High-performance liquid chromatography study of gatifloxacin and sparfloxacin using erythrosine as post-column resonance Rayleigh scattering reagent and mechanism study.

Herein, a highly selective high-performance liquid chromatography (HPLC) coupled with resonance Rayleigh scattering (RRS) method was developed to detect gatifloxacin (GFLX) and sparfloxacin (SPLX). GFLX and SPLX were first separated by HPLC, then, in pH 4.4 Britton-Robinson (BR) buffer medium, protonic quaternary ammonia cation of GFLX and SPLX reacted with erythrosine (ERY) to form 1:1 ion-association complexes, which resulted in a significant enhancement of RRS signal. The experimental conditions of HPLC and post-column RRS have been investigated, including detection wavelength, flow rate, pH, reacting tube length and reaction temperature. Reaction mechanism were studied in detail by calculating the distribution fraction. The maximum RRS signals for GFLX and SPLX were recorded at λex  = λem  = 330 nm. The detection limits were 3.8 ng ml-1 for GFLX and 17.5 ng ml-1 for SPLX at a signal-to-noise ratio of 3. The developed method was successfully applied to the determination of GFLX and SPLX in water samples. Recoveries from spiked water samples were 97.56-98.85%.