Abstract

A LC-MS/MS technique separated the bovine and mouse brain gangliosides monosialotetrahexosylgangliosides (GM1), disialotetrahexosylgangliosides (GD1a), trisialotetrahexosylgangliosides (GT1b) and tetrasialotetrahexosylgangliosides (GQ1b) using a phenyl-hexyl HPLC column and employing a linear methanol gradient in water, which is 0.028% in ammonium hydroxide. The gangliosides were separated according to sialic acid class, and within a particular class, gangliosides having different ceramide carbon chain lengths were also separated. All gangliosides of a particular sialic acid class eluted in characteristic retention time windows in the order of GQ1b, (earliest), GT1b, GD1a, and GM1 (latest). Within each specific retention time window for a particular ganglioside class, gangliosides were separated in the order of increasing ceramide carbon chain length. The phenyl-hexyl column separation of gangliosides is advantageous over established hydrophilic interaction and conventional reversed-phase chromatography techniques, in that the former separates gangliosides according to sialic acid class but not ceramide composition and the latter distributes all the sialic acid ganglioside classes throughout the entire chromatogram. The mechanism of separation of the ganglioside sialic acid classes is proposed to be a π-electron repulsion of negatively-charged gangliosides by the column phenyl moiety.

摘要

LC-MS / MS技术通过使用HPLC-HPLC和液相色谱分离的方法，通过使用HPLC-HPLC分离的牛和小鼠脑神经节苷脂单唾液酸四己糖基神经节苷脂（GD1a），二唾液酸四己糖基神经节苷脂（GT1b）和四唾液酸四己糖基神经节苷脂（GQ1b）在氢氧化铵中为0.028％。根据唾液酸类别分离神经节苷脂，并且在特定类别中，还分离具有不同神经酰胺碳链长度的神经节苷脂。特定唾液酸类的所有神经节苷脂在特征性保留时间窗口中按GQ1b（最早），GT1b，GD1a和GM1（最新）的顺序洗脱。在特定神经节苷脂类别的每个特定保留时间窗口内，神经节苷脂以神经酰胺碳链长度增加的顺序分离。神经节苷脂的苯基-己基柱分离优于已建立的亲水相互作用和常规的反相色谱技术，因为前者根据唾液酸类别分离神经节苷脂，而不是神经酰胺成分，而后者则将所有唾液酸神经节苷脂类别分布在整个区域色谱。认为神经节苷脂唾液酸类的分离机理是通过柱苯基部分使带负电的神经节苷脂的π电子排斥。因为前者根据唾液酸类别分离神经节苷脂而不是神经酰胺成分，而后者根据整个色谱图分配所有唾液酸神经节苷脂类别。认为神经节苷脂唾液酸类的分离机理是通过柱苯基部分使带负电的神经节苷脂的π电子排斥。因为前者根据唾液酸类别分离神经节苷脂而不是神经酰胺成分，而后者根据整个色谱图分配所有唾液酸神经节苷脂类别。认为神经节苷脂唾液酸类的分离机理是通过柱苯基部分使带负电的神经节苷脂的π电子排斥。