title: High throughput assay for the determination of piperaquine in plasma.
abstract: 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.affiliation: Faculty of Tropical Medicine, Mahidol University, Bangkok 10400, Thailand. niklas@tropmedres.ac
authors.forename: N
authors.initials: N
authors.lastname: Lindegårdh
authors.forename: N J
authors.initials: NJ
authors.lastname: White
authors.forename: N P J
authors.initials: NP
authors.lastname: Day
mesh_terms: D000818:Animals; D000962:Antimalarials / Q000032:analysis* / Q000097:blood*; D002138:Calibration; D002626:Chemistry, Pharmaceutical / Q000379:methods*; D002845:Chromatography; D002853:Chromatography, Liquid; D004305:Dose-Response Relationship, Drug; D004345:Drug Industry / Q000379:methods*; D016903:Drug Monitoring / Q000379:methods*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008956:Models, Chemical; D011804:Quinolines / Q000032:analysis* / Q000097:blood*; D015203:Reproducibility of Results; D013997:Time Factors; D014466:Ultraviolet Rays; D010879:Piperazines
chemical_list: D000962:Antimalarials; D011804:Quinolines; C034759:piperaquine; D010879:Piperazines