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高效液相色谱法测定人血浆中的儿茶酚胺:荧光检测新方法与电化学检测既定方法的比较

Determination of catecholamines in human plasma by high-performance liquid chromatography: comparison between a new method with fluorescence detection and an established method with electrochemical detection.

作者信息

van der Hoorn F A, Boomsma F, Man in 't Veld A J, Schalekamp M A

机构信息

Department of Internal Medicine I, University Hospital Dijkzigt, Erasmus University, Rotterdam, The Netherlands.

出版信息

J Chromatogr. 1989 Jan 27;487(1):17-28. doi: 10.1016/s0378-4347(00)83003-0.

Abstract

We report a sensitive fluorimetric method, in which catecholamines are concentrated from plasma by liquid-liquid extraction and derivatized with the selective fluorescent agent 1,2-diphenylethyl-enediamine prior to chromatography. Optimal conditions for extraction, derivatization and chromatography were investigated. With alpha-methylnorepinephrine as internal standard, the chromatographic separations are complete within 6 min. Limits of detection are 0.3 pg for norepinephrine and epinephrine and 0.5 pg for dopamine. Coefficients of variation are low (3-7%). Comparison of plasma catecholamine values determined with this method and with an established method with electrochemical detection (n = 135) shows good correlation (r = 0.94-1.00), and regression lines are close to lines of identity.

摘要

我们报告了一种灵敏的荧光分析法,该方法中,儿茶酚胺通过液-液萃取从血浆中富集,并在色谱分析前用选择性荧光剂1,2-二苯基乙二胺进行衍生化。研究了萃取、衍生化和色谱分析的最佳条件。以α-甲基去甲肾上腺素作为内标,6分钟内可完成色谱分离。去甲肾上腺素和肾上腺素的检测限为0.3皮克,多巴胺的检测限为0.5皮克。变异系数较低(3-7%)。用该方法与已建立的电化学检测方法测定血浆儿茶酚胺值(n = 135)进行比较,显示出良好的相关性(r = 0.94-1.00),回归线接近恒等线。

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