Výhody a nevýhody zvýšené teploty při separaci peptidů na obrácených fázích

• Main Author: Šemlej, Tomáš
• Other Authors: Lenčo, Juraj
• Format: Dissertation
• Language: Czech
• Published: 2019
• Online Access: http://www.nusl.cz/ntk/nusl-397426

Charles University, Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Tomáš Šemlej Supervisor: PharmDr. Juraj Lenčo, Ph.D. Title of diploma thesis: Pros and cons of using elevated temperature for peptide separation using reversed phases Nowadays, powerful mass spectrometers are available for bottom-up LC-MS proteomic analyses. They allow to identify up to several thousand proteins in one bottom-up analysis. It has been recently demonstrated, however, that their capabilities cannot be fully exploited without significant improvements in peptide separation. One of the easiest, but often neglected, ways to enhance peptide separation efficiency is to increase column temperature. Elevated column temperature may contribute to more efficient separation especially by increasing low diffusivity of peptides. On the other hand, stability of both a stationary phase and peptides limits increasing column temperature to values affording maximum effect on separation efficiency. In this thesis, we explored the effect of increased column temperature on peptide separation efficiency using columns packed either with superficially porous particles or with fully porous particles. Elevated column temperature narrowed peaks for peptides eluted from fully porous particles whereas peak...