TY - JOUR
T1 - Simultaneous determination of gemcitabine and its main metabolite, dFdU, in plasma of patients with advanced non-small-cell lung cancer by high-performance liquid chromatography-tandem mass spectrometry
AU - Marangon, Elena
AU - Sala, Federica
AU - Caffo, Orazio
AU - Galligioni, Enzo
AU - D'Incalci, Maurizio
AU - Zucchetti, Massimo
PY - 2008/2
Y1 - 2008/2
N2 - Gemcitabine, 2′,2′-difluoro-2′-deoxycytidine (dFdC) is a pyrimidine antimetabolite employed against several human malignancies. It undergoes intracellular activation to the pharmacologically active triphosphate form (dFdCTP) and metabolic inactivation to the metabolite 2′,2′- difluorodeoxyuridine (dFdU). In order to investigate the human plasma pharmacokinetics of dFdC and dFdU, we developed and validated an HPLC-MS/MS method, adding 2′-deoxycytidine as internal standard and simply precipitating the protein with acetonitrile. The method requires a small sample (125 μl), and it is rapid and selective, allowing good resolution of peaks from the plasma matrix in only 7 min. It is sensitive, precise and accurate, with overall precision, expressed as CV%, always less than 10.0% for both analytes and high recovery: ≥80%. The limits of detection for dFdC and dFdU were 0.1 and 1.1 ng/ml, but considering the high concentrations in the plasma of patients investigated, we set the limit of quantitation at 20 ng/ml (0.08 μM) for dFdC and 250 ng/ml for dFdU, and validated the assay up to the dFdC concentration of 6.0 μg/ml (22.8 μM). The method was successfully used to study the drug pharmacokinetics in patients with advanced non-small-cell lung cancer in a phase II trial with gemcitabine administered as a fixed dose-rate infusion.
AB - Gemcitabine, 2′,2′-difluoro-2′-deoxycytidine (dFdC) is a pyrimidine antimetabolite employed against several human malignancies. It undergoes intracellular activation to the pharmacologically active triphosphate form (dFdCTP) and metabolic inactivation to the metabolite 2′,2′- difluorodeoxyuridine (dFdU). In order to investigate the human plasma pharmacokinetics of dFdC and dFdU, we developed and validated an HPLC-MS/MS method, adding 2′-deoxycytidine as internal standard and simply precipitating the protein with acetonitrile. The method requires a small sample (125 μl), and it is rapid and selective, allowing good resolution of peaks from the plasma matrix in only 7 min. It is sensitive, precise and accurate, with overall precision, expressed as CV%, always less than 10.0% for both analytes and high recovery: ≥80%. The limits of detection for dFdC and dFdU were 0.1 and 1.1 ng/ml, but considering the high concentrations in the plasma of patients investigated, we set the limit of quantitation at 20 ng/ml (0.08 μM) for dFdC and 250 ng/ml for dFdU, and validated the assay up to the dFdC concentration of 6.0 μg/ml (22.8 μM). The method was successfully used to study the drug pharmacokinetics in patients with advanced non-small-cell lung cancer in a phase II trial with gemcitabine administered as a fixed dose-rate infusion.
KW - Gemcitabine
KW - HPLC-MS/MS
KW - Human pharmacokinetics
KW - Non-small-cell lung cancer
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U2 - 10.1002/jms.1293
DO - 10.1002/jms.1293
M3 - Article
C2 - 17941128
AN - SCOPUS:39749135507
VL - 43
SP - 216
EP - 223
JO - Biomedical Mass Spectrometry
JF - Biomedical Mass Spectrometry
SN - 1076-5174
IS - 2
ER -