Size-exclusion chromatography performed in capillaries - Studies by liquidchromatography mass spectrometry

Miniaturization of the chromatographic column led to increased sensitivity and shortened time of analysis. In our work we applied 300 mu m LD. capilla ries packed with a novel stationary phase Superdex Peptide for the size-exc lusion chromatography, capable of separating molecules within the mass rang e of 0.1-7 kDa. Here we proved that such capillary columns can operate effe ctively at high sensitivity. Several peptide mixtures were efficiently chro matographed and analyzed on line with electrospray ionization mass spectrom etry as a detection technique. A CNBr peptide map, derived from human globi n alpha subunit, was effectively separated using this method. These fragmen ts are difficult to elute from the reversed-phase column at low pH, therefo re, such approach can be considered as a complementary to other separation techniques, in particular for analyzing hydrophobic components and complex mixtures. (C) 1999 Elsevier Science B.V. All rights reserved.