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利用深紫外本征荧光和质谱法对活性香蕉成分进行芯片电泳及无标记检测。

Chip electrophoresis of active banana ingredients with label-free detection utilizing deep UV native fluorescence and mass spectrometry.

机构信息

Institute of Analytical Chemistry, University of Leipzig, Johannisallee 29, 04103 Leipzig, Germany.

出版信息

Anal Bioanal Chem. 2011 Feb;399(5):1853-7. doi: 10.1007/s00216-010-4557-z. Epub 2010 Dec 22.

Abstract

In the present work, we report on a rapid and straightforward approach for the determination of biologically active compounds in bananas applying microchip electrophoresis (MCE). For this purpose, we applied label-free detection utilizing deep UV fluorescence detection with excitation at 266 nm. Using this approach, we could identify dopamine and serotonin, their precursors tryptophan and tyrosine and also the isoquinoline alkaloid salsolinol in less than 1 min. In bananas, after 10 days of ripening, we additionally found the compound levodopa which is a metabolite of the tyrosine pathway. Quantitative analysis of extracts by external calibration revealed concentrations of serotonin, tryptophan, and tyrosine from 2.7 to 7.6 μg/mL with relative standard deviations of less than 3.5%. The corresponding calibration plots showed good linearity with correlation coefficients higher than 0.985. For reliable peak assignment, the compounds were also analyzed by coupling chip electrophoresis with mass spectrometry. This paper demonstrates exemplarily the applicability of MCE with native fluorescence detection for rapid analysis of natural compounds in fruits and reveals the potential of chip-based separation systems for the analysis of complex mixtures.

摘要

在本工作中,我们报告了一种快速而直接的方法,用于应用微芯片电泳(MCE)测定香蕉中的生物活性化合物。为此,我们应用了无标记检测,利用 266nm 的深紫外荧光检测进行激发。使用这种方法,我们可以在不到 1 分钟的时间内鉴定出多巴胺和血清素、它们的前体色氨酸和酪氨酸,以及异喹啉生物碱萨索林醇。在成熟 10 天后的香蕉中,我们还发现了左旋多巴,它是酪氨酸途径的代谢产物。通过外部校准对提取物进行定量分析,发现血清素、色氨酸和酪氨酸的浓度为 2.7 至 7.6μg/mL,相对标准偏差小于 3.5%。相应的校准曲线具有良好的线性,相关系数高于 0.985。为了可靠地进行峰分配,还通过与质谱法耦合的芯片电泳对化合物进行了分析。本文以实例证明了 MCE 与天然荧光检测相结合在快速分析水果中的天然化合物方面的适用性,并揭示了基于芯片的分离系统在分析复杂混合物方面的潜力。

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