplate formats throughout the whole procedure, and easily enables a throughput of 192 samples a day using a single LC system. Buffer (pH 2.0; 0.05 M) containing internal standard was added to 0.25 mL plasma in a 96-wellplate (2 mL wells). The samples were extracted on a MPC solid phase extraction deep well 96-wellplate (3M Empore). Piperaquine and internal standard were analysed by liquid chromatography with UV detection on a Chromolith Performance (100 mm x 4.6 mm) column with a mobile phase containing acetonitrile-phosphate buffer (pH 2.5; 0.1 M) (8:92, v/v) at a flow rate of 3.0 mL/min. The within-day precisions for piperaquine were 3.3 and 2.3% at 40 and 1250 ng/mL, respectively. The between-day precisions for piperaquine were 5.8 and 1.3% at 40 and 1250 ng/mL, respectively. The total assay precisions using 29 replicates over 5 days were 6.7, 4.5 and 2.7% at 40, 200 and 1250 ng/mL, respectively. The lower limit of quantification (LLOQ) and the limit of detection (LOD) were 10 and 5 ng/mL, respectively using 0.25 mL plasma. Using 1 mL of plasma, it was possible to decrease LLOQ and LOD to 2.5 and 1.25 ng/mL, respectively.

Authors: Lindeg?rdh, N; White, N; Day, N

• Publication status: Published
• Journal: Journal of pharmaceutical and biomedical analysis
• Volume: 39
• Issue: 3-4
• Pages: 601-605
• Publication date: 2005-09-01
• DOI: 10.1016/j.jpba.2005.03.031
• EISSN: 1873-264X
• ISSN: 0731-7085
• Language: English
• Keywords: Chromatography, Liquid; Drug Monitoring; Dose-Response Relationship, Drug; Chemistry, Pharmaceutical; Antimalarials; Hydrogen-Ion Concentration; Models, Chemical; Time Factors; Quinolines; Calibration; Ultraviolet Rays; Animals; Chromatography; Humans; Drug Industry; Reproducibility of Results