Journal
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
Volume 988, Issue -, Pages 53-58Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jchromb.2015.02.025
Keywords
Indole-3-acetic acid; Liquid chromatography-tandem mass spectrometry; IAA biosynthesis; Tryptophan
Funding
- Tzu University [TCMRC-P-102016]
- National Science Council of Taiwan [NSC 99-2113-M-320-002-MY2]
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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 C-18 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 mu M, and the lower limits of quantification ranged from 0.05 to 2 mu M. 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. (C) 2015 Elsevier B.V. All rights reserved.
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