title: High efficiency, high temperature separations on silica based monolithic columns.
abstract: The effect of temperature on separation using reversed-phase monolithic columns has been investigated using a nano-LC pumping system for gradient separation of tryptic peptides with MS detection. A goal of this study was to find optimal conditions for high-speed separations. The chromatographic performance of the columns was evaluated by peak capacity and peak capacity per time unit. Column lengths ranging from 20 to 100 cm and intermediate gradient times from 10 to 30 min were investigated to assess the potential of these columns in a final step separation, e.g. after fractionation or specific sample preparation. Flow rates from 250 to 2000 nL/min and temperatures from 20 to 120°C were investigated. Temperature had a significant effect on fast separations, and a flow rate of 2000 nL/min and a temperature of 80°C gave the highest peak capacity per time unit. These settings produced 70% more protein identifications in a biological sample compared to a conventional packed column. Alternatively, an equal amount of protein identifications was obtained with a 40% reduction in run time compared to the conventional packed column.
authors: Department of Chemistry, University of Oslo, Oslo, Norway. magnus.rogeberg@kjemi.uio.no
authors: Magnus
authors: M
authors: Rogeberg
authors: Steven Ray
authors: SR
authors: Wilson
authors: Helle
authors: H
authors: Malerod
authors: Elsa
authors: E
authors: Lundanes
authors: Nobuo
authors: N
authors: Tanaka
authors: Tyge
authors: T
authors: Greibrokk
mesh_terms: D000818:Animals; D002853:Chromatography, Liquid / Q000295:instrumentation* / Q000379:methods; D006358:Hot Temperature; D008099:Liver / Q000737:chemistry; D008855:Microscopy, Electron, Scanning; D010455:Peptides / Q000737:chemistry / Q000302:isolation & purification; D011506:Proteins / Q000737:chemistry / Q000302:isolation & purification; D051381:Rats; D012822:Silicon Dioxide / Q000737:chemistry*; D013696:Temperature
chemical_list: D010455:Peptides; D011506:Proteins; D012822:Silicon Dioxide