Article ID Journal Published Year Pages File Type
1213046 Journal of Chromatography B 2015 6 Pages PDF
Abstract

•Simultaneous determination of six intermediates in IAA biosynthesis by LC–MS/MS.•Application in Pseudomonas putida with known IAA biosynthesis pathway.•A simple, rapid, and reliable assay for understanding the IAA biosynthesis pathway.

Indole-3-acetic acid (IAA) is produced from tryptophan through five synthesis pathways. A comprehensive method for the quantification of IAA and biosynthesis-related intermediates in a culture medium was developed. Sample preparation was simple with protein precipitation. The analytes were separated on a superficially porous C18 silica column and detected by electrospray ionization-tandem mass spectrometry in the positive ion multiple reaction monitoring mode. The limit of detection was 0.05 μM, and the lower limits of quantification ranged from 0.05 to 2 μM. The intra-day and inter-day precision and accuracy were less than 13.96%. Ion suppression was observed, and the deuterated internal standards were used to compensate for the matrix effect. The method was applied to analyze changes in tryptophan catabolism in a culture medium of Pseudomonas putida. The proposed method is robust and suitable for the systematic profiling of IAA biosynthesis in culture supernatant.

Related Topics
Physical Sciences and Engineering Chemistry Analytical Chemistry
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