title: Microdialysate analysis of monoamine neurotransmitters--a versatile and sensitive LC-MS/MS method.
abstract: We have developed and validated a sensitive method for the simultaneous determination of some monoamine neurotransmitters like dopamine (DA), norepinephrine (NE) and serotonin (5-HT) in rat brain microdialysate using high-performance liquid chromatography coupled to tandem mass spectrometry (LC/MS/MS). Sensitivity enhancement has been achieved by amine derivatization with the reagent (5-N-succinimidoxy-5-oxopentyl)triphenylphosphonium bromide (SPTPP) under mild conditions. The use of the selected reaction monitoring (SRM) mode has allowed detection of the analytes at a concentration of 30 pM (lower limit of quantification, LLOQ, signal-to-noise ratio higher than 5) with an accuracy of ≤3.80% and a precision of ±7.39 (%CV) for all neurotransmitters. Derivatization improves resolution and chromatographic retention times (3 min) by lipophilization. Linearity has been good (R>0.99) over a large concentration range (30-50,000 pM). The intra and inter-batch accuracy and precision were not greater than 4.8% and 6.4%, respectively. Therefore, the method was successfully applied for monitoring the concentration changes of neurotransmitters in microdialysis samples deriving from striatum rat brain region after amphetamine administration (3 mg kg(-1), i.p.).
authors.affiliation: Merz Pharmaceuticals GmbH, Department of Biological Analytics, Frankfurt am Main, Germany. sergio.greco@merz.de
authors.forename: S
authors.initials: S
authors.lastname: Greco
authors.forename: W
authors.initials: W
authors.lastname: Danysz
authors.forename: A
authors.initials: A
authors.lastname: Zivkovic
authors.forename: R
authors.initials: R
authors.lastname: Gross
authors.forename: H
authors.initials: H
authors.lastname: Stark
mesh_terms: D000818:Animals; D015306:Biogenic Monoamines / Q000032:analysis* / Q000737:chemistry; D001921:Brain / Q000166:cytology; D002853:Chromatography, Liquid / Q000379:methods*; D008297:Male; D017551:Microdialysis / Q000379:methods*; D018377:Neurotransmitter Agents / Q000032:analysis* / Q000737:chemistry; D009943:Organophosphorus Compounds / Q000737:chemistry; D051381:Rats; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results; D013388:Succinimides / Q000737:chemistry; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: C550832:(5-N-succinimidoxy-5-oxopentyl)triphenylphosphonium; D015306:Biogenic Monoamines; D018377:Neurotransmitter Agents; D009943:Organophosphorus Compounds; D013388:Succinimides