The dynamics of a novel method coupling high-performance liquid chromatography (HPLC) with resonance Rayleigh scattering (RRS) is presented in the paper. The method was employed in the detection of fluoroquinolones (FQs) of urine samples from healthy human beings, by forming ion-association complexes between the components separated from HPLC and [Ce(OH)(3) ](+) as the molecular recognition probe. The RRS signal was measured at λ(ex) =λ(em) =365 nm. It was applied to detect three FQs and obtained satisfactory results. The RRS spectral characteristics of the analytes and the kinetics of flow rate, proportion of organic phase, reaction time and the aggregation level of ion-association complexes were investigated, which provided a new basis for the development of the hyphenated techniques. It was established that the presence of HPLC-RRS would open up a new field in the determination of analytes in the absence of UV or fluorescence.