采用高效液相色谱-荧光检测法测定大鼠脑组织中色氨酸羟化酶的活性。
Assay for tryptophan hydroxylase activity in rat brain by high-performance liquid chromatography with fluorescence detection.
作者信息
Iizuka R, Ishida J, Yoshitake T, Nakamura M, Yamaguchi M
机构信息
Chemical Biotesting Center, Chemical Inspection and Testing Institute, Oita, Japan.
出版信息
Biol Pharm Bull. 1996 May;19(5):762-4. doi: 10.1248/bpb.19.762.
A highly sensitive and selective liquid chromatographic (HPLC) method with post-column fluorescence detection has been developed for the determination of tryptophan 5-monooxygenase activity in rat brain tissue homogenate. 5-Hydroxytryptophan, formed enzymatically from tryptophan (incubation time, 20 min), is extracted with perchloric acid and determined by HPLC. Detection is performed fluorometrically after post-column derivatization by a reaction with benzylamine in the presence of potassium hexacyanoferrate (III). The lower limit of detection for 5-hydroxytryptophan formed enzymatically is 100 fmol at a signal-to-noise ratio of three.
已开发出一种具有柱后荧光检测功能的高灵敏度和高选择性液相色谱（HPLC）方法，用于测定大鼠脑组织匀浆中色氨酸5-单加氧酶的活性。由色氨酸酶促形成的5-羟色氨酸（孵育时间为20分钟）用高氯酸提取，并用HPLC测定。在柱后通过与苄胺在六氰合铁（III）酸钾存在下反应进行衍生化后，进行荧光检测。酶促形成的5-羟色氨酸的检测下限为100 fmol，信噪比为3。
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