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09월 03일 23시 이후 : 초록수정 불가능, 일정확인 및 검색만 가능
제122회 대한화학회 학술발표회, 총회 및 기기전시회 안내
“Hidden” isocratic elution on a dual online reverse-phase liquid chromatography to increase the utilization of LC-MS/MS experiment time for highly efficient proteomic analysis
2018년 8월 30일 16시 22분 44초
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10월 19일 (금요일) 11:00~12:30
, Jeonghei Choi
, Hangyeore Lee
, Sang-Won Lee
Korea University, Korea
Department of Chemistry, Korea University, Korea
Reverse-phase liquid chromatography (RPLC) is a method of choice in current proteomics and generally employs a hydrophobic gradient of a mobile phase to efficiently elute peptides. However, in this method, the hydrophilic peptides with little retention to the hydrophobic stationary phase tend to elute at the same time at early gradient, suffering from ion suppression due to the co-eluting peptides. Isocratic elution offers an advantage of separating the low retaining hydrophilic peptides while it is generally not suitable for eluting hydrophobic peptides. In this work, we developed a new separation scheme in which an isocratic elution prior to gradient elution was used and the isocratic elution is “hidden” in the LC-MS/MS experiments time so that the entire experiment time remains the same as if there is only gradient elution being employed. This “hidden” isocratic elution was accomplished by performing the back-flushing sample injection and isocratic elution on one column while RP gradient separation is being executed on the other column on a dual online reverse-phase liquid chromatography (DO-RPLC) system. The duration of the isocratic elution was determined so that the first peptide peak eluting from the column was to be measured by a mass spectrometer, effectively removing lead time in LC-MS/MS measurement. The “hidden” isocratic elution prior to gradient elution on a DO-RPLC system was demonstrated to provide maximum utilization of LC-MS/MS experiment times while accomplishing increased identification of hydrophilic peptides.
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