title: An improved microbore UHPLC method with electrochemical detection for the simultaneous determination of low monoamine levels in in vivo brain microdialysis samples.
abstract: The simultaneous determination of the monoamines dopamine (DA), noradrenaline (NA) and serotonin (5-HT) in in vivo microdialysis samples remains challenging because of the low extracellular neurotransmitter levels in different brain regions, specific sample characteristics, and the quest for high temporal resolution and a multi-target strategy in neuropharmacological research. A fast and sensitive microbore (1.0mm i.d. column) UHPLC method coupled to electrochemical detection (ECD) is developed by means of design of experiments with the emphasis on sufficient retention of NA within an acceptable total analysis time. Indeed, NA is the earliest eluting compound and often interferes with the broad solvent front originating from the sample matrix. The sensitive UHPLC-ECD assay (LLOQ of 100pM for NA and 150pM for DA and 5-HT) with an analysis time of 8min for standard solutions and 20min for in vivo microdialysis samples originating from rat hippocampus, prefrontal cortex and striatum, is validated applying accuracy profiles. The combination of in vivo microdialysis and microbore UHPLC-ECD has shown to be particularly suitable for future contributions to neuropharmacological research on the monoaminergic system.
authors.affiliation: Department of Pharmaceutical Chemistry and Drug Analysis, Center for Neurosciences (C4N), Vrije Universiteit Brussel (VUB), Laarbeeklaan 103, 1090 Brussels, Belgium. Electronic address: Jolien.Van.Schoors@vub.ac.be.
authors.forename: Jolien
authors.initials: J
authors.lastname: Van Schoors
authors.affiliation: Department of Analytical Chemistry and Pharmaceutical Technology, Vrije Universiteit Brussel (VUB), Laarbeeklaan 103, 1090 Brussels, Belgium. Electronic address: Johan.Viaene@vub.ac.be.
authors.forename: Johan
authors.initials: J
authors.lastname: Viaene
authors.affiliation: Department of Pharmaceutical Chemistry and Drug Analysis, Center for Neurosciences (C4N), Vrije Universiteit Brussel (VUB), Laarbeeklaan 103, 1090 Brussels, Belgium. Electronic address: Yannick.Van.Wanseele@vub.ac.be.
authors.forename: Yannick
authors.initials: Y
authors.lastname: Van Wanseele
authors.affiliation: Department of Pharmaceutical Chemistry and Drug Analysis, Center for Neurosciences (C4N), Vrije Universiteit Brussel (VUB), Laarbeeklaan 103, 1090 Brussels, Belgium. Electronic address: Ilse.Smolders@vub.ac.be.
authors.forename: Ilse
authors.initials: I
authors.lastname: Smolders
authors.affiliation: Department of Analytical Chemistry and Pharmaceutical Technology, Vrije Universiteit Brussel (VUB), Laarbeeklaan 103, 1090 Brussels, Belgium; Laboratory of Instrumental Analysis and Bioelectrochemistry, Université Libre de Bruxelles (ULB), Boulevard du Triomphe, 1050 Brussels, Belgium. Electronic address: Bieke.Dejaegher@vub.ac.be.
authors.forename: Bieke
authors.initials: B
authors.lastname: Dejaegher
authors.affiliation: Department of Analytical Chemistry and Pharmaceutical Technology, Vrije Universiteit Brussel (VUB), Laarbeeklaan 103, 1090 Brussels, Belgium. Electronic address: Yvan.Vander.Heyden@vub.ac.be.
authors.forename: Yvan
authors.initials: Y
authors.lastname: Vander Heyden
authors.affiliation: Department of Pharmaceutical Chemistry and Drug Analysis, Center for Neurosciences (C4N), Vrije Universiteit Brussel (VUB), Laarbeeklaan 103, 1090 Brussels, Belgium. Electronic address: aveeckha@vub.ac.be.
authors.forename: Ann
authors.initials: A
authors.lastname: Van Eeckhaut
mesh_terms: D000818:Animals; D015306:Biogenic Monoamines / Q000032:analysis* / Q000378:metabolism; D001921:Brain / Q000378:metabolism*; D002138:Calibration; D002853:Chromatography, Liquid / Q000295:instrumentation / Q000379:methods*; D055664:Electrochemical Techniques / Q000295:instrumentation / Q000379:methods*; D006624:Hippocampus / Q000378:metabolism; D057230:Limit of Detection; D008297:Male; D017551:Microdialysis / Q000295:instrumentation / Q000379:methods*; D017208:Rats, Wistar; D015203:Reproducibility of Results
chemical_list: D015306:Biogenic Monoamines
keywords: Accuracy profiles; Design of experiments; Electrochemical detection; In vivo microdialysis; Monoamine neurotransmitters; UHPLC