Quantitation of the enantiomers of ofloxacin by capillary electrophoresis in the parts per billion concentration range for in vitro drug absorption studies

Ofloxacin, a chiral fluoroquinolone, possesses two optical isomers. The antibacterial activity of S-(−)-ofloxacin is reported to be 8–128 times higher than that of R-(+)-ofloxacin. A capillary zone electrophoresis method has been developed to quantify the enantiomers of ofloxacin in high diluted samples (20–700 ng/ml for each enantiomer). After fluid–fluid extraction of ofloxacin from physiological solution electrokinetic injection was employed to improve the sensitivity. The method was optimised using a central composite design. Four experimental factors were investigated: the background electrolyte concentration, the methyl-β-cyclodextrin concentration, the buffer pH and the temperature. The amount migrated into the capillary, determined by the peak area, the resolution between the ofloxacin enantiomers, the migration time and the generated current were evaluated as responses. The quantification limit is 11.4 ng/ml for S-ofloxacin and 10.8 ng/ml for R-ofloxacin. The method has shown good validation data in terms of precision and recovery rate.