HIGH-SPEED HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY OF PEPTIDES AND PROTEINS
HIGH-SPEED HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY OF PEPTIDES AND PROTEINS
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DOI：
10.1016/0021-9673(94)01254-c
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发表时间：
1995-06-23
影响因子：
4.1
通讯作者：
HORVATH, C
中科院分区：
文献类型：
--
作者：
CHEN, H;HORVATH, C
在过去的三十年里，HPLC这个名字已经成为了高速液相色谱的代名词，在过去的十年里，我们经历了分析速度的急剧提高，特别是在生物大分子（如蛋白质）的分离方面。色谱柱技术具有坚实的色谱理论基础，是该领域取得进展的主要原因。最近的研究表明，用双分散或双峰型的微孔或巨孔固定相填充的柱有助于在流动相和固定相之间快速传质，从而可以在很短的时间内提供高分辨率的分离。这表明高效液相色谱法有潜力成为实时在线监测生物技术过程的主要分析技术。进一步提高分离速度来自于高温的使用。温度在HPLC中的作用在很大程度上被忽视，大多数商用仪器没有配备适当的温度控制。然而，本文提出的结果强烈表明，升高的柱温可能会在大分子的HPLC中发现越来越多的用途。在这样的分析应用中，温度编程也可以发挥主要作用，提供低热容量的柱，如填充熔融二氧化硅毛细管，在HPLC中获得更广泛的应用。
Over the last thirty years the name HPLC has been synonymous with high-speed liquid chromatography and during the last ten years we have experienced a dramatic increase in the speed of analysis particularly as far as the separation of biological macromolecules, such as proteins, is concerned. With a solid grounding in the chromatographic theories, column technology has been mainly responsible for the advances in this field. Recent development shows that columns packed with micropellicular or gigaporous stationary phases of the bidisperse or the bimodal type facilitate rapid mass transfer between the mobile and stationary phases and thus can deliver high resolution separations in a very short time. This suggests that HPLC has the potential to be the prime analytical technique for on-line monitoring of biotechnological processes in real time. Further enhancement of the speed of separation comes from the use of elevated temperatures. The role of temperature in HPLC has largely been ignored and most commercial instruments are not equipped with appropriate temperature control. Results presented here strongly suggest, however, that elevated column temperature may find increasing use in the HPLC of large molecules. In such analytical applications temperature programming may also play a major role provided columns with low heat capacity, such as packed fused-silica capillaries, gain wider employment in HPLC.