AbstractIn this work, the development of two solid‐phase extraction procedures (off‐line and on‐line formats) for the identification and quantification of several (fluoro)quinolones in hospital sewage water by HPLC‐UV is described. Both procedures are based on the use of C18 and anion exchange (SAX) sorbents for the preconcentration and clean‐up steps, respectively, and all variables influencing both steps were optimised. In the off‐line format, after its pH was adjusted to 2.5, sample was preconcentrated on a C18 cartridge and eluted with 4 mL of methanol/ammonia (94/6). The methanolic extract must be diluted up to 10 mL with water to allow quantitative retention of the analytes on the SAX cartridge. In the on‐line format, the addition of 2.5% of NH4Cl to the sewage water sample (pH = 2.5) was necessary to increase the breakthrough volumes of the analytes in the C18 precolumn. Quantitative transfer of the (fluoro)quinolones from the C18 precolumn to the SAX precolumn was accomplished by pumping 2 mL of a mixture methanol/water (40/60, pH = 9.2) at 2 mL min–1. Elution of the analytes from the SAX precolumn by means of the chromatographic mobile phase required the inclusion of an additional isocratic step at the beginning of the gradient program. Both off‐line and on‐line solid phase extraction procedures coupled to HPLC‐UV were applied to the analysis of a sewage water sample collected in the sewer system at the output of the St Dimphna Hospital (Geel, Belgium). The fluoroquinolone ciprofloxacin was found in this sample and quantified at 5.8 ± 0.4 μg L–1 (off‐line method) and 5.6 ± 0.5 μg L–1 (on‐line method). The analysis of spiked samples containing the seven (fluoro)quinolones studied provided quantitative recoveries in all cases with low RSD values (from 6 to 12%), and all the analytes could be identified by means of their UV spectra with match factors varying from 950 to 985 depending on the (fluoro)quinolone.