Determination of enzyme activity by high-performance liquid chromatography

The application of high-performance liquid chromatography (HPLC) in the study of enzymatic reactions is reviewed. The rationale for using HPLC is given and whether the components of the reaction mixture should be derivatized prior to or after HPLC. An alphabetical list of enzymes assayed by HPLC is given. Substrate and product are included as well the derivatization reagent, detection method and biological matrix. Specific examples of these assays in a complex biological matrix viz. faeces are given. Future prospects are the detection of new enzymes using synthetic substrates and implementation of mass spectrometry to elucidate enzyme specificities.