title: Stable isotope dilution ultra-high performance liquid chromatography-tandem mass spectrometry quantitative profiling of tryptophan-related neuroactive substances in human serum and cerebrospinal fluid.
abstract: Many compounds related to L-tryptophan (L-TRP) have interesting biological or pharmacological activity, and their abnormal neurotransmission seems to be linked to a wide range of neurodegenerative and psychiatric diseases. A high-throughput method based on ultra-high performance liquid chromatography connected to electrospray tandem mass spectrometry (UHPLC-ESI-MS/MS) was developed for the quantitative analysis of L-TRP and 16 of its metabolites in human serum and cerebrospinal fluid (CSF), representing both major and minor routes of L-TRP catabolism. The combination of a fast LC gradient with selective tandem mass spectrometry enabled accurate analysis of almost 100 samples in 24h. The standard isotope dilution method was used for quantitative determination. The method's lower limits of quantification for serum and cerebrospinal fluid ranged from 0.05 to 15nmol/L and 0.3 to 45nmol/L, respectively. Analytical recoveries ranged from 10.4 to 218.1% for serum and 22.1 to 370.0% for CSF. The method's accuracy ranged from 82.4 to 128.5% for serum matrix and 90.7 to 127.7% for CSF matrix. All intra- and inter-day coefficients of variation were below 15%. These results demonstrate that the new method is capable of quantifying endogenous serum and CSF levels of a heterogeneous group of compounds spanning a wide range of concentrations. The method was used to determine the physiological levels of target analytes in serum and CSF samples from 18 individuals, demonstrating its reliability and potential usefulness in large-scale epidemiological studies.
authors.affiliation: Laboratory of Growth Regulators, Centre of the Region Haná for Biotechnological and Agricultural Research, Faculty of Science of Palacký University & Institute of Experimental Botany CAS, Šlechtitelů 27, CZ-783 71 Olomouc, Czech Republic.
authors.forename: Eva
authors.initials: E
authors.lastname: Hényková
authors.affiliation: Department of Neurology, University Hospital in Olomouc, I. P. Pavlova 6, CZ-775 20 Olomouc, Czech Republic.
authors.forename: Hana Přikrylová
authors.initials: HP
authors.lastname: Vránová
authors.affiliation: Laboratory of Growth Regulators, Centre of the Region Haná for Biotechnological and Agricultural Research, Faculty of Science of Palacký University & Institute of Experimental Botany CAS, Šlechtitelů 27, CZ-783 71 Olomouc, Czech Republic.
authors.forename: Petra
authors.initials: P
authors.lastname: Amakorová
authors.affiliation: Department of Chemical Biology and Genetics, Centre of the Region Haná for Biotechnological and Agricultural Research, Faculty of Science of Palacký University, Šlechtitelů 27, CZ-783 71 Olomouc, Czech Republic.
authors.forename: Tomáš
authors.initials: T
authors.lastname: Pospíšil
authors.affiliation: Laboratory of Growth Regulators, Centre of the Region Haná for Biotechnological and Agricultural Research, Faculty of Science of Palacký University & Institute of Experimental Botany CAS, Šlechtitelů 27, CZ-783 71 Olomouc, Czech Republic; Department of Biology, Faculty of Medicine and Dentistry, Palacký University, Hněvotínská 3, CZ-775 15 Olomouc, Czech Republic.
authors.forename: Asta
authors.initials: A
authors.lastname: Žukauskaitė
authors.affiliation: Laboratory of Growth Regulators, Centre of the Region Haná for Biotechnological and Agricultural Research, Faculty of Science of Palacký University & Institute of Experimental Botany CAS, Šlechtitelů 27, CZ-783 71 Olomouc, Czech Republic.
authors.forename: Magdaléna
authors.initials: M
authors.lastname: Vlčková
authors.affiliation: Laboratory of Growth Regulators, Centre of the Region Haná for Biotechnological and Agricultural Research, Faculty of Science of Palacký University & Institute of Experimental Botany CAS, Šlechtitelů 27, CZ-783 71 Olomouc, Czech Republic.
authors.forename: Lubor
authors.initials: L
authors.lastname: Urbánek
authors.affiliation: Laboratory of Growth Regulators, Centre of the Region Haná for Biotechnological and Agricultural Research, Faculty of Science of Palacký University & Institute of Experimental Botany CAS, Šlechtitelů 27, CZ-783 71 Olomouc, Czech Republic. Electronic address: novako@ueb.cas.cz.
authors.forename: Ondřej
authors.initials: O
authors.lastname: Novák
authors.affiliation: Department of Neurology, University Hospital in Olomouc, I. P. Pavlova 6, CZ-775 20 Olomouc, Czech Republic.
authors.forename: Jan
authors.initials: J
authors.lastname: Mareš
authors.affiliation: Department of Neurology, University Hospital in Olomouc, I. P. Pavlova 6, CZ-775 20 Olomouc, Czech Republic.
authors.forename: Petr
authors.initials: P
authors.lastname: Kaňovský
authors.affiliation: Laboratory of Growth Regulators, Centre of the Region Haná for Biotechnological and Agricultural Research, Faculty of Science of Palacký University & Institute of Experimental Botany CAS, Šlechtitelů 27, CZ-783 71 Olomouc, Czech Republic. Electronic address: miroslav.strnad@upol.cz.
authors.forename: Miroslav
authors.initials: M
authors.lastname: Strnad
mesh_terms: D001774:Blood Chemical Analysis / Q000379:methods*; D002623:Chemistry Techniques, Analytical / Q000379:methods*; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D007201:Indicator Dilution Techniques; D007554:Isotopes; D008297:Male; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry; D014364:Tryptophan / Q000097:blood* / Q000134:cerebrospinal fluid*
chemical_list: D007554:Isotopes; D014364:Tryptophan
keywords: Cerebrospinal fluid; Neurochemicals; Serum; Tryptophan metabolism; UHPLC–MS/MS