A simple and rapid SPE-HPLC-DAD method was developed for the determination of 5-fluorouracil (5-FU) in surface samples. A C18 column (250 x 4.6 mm i.d., 5 µm) and a similar pre-column were
used for the separation at 25 ºC, using 20 mM ammonium acetate buffer solution pH 4.7: methanol (95:5,
v/v) as mobile phase at a flow rate of 1.2 mL/min. Under optimal conditions, the linearity was 0.9993, in a
range of 25-100 µg/mL. The limits of detection and quantification were 5 and 25 µg/mL, respectively. The
relative standard deviation (%) was below 15 % for the evaluation of precision and the mean recovery was
77 %. The extracting procedure followed HPLC analysis showed their applicability in order to examine 5-
FU in surfaces samples. Moreover, it could be suggested that the developed method is an alternative in the
monitoring of the occupational exposure to antineoplastic agents, once the analyte in question is considered an indicator for this purpose.