Development of new liquid chromatography techniques for mass spectrometric analysis of metabolites and proteins.

摘要

Three liquid chromatography (LC) techniques are developed to combine with mass spectrometry (MS) for proteomics and metabolomics applications. The first technique involves the use of a bifunctional LC column with surface chemistry including an ionic group and a long chain hydrophobic group for multidimensional separation. This new single-column multidimensional LC technique is combined with electrospray ionization (ESI) MS for amino acid analysis in human urine samples. In the second technique, bacterial proteins are digested by trypsin and the digested peptides are injected into LC-ESI MS/MS with a separation window of as short as 2.5 min for identification of specific bacterial proteins which leads to rapid bacterial identification. Finally, a chromatofocusing method with a pH range from 3 to 12 has been developed and the applicability of this technique is demonstrated in milk and serum proteome analysis.