Kamerling Johannis P, Gerwig Gerrit J
Bijvoet Center, Department of Bio-Organic Chemistry, Utrecht University, Utrecht, The Netherlands.
Methods Mol Biol. 2006;347:69-91. doi: 10.1385/1-59745-167-3:69.
Over the years several methodologies have been developed for the structural analysis of naturally occurring sialic acids (Sias), a family with more than 62 members. Currently there are two primary instrumental approaches: analysis of volatile Sia derivatives by gas-liquid chromatography (GLC) combined with electron-impact mass spectrometry (EI/MS), and analysis of fluorescently labeled Sias by high-performance liquid chromatography (HPLC) eventually coupled with electrospray mass spectrometry (ESI/MS). This chapter presents both approaches in detail. The volatile Sia derivatives are comprised of trimethylsilylated methyl ester derivatives, heptafluorobutylated methyl ester derivatives, or pertrimethylsilylated derivatives. The fluorescent Sia derivatives are prepared by reaction with 1,2-diamino-4,5-methylenedioxybenzene. For the identification of the different Sia derivatives, detailed GLC, HPLC, EI/MS, and ESI/MS data are included.
多年来，已经开发出几种方法用于天然存在的唾液酸（Sias）的结构分析，唾液酸家族有60多个成员。目前有两种主要的仪器分析方法：通过气液色谱法（GLC）结合电子轰击质谱法（EI/MS）分析挥发性唾液酸衍生物，以及通过高效液相色谱法（HPLC）最终结合电喷雾质谱法（ESI/MS）分析荧光标记的唾液酸。本章详细介绍了这两种方法。挥发性唾液酸衍生物包括三甲基硅烷基化甲酯衍生物、七氟丁酰化甲酯衍生物或全三甲基硅烷基化衍生物。荧光唾液酸衍生物是通过与1,2-二氨基-4,5-亚甲二氧基苯反应制备的。为了鉴定不同的唾液酸衍生物，文中包含了详细的GLC、HPLC、EI/MS和ESI/MS数据。
