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用于生物胺快速分析的集成芯片衍生化和电泳技术。

Integrated on-chip derivatization and electrophoresis for the rapid analysis of biogenic amines.

作者信息

Beard Nigel P, Edel Joshua B, deMello Andrew J

机构信息

Department of Chemistry, Imperial College London, Exhibition Road, South Kensington, London, UK.

出版信息

Electrophoresis. 2004 Jul;25(14):2363-73. doi: 10.1002/elps.200305919.

Abstract

We demonstrate the monolithic integration of a chemical reactor with a capillary electrophoresis device for the rapid and sensitive analysis of biogenic amines. Fluorescein isothiocyanate (FITC) is widely employed for the analysis of amino-group containing analytes. However, the slow reaction kinetics hinders the use of this dye for on-chip labeling applications. Other alternatives are available such as o-phthaldehyde (OPA), however, the inferior photophysical properties and the UV lambdamax present difficulties when using common excitation sources leading to a disparity in sensitivity. Consequently, we present for the first time the use of dichlorotriazine fluorescein (DTAF) as a superior in situ derivatizing agent for biogenic amines in microfluidic devices. The developed microdevice employs both hydrodynamic and electroosmotic flow, facilitating the creation of a polymeric microchip to perform both precolumn derivatization and electrophoretic analysis. The favorable photophysical properties of the DTAF and its fast reaction kinetics provide detection limits down to 1 nM and total analysis times (including on-chip mixing and reaction) of <60 s. The detection limits are two orders of magnitude lower than current limits obtained with both FITC and OPA. The optimized microdevice is also employed to probe biogenic amines in real samples.

摘要

我们展示了一种化学反应器与毛细管电泳装置的单片集成,用于快速、灵敏地分析生物胺。异硫氰酸荧光素(FITC)被广泛用于分析含氨基的分析物。然而,缓慢的反应动力学阻碍了这种染料在芯片标记应用中的使用。还有其他替代方法,如邻苯二甲醛(OPA),然而,其较差的光物理性质和紫外最大吸收波长在使用普通激发源时存在困难,导致灵敏度存在差异。因此,我们首次提出使用二氯三嗪荧光素(DTAF)作为微流控装置中生物胺的一种优越的原位衍生剂。所开发的微装置采用了流体动力学和电渗流,便于制造一种聚合物微芯片,以进行柱前衍生和电泳分析。DTAF良好的光物理性质及其快速的反应动力学使得检测限低至1 nM,总分析时间(包括芯片上的混合和反应)小于60秒。检测限比使用FITC和OPA获得的当前限值低两个数量级。优化后的微装置还用于检测实际样品中的生物胺。

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