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首页> 外文期刊>Journal of chromatography, A: Including electrophoresis and other separation methods >Development of a new in-line coupling of a miniaturized boronate affinity monolithic column with reversed-phase silica monolithic capillary column for analysis of cis-diol-containing nucleoside compounds
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Development of a new in-line coupling of a miniaturized boronate affinity monolithic column with reversed-phase silica monolithic capillary column for analysis of cis-diol-containing nucleoside compounds

机译：用反相二氧化硅整体毛细管进行小型化硼酸盐亲和整体柱的新型型硼酸酯整体圆柱形的开发，用于分析含CIS-DIOL的核苷化合物

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摘要

In-line coupling of capillary columns is an effective means for achieving miniaturized and automated separation methods. The use of multimodal column designed to allow the direct integration of a sample preparation step to the separation column is one example. Herein we propose a novel in-line coupling at the capillary scale between a boronate affinity capillary column (mu BAMC unit) and a reversed-phase separation column. This has been made possible due to the elaboration of a new and efficient mu BAMC unit. A thiol-activated silica monolithic capillary column was functionalized through thiol-ene photoclick reaction. This simple and fast reaction allows to prepare stable mu BAMC units having grafting densities of 1.93 +/- 0.17 nmol cm(-1). This grafting strategy increases the surface density by a factor 4 compared to our previous strategies and opens the frame to in-line coupling with reversed-phase capillary column. Proof of concept of the in-line coupling was done by coupling a 1-cm length mu BAMC unit to a 7-cm length reversed phase capillary column. The conditions of loading, elution and separation were optimized for cis-diol nucleosides analysis (uridine, cytidine, adenosine, guanosine). A loading volume (at pH 8.5) of up to 21 hold volume (i.e 1 mu l) of the mu BAMC unit can be loaded without sample breakthrough. For the least retained nucleoside (uridine) a limit of detection of 50 ng mL(-1) was estimated. Elution and full separation of the four nucleosides was triggered by flushing the multimodal column with an acetic acid (50 mM) / methanol (98/2, v/v) mobile phase. (C) 2019 Elsevier B.V. All rights reserved.
机译：毛细管柱的在线偶联是实现小型化和自动分离方法的有效手段。设计用于允许将样品制备步骤直接整合到分离柱的多模式柱是一个示例。在此，我们在硼酸盐亲和毛细管柱（MU BAMC单元）和反相分离塔之间的毛细管尺度上提出了一种新的在线偶联。由于阐述了一种新的和高效的MU BAMC单元，这是可能的。通过硫醇-NEE光点燃反应官能化硫醇活化二氧化硅整体毛细管。这种简单且快速的反应允许制备具有1.93 +/- 0.17nmol cm（-1）的接枝密度的稳定MU BAMC单元。与我们以前的策略相比，该接枝策略将表面密度增加了因子4，并与反相毛细管柱进行框架到线耦合。通过将1cm长度Mu Bamc单元耦合到7cm长度反相毛细管柱来完成内联耦合的概念证明。针对顺式二醇核苷分析（尿苷，胞苷，腺苷，鸟苷）优化了装载，洗脱和分离的条件。可以在没有样品突破的情况下加载MU BAMC单元的最多21个保持体积（即1μl）的负载量（在pH 8.5）中。对于最小保留的核苷（尿苷），估计了50ng ml（-1）的检测限。通过用乙酸（50mM）/甲醇（98/2，v / v）流动相冲洗多模柱，触发四种核苷的洗脱和完全分离。 （c）2019 Elsevier B.v.保留所有权利。

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