微分離管高效液相層析技術及其應用

Microcolumn HPLC and Its Applications

Jean-Pierre Chervet ； 吳玉琛(Yuh-Shen Wu) ； 石瑩(Ying Shih)

《化學》 54卷3期 (1996/09) Pp. 71-89

https://doi.org/10.6623/chem.1996050

摘要

對於微量樣本的分析技術的成長，與分析技術要求增進靈敏度，快速液相層析分析技術，以及最小量的環境污染及低價的分析技術都是尋求或發展微分離管層析技術的趟動力。微分離管層析技術結合電子噴灑質譜術、蛋白質研究、生化或生化高分子分析，法醫學研究或神經科學，都產生許多有趣的微分離管或毛細管高效液相層應用研究領域，甚至毫微高效液相層析技術也會在不久的將來被引進。在本文中除了回顧微分離管層析技術之原理及定義的始末，詳細的描述儀器的需求，幫浦的規格、注射器、管柱、連接管等等，實際的圖譜也用來顯示或驗證微管層析上的理論重點，並從大量的文獻報導中探討最常用的偵測器，包括紫外光偵測器、螢光偵測器、電化學偵測器及電子噴灑質譜儀的連線技銜。最後對於能達成最佳化之微分離管高效率相層析儀器需求作一簡要說明。

並列摘要

The growing interest in the analysis of minute samples and the permanent need for increased sensitivities, better separation efficiencies and faster analysis in almost any field of liquid chromatography, combined with the increasing demand for analytical techniques with minimal environmental hazards and lower cost of operation, are some of the driving forces towards microcolumn HPLC. Coupling of liquid chromatography with electrospray mass spectrometry (ESIMS) and MALDI-TOFIMS, protein/peptide research, bio- and biopolymer analysis, forensic applications, neuroscience, combinatorial chemistry, etc., are some of the fields where micro- and capillary HPLC are used routinely and where new techniques such as nano HPLC will emerge very rapidly. In this review some definitions and the theoretical basis of microcolumn HPLC are reviewed. A detailed description of the instrumental requirements including specifications for pumps, injectors, columns, connecting tubing, detectors, etc ., are given. Practical examples based on authentic chromatograms are used to demonstrate and underlay the theoretical principles. From the large variety of detection principles adapted for microcolumn HPLC the most frequently used, such as UV absorbance, fluorescence, electrochemical and ESIMS are, discussed in detail. Finally, the instrumental requirements for micro-, capillary and nano HPLC are summarized and suggestions are given for optimal system configurations.