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Validated assay for the simultaneous determination of the anti‐cancer agent gemcitabine and its metabolite 2′,2′‐difluorodeoxyuridine in human plasma by high‐performance liquid chromatography with tandem mass spectrometry
Author(s) -
Vainchtein Liia D.,
Rosing Hilde,
Thijssen Bas,
Schellens Jan H. M.,
Beijnen Jos H.
Publication year - 2007
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.3096
Subject(s) - chemistry , chromatography , ammonium acetate , metabolite , high performance liquid chromatography , tandem mass spectrometry , mass spectrometry , electrospray ionization , acetonitrile , liquid chromatography–mass spectrometry , selected reaction monitoring , extraction (chemistry) , biochemistry
Abstract A sensitive and specific high‐performance liquid chromatography/tandem mass spectrometry (HPLC/MS/MS) assay for the quantitative determination of gemcitabine (dFdC) and its metabolite 2′,2′‐difluorodeoxyuridine (dFdU) is presented. A 200‐µL aliquot of human plasma was spiked with a mixture of internal standards, didanosine, lamivudine and fludarabine, and extracted using solid‐phase extraction. Dried extracts were reconstituted in 1 mM ammonium acetate/acetonitrile (97:3, v/v) and 10‐µL volumes were injected onto the HPLC system. Separation was achieved on a 150 × 2.1 mm C18 bonded phase endcapped with polar groups (Synergi Hydro‐RP column) using an eluent composed of 1 mM ammonium acetate (pH 6.8)/acetonitrile (94:6, v/v). Detection was performed by positive ion electrospray ionization followed by MS/MS. The assay quantifies a range from 0.5 to 1000 ng/mL for gemcitabine and from 5 to 10 000 ng/mL for dFdU using 200 µL of human plasma sample. Validation results demonstrate that gemcitabine and dFdU concentrations can be accurately and precisely quantified in human plasma. This assay is used to support clinical pharmacologic studies with gemcitabine. Copyright © 2007 John Wiley & Sons, Ltd.