SEPARATION OF NANOLITER SAMPLES OF BIOLOGICAL AMINES BY A COMPREHENSIVE 2-DIMENSIONAL MICROCOLUMN LIQUID-CHROMATOGRAPHY SYSTEM
SEPARATION OF NANOLITER SAMPLES OF BIOLOGICAL AMINES BY A COMPREHENSIVE 2-DIMENSIONAL MICROCOLUMN LIQUID-CHROMATOGRAPHY SYSTEM
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DOI：
10.1021/ac00114a026
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发表时间：
1995-09-15
影响因子：
7.4
通讯作者：
JORGENSON, JW
中科院分区：
文献类型：
--
作者：
HOLLAND, HA;JORGENSON, JW
提出了一种基于由阴离子交换色谱提供的电荷分离机制与由反相色谱提供的疏水分离机制的组合的二维液相色谱系统。将90 cm长的阴离子交换微柱与3 cm长的反相微柱偶联。两个微柱通过两个电子控制阀连接。阴离子交换微柱的流出物收集在样品环中，然后浓缩到反相微柱的顶部。由标记的胺组成的分析物峰从第二柱中分离出来，并通过激光诱导荧光检测。整个操作由个人电脑控制。该系统的分辨率证明了从猪甲状腺球蛋白的胰蛋白酶消化获得的肽的二维色谱图。该系统的灵敏度证明了一个单一的牛嗜铬细胞的内容物的二维色谱图。
A two-dimensional liquid chromatography system based on the combination of a charge separation mechanism, provided by anion exchange chromatography, with a hydrophobic separation mechanism, provided by reversed phase chromatography, is presented. A 90 cm long anion exchange microcolumn is coupled to a 3 cm long reversed phase microcolumn. Both microcolumns are interfaced by two electronically controlled valves. Effluent from the anion exchange microcolumn is collected in a sample loop and then concentrated onto the head of the reversed phase microcolumn. Analyte peaks consisting of tagged amines elute from the second column and are detected by laser-induced fluorescence. The entire operation is controlled by a personal computer. The resolving power of this system is demonstrated with a two-dimensional chromatogram of the peptides obtained from a tryptic digest of porcine thyroglobulin. The sensitivity of the system is demonstrated with a two-dimensional chromatogram of the contents of a single bovine chromaffin cell.