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用于毛细管电泳检测吸烟者尿液中肾上腺素的高灵敏度三价铜螯合物-鲁米诺化学发光体系。

Highly sensitive trivalent copper chelate-luminol chemiluminescence system for capillary electrophoresis detection of epinephrine in the urine of smoker.

机构信息

Key Laboratory of Luminescence and Real-Time Analysis (Ministry of Education), College of Pharmaceutical Sciences, Southwest University, Chongqing 400716, China.

出版信息

J Chromatogr B Analyt Technol Biomed Life Sci. 2012 Dec 12;911:1-5. doi: 10.1016/j.jchromb.2012.10.017. Epub 2012 Oct 23.

Abstract

Epinephrine (EP) is one of the most important neurotransmitters and hormones. Some previous literatures show that there is a close relation between its release and smoking. To compare the levels of EP in urines of smokers and nonsmokers, a sensitive chemiluminescence (CL) system, luminol-diperiodatocuprate (III) (K(5)[Cu(HIO(6))(2)], DPC), has been developed and validated for the determination of EP after CE separation. The DPC-luminol-EP CL reaction showed very intensive emission and fast kinetic characteristics, thus led to a high sensitivity in the flow-through detection mode for capillary electrophoresis. With the peak height as a quantitative parameter, the relative CL intensity was linear with the EP concentration in the range of 2.0-400ng/mL, with a limit of detection of 0.82ng/mL (S/N=3). The reproducibility was assessed by intra- and inter-day relative standard deviations (RSDs) for 11 replicate determinations of EP standard samples at low, medium and high concentrations. The intra- and inter-day RSDs for CL signals were 5.5%-6.6% and 6.1%-7.5%, respectively, and those for migration times were 3.4%-5.8% and 4.3%-6.3%, respectively. The presented method was successfully applied to the determination of EP in EP injection and urine samples of smokers and nonsmokers. The recovery test results for urine samples ranged from 86.5 to 112.0%, which demonstrated the reliability of this method. The results for urine sample detection indicate that the average level of EP in the urines of the smoker group is obviously higher than that in the urines of the nonsmoker group, which may demonstrate that smoking can stimulate the release of EP in human body.

摘要

肾上腺素(EP)是最重要的神经递质和激素之一。一些先前的文献表明,它的释放与吸烟之间存在密切关系。为了比较吸烟者和不吸烟者尿液中 EP 的水平,开发并验证了一种灵敏的化学发光(CL)系统,即鲁米诺-双过碘酸盐铜(III)(K(5)[Cu(HIO(6))(2)],DPC),用于 CE 分离后 EP 的测定。DPC-鲁米诺-EP CL 反应具有非常强烈的发射和快速的动力学特性,因此在毛细管电泳的流通检测模式下具有很高的灵敏度。以峰高为定量参数,相对 CL 强度与 EP 浓度在 2.0-400ng/mL 范围内呈线性关系,检测限为 0.82ng/mL(S/N=3)。通过在低、中、高浓度下对 EP 标准样品进行 11 次重复测定,评估了日内和日间重现性。CL 信号的日内和日间 RSD 分别为 5.5%-6.6%和 6.1%-7.5%,迁移时间的日内和日间 RSD 分别为 3.4%-5.8%和 4.3%-6.3%。该方法成功应用于 EP 注射液和吸烟者和不吸烟者尿液中 EP 的测定。尿样的回收率试验结果在 86.5%至 112.0%之间,证明了该方法的可靠性。尿样检测结果表明,吸烟者尿液中 EP 的平均水平明显高于不吸烟者尿液中的 EP 水平,这可能表明吸烟会刺激人体释放 EP。

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