Nissinen E, Männistö P
Anal Biochem. 1984 Feb;137(1):69-73. doi: 10.1016/0003-2697(84)90348-8.
A rapid and simple assay for catechol-O-methyltransferase (COMT) activity by high-performance liquid chromatography with electrochemical detection is described. The method is based on the measurement of 3-O- and 4-O-methylated products (3-methoxy-4-hydroxy- and 4-methoxy-3-hydroxybenzylamine, respectively) of the substrate 3,4-dihydroxybenzylamine. These compounds are determined from the incubation mixture after removal of protein injecting aliquot into the liquid chromatograph. The detection limit is 1 pmol of product. This method is very suitable for screening of COMT activity as well as for determining the meta/para product ratios.
描述了一种通过高效液相色谱-电化学检测快速简便地测定儿茶酚-O-甲基转移酶（COMT）活性的方法。该方法基于对底物3,4-二羟基苄胺的3-O-和4-O-甲基化产物（分别为3-甲氧基-4-羟基和4-甲氧基-3-羟基苄胺）的测量。在将等分试样注入液相色谱仪之前，通过去除蛋白质从孵育混合物中测定这些化合物。检测限为1 pmol产物。该方法非常适合筛选COMT活性以及确定间位/对位产物比率。
