Lee M, Nohta H, Ohkura Y, Yoo B
J Chromatogr. 1985 Dec 4;348(2):407-15. doi: 10.1016/s0021-9673(01)92479-3.
A highly sensitive assay method for phenylethanolamine N-methyltransferase in rat adrenal medulla and brain is described which employs high-performance liquid chromatography with fluorescence detection. Epinephrine formed enzymatically from the substrate norepinephrine and isoproterenol (internal standard), after chromatography on a small cartridge of a cation exchanger, Toyopak SP, are converted into the corresponding fluorescent compounds by reaction with 1,2-diphenylethylenediamine, a selective fluorescence derivatization reagent for catechol compounds. The derivatives are separated by reversed-phase chromatography on TSK gel ODS-120T. The detection limit for epinephrine formed enzymatically is 0.66 pmol per assay tube.
本文描述了一种用于测定大鼠肾上腺髓质和脑组织中苯乙醇胺N-甲基转移酶的高灵敏度检测方法，该方法采用带荧光检测的高效液相色谱法。由底物去甲肾上腺素和异丙肾上腺素（内标）酶促形成的肾上腺素，在阳离子交换剂Toyopak SP的小柱上进行色谱分离后，通过与1,2-二苯基乙二胺反应转化为相应的荧光化合物，1,2-二苯基乙二胺是一种用于儿茶酚化合物的选择性荧光衍生试剂。衍生物在TSK凝胶ODS-120T上通过反相色谱法分离。酶促形成的肾上腺素的检测限为每个测定管0.66皮摩尔。
