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HPLC Linked Electrospray Tandem Mass Spectrometry: A Rapid and Reliable Method to Analyse Indole‐3‐Acetic Acid Metabolism in Bacteria
Author(s) -
Prinsen Els,
Van Dongen Walter,
Esmans Eddy L.,
Van Onckelen Henri A.
Publication year - 1997
Publication title -
journal of mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.475
H-Index - 121
eISSN - 1096-9888
pISSN - 1076-5174
DOI - 10.1002/(sici)1096-9888(199701)32:1<12::aid-jms444>3.0.co;2-7
Subject(s) - chemistry , indole test , chromatography , tryptamine , indole 3 acetic acid , tryptophan , tandem mass spectrometry , isatin , mass spectrometry , selected reaction monitoring , organic chemistry , amino acid , biochemistry , auxin , gene
Electrospray tandem mass spectrometry in combination with high‐performance liquid chromatography can be used for the simultaneous detection and quantification of indole‐3‐acetic acid, indole‐3‐methanol, tryptophan, indole‐3‐acetamide, indole‐3‐ethanol, tryptamine, indole‐3‐acetaldehyde, indole‐3‐acetonitrile, indole‐3‐aldehyde, indole‐3‐lactic acid, indole‐3‐pyruvate and anthranilate on an individual basis. Although methylation is not an absolute prerequisite for analysis of most carboxyl‐type indole compounds, samples were methylated before analysis and this improved the detection limits for tryptophan, indole‐3‐acetic acid, anthranilate and indole‐3‐lactate up to 1000‐fold. Moreover, owing to this methylation no ion suppression conditions, necessary for chromatography of organic acids, have to be used and stability of indole‐3‐pyruvate was achieved. A detection limit of 100 fmol indole‐3‐acetic acid methyl ester injected on‐column was obtained under multiple reaction monitoring conditions. In view of the analysis of a large number of samples, the chromatographic conditions selected are a compromise between speed of analysis and resolution. © 1997 by John Wiley & Sons, Ltd.