A capillary electrophoresis (CE) and a reversed phase high-performance liquid chromatography (RP-HPLC) method with UV detection have been developed for florfenicol analysis in plasma samples. The suitabilities of both methods for quantitative determination of florfenicol were approved through validation specification, such as linearity, precision, selectivity, accuracy, limit of detection and quantification. The capillary electrophoresis (CE) and high-performance liquid chromatography (HPLC) assay were compared by analyzing a series of plasma samples containing florfenicol in different concentrations using the two methods. The extraction procedure is simple and no gradient elution or derivatization is required. Furthermore, the analysis time of the CE method is two times shorter than the respective parameter in HPLC and solvent consumptions is considerably lower. The calibration curve were linear to at least 0.05-10 microg/ml (r = 0.9998) and 0.1-10 microg/ml (r = 0.9998) for CE and HPLC, respectively. The separation efficiency are good for both methods. The detection limits for florfenicol were 0.015 microg/ml with CE and 0.03 microg/ml with HPLC and CE method gave lower value, even though UV detector was applied in the both cases. The both methods were selective, robust and reliable quantification of florfenicol and can be useful for clinical and biomedical investigations.