High throughput assay for the determination of piperaquine in plasma

J Pharm Biomed Anal. 2005 Sep 15;39(3-4):601-5. doi: 10.1016/j.jpba.2005.03.031.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antimalarials / analysis*
  • Antimalarials / blood*
  • Calibration
  • Chemistry, Pharmaceutical / methods*
  • Chromatography
  • Chromatography, Liquid
  • Dose-Response Relationship, Drug
  • Drug Industry / methods*
  • Drug Monitoring / methods*
  • Humans
  • Hydrogen-Ion Concentration
  • Models, Chemical
  • Quinolines / analysis*
  • Quinolines / blood*
  • Reproducibility of Results
  • Time Factors
  • Ultraviolet Rays

Substances

  • Antimalarials
  • Quinolines
  • piperaquine