| Article ID | Journal | Published Year | Pages | File Type |
|---|---|---|---|---|
| 1965149 | Clinica Chimica Acta | 2016 | 8 Pages |
•Liquid chromatography–mass spectrometry used to monitor neurotransmitter and metabolite levels•Protein precipitation as sample preparation in rat brain matrix•Method validated according to the recommendations of regulatory agencies•Rapid quantification of neurotransmitters and metabolites to diagnose neurological diseases•Application of the method to the murine Nrf2 depression model
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 (R2 > 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.
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