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Quantitative analysis of EO9 (apaziquone) and its metabolite EO5a in human plasma by high-performance liquid chromatography under basic conditions coupled to electrospray tandem mass spectrometry

✍ Scribed by Liia D. Vainchtein; Hilde Rosing; Dorla Mirejovsky; Van Huynh; Luigi Lenaz; Michel J. X. Hillebrand; Jan H. M. Schellens; Jos H. Beijnen

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

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✦ Synopsis

Abstract

A sensitive and specific LC‐MS/MS assay for the quantitative determination of EO9 and its metabolite EO5a is presented. A 200‐µl human plasma aliquot was spiked with a mixture of deuterated internal standards EO9‐d~3~ and EO5a‐d~4~ and extracted with 1.25 ml ethyl acetate. Dried extracts were reconstituted in 0.1 M ammonium acetate—methanol (7 : 3, v/v) and 25 µl‐volumes were injected into the HPLC system. Separation was achieved on a 150 × 2.1 mm C18 column using an alkaline eluent (1 mM ammonium hydroxide—methanol (gradient system)). Detection was performed by positive ion electrospray followed by tandem mass spectrometry. The assay quantifies a range from 5 to 2500 ng/ml for EO9 and from 10 to 2500 ng/ml EO5a using 200 µl of human plasma samples. Validation results demonstrate that EO9 and EO5a concentrations can be accurately and precisely quantified in human plasma. This assay will be used to support clinical pharmacologic studies with EO9. Copyright © 2006 John Wiley & Sons, Ltd.

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