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09월 03일 23시 이후 : 초록수정 불가능, 일정확인 및 검색만 가능
대한화학회 제122회 학술발표회 및 총회
LC/MS/MS based Construction of Isomer-Specific and Region-Specific Mouse Brain Ganglioside Library
2018년 8월 16일 18시 16분 30초
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10월 19일 (금요일) 11:00~12:30
, Hyun Joo An
Graduate School of Analytical Science and Technology, Chungnam National University, Korea
Gangliosides are anionic glycosphingolipids containing one to several sialic acid residues. Although they play an important role in neuro-biological functions including synaptic plasticity and memory formation, there have been only few studies in ganglioside due to their structural complexity and the lack of effective analytical methods. In particular, isomeric investigation of gangliosides is necessarily required in order to understand the ganglioside biology with the consideration of its biosynthesis however, isomer separation based on reversed-phase (RP) chromatography is highly challengeable. In this study, for the purpose of construction of localized mouse brain ganglioside library, we examined ganglioside from anatomically dissected nine mouse brain regions by isomer-specific LC/MS and LC/MS/MS. Briefly, gangliosides were extracted by modified Folch method with chloroform and methanol, followed by purification and enrichment by C18-SPE. Then they were identified and quantified by UHPLC (C18 column) QTOF MS. Ganglioside isomers were completely separated depending on their glycan traits on a C18 column by addition of formic acid in the mobile phase. Furthermore, tandem MS analysis was performed to confirm the identification based on diagnostic fragment ions. Approximately, isomeric structures of 40 ganglioside compositions were elucidated including O-acetylated species. Through LC-MS/MS based isomeric investigation of gangliosides, region-specific mouse brain ganglioside biosynthetic pathway including 0-, a-, b- and c- series could be suggested. Interestingly, major ganglioside were distributed with distinguished qualitative and quantitative pattern for each nine brain regions. Constructed mouse brain ganglioside library is going to be used for monitoring alteration in KO mouse models including glycosylation transferase KO mice.
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