title: Determination of Serotonin and Dopamine Metabolites in Human Brain Microdialysis and Cerebrospinal Fluid Samples by UPLC-MS/MS: Discovery of Intact Glucuronide and Sulfate Conjugates.
abstract: An UPLC-MS/MS method was developed for the determination of serotonin (5-HT), dopamine (DA), their phase I metabolites 5-HIAA, DOPAC and HVA, and their sulfate and glucuronide conjugates in human brain microdialysis samples obtained from two patients with acute brain injuries, ventricular cerebrospinal fluid (CSF) samples obtained from four patients with obstructive hydrocephalus, and a lumbar CSF sample pooled mainly from patients undergoing spinal anesthesia in preparation for orthopedic surgery. The method was validated by determining the limits of detection and quantification, linearity, repeatability and specificity. The direct method enabled the analysis of the intact phase II metabolites of 5-HT and DA, without hydrolysis of the conjugates. The method also enabled the analysis of the regioisomers of the conjugates, and several intact glucuronide and sulfate conjugates were identified and quantified for the first time in the human brain microdialysis and CSF samples. We were able to show the presence of 5-HIAA sulfate, and that dopamine-3-O-sulfate predominates over dopamine-4-O-sulfate in the human brain. The quantitative results suggest that sulfonation is a more important phase II metabolism pathway than glucuronidation in the human brain.
authors.affiliation: Division of Pharmaceutical Chemistry, Faculty of Pharmacy, University of Helsinki, Helsinki, Finland.
authors.forename: Tina
authors.initials: T
authors.lastname: Suominen
authors.forename: Päivi
authors.initials: P
authors.lastname: Uutela
authors.forename: Raimo A
authors.initials: RA
authors.lastname: Ketola
authors.forename: Jonas
authors.initials: J
authors.lastname: Bergquist
authors.forename: Lars
authors.initials: L
authors.lastname: Hillered
authors.forename: Moshe
authors.initials: M
authors.lastname: Finel
authors.forename: Hongbo
authors.initials: H
authors.lastname: Zhang
authors.forename: Aki
authors.initials: A
authors.lastname: Laakso
authors.forename: Risto
authors.initials: R
authors.lastname: Kostiainen
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D015415:Biomarkers / Q000134:cerebrospinal fluid; D001927:Brain Diseases / Q000378:metabolism; D002853:Chromatography, Liquid / Q000379:methods*; D004298:Dopamine / Q000378:metabolism*; D005260:Female; D020719:Glucuronides / Q000378:metabolism; D006801:Humans; D008297:Male; D017551:Microdialysis; D008875:Middle Aged; D000072161:Orthopedic Surgeons; D015203:Reproducibility of Results; D012701:Serotonin / Q000378:metabolism*; D013129:Spinal Puncture; D013431:Sulfates / Q000378:metabolism; D053719:Tandem Mass Spectrometry / Q000379:methods*; D055815:Young Adult
chemical_list: D015415:Biomarkers; D020719:Glucuronides; D013431:Sulfates; D012701:Serotonin; D004298:Dopamine