Department of Analytical Chemistry, Faculty of Science, Charles University, Hlavova 8, 128 43 Prague 2, Czech Republic.
Department of Analytical Chemistry, Faculty of Science, Charles University, Hlavova 8, 128 43 Prague 2, Czech Republic.
J Chromatogr B Analyt Technol Biomed Life Sci. 2020 Aug 1;1150:122196. doi: 10.1016/j.jchromb.2020.122196. Epub 2020 May 28.
The composition of a sample solvent has a crucial impact on separations in hydrophilic interaction liquid chromatography (HILIC). In this short communication, we studied the effect of an organic modifier in the sample solvent on the solubility of different tryptic glycopeptides of hemopexin and haptoglobin proteins. The results showed that the solubility of glycopeptides in solvents with a high acetonitrile content depends on the type of attached N-glycan. We observed lower solubility in larger glycans attached to the same peptide backbone, and we demonstrated that glycopeptides containing sialic acids precipitate more readily than those without sialic acid. Therefore, the sample solvent composition in HILIC must be carefully optimized for accurate quantitative data collection and for adequate separation.
样品溶剂的组成对亲水相互作用液相色谱（HILIC）中的分离有至关重要的影响。在本简短通讯中，我们研究了样品溶剂中有机改性剂对血色素结合蛋白和触珠蛋白的不同胰蛋白酶糖肽溶解度的影响。结果表明，在高乙腈含量的溶剂中糖肽的溶解度取决于连接的 N-聚糖的类型。我们观察到连接到同一肽主链上的较大聚糖的溶解度较低，并且我们证明含有唾液酸的糖肽比不含唾液酸的糖肽更容易沉淀。因此，HILIC 中的样品溶剂组成必须仔细优化，以获得准确的定量数据采集和充分的分离。
