Beijing Key Laboratory for Green Catalysis and Separation, Department of Chemistry and Chemical Engineering, Beijing University of Technology, Beijing 100124, PR China.
Beijing Key Laboratory for Green Catalysis and Separation, Department of Chemistry and Chemical Engineering, Beijing University of Technology, Beijing 100124, PR China.
Talanta. 2019 Jan 15;192:360-367. doi: 10.1016/j.talanta.2018.09.025. Epub 2018 Sep 11.
A novel composite monolithic column based on graphene oxide-trimethyl-2-methacroyloxyethylammonium chloride-titania (GO-META-TiO) was developed for the enrichment of phosphopeptides. META was proposed as a "bridge" to connect GO and TiO species to prepare GO-META-TiO composite. This high surface area composite (surface area = 196.93 m g) was fixed in the monolithic column via an in situ UV polymerization process. In-tube solid phase microextraction (IT-SPME) using this composite was coupled with matrix-assisted laser desorption ionization time-of-flight-mass spectrometry (MALDI-TOF-MS) for the enrichment and detection of phosphorylated peptides from a digestion mixture of α-casein, β-casein, and bovine serum albumin (BSA) in molar ratios of 1:1:1, 1:1:10, and 1:1:100. The key factors affecting the IT-SPME of the phosphopeptides, such as the elution solution concentrations, the extraction flow rate, and the elution flow rate were comprehensively investigated. For further demonstration, this method was employed for the enrichment and detection of phosphorylated peptides from digested chicken egg white. The obtained results indicated that the GO-META-TiO composite monolithic column rapidly and efficiently captured the phosphopeptides present in these complex biological samples, even in the 10 fmol β-casein tryptic digest. We therefore propose that the reported GO-META-TiO composite monolithic column possesses a suitable affinity for the selective extraction of phosphopeptides from biological samples. This method paves a way in extending the application of nanomaterials.
一种新型的基于氧化石墨烯-三甲基-2-甲丙烯基氯化铵-二氧化钛（GO-META-TiO）的整体式复合柱被开发用于富集磷酸肽。META 被提议作为连接 GO 和 TiO 物种的“桥梁”，以制备 GO-META-TiO 复合材料。这种高表面积的复合材料（表面积=196.93m²/g）通过原位 UV 聚合过程固定在整体式柱中。使用这种复合材料的管内固相微萃取（IT-SPME）与基质辅助激光解吸电离飞行时间质谱（MALDI-TOF-MS）相结合，用于从α-酪蛋白、β-酪蛋白和牛血清白蛋白（BSA）的消化混合物中富集和检测磷酸化肽，摩尔比为 1:1:1、1:1:10 和 1:1:100。综合考察了影响 IT-SPME 磷酸肽的关键因素，如洗脱液浓度、萃取流速和洗脱流速等。为了进一步验证，该方法用于从鸡蛋白消化液中富集和检测磷酸化肽。结果表明，GO-META-TiO 整体式复合柱能够快速有效地捕获这些复杂生物样品中存在的磷酸肽，即使在 10 fmol 的β-酪蛋白胰蛋白酶消化物中也是如此。因此，我们提出所报道的 GO-META-TiO 整体式复合柱对从生物样品中选择性提取磷酸肽具有合适的亲和力。该方法为纳米材料的应用拓展提供了一种途径。
