Structural characterization of human milk oligosaccharides using ultrahigh performance liquid chromatography–helium charge transfer dissociation mass spectrometry
Structural characterization of human milk oligosaccharides using ultrahigh performance liquid chromatography–helium charge transfer dissociation mass spectrometry
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超高效液相色谱-氦电荷转移解离质谱法表征母乳寡糖的结构
DOI：
10.1093/glycob/cwac010
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发表时间：
2022
期刊：
影响因子：
4.3
通讯作者：
Jackson, Glen P
中科院分区：
文献类型：
--
作者：
Mendis, Praneeth M;Jackson, Glen P
采用氦电荷转移解离质谱（He-CTD-MS）和高效液相色谱（UHPLC）联用技术对酸性和中性人乳低聚糖（HMOs）进行分析。研究的重点是单糖序列的鉴定，分支模式，唾液酸化/岩藻糖基化的安排，并在混合物中的异构寡糖的分化。初始研究首先优化了UHPLC分离条件和He-CTD-MS条件。结果表明，He-CTD与UHPLC时间尺度兼容，并从还原和非还原末端提供明确的糖苷和交叉环裂解，这通常不可能使用碰撞诱导解离。He-CTD产生信息片段，包括0，3 Anion和0，4 Anion，其已通过电子转移解离、电子脱离解离和紫外光解离（UVPD）观察到，并且对于区分唾液酸-N-四糖HMO中存在的α-2，3-与α-2，6-连接的唾液酸（Neu 5Ac）残基至关重要。除了连接位置，He-CTD还能够通过提供0，2An，0，2Xn和1，5An型的独特，明确的交叉环裂解来区分唾液酸乳糖-N-四糖和乳糖-N-岩藻五糖结构的结构异构体，同时保留大部分不稳定的Neu 5Ac和岩藻糖基团。
The combination of helium charge transfer dissociation mass spectrometry (He–CTD–MS) with ultrahigh performance liquid chromatography (UHPLC) is presented for the analysis of a complex mixture of acidic and neutral human milk oligosaccharides (HMOs). The research focuses on the identification of the monosaccharide sequence, the branching patterns, the sialylation/fucosylation arrangements, and the differentiation of isomeric oligosaccharides in the mixture. Initial studies first optimized the conditions for the UHPLC separation and the He–CTD–MS conditions. Results demonstrate that He–CTD is compatible with UHPLC timescales and provides unambiguous glycosidic and cross-ring cleavages from both the reducing and the nonreducing ends, which is not typically possible using collision-induced dissociation. He–CTD produces informative fragments, including0,3Anand0,4Anions, which have been observed with electron transfer dissociation, electron detachment dissociation, and ultraviolet photodissociation (UVPD) and are crucial for differentiating the α-2,3- versus α-2,6-linked sialic acid (Neu5Ac) residues present among sialyllacto-N-tetraose HMOs. In addition to the linkage positions, He–CTD is able to differentiate structural isomers for both sialyllacto-N-tetraoses and lacto-N-fucopentaoses structures by providing unique, unambiguous cross-ring cleavages of types0,2An,0,2Xn, and1,5Anwhile preserving most of the labile Neu5Ac and fucose groups.