Open-Tubular Capillary Electrochromatography with an On-Line Ion Trap Storage/Reflectron Time-of-Flight Mass Detector for Ultrafast Peptide Mixture Analysis

Abstract

In this work, a novel open-tubular column (OTC) capillary electrochromatography (CEC) system has been coupled to an on-line ion trap storage/reflectron time-of-flight mass spectrometer for ultrafast peptide mixture analysis. Reversed-phase OTCs prepared by the sol−gel process were coated with an amine, which greatly enhanced the electroosmotic flow in an acidic buffer solution and considerably reduced nonspecific adsorption between the peptides and the column wall. A six-peptide mixture could be separated to baseline within 3 min on this system. A full mass range detection speed of 8 Hz was used in all these experiments, which was sufficiently rapid to maintain the high efficiency of ultrafast separations. Because of the high duty cycle of the mass spectrometer and the column path length-independent concentration-sensitive feature of the electrospray ionization process, high-quality total ion chromatograms could be obtained with injections of only 1−2 fmol of peptide samples. A concentration limit of detection of 1 × 10^-6 M was also achieved due to the preconcentration capability of CEC. In addition, a novel gradient CEC device was demonstrated which did not result in a pressure-driven flow. A tryptic digest of horse heart myoglobin was successfully separated on the gradient CEC system within 6 min. The use of the mass spectrometer increased the resolving power of this system by clearly identifying coeluting components.