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采用柱端安培检测法的毛细管电泳法检测尿中的8-羟基-2'-脱氧鸟苷。

Capillary electrophoresis with end-column amperometric detection of urinary 8-hydroxy-2'-deoxyguanosine.

作者信息

Mei Su-Rong, Yao Qing-Hong, Cai Ling-Suang, Xing Jun, Xu Guo-Wang, Wu Cai-Ying

机构信息

National Chromatographic R & A Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, China.

出版信息

Electrophoresis. 2003 May;24(9):1411-5. doi: 10.1002/elps.200390181.

Abstract

Urinary 8-hydroxy-2'-deoxyguanosine (8OHdG) is an excellent marker of oxidative DNA damage. Until now, urinary 8OHdG has been measured by high-performance liquid chromatography with electrochemical detection. A simple and sensitive method for the analysis of urinary 8OHdG by capillary electrophoresis with end-column amperometric detection has been developed in our laboratory. A single-step solid-phase extraction procedure was optimized and used for extracting 8OHdG from human urine. To improve the sensitivity of this method, a new focusing technique based on a dynamic pH junction was used. The limit of detection was 20 nM (signal-to-noise ratio S/N = 3), the linear range was 50 nM-10 microM, and the correlation coefficient was better than 0.999. The relative standard deviation (RSD) was found to be 0.57% for migration time, and 4.79% for peak current. To show the usefulness of the method, the urinary concentration of 8OHdG in nine healthy persons and ten cancer patients was determined. The urinary concentration of 8OHdG in cancer patients was significantly higher than that in healthy persons.

摘要

尿8-羟基-2'-脱氧鸟苷(8OHdG)是氧化DNA损伤的一个优秀标志物。到目前为止,尿8OHdG一直通过高效液相色谱-电化学检测法进行测定。我们实验室已开发出一种采用柱端安培检测的毛细管电泳法来分析尿8OHdG的简单且灵敏的方法。优化了一种单步固相萃取程序并用于从人尿中提取8OHdG。为提高该方法的灵敏度,采用了一种基于动态pH连接的新型聚焦技术。检测限为20 nM(信噪比S/N = 3),线性范围为50 nM - 10 μM,相关系数优于0.999。迁移时间的相对标准偏差(RSD)为0.57%,峰电流的相对标准偏差为4.79%。为证明该方法的实用性,测定了9名健康人和10名癌症患者尿中8OHdG的浓度。癌症患者尿中8OHdG的浓度显著高于健康人。

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