A modified chromatographic process tvas used to determine the enantiomeric ratio (E) in enantioselective: biocatalysis. Theoretical results indicated that the established relationship, E = ln(l-x(S))/ln(l-x(R)), was valid for this modified operation, in addition, the Candida cylindracea Lipase-catalyzed enantioselective esterification of racemic naproxen was selected to verify experimentally the effectiveness of the proposed method. The ease of operation and simplicity of data analysis of the proposed method makes it a promising alternative for determining the enantioselectivity of enzymatic chiral resolution.