title: Quantitative profiling of tryptophan metabolites in serum, urine, and cell culture supernatants by liquid chromatography-tandem mass spectrometry.
abstract: A sensitive, selective, and comprehensive method for the quantitative determination of tryptophan and 18 of its key metabolites in serum, urine, and cell culture supernatants was developed. The analytes were separated on a C18 silica column by reversed-phase liquid chromatography and detected by electrospray ionization tandem mass spectrometry in positive ion multiple reaction monitoring (MRM) mode, except for indoxyl sulfate which was measured in negative ion MRM mode in a separate run. The limits of detection and lower limits of quantification were in the range of 0.1-50 and 0.5-100 nM, respectively. Fully (13)C isotope-labeled and deuterated internal standards were used to achieve accurate quantification. The applicability of the method to analyze serum, urine, and cell culture supernatants was demonstrated by recovery experiments and the evaluation of matrix effects. Precision for the analysis of serum, urine, and cell culture supernatants ranged between 1.3% and 16.0%, 1.5% and 13.5%, and 1.0% and 17.4%, respectively. The method was applied to analyze changes in tryptophan metabolism in cell culture supernatants from IFN-γ-treated monocytes and immature or mature dendritic cells.
authors: Institute of Functional Genomics, University of Regensburg, Regensburg, Germany.
authors: Wentao
authors: W
authors: Zhu
authors: Axel P
authors: AP
authors: Stevens
authors: Katja
authors: K
authors: Dettmer
authors: Eva
authors: E
authors: Gottfried
authors: Sabine
authors: S
authors: Hoves
authors: Marina
authors: M
authors: Kreutz
authors: Ernst
authors: E
authors: Holler
authors: André B
authors: AB
authors: Canelas
authors: Ido
authors: I
authors: Kema
authors: Peter J
authors: PJ
authors: Oefner
mesh_terms: D002460:Cell Line; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003470:Culture Media / Q000737:chemistry; D003713:Dendritic Cells / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D009000:Monocytes / Q000737:chemistry / Q000378:metabolism; D021241:Spectrometry, Mass, Electrospray Ionization; D053719:Tandem Mass Spectrometry / Q000379:methods*; D014364:Tryptophan / Q000097:blood* / Q000378:metabolism / Q000652:urine*
chemical_list: D003470:Culture Media; D014364:Tryptophan