Lattova Erika, Perreault Hélène
Chemistry Department, University of Manitoba, 144 Dysart Road, Manitoba, Winnipeg, Canada R3T 2N2.
J Chromatogr B Analyt Technol Biomed Life Sci. 2003 Aug 5;793(1):167-79. doi: 10.1016/s1570-0232(03)00374-x.
A well-known reaction of carbonyl compounds with phenylhydrazine has been applied to saccharides, providing increased sensitivity for mass spectrometric (MS) and ultraviolet (UV) detection during high-performance liquid chromatographic (HPLC) separations. After a simple derivatization procedure for 1 h at 70 degrees C and purification of the reaction mixture from excess reagent by extraction, the sugar derivatives were characterized by direct injection or on-line HPLC/electrospray ionization (ESI) and by matrix-assisted laser desorption/ionization (MALDI) MS. Because no salts are used or produced upon reaction, this procedure is very simple and suitable for the tagging of saccharides. The reaction allows for on-target derivatization and products are very stable. The derivatization procedure has been applied to commercially-obtained small saccharides and standard N-linked oligosaccharides. Lastly, hen ovalbumin N-glycans were detached enzymatically and characterized by MALDI-MS as their phenylhydrazone derivatives.
羰基化合物与苯肼的一种著名反应已应用于糖类，在高效液相色谱（HPLC）分离过程中提高了质谱（MS）和紫外（UV）检测的灵敏度。在70℃下进行1小时的简单衍生化程序，并通过萃取从过量试剂中纯化反应混合物后，通过直接进样或在线HPLC/电喷雾电离（ESI）以及基质辅助激光解吸/电离（MALDI）MS对糖衍生物进行表征。由于反应过程中不使用或产生盐，该程序非常简单，适用于糖类的标记。该反应允许进行靶向衍生化，且产物非常稳定。该衍生化程序已应用于商业获得的小糖类和标准N-连接寡糖。最后，从鸡卵清蛋白中酶解下N-聚糖，并通过MALDI-MS将其作为苯腙衍生物进行表征。
