TY - JOUR
T1 - Concurrent quantification of tryptophan and its major metabolites
AU - Lesniak, Wojciech G.
AU - Jyoti, Amar
AU - Mishra, Manoj K.
AU - Louissaint, Nicolette
AU - Romero, Roberto
AU - Chugani, Diane C.
AU - Kannan, Sujatha
AU - Kannan, Rangaramanujam M.
N1 - Funding Information:
This research was supported in part by the Perinatology Research Branch, Division of Intramural Research, Eunice Kennedy Shriver National Institute of Child Health and Human Development (NICHD), NIH, and by R01HD069562, NICHD (S.K.).
PY - 2013
Y1 - 2013
N2 - 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.
AB - 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.
KW - HPLC
KW - Kynurenic acid
KW - Kynurenine pathway
KW - Serotonin
KW - Tryptophan metabolites
UR - http://www.scopus.com/inward/record.url?scp=84886066556&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84886066556&partnerID=8YFLogxK
U2 - 10.1016/j.ab.2013.09.001
DO - 10.1016/j.ab.2013.09.001
M3 - Article
C2 - 24036037
AN - SCOPUS:84886066556
SN - 0003-2697
VL - 443
SP - 222
EP - 231
JO - Analytical biochemistry
JF - Analytical biochemistry
IS - 2
ER -