Profiling of basic amino acids and polyamines in microbial culture supernatants by electrospray mass spectrometry.
As part of our efforts to investigate the biosynthesis of proferrioxamines in Erwinia amylovora, it was of interest to develop a methodology with which a large number of basic amino acids, di- and polyamines, and C- and N-hydroxy derivatives thereof could be monitored simultaneously. Towards this end, the on-line coupling of electrospray mass spectrometry with ion chromatography or reversed-phase chromatography was explored. Tandem mass spectrometry was found to be an excellent method for obtaining relevant structural information on underivatized polyamines and basic amino acid, including N-hydroxy derivatives. However, the coupling of ion chromatography with electrospray mass spectrometry for on-line extraction and separation of the underivatized compounds is hampered by compatibility problems. For most target compounds, on-line reversed-phase liquid chromatography/electrospray mass spectrometry was the method of choice but required derivatization and in this respect dansylation was preferred over benzoylation. Neither derivatization method is suitable for N-hydroxy compounds, which are base-sensitive.
Beckman Research Institute of the City of Hope, Duarte, California 011010-0269.
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Feistner
D000596:Amino Acids / Q000032:analysis*; D002852:Chromatography, Ion Exchange; D003470:Culture Media; D003619:Dansyl Compounds / Q000737:chemistry; D004885:Erwinia / Q000737:chemistry*; D013058:Mass Spectrometry / Q000379:methods*; D011073:Polyamines / Q000032:analysis*
D000596:Amino Acids; D003470:Culture Media; D003619:Dansyl Compounds; D011073:Polyamines