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Simultaneous quantification of fludarabine and cyclophosphamide in human plasma by high‐performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry
Author(s) -
Silvertand L. H. H.,
Vazvaei F.,
Weigl P.,
Rosing H.,
Hillebrand M. J. X.,
van Maanen M. J.,
Beijnen J. H.
Publication year - 2005
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.2242
Subject(s) - chemistry , chromatography , protein precipitation , electrospray ionization , selected reaction monitoring , triple quadrupole mass spectrometer , tandem mass spectrometry , mass spectrometry , liquid chromatography–mass spectrometry , electrospray , fludarabine , high performance liquid chromatography , analytical chemistry (journal) , cyclophosphamide , medicine , surgery , chemotherapy
Fludarabine and cyclophosphamide are anticancer agents mainly used in the treatment of hematologic malignancies. We have developed and validated an assay using high‐performance liquid chromatography (HPLC) coupled with electrospray ionization tandem mass spectrometry for the quantification of fludarabine in combination with cyclophosphamide in human heparin and human EDTA plasma. Sample pre‐treatment consisted of a protein precipitation with cold acetonitrile (−20°C) using 250 µL of plasma. Separation was performed on an Extend C18 column (150 × 2.1 mm i.d.; 5 µm) with a stepwise gradient using 1 mM ammonia solution and acetonitrile at a flow rate of 400 µL/min. The analytical run time was 12 min. The triple quadrupole mass spectrometer was operated in the positive ion mode and multiple reaction monitoring was used for drug quantification. The method was validated over a concentration range of 1 to 100 ng/mL for fludarabine and cyclophosphamide in human heparin and human EDTA plasma. The coefficients of variation were <13.9% for inter‐ and intra‐day precisions. Mean accuracies were also within the designated limits (±15%). The analytes were stable in plasma, processed extracts and in stock solution under all relevant conditions. Copyright © 2005 John Wiley & Sons, Ltd.