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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
- Source :
- Clinica Chimica Acta, 453, 174-181. Elsevier
- Publication Year :
- 2016
-
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), gamma-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 1-50 ng/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.
- Subjects :
- Male
0301 basic medicine
Time Factors
NF-E2-Related Factor 2
Formic acid
Metabolite
Electrospray ionization
Clinical Biochemistry
Rat brain
Mass spectrometry
Hippocampus
01 natural sciences
Biochemistry
High-performance liquid chromatography
Gene Knockout Techniques
Mice
03 medical and health sciences
chemistry.chemical_compound
Tandem Mass Spectrometry
Liquid chromatography–mass spectrometry
Metabolites
Animals
Protein precipitation
Neurotransmitter Agents
Chromatography
Depression
Chemistry
010401 analytical chemistry
Biochemistry (medical)
General Medicine
Glutamic acid
Neurotransmitters
Rats
0104 chemical sciences
Disease Models, Animal
030104 developmental biology
Liquid chromatography-tandem mass spectrometry
Calibration
Linear Models
Blood Chemical Analysis
Chromatography, Liquid
Subjects
Details
- Language :
- English
- ISSN :
- 00098981
- Database :
- OpenAIRE
- Journal :
- Clinica Chimica Acta, 453, 174-181. Elsevier
- Accession number :
- edsair.doi.dedup.....7b13d2672c31b8a991c6fd4d6a7c0cbd