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Quantification of fudosteine in human plasma by high-performance liquid chromatography-electrospray ionization mass spectrometry employing precolumn derivatization with 9-fluorenylmethyl chloroformate

✍ Scribed by Fengguo Xu; Zunjian Zhang; Haoyan Jiao; Yuan Tian; Beibei Zhang; Yun Chen

Publisher
John Wiley and Sons
Year
2006
Tongue
English
Weight
148 KB
Volume
41
Category
Article
ISSN
1076-5174

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✦ Synopsis

Abstract

This paper describes a novel method for the sensitive and selective determination of fudosteine in human plasma. The method involves a derivatization step with 9‐fluorenylmethyl chloroformate (FMOC‐Cl) in borate buffer and detection based on high‐performance liquid chromatography‐electrospray ionization mass spectrometry (LC/ESI/MS). After acetonitrile‐induced protein precipitation of plasma samples, fudosteine was derivatized with FMOC‐Cl, then extracted by ethyl acetate, evaporated, reconstituted and injected using an LC/ESI/MS instrument. Separation was achieved using an ODS column and isocratic elution. Excellent linearity was obtained for the entire calibration range from 0.05 to 20 µg/ml. Validation assays of the lower limit of quantification (LLOQ) as well as for the intra‐ and inter‐batch precision and accuracy met the international acceptance criteria for bioanalytical method validation. Using the developed analytical method, fudosteine could be detected for the first time in human plasma with a low limit of detection (LLOD) of 0.03 µg/ml. The proposed method has been successfully applied to study the pharmacokinetics of fudosteine in healthy Chinese volunteers after single and multiple oral administration. Copyright © 2006 John Wiley & Sons, Ltd.

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