Simultaneous separation and detection of three organomercury species, namely methylmercury (MeHg), ethylmercury (EtHg), and phenylmercury (PhHg), was performed by using capillary electrophoresis (CE) with UV detection. Pre-column derivatization with thiosalicylic acid and on-line salt-induced stacking significantly improved the detection performance. Buffer pH, ion strength, and additive were optimized for CE separation, concentration of NaCl in sample solution and injection time were optimized for on-line stacking. The limits of detection were 76.9, 83.0 and 76.4 μg/L for PhHg, EtHg and MeHg, respectively. The developed method was validated by certified reference material and liquid chromatography-atomic fluorescence spectroscopy, which suggests this method could be useful in the speciation of organomercury compounds in biological samples.