期刊
JOURNAL OF CHROMATOGRAPHY A
卷 1614, 期 -, 页码 -出版社
ELSEVIER
DOI: 10.1016/j.chroma.2019.460650
关键词
HILIC; Retention modelling; Bottom-up proteomics; Mass spectrometry
资金
- Horizon 2020 - Excellent Science - European Research Council (ERC) [694151]
- European Research Council (ERC) [694151] Funding Source: European Research Council (ERC)
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%. (C) 2019 The Authors. Published by Elsevier B.V.
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