Utilization of hydralazine as a reactive matrix for enhanced detection and on-MALDI-target derivatization of saccharides
Utilization of hydralazine as a reactive matrix for enhanced detection and on-MALDI-target derivatization of saccharides
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肼苯哒嗪作为反应基质用于增强检测和靶向MALDI衍生化的用途
DOI：
10.1016/j.cclet.2023.108885
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发表时间：
2023-08
影响因子：
9.1
通讯作者：
Dan Ouyang;Huan Huang;Yanting He;Jiajing Chen;Jiali Lin;Zhuling Chen;Zongwei Cai;Zian Lin
中科院分区：
文献类型：
--
作者：
Dan Ouyang;Huan Huang;Yanting He;Jiajing Chen;Jiali Lin;Zhuling Chen;Zongwei Cai;Zian Lin
糖类是生命中普遍存在的一种重要分子。然而，基质辅助激光解吸/电离质谱 (MALDI-MS) 糖类分析的应用受到电离效率低和唾液酸组分不稳定的限制。基于非还原胺化的衍生策略提供了一个很好的解决方案，然而，这通常非常耗时，并且可能由于去除过量的衍生试剂而导致样品损失。在此，肼屈嗪（HZN）被用作反应基质，直接在 MALDI 靶标上标记还原糖，从而消除了繁琐的样品制备并避免了样品损失。经过优化，在正负模式下均实现了中性和酸性糖类的有效且可重复的 MALDI 靶点衍生化。与2,5-二羟基苯甲酸（DHB）和9-氨基吖啶（9-AA）相比，HZN提高了还原糖的检测灵敏度，并在MS/MS分析中提供了更丰富的碎片离子。此外，还分别从卵清蛋白（OVA）和牛胎球蛋白中鉴定出26种中性聚糖和5种唾液酸聚糖。结合统计模型，该策略可用于区分和预测6个品牌啤酒的样本，并区分2种啤酒发酵模式。此外，应用HZN对尿样中的葡萄糖进行定量分析，得到的糖尿病患者尿糖浓度与临床检测结果一致，显示出对复杂样品中还原糖进行定性定量分析的潜力。
Saccharides are a sort of ubiquitous and vital molecules within the whole life. However, the application of saccharides analysis with matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) is restricted by their low ionization efficiency and the instability of the sialic acid fraction. Derivatization strategy based on nonreductive amination provides a good solution, however, this is often time consuming and may result in sample loss due to removal of excessive derivatization reagents. Herein, hydralazine (HZN) was utilized as a reactive matrix for labeling reducing saccharides directly on MALDI target which eliminated tedious sample preparation and avoided sample loss. After optimization, effective and reproducible on-MALDI-target derivatization of neutral and acidic saccharides was achieved in both positive and negative modes. Compared with 2,5-dihydroxybenzoic acid (DHB) and 9-aminoacridine (9-AA), HZN improved the detection sensitivity of reducing saccharides and provided more abundant fragment ions in MS/MS analysis. Moreover, 26 kinds of neutral glycans and 5 kinds of sialic glycans were identified from ovalbumin (OVA) and bovine fetuin, respectively. Combined with the statistical models, this strategy could be used to distinguish and predict samples of 6 brands of beer, and discriminate 2 kinds of beer fermentation modes. In addition, HZN was applied for quantitative analysis of glucose in urine samples, and the obtained urine glucose concentrations of diabetic patients were consistent with the clinical test results, showing the potential of qualitative and quantitative analysis of reducing saccharides in complex samples.