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首页> 外文期刊>CHROMATOGRAPHY >Online Cleanup Liquid Chromatography for the Analysis of Glycoprotein-Derived Oligosaccharides Labeled with 7-Amino-4-methylcoumarin
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Online Cleanup Liquid Chromatography for the Analysis of Glycoprotein-Derived Oligosaccharides Labeled with 7-Amino-4-methylcoumarin

机译：在线纯化液相色谱法，分析标记有7-氨基-4-甲基香豆素的糖蛋白衍生寡糖

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In HPLC analysis of glycoprotein glycans, oligosaccharides released from glycoproteins are often derivatized with fluorescent tags for enhancing sensitivity and selectivity. However, existing analysis methods require multiple steps for removing excess reagents and are time consuming and sometime spoil the reliability in quantitative determination. Here, we developed an online sample cleanup procedure for the analysis of 7-amino-4-methylcoumarin (AMC)-labeled oligosaccharides. An NH_(2)-bonded silica packed short column (1 mm I.D. x 15 mm) and a Waters HLB~(TM)-packed column (4 mm I.D. x 5 cm) were used for online cleanup HPLC analysis with Amide80 and ODS columns, respectively. Optimized conditions enabled the direct injection of the diluted solution of the reaction mixture onto the chromatographic system without any prior removal of the excess labeling reagents. The method was successfully applied to the analysis of N -linked glycans released from some glycoprotein samples.
机译：在糖蛋白聚糖的HPLC分析中，从糖蛋白释放的寡糖通常用荧光标签衍生化，以增强灵敏度和选择性。但是，现有的分析方法需要多个步骤才能去除多余的试剂，并且很耗时，并且有时会破坏定量测定的可靠性。在这里，我们开发了一种在线样品净化程序，用于分析7-氨基-4-甲基香豆素（AMC）标记的寡糖。使用NH_（2）键合的硅胶填充短柱（1 mm内径x 15毫米）和Waters HLB〜TM填充柱（4 mm内径x 5厘米）进行在线纯化，使用Amide80和ODS色谱柱进行HPLC分析， 分别。优化的条件可以将反应混合物的稀释溶液直接注入色谱系统，而无需事先去除过量的标记试剂。该方法已成功地用于分析从某些糖蛋白样品中释放的N-连接的聚糖。

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