title: Simultaneous determination of 8 neurotransmitters and their metabolite levels in rat brain using liquid chromatography in tandem with mass spectrometry: Application to the murine Nrf2 model of depression.
abstract: Analysis of neurotransmitters and their metabolites is useful for the diagnosis of central nervous system diseases. A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method with protein precipitation was developed to monitor levels of adrenaline (AD), noradrenaline (NA), glutamic acid (Glu), γ-aminobutyric acid (GABA), dopamine (DA), 5-hydroxytryptamine (5-HT), 5-hydroxyindole acetic acid (5-HIAA), and 3-methoxy-4-hydroxyphenylglycol (MHPG) in rat brain tissue. Isoprenaline was used as an internal standard (IS). Neurotransmitters and metabolites were eluted with a reverse phase column under gradient conditions through a mobile phase consisting of 0.2% formic acid water solution/acetonitrile. The compounds were detected and quantified by LC-MS/MS with positive or negative electrospray ionization, which operates in multiple-reaction monitoring mode. The method was linear or polynomial (R(2)>0.99) for AD, NA, Glu, GABA, DA, 5-HT, 5-HIAA, and MHPG in the range of 0.25-200, 0.5-200, 250-20,000, 250-20,000, 0.25-200, 10-3000, 1-50, and 1-50ng/mL, respectively. The validation assays for accuracy and precision, matrix effect, extraction recovery, stability and carry-over of the samples for neurotransmitters and metabolites were consistent with the requirements of regulatory agencies. The method enables rapid quantification of neurotransmitters and their metabolites and has been applied in the nuclear factor (erythroid 2-derived)-like 2 (Nrf2) knockout mouse model of depression.
authors.affiliation: Instituto-Fundación Teófilo Hernando, Universidad Autónoma de Madrid, Madrid, Spain; Departamento de Farmacología y Terapéutica, Facultad de Medicina, Universidad Autónoma de Madrid, Madrid, Spain; Servicio de Farmacología Clínica, Instituto de Investigación Sanitaria, Hospital Universitario de la Princesa, Universidad Autónoma de Madrid, Madrid, Spain.
authors.forename: Aneta
authors.initials: A
authors.lastname: Wojnicz
authors.affiliation: Departamento de Farmacología y Terapéutica, Facultad de Medicina, Universidad Autónoma de Madrid, Madrid, Spain.
authors.forename: José
authors.initials: J
authors.lastname: Avendaño Ortiz
authors.affiliation: Departamento de Farmacología y Terapéutica, Facultad de Medicina, Universidad Autónoma de Madrid, Madrid, Spain; Department of Pharmacology, CARIM, and Maastricht Institute for Advanced Studies, Maastricht University, The Netherlands.
authors.forename: Ana I
authors.initials: AI
authors.lastname: Casas
authors.affiliation: Departamento de Farmacología y Terapéutica, Facultad de Medicina, Universidad Autónoma de Madrid, Madrid, Spain; Center of Biological Sciences, Universidade Federal de Santa Catarina, Campus Universitário, Trindade, Florianópolis, Brazil.
authors.forename: Andiara E
authors.initials: AE
authors.lastname: Freitas
authors.affiliation: Instituto-Fundación Teófilo Hernando, Universidad Autónoma de Madrid, Madrid, Spain; Departamento de Farmacología y Terapéutica, Facultad de Medicina, Universidad Autónoma de Madrid, Madrid, Spain.
authors.forename: Manuela
authors.initials: M
authors.lastname: G López
authors.affiliation: Instituto-Fundación Teófilo Hernando, Universidad Autónoma de Madrid, Madrid, Spain; Servicio de Farmacología Clínica, Instituto de Investigación Sanitaria, Hospital Universitario de la Princesa, Universidad Autónoma de Madrid, Madrid, Spain. Electronic address: anaruiznuno@yahoo.es.
authors.forename: Ana
authors.initials: A
authors.lastname: Ruiz-Nuño
mesh_terms: D000818:Animals; D001774:Blood Chemical Analysis / Q000379:methods*; D002138:Calibration; D002853:Chromatography, Liquid; D003863:Depression / Q000097:blood* / Q000235:genetics; D004195:Disease Models, Animal; D055786:Gene Knockout Techniques; D006624:Hippocampus / Q000378:metabolism; D016014:Linear Models; D008297:Male; D051379:Mice; D051267:NF-E2-Related Factor 2 / Q000172:deficiency* / Q000235:genetics*; D018377:Neurotransmitter Agents / Q000097:blood* / Q000378:metabolism*; D051381:Rats; D053719:Tandem Mass Spectrometry; D013997:Time Factors
chemical_list: D051267:NF-E2-Related Factor 2; D018377:Neurotransmitter Agents; C495636:Nfe2l2 protein, mouse
keywords: Liquid chromatography–tandem mass spectrometry; Metabolites; Neurotransmitters; Protein precipitation; Rat brain