121st General Meeting of the KCS

Type Poster Presentation
Area Analytical Chemistry
Room No. Event Hall
Time 4월 19일 (목요일) 11:00~12:30
Code ANAL.P-288
Subject “Hidden” isocratic elution prior to gradient elution increases the utilization of LC-MS/MS experiment time for efficient proteomic analyses
Authors Do-Woon Nam, Sang-Won Lee*, Hangyeore Lee, Jeonghei Choi
Department of Chemistry, Korea University, Korea
Abstract In advanced proteomics, liquid chromatography - tandem mass spectrometry (LC-MS/MS) is commonly used to characterize proteins and proteomes and, moreover, to discover candidate biomarkers. To do so, complex peptide mixture should be effectively resolved by high-resolving power LC with selected separation method. Reverse-Phase Liquid Chromatography (RPLC) which is generally used in proteomics can be subdivided into isocratic and gradient from elution type. Isocratic elution method have the advantage of separating hydrophilic peptide while gradient elution method have merit of separating hydrophobic peptide. We employed a new separation method by combining isocratic elution method and gradient elution method and applied it to dual online-ultrahigh pressure liquid chromatography (DO-UHPLC) system. DO-UHPLC system has two time bases which allows one pump to perform gradient elution while another pump can simultaneously carry out equilibration, injection, and isocratic elution. When injected sample finished isocratic elution by one pump, it instantly switched into gradient elution pump and start analyzing. Consequently this method increase total elution time during the same analysis time and hydrophilic peptides in sample mixture can be separated. Thus, this new mixed separation method allows us to exploit overall separation space while using almost 100% LC-MS/MS duty cycle.
E-mail dowoon0517@korea.ac.kr