Abstract: The distribution behavior of ofloxacin enantiomers was examined in the aqueous and organic solvent of a two-phase system containing a chiral selector L-tartrate， and extraction equilibrium time and effect of pH , organic solvents and length of alkyl chain of L-tartrates on the partition coefficients and enantioselectivity of RS-ofloxacins, were investigated. The L-tartaric acid esters formed more stable complexes with R-ofloxacin than that with S-ofloxacin.The partition coefficients and enantioselectivity of RS-ofloxacins increased with increasing length of alkyl chain of L-tartrates.Solvents showed a large influence on enantioselectivity and partition coefficients.Optimum pH was about 7 for separation of RS-ofloxacins by extraction.