Department of Chemistry, University of British Columbia, Vancouver, British Columbia, Canada.
Anal Chem. 2012 Oct 16;84(20):8756-62. doi: 10.1021/ac301889k. Epub 2012 Sep 26.
O-acetylated N-glycans from fish serum of Atlantic salmon (Salmo salar) are characterized by capillary electrophoresis (CE) in conjunction with both laser-induced fluorescence (LIF) and mass spectrometry (MS) detection methods. Glycans derivatized with negatively charged fluorescent label 8-aminopyrene-1,2,6-trisulfonate (APTS) were separated to obtain a CE-LIF profile of the complex glycan mixture, and the profile concurs with that obtained by using electrospray mass spectrometry. The identity of the APTS-labeled glycans was confirmed by CE-MS. The same glycans can be identified also in their native state by CE-MS without derivatization. The structural variations of O-acetylated sialic acid isomers in fish serum glycans are investigated by CE-MS/MS. Selected ion monitoring provided useful structural information of the underivatized glycans from fragmentation spectra. New complex fish serum glycans that are not reported previously were observed and characterized. These methods may be useful not only for the characterization of acetylation of complex glycans but also to study other types of glycan modifications, as well as to allow determination of overall glycan composition in glycoproteins.
采用毛细管电泳（CE）与激光诱导荧光（LIF）和质谱（MS）检测方法联用，对来自大西洋三文鱼（Salmo salar）血清的 O-乙酰化 N-聚糖进行了表征。经带负电荷的荧光标记物 8-氨基芘-1,2,6-三磺酸（APTS）衍生化的聚糖得以分离，得到了复杂糖混合物的 CE-LIF 图谱，该图谱与采用电喷雾质谱获得的图谱一致。通过 CE-MS 确认了 APTS 标记聚糖的身份。同样的聚糖也可以通过 CE-MS 在未经衍生化的情况下以其天然状态进行鉴定。通过 CE-MS/MS 研究了鱼血清聚糖中 O-乙酰化唾液酸异构体的结构变化。选择离子监测为来自碎片谱的未衍生化聚糖提供了有用的结构信息。观察并表征了先前未报道的新型复杂鱼血清聚糖。这些方法不仅可用于表征复杂聚糖的乙酰化，还可用于研究其他类型的聚糖修饰，以及允许确定糖蛋白中的总体聚糖组成。
