title: Determination of monoamine neurotransmitters in zebrafish (Danio rerio) by gas chromatography coupled to mass spectrometry with a two-step derivatization.
abstract: A sensitive analytical method for the determination of monoamine neurotransmitters (MNTs) in zebrafish larvae was developed using gas chromatography coupled to mass spectrometry. Six MNTs were selected as target compounds for neurotoxicity testing. MNTs underwent a two-step derivatization with hexamethyldisilazane (HDMS) for O-silylation followed by N-methyl-bis-heptafluorobutyramide (MBHFBA) for N-perfluoroacylation. Derivatization conditions were optimized by an experimental design approach. Method validation showed linear calibration curves (r2> 0.9976) in the range of 1-100 ng for all the compounds. The recovery rates were between 92 and 119%. The method was repeatable and reproducible with relative standard deviations (RSD) in the range of 2.5-9.3% for intra-day and 4.8-12% for inter-day variation. The limits of detection and the limits of quantitation were 0.4-0.8 and 1.2-2.7 ng/mL, respectively. The method was successfully applied to detect and quantify trace levels of MNTs in 5-day-old zebrafish larvae that were exposed to low concentrations of neurotoxic chemicals such as pesticides and methylmercury. Although visual malformations were not detected, the MNT levels varied significantly during early zebrafish development. These results show that exposure to neurotoxic chemicals can alter neurotransmitter levels and thereby may influence early brain development. Graphical abstract ᅟ.
authors.affiliation: Institute for Environmental Studies, VU University Amsterdam, De Boelelaan 1087, 1081 HV, Amsterdam, The Netherlands. alvaroarse@gmail.com.|Botanical Institute, Botanic Garden, University of Castilla-La Mancha, University Campus, 02071, Albacete, Spain. alvaroarse@gmail.com.
authors.forename: Alvaro
authors.initials: A
authors.lastname: Aragon
authors.affiliation: Institute for Environmental Studies, VU University Amsterdam, De Boelelaan 1087, 1081 HV, Amsterdam, The Netherlands.
authors.forename: Jessica
authors.initials: J
authors.lastname: Legradi
authors.affiliation: Institute for Environmental Studies, VU University Amsterdam, De Boelelaan 1087, 1081 HV, Amsterdam, The Netherlands.|Department of Analytical Chemistry, Institute of Fine Chemistry and Nanochemistry, University of Cordoba, Marie Curie Building (Annex), Campus of Rabanales, Cordoba, 14071, Spain.
authors.forename: Ana
authors.initials: A
authors.lastname: Ballesteros-Gómez
authors.affiliation: Institute for Environmental Studies, VU University Amsterdam, De Boelelaan 1087, 1081 HV, Amsterdam, The Netherlands.|Institute for Environment, Health and Societies, Brunel University London, Uxbridge, Middlesex, UB8 3PH, UK.
authors.forename: Juliette
authors.initials: J
authors.lastname: Legler
authors.affiliation: Institute for Environmental Studies, VU University Amsterdam, De Boelelaan 1087, 1081 HV, Amsterdam, The Netherlands.
authors.forename: Martin
authors.initials: M
authors.lastname: van Velzen
authors.affiliation: Institute for Environmental Studies, VU University Amsterdam, De Boelelaan 1087, 1081 HV, Amsterdam, The Netherlands.
authors.forename: Jacob
authors.initials: J
authors.lastname: de Boer
authors.affiliation: Institute for Environmental Studies, VU University Amsterdam, De Boelelaan 1087, 1081 HV, Amsterdam, The Netherlands.
authors.forename: Pim
authors.initials: P
authors.lastname: Leonards
mesh_terms: D000818:Animals; D015306:Biogenic Monoamines / Q000378:metabolism*; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods*; D007814:Larva / Q000187:drug effects* / Q000378:metabolism*; D009498:Neurotoxins / Q000008:administration & dosage*; D018377:Neurotransmitter Agents / Q000378:metabolism*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D015027:Zebrafish / Q000378:metabolism*
chemical_list: D015306:Biogenic Monoamines; D009498:Neurotoxins; D018377:Neurotransmitter Agents
keywords: Derivatization; Design of experiments; GC-MS; Monoamine neurotransmitters; Neurotoxic chemical exposure; Zebrafish