Davies M J, Hounsell E F
Department of Biochemistry and Molecular Biology, University College London, UK.
Biomed Chromatogr. 1996 Nov-Dec;10(6):285-9. doi: 10.1002/(SICI)1099-0801(199611)10:6<285::AID-BMC616>3.0.CO;2-W.
In attempting to analyse complex carbohydrates by HPLC or capillary electrophoresis (CE) the chromatographer is faced with many challenges. The diversity of oligosaccharide structures described the date and their poor resolution on many chromatographic systems present a significant challenge to the chromatographer. In addition to this, the non-chromogenic nature of most carbohydrates greatly decreases the sensitivity of UV detection and refractive index is not an option for sensitive analysis. Over the last five years there have been significant advances in separation and detection methods for carbohydrates, and in this paper we will attempt to describe the most significant advances and highlight potential future developments. The detection of 200-300 fmol (2-3 x 10(-3) mol) of oligosaccharides can now be readily achieved using electrochemical detection or fluorescence detection (after labelling) with conventional HPLC columns and detectors. Miniaturized separation systems have greatly increased sensitivities with a CE-laser induced fluorescence instrument detecting as little as 100 yoctomoles (1 x 10(-22) mol).
在尝试通过高效液相色谱法（HPLC）或毛细管电泳（CE）分析复杂碳水化合物时，色谱工作者面临诸多挑战。已描述的寡糖结构的多样性以及它们在许多色谱系统上的低分辨率给色谱工作者带来了重大挑战。除此之外，大多数碳水化合物的非发色性质极大地降低了紫外检测的灵敏度，而折光指数不适用于灵敏分析。在过去五年中，碳水化合物的分离和检测方法取得了重大进展，在本文中，我们将试图描述最显著的进展并突出潜在的未来发展。现在，使用传统的HPLC柱和检测器，通过电化学检测或荧光检测（标记后）能够轻松实现对200 - 300飞摩尔（2 - 3×10⁻³摩尔）寡糖的检测。小型化分离系统极大地提高了灵敏度，一台CE - 激光诱导荧光仪器能够检测低至100尧摩尔（1×10⁻²²摩尔）的物质。
