Mashige F, Takai N, Matsushima Y, Ito A, Takano M, Tsuchiya E, Kanazawa H, Nagata Y, Shinozuka N, Wada H
Department of Laboratory Medicine, Faculty of Medicine, University of Tokyo.
Rinsho Byori. 1994 Jun;42(6):591-9.
We have developed two HPLC systems for the simultaneous determination of CA, 5-HT and their precursors and metabolites in human body fluid. One system consisted of two reversed-phase columns, a column switching device, a pair of electrodes for elimination of interferents and two sets of new electrochemical detectors with four electrodes. Using this system, adequate separation of peaks for 17 kinds of CA, 5-HT and their related metabolites in a standard solution required only 17 min. NE, MHPG, DOPAC, 5-HIAA, HVA and TRP levels in cerebrospinal fluid (CSF) were determined without any pretreatment of the CSF samples. The concentrations of DOPAC, 5-HIAA and HVA in CSF were significantly lower in Alzheimer's disease, vascular dementia and Parkinson's disease than in the controls. The other system was developed for determination of CA acidic metabolites and 5-HIAA in human urine. This system consisted of a mixed-mode column (C18/anion) and 8-channel electrochemical detector with isocratic elution of citrate buffer. Detection limits, precision and analytical recoveries of this method were satisfactory for clinical use.
我们开发了两种高效液相色谱系统,用于同时测定人体体液中的儿茶酚胺(CA)、5-羟色胺(5-HT)及其前体和代谢物。一种系统由两根反相柱、一个柱切换装置、一对用于消除干扰物的电极以及两套带有四个电极的新型电化学检测器组成。使用该系统,标准溶液中17种CA、5-HT及其相关代谢物的峰得到充分分离仅需17分钟。脑脊液(CSF)中的去甲肾上腺素(NE)、3-甲氧基-4-羟基苯乙二醇(MHPG)、3,4-二羟基苯乙酸(DOPAC)、5-羟吲哚乙酸(5-HIAA)、高香草酸(HVA)和色氨酸(TRP)水平在脑脊液样本未经任何预处理的情况下即可测定。在阿尔茨海默病、血管性痴呆和帕金森病患者的脑脊液中,DOPAC、5-HIAA和HVA的浓度显著低于对照组。另一种系统是为测定人体尿液中的CA酸性代谢物和5-HIAA而开发的。该系统由一个混合模式柱(C18/阴离子)和一个8通道电化学检测器组成,采用柠檬酸盐缓冲液等度洗脱。该方法的检测限、精密度和分析回收率对于临床应用来说是令人满意的。