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亲水作用液相色谱(HILIC)近年来引起了越来越多的关注,然而用于亲水液相色谱的固定相种类仍然有限,需要开发更多的固定相和分离分析方法。我们课题组开发和评价了基于硅胶基质的叠氮基固定相。首先,采用核苷作为测试样品对固定相的保留性质进行了评价,结果表明核苷在叠氮基固定相上的保留符合亲水液相色谱的典型特征。其次,比较了亲水模式下流动相中两种缓冲盐(碳酸氢铵和甲酸铵)和酸添加剂对叠氮基固定相糖肽富集选择性的影响。结果表明当流动相中不加缓冲盐并且调成酸性时,糖肽富集选择性更高,在优化富集条件后,从辣根过氧化物酶(HRP)中富集得到9条糖肽信号,所开发的叠氮基固定相具有在HILIC模式下使用的广阔前景。
Hydrophilic interaction liquid chromatography (HILIC) has attracted more and more attention in recent years. However, the types of stationary phases used in hydrophilic liquid chromatography are still limited, and more stationary phases and separation methods need to be developed. Our team developed and evaluated azide-based silica-based stationary phases. First, the retention properties of the stationary phase using nucleosides as test samples were evaluated and the results show that the retention of nucleosides on the azido stationary phase conforms to the typical characteristics of hydrophilic liquid chromatography. Secondly, the effects of two buffer salts (ammonium bicarbonate and ammonium formate) and acid additives in the mobile phase on the enrichment selectivity of glycopeptides in azido phase were compared. The results showed that when the mobile phase without buffer salt and tune into acidic, glycopeptide enrichment selectivity higher, the optimal enrichment conditions, horseradish peroxidase (HRP) enrichment obtained nine glycopeptides Signal, the developed azide-based stationary phase has broad prospects for use in HILIC mode.