Determination of S-catechol-O-methyltransferase Activity in rat tissues using High-Performance Liquid Chromatography with Electrochemical detection

摘要

A high-performance liquid chromatography with electrochemical detection (HPLC-ECD) method was developed determine the activity of S-catechol-O-methyltransferase (COMT) in rat tissues including the liver, kidney, lung,spleen and brain. The 3,4-dihydroxybenzoic acid was used as the substrate for SCOMT in rat tissues. After incubated with different tissues, the product 4-hydroxy-3-methoxybenzoic acid was formed from the substrate and separated to measure the COMT activity. The analysis was performed on a Agilent C8 column (5μm, 150×4.6mm) at 550mV potential. The mobile phase was a mixture of methanol and 0.1MNa2HPO4(pH=3.3) containing 0.13mM ethylenediamine tetraacetic acid (v/v, 14/86) with a flow rate of 1ml/min. This approach was used to study S-COMT activity vitro from various tissues.