Development and validation of a high-throughput zwitterionic hydrophilic interaction liquid chromatography solid-phase extraction–liquid chromatography–tandem mass spectrometry method fo

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• 作者：N. Lindegardh ; W. Hanpithakpong ; A. Phakdeeraj ; P. Singhasivanon ; J. Farrar ; T.T. Hien ; N.J. White ; N.P.J. Day
• 关键词：Avian influenza ; Birdflu ; High-throughput ; HILIC ; Ion suppression ; Liquid chromatography/tandem mass spectrometry (LC/MS/MS) ; Peramivir ; Solid phase extraction ; Stable isotope labeled (SIL) internal standard
• 刊名：Journal of Chromatography A
• 出版年：2008
• 出版时间：26 December 2008
• 年：2008
• 卷：1215
• 期：1-2
• 页码：145-151
• 全文大小：755 K

文摘

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.