title: Analytical approach to determine biogenic amines in urine using microextraction in packed syringe and liquid chromatography coupled to electrochemical detection.
abstract: The goal of this work was to develop and validate an analytical method for the detection and quantification of the biogenic amines serotonin (5-HT), dopamine (DA) and norepinephrine (NE), using microextraction in packed syringe (MEPS) and liquid chromatography coupled to electrochemical detection (HPLC-ED) in urine. The method was validated according to internationally accepted guidelines from the Food and Drug Administration. Linearity was established between 50 and 1000 ng/mL for 5-HT and between 5 and 1000 ng/mL for DA and NE, with determination coefficients (R(2)) >0.99 for all compounds. The limits of quantification and detection were respectively 50 and 20 ng/mL for 5-HT, and 5 and 2 ng/mL for DA and NE. Within- and between-run precision ranged from 0.84 to 9.41%, while accuracy ranged from 0.79 to 12.76% for all compounds. The intermediate precision and accuracy were 1.50-8.36 and 0.54-13.51%, respectively. The method was found suitable for clinical routine analysis of the studied compounds, using a sample volume of 0.5 mL. This is the first study employing a commercially available MEPS column for the simultaneous detection and quantification of 5-HT, DA and NE in urine by coulometric detection.
authors.affiliation: CICS-UBI, Centro de Investigação em Ciências da Saúde, Universidade da Beira Interior, Av. Infante D. Henrique, 6201-556 Covilhã, Portugal.
authors.forename: David
authors.initials: D
authors.lastname: Oppolzer
authors.forename: Ivo
authors.initials: I
authors.lastname: Moreno
authors.forename: Beatriz
authors.initials: B
authors.lastname: da Fonseca
authors.forename: Luís
authors.initials: L
authors.lastname: Passarinha
authors.forename: Mário
authors.initials: M
authors.lastname: Barroso
authors.forename: Suzel
authors.initials: S
authors.lastname: Costa
authors.forename: João António
authors.initials: JA
authors.lastname: Queiroz
authors.forename: Eugenia
authors.initials: E
authors.lastname: Gallardo
mesh_terms: D001679:Biogenic Amines / Q000302:isolation & purification / Q000652:urine*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D055664:Electrochemical Techniques / Q000295:instrumentation / Q000379:methods; D006801:Humans; D016014:Linear Models; D059627:Liquid Phase Microextraction / Q000295:instrumentation / Q000379:methods*; D000432:Methanol / Q000737:chemistry; D015203:Reproducibility of Results; D012107:Research Design; D012680:Sensitivity and Specificity
chemical_list: D001679:Biogenic Amines; D000432:Methanol