期刊
JOURNAL OF CHROMATOGRAPHY A
卷 1215, 期 1-2, 页码 145-151出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.chroma.2008.11.009
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
资金
- US National Institute of Allergy and Infectious Diseases [NIH N01-AO-00042]
- Wellcome Trust of Great Britain [077166/Z/05/Z]
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 I ng/mL. and the upper limit of quantification was 50,000 ng/mL The within-clay and between-clay 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. (C) 2008 Elsevier B.V. All rights reserved.
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