Development and validation of a high-throughput zwitterionic hydrophilic interaction liquid chromatography solid-phase extraction-liquid chromatography-tandem mass spectrometry method for determination of the anti-influenza drug peramivir in plasma.

Journal article

An assay for the analysis for the quantification of the anti-influenza drug peramivir in human plasma using high-throughput zwitterionic (ZIC) hydrophilic interaction liquid chromatography (HILIC) solid-phase extraction (SPE) in a 96-wellplate format and liquid chromatography coupled to positive tandem mass spectroscopy has been developed and validated. The ZIC-HILIC SPE efficiently removed sources of interference present in the supernatant after protein precipitation of plasma proteins. The main advantage of the ZIC-HILIC SPE sample preparation step was that it allowed load and elution conditions to be optimised to extract only peramivir and minimize co-extraction of lipophilic phospholipids. The method was validated according to published US Food and Drugs Administration guidelines and showed excellent performance. The assay was validated over two calibration ranges (0.952-500 and 50-50,000 ng/mL) to support analysis of peramivir after intra-venous administration. The lower limit of quantification for peramivir in plasma was 1 ng/mL and the upper limit of quantification was 50,000 ng/mL. The within-day and between-day precisions expressed as RSD, were lower than 8% at all tested quality control concentrations and below 11% at the lower limit of quantification. Validation of over-curve samples ensured that it would be possible with dilution if samples went outside the calibration range.

Authors: Lindegardh, N; Hanpithakpong, W; Phakdeeraj, A; Singhasivanon, P; Farrar, J

• Publication status: Published
• Journal: Journal of chromatography. A
• Volume: 1215
• Issue: 1-2
• Pages: 145-151
• Publication date: 2008-12-01
• DOI: 10.1016/j.chroma.2008.11.009
• EISSN: 1873-3778
• ISSN: 0021-9673
• Language: English
• Keywords: Humans; Tandem Mass Spectrometry; Reference Standards; Guanidines; Reproducibility of Results; Chromatography, Liquid; Cyclopentanes; Antiviral Agents