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A new molecularly imprinted polymer, prepared following a non-covalent approach, was synthesised using enrofloxacin as a template molecule. The imprinting effect of the polymer was verified by chromatographic evaluation and, interestingly, this evaluation also revealed that the imprinted polymer showed a high degree of cross-reactivity for ciprofloxacin, the major metabolite of enrofloxacin. The molecularly imprinted polymer was then applied as a selective sorbent in a two-step solid-phase extraction method focussing upon complex biological matrices, specifically human urine and pig liver. This two-step solid-phase extraction protocol, in which a commercial Oasis HLB cartridge and a molecularly imprinted solid-phase extraction cartridge were combined, allowed enrofloxacin and ciprofloxacin to be determined by liquid chromatography coupled to a UV detector at levels below the maximum residue limits established by the European Union. The quantification and detection limits in tissue samples of enrofloxacin and ciprofloxacin were established at 50 mu g kg(-1) and 30 mu g kg(-1), respectively. (c) 2006 Elsevier B.V. All rights reserved.