Román David Arráez, Carretero Antonio Segura, Blanco Carmen Cruces, Gutiérrez Alberto Fernández
Department of Analytical Chemistry, Faculty of Sciences, University of Granada, Spain.
Biomed Chromatogr. 2004 Sep;18(7):422-6. doi: 10.1002/bmc.330.
In this work, a sub-minute and sensitive capillary electrophoresis with laser-induced fluorescence (CE-LIF) method was developed for the analysis and quantitation of the neurotransmitter 5-hydroxytryptamine (5-HT) or serotonin in urine. The method involves precolumn derivatization with fluorescein isothiocyanate isomer I (FITC) using an excitation light from an argon ion laser of 488 nm and a 520 nm band pass emission filter. Different variables that affect derivatization (pH, FITC concentration, reaction time and temperature) and separation (buffer concentration, pH, applied voltage and injection time) were studied. The linear dynamic range obtained was between 0 and 188 nM with a detection limit of 16 nm with a RSD between 2 and 9%. The applicability of the proposed method was demonstrated by analysis of 5-HT in human urine, establishing a concentration of 57 nM in control urine. The method was validated by standard-addition methodology.
在本研究中，开发了一种用于分析和定量尿液中神经递质5-羟色胺（5-HT）或血清素的亚分钟级灵敏毛细管电泳-激光诱导荧光（CE-LIF）方法。该方法包括使用来自氩离子激光器的488 nm激发光和520 nm带通发射滤光片，用异硫氰酸荧光素异构体I（FITC）进行柱前衍生化。研究了影响衍生化（pH、FITC浓度、反应时间和温度）和分离（缓冲液浓度、pH、施加电压和进样时间）的不同变量。获得的线性动态范围为0至188 nM，检测限为16 nM，相对标准偏差为2%至9%。通过分析人尿液中的5-HT证明了该方法的适用性，在对照尿液中确定浓度为57 nM。该方法通过标准加入法进行了验证。
