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Capillary liquid chromatography-fast atom bombardment mass spectrometry using a high-resolving cation exchanger,
Y M Li1, P Brostedt, S Hjertén
1Department of Clinical Neuroscience, Karolinska Institute, Stockholm, Sweden.
Abstract:
Hyphenated mass spectrometric techniques such as LC-MS are advantageous over standard MS methods, because they provide increased sensitivity and minimize signal suppression by other compounds present in the reaction mixture. Recently, we have introduced so-called continuous beds, and applied this technique to prepare a 0.32 mm I.D. cation-exchange capillary column, in order to separate the reaction product substance P(1-7) after proteolytic cleavage of substance P by an endopeptidase recovered from human cerebrospinal fluid. The use of a volatile buffer for elution provides very good flow stability. Ion-exchange microcolumns may be particularly useful for the separation of those peptides that co-elute in reversed-phase chromatography because the separation mechanisms of these two methods are different.