Pasas Stephanie A, Lacher Nathan A, Davies Malonne I, Lunte Susan M
Department of Pharmaceutical Chemistry, University of Kansas, Lawrence, KS 66047, USA.
Electrophoresis. 2002 Mar;23(5):759-66. doi: 10.1002/1522-2683(200203)23:5<759::AID-ELPS759>3.0.CO;2-4.
A method based on capillary electrophoresis (CE) with electrochemical (EC) detection for the determination of both total homocysteine (tHcy) and protein-bound homocysteine (pbHcy) in plasma is described. Both end-column and off-column amperometric detection were investigated. Off-column detection resulted in a more sensitive assay for the determination of homocysteine (Hcy). The detection limit for homocysteine was 500 nM using off-column EC detection and the response was linear over the range 1-100 microM. Therefore, this assay is appropriate for the quantification of Hcy over the physiological concentration ranges found in all disease states. Methodologies for the determination of tHcy and pbHcy in human plasma were investigated and optimized and the concentrations of both pbHcy and tHcy in plasma obtained from a healthy individual were determined to be 2.79+/-0.31 nuM (n = 4) and 3.37+/-0.15 microM (n = 3), respectively. The methodology was then transferred to a microchip CE-EC format and Hcy and reduced glutathione (GSH) were detected. Future work will focus on the development of ancillary methodologies to identify the other forms of Hcy in vivo.
本文描述了一种基于毛细管电泳（CE）结合电化学（EC）检测的方法，用于测定血浆中的总同型半胱氨酸（tHcy）和蛋白结合型同型半胱氨酸（pbHcy）。研究了柱端和柱外安培检测。柱外检测对同型半胱氨酸（Hcy）的测定产生了更灵敏的分析方法。使用柱外EC检测时，同型半胱氨酸的检测限为500 nM，响应在1-100 microM范围内呈线性。因此，该分析方法适用于在所有疾病状态下生理浓度范围内同型半胱氨酸的定量。研究并优化了测定人血浆中tHcy和pbHcy的方法，确定从一名健康个体获得的血浆中pbHcy和tHcy的浓度分别为2.79±0.31μM（n = 4）和3.37±0.15 microM（n = 3）。然后将该方法转移到微芯片CE-EC形式，并检测了Hcy和还原型谷胱甘肽（GSH）。未来的工作将集中在开发辅助方法以鉴定体内其他形式的Hcy。
