title: Liquid chromatography-tandem mass spectrometry method for determination of panel of neurotransmitters in cerebrospinal fluid from the rat model for tauopathy.
abstract: Alzheimer's disease (AD) is still being recognized today as an unmet medical need. Currently, there is no cure and early preclinical diagnostic assay available for AD. Therefore much attention is now being directed at the development of novel methods for quantitative determination of AD biomarkers in the cerebrospinal fluid (CSF). Here, we describe the liquid chromatography-tandem mass spectrometry method for determination of 5-hydroxytryptamine (SER), 5-hydroxyindoleacetic acid (5-HIAA), homovanilic acid (HVA), noradrenaline (NADR), adrenaline (ADR), dopamine (DA), glutamic acid (Glu), γ-aminobutyric acid (GABA), 3,4-dihydroxyphenylacetic acid (DOPAC) and histamine (HIS) in cerebrospinal fluid (CSF) from the rat model for human tauopathy. The benzoyl chloride was used as pre-column derivatization reagents. Neurotransmitters and metabolites were analysed on ultra performance liquid chromatography (UPLC) on C18 column in combination with tandem mass spectrometry. The method is simple, highly sensitive and showed excellent linearity with regression coefficients higher than 0.99. The accuracy was in a range of 93-113% for all analytes. The inter-day precision (n=5 days), expressed as %RSD, was in a range 2-10% for all analytes. Using this method we detected significant changes of CSF levels of two important neurotransmitters/metabolites, ADR and 5-HIAA, which correlates with progression of neurodegeneration in our animal model.
authors.affiliation: Institute of Neuroimmunology, Slovak Academy of Sciences, Dubravska cesta 9, 84510 Bratislava, Slovak Republic; AXON Neuroscience SE, Dvorakovo nabrezie 10, 811 02 Bratislava, Slovak Republic.
authors.forename: Andrej
authors.initials: A
authors.lastname: Kovac
authors.affiliation: Institute of Neuroimmunology, Slovak Academy of Sciences, Dubravska cesta 9, 84510 Bratislava, Slovak Republic.
authors.forename: Zuzana
authors.initials: Z
authors.lastname: Somikova
authors.affiliation: Institute of Neuroimmunology, Slovak Academy of Sciences, Dubravska cesta 9, 84510 Bratislava, Slovak Republic; AXON Neuroscience SE, Dvorakovo nabrezie 10, 811 02 Bratislava, Slovak Republic.
authors.forename: Norbert
authors.initials: N
authors.lastname: Zilka
authors.affiliation: Institute of Neuroimmunology, Slovak Academy of Sciences, Dubravska cesta 9, 84510 Bratislava, Slovak Republic; AXON Neuroscience SE, Dvorakovo nabrezie 10, 811 02 Bratislava, Slovak Republic. Electronic address: michal.novak@savba.sk.
authors.forename: Michal
authors.initials: M
authors.lastname: Novak
mesh_terms: D000818:Animals; D002853:Chromatography, Liquid / Q000379:methods*; D004195:Disease Models, Animal; D057230:Limit of Detection; D018377:Neurotransmitter Agents / Q000134:cerebrospinal fluid*; D051381:Rats; D055647:Rats, Transgenic; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods*; D024801:Tauopathies / Q000134:cerebrospinal fluid*
chemical_list: D018377:Neurotransmitter Agents
keywords: 3,4-dihydroxyphenylacetic acid; 5-HIAA; 5-hydroxyindoleacetic acid; 5-hydroxytryptamine; AD; ADR; Alzheimer's disease; CSF; Cerebrospinal fluid; DA; DOPAC; GABA; Glu; HIS; HPLC; HVA; IS; LOD; LOQ; NADR; NT; Neurotransmitters; Rat model; SER; Tau protein; UPLC; UPLC/MS; adrenaline; cerebrospinal fluid; dopamine; glutamic acid; high-performance liquid chromatography; histamine; homovanilic acid; internal standard; limit of detection; limit of quantification; neurotransmitters; noradrenaline; ultra-performance liquid chromatography; γ-aminobutyric acid