Faculty of Pharmaceutical Sciences, Fukuoka University, 8-19-1 Nanakuma, Johnan, Fukuoka 814-0180, Japan.
Rapid Commun Mass Spectrom. 2010 Oct 15;24(19):2868-74. doi: 10.1002/rcm.4710.
We have developed a novel method for selective and sensitive analysis of sialic acids (N-acetylneuraminic, N-glycolylneuraminic, and 2-keto-3-deoxy-D-glycero-D-galactonononic acid) utilizing liquid chromatography/tandem mass spectrometry (LC/MS/MS) combined with a fluorous derivatization technique. In this method, the carboxylic groups in the sialic acids are derivatized via amidation with heptadecafluoroundecylamine, a commercially available perfluoroalkylamine reagent. This reaction proceeds rapidly and readily at room temperature in the presence of a condensation reagent. Subsequently, the derivatives are retained specifically on an LC column with a perfluoroalkyl stationary phase by means of a fluorophilic or 'fluorous' interaction, and detected by positive electrospray ionization MS/MS. The detection limits of the examined sialic acids are in the range of 60-750 amol on column. We show that the proposed method can be used to analyze trace amounts of sialic acids in biological samples.
我们开发了一种新的方法，利用液相色谱/串联质谱（LC/MS/MS）结合氟衍生化技术，对唾液酸（N-乙酰神经氨酸、N-羟乙酰神经氨酸和 2-酮基-3-脱氧-D-甘油-D-半乳糖壬酮酸）进行选择性和灵敏的分析。在该方法中，通过与十七氟十一烷基胺（一种市售的全氟烷基胺试剂）进行酰胺化反应，使唾液酸中的羧基衍生化。在缩合剂存在的情况下，该反应在室温下迅速且容易进行。随后，通过亲氟或“氟”相互作用，衍生物特异性地保留在具有全氟烷基固定相的 LC 柱上，并通过正电喷雾电离 MS/MS 进行检测。所检测的唾液酸的检测限在柱上的范围为 60-750 amol。我们表明，所提出的方法可用于分析生物样品中痕量的唾液酸。
