Van 't Hoff Institute for Molecular Sciences, Science Park 904, 1098 XH Amsterdam, the Netherlands; Centre for Analytical Science Amsterdam, Science Park 904, 1098 XH Amsterdam, the Netherlands.
Van 't Hoff Institute for Molecular Sciences, Science Park 904, 1098 XH Amsterdam, the Netherlands.
J Chromatogr A. 2020 Mar 15;1614:460650. doi: 10.1016/j.chroma.2019.460650. Epub 2019 Oct 23.
The applicability of models to describe peptide retention in hydrophilic interaction liquid chromatography (HILIC) was investigated. A tryptic digest of bovine-serum-albumin (BSA) was used as a test sample. Several different models were considered, including adsorption, mixed-mode, exponential, quadratic and Neue-Kuss models. Gradient separations were performed on three different HILIC stationary-phases under three different mobile-phase conditions to obtain model parameters. Methods to track peaks for specific peptides across different chromatograms are shown to be essential. The optimal mobile-phase additive for the separation of BSA digest on each of the three columns was selected by considering the retention window, peak width and peak intensity with mass-spectrometric detection. The performance of the models was investigated using the Akaike information criterion (AIC) to measure the goodness-of-fit and evaluated using prediction errors. The F-test for regression was applied to support model selection. RPLC separations of the same sample were used to test the models. The adsorption model showed the best performance for all the HILIC columns investigated and the lowest prediction errors for two of the three columns. In most cases prediction errors were within 1%.
考察了模型在描述亲水性相互作用液相色谱（HILIC）中肽保留的适用性。牛血清白蛋白（BSA）的胰蛋白酶消化物被用作测试样品。考虑了吸附、混合模式、指数、二次和Neue-Kuss 模型等几种不同的模型。在三种不同的 HILIC 固定相上进行梯度分离，以获得模型参数。显示出跟踪特定肽在不同色谱图中峰的方法是必不可少的。通过考虑保留窗口、峰宽和峰强度以及质谱检测，选择了用于分离三种柱子上 BSA 消化物的最佳流动相添加剂。使用赤池信息量准则（AIC）评估模型的性能，以衡量拟合优度，并使用预测误差进行评估。应用回归 F 检验支持模型选择。使用相同样品的 RPLC 分离来测试模型。吸附模型在研究的所有 HILIC 柱中表现出最佳性能，并且对于其中两个柱的预测误差最低。在大多数情况下，预测误差在 1%以内。
