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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
Author(s) -
Vainchtein Liia D.,
Rosing Hilde,
Mirejovsky Dorla,
Huynh Van,
Lenaz Luigi,
Hillebrand Michel J. X.,
Schellens Jan H. M.,
Beijnen Jos H.
Publication year - 2006
Publication title -
journal of mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.475
H-Index - 121
eISSN - 1096-9888
pISSN - 1076-5174
DOI - 10.1002/jms.1086
Subject(s) - chemistry , chromatography , metabolite , tandem mass spectrometry , electrospray , mass spectrometry , ammonium acetate , quantitative analysis (chemistry) , high performance liquid chromatography , selected reaction monitoring , biochemistry
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 m M 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.