The enantiomeric separation of ketoprofen in three mobile phase systems on an ovomucoid (OVM) chiral stationary phase (CSP) column was investigated. Suitable chromatographic conditions for simultaneous separation of the two pairs of enantiomers of ketoprofen and its methyl ester were adopted. All of the four compounds occuring in the process of enzymatic stereoselective hydrolysis or esterification can be simultaneously separated.