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Anal Biochem. 2013 Dec 15;443(2):222-31. doi: 10.1016/j.ab.2013.09.001. Epub 2013 Sep 11.

Concurrent quantification of tryptophan and its major metabolites.

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  • 1Department of Chemical Engineering and Material Science, Children's Hospital of Michigan, Wayne State University, Detroit, MI 48202, USA; The Center for Nanomedicine, Department of Ophthalmology, Johns Hopkins University School of Medicine, Baltimore, MD 21287, USA; Perinatology Research Branch, National Institute of Child Health and Human Development, NIH, DHHS, Detroit, MI 48201, USA; Perinatology Research Branch, National Institute of Child Health and Human Development, NIH, DHHS, Bethesda, MD 20892, USA.

Abstract

An imbalance in tryptophan (TRP) metabolites is associated with several neurological and inflammatory disorders. Therefore, analytical methods allowing for simultaneous quantification of TRP and its major metabolites would be highly desirable, and may be valuable as potential biomarkers. We have developed a HPLC method for concurrent quantitative determination of tryptophan, serotonin, 5-hydroxyindoleacetic acid, kynurenine, and kynurenic acid in tissue and fluids. The method utilizes the intrinsic spectroscopic properties of TRP and its metabolites that enable UV absorbance and fluorescence detection by HPLC, without additional labeling. The origin of the peaks related to analytes of interest was confirmed by UV-Vis spectral patterns using a PDA detector and mass spectrometry. The developed methods were validated in rabbit fetal brain and amniotic fluid at gestational day 29. Results are in excellent agreement with those reported in the literature for the same regions. This method allows for rapid quantification of tryptophan and four of its major metabolites concurrently. A change in the relative ratios of these metabolites can provide important insights in predicting the presence and progression of neuroinflammation in disorders such as cerebral palsy, autism, multiple sclerosis, Alzheimer disease, and schizophrenia.

Copyright © 2013 Elsevier Inc. All rights reserved.

KEYWORDS:

HPLC, Kynurenic acid, Kynurenine pathway, Serotonin, Tryptophan metabolites

PMID:
24036037
[PubMed - in process]
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