Abstract
A gas chromatography-mass spectrometry (GC-MS) method for the simultaneous determination of 5′-deoxy-5-fluorouridine (doxifluridine, 5′-dFUR), 5-fluorouracil (5-FU) and its main catabolite 5,6-dihydro-5-fluorouracil (5-FUH 2) in human plasma has been developed. Sample preparation consisted of protein precipitation with ammonium sulphate followed by analyte extraction with ethyl acetate/isopropanol ( 90 10 , v/v) mixture. Extracts were then analysed by GC-MS in positive electron impact mode after derivatization with N,O-bis(trimethylsilyl)trifluoroacetamide/pyridine ( 1 1 , v/v). The trimethylsilyl (TMS) derivative of 5′-dFUR was proved to be stable so that no interference in 5-FU determination was observed. Mass spectra of TMS derivatives of 5′-dFUR, 5-FU and 5-FUH 2 are discussed. Analyte response was linear over two decades of concentration and detection limits were typically 20 ng/ml of plasma.
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