title: A Collaborative Evaluation of LC-MS/MS Based Methods for BMAA Analysis: Soluble Bound BMAA Found to Be an Important Fraction.
abstract: Exposure to β-N-methylamino-l-alanine (BMAA) might be linked to the incidence of amyotrophic lateral sclerosis, Alzheimer's disease and Parkinson's disease. Analytical chemistry plays a crucial role in determining human BMAA exposure and the associated health risk, but the performance of various analytical methods currently employed is rarely compared. A CYANOCOST initiated workshop was organized aimed at training scientists in BMAA analysis, creating mutual understanding and paving the way towards interlaboratory comparison exercises. During this workshop, we tested different methods (extraction followed by derivatization and liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) analysis, or directly followed by LC-MS/MS analysis) for trueness and intermediate precision. We adapted three workup methods for the underivatized analysis of animal, brain and cyanobacterial samples. Based on recovery of the internal standard D₃BMAA, the underivatized methods were accurate (mean recovery 80%) and precise (mean relative standard deviation 10%), except for the cyanobacterium Leptolyngbya. However, total BMAA concentrations in the positive controls (cycad seeds) showed higher variation (relative standard deviation 21%-32%), implying that D₃BMAA was not a good indicator for the release of BMAA from bound forms. Significant losses occurred during workup for the derivatized method, resulting in low recovery (<10%). Most BMAA was found in a trichloroacetic acid soluble, bound form and we recommend including this fraction during analysis.
authors.affiliation: Aquatic Ecology &amp; Water Quality Management Group, Wageningen University, P.O. Box 47, Wageningen 6700 DD, The Netherlands. els.faassen@wur.nl.
authors.forename: Elisabeth J
authors.initials: EJ
authors.lastname: Faassen
authors.affiliation: Department of Environmental Science and Technology, Cyprus University of Technology, 3036 Lemesos, Cyprus. maria.antoniou@cut.ac.cy.
authors.forename: Maria G
authors.initials: MG
authors.lastname: Antoniou
authors.affiliation: Aquatic Ecology &amp; Water Quality Management Group, Wageningen University, P.O. Box 47, Wageningen 6700 DD, The Netherlands. wendy.beekman-lukassen@wur.nl.
authors.forename: Wendy
authors.initials: W
authors.lastname: Beekman-Lukassen
authors.affiliation: Faculty of Science, RECETOX, Masaryk University, Kamenice 5, 625 00 Brno, Czech Republic. blahova@recetox.muni.cz.
authors.forename: Lucie
authors.initials: L
authors.lastname: Blahova
authors.affiliation: Saint-Petersburg Scientific-Research Centre for Ecological Safety, Russian Academy of Sciences, 18, Korpusnaya street, St. Petersburg 197110, Russia. s3561389@ya.ru.
authors.forename: Ekaterina
authors.initials: E
authors.lastname: Chernova
authors.affiliation: Laboratory of Catalytic-Photocatalytic Processes and Environmental Analysis, Institute of Nanoscience &amp; Nanotechnology, National Center for Scientific Research "Demokritos", Patriarchou Grigoriou &amp; Neapoleos, 15310 Agia Paraskevi, Athens, Greece. c.christoforidis@inn.demokritos.gr.
authors.forename: Christophoros
authors.initials: C
authors.lastname: Christophoridis
authors.affiliation: Department of Analytical, Bioanalytical Sciences and Miniaturization (LSABM), UMR CBI 8231 ESPCI ParisTech/CNRS, PSL Research University, ESPCI ParisTech, 75005 Paris, France. audrey.combes@espci.fr.
authors.forename: Audrey
authors.initials: A
authors.lastname: Combes
authors.affiliation: Pharmacy &amp; Life Sciences, Robert Gordon University, Aberdeen AB10 7GJ, UK. c.edwards@rgu.ac.uk.
authors.forename: Christine
authors.initials: C
authors.lastname: Edwards
authors.affiliation: Drinking-Water Resources and Water Treatment, Federal Environment Agency, Schichauweg 58, 12307 Berlin, Germany. jutta.fastner@uba.de.
authors.forename: Jutta
authors.initials: J
authors.lastname: Fastner
authors.affiliation: Alterra, P.O. Box 47, Wageningen 6700 DD, The Netherlands. joop.harmsen@wur.nl.
authors.forename: Joop
authors.initials: J
authors.lastname: Harmsen
authors.affiliation: Laboratory of Catalytic-Photocatalytic Processes and Environmental Analysis, Institute of Nanoscience &amp; Nanotechnology, National Center for Scientific Research "Demokritos", Patriarchou Grigoriou &amp; Neapoleos, 15310 Agia Paraskevi, Athens, Greece. a.hiskia@inn.demokritos.gr.
authors.forename: Anastasia
authors.initials: A
authors.lastname: Hiskia
authors.affiliation: Department of Environmental Science and Analytical Chemistry, Stockholm University, SE-10691 Stockholm, Sweden. leopold.ilag@aces.su.se.
authors.forename: Leopold L
authors.initials: LL
authors.lastname: Ilag
authors.affiliation: Water Quality Department, Division of Quality, Research and Development (R&amp;D), Athens Water Supply and Sewerage Company (EYDAP SA), 156 Oropou str., 11146 Athens, Greece. kaloudis@eydap.gr.
authors.forename: Triantafyllos
authors.initials: T
authors.lastname: Kaloudis
authors.affiliation: Institute for Pathological Physiology, School of Medicine, University of Belgrade, 11000 Belgrade, Serbia. slopicic@med.bg.ac.rs.
authors.forename: Srdjan
authors.initials: S
authors.lastname: Lopicic
authors.affiliation: Aquatic Ecology &amp; Water Quality Management Group, Wageningen University, P.O. Box 47, Wageningen 6700 DD, The Netherlands. miquel.lurling@wur.nl.|NIOO-KNAW, Droevendaalsesteeg 10, Wageningen 6708 PB, The Netherlands. miquel.lurling@wur.nl.
authors.forename: Miquel
authors.initials: M
authors.lastname: Lürling
authors.affiliation: Department of Marine Biotechnology, University of Gdansk, Al. Marszalka Pilsudskiego 46, Gdynia 81-378, Poland. biohm@ug.edu.pl.
authors.forename: Hanna
authors.initials: H
authors.lastname: Mazur-Marzec
authors.affiliation: Biochemistry, Faculty of Science and Engineering, Åbo Akademi University, Tykistökatu 6A 3rd Floor, Turku 20520, Finland. jussi.meriluoto@abo.fi.
authors.forename: Jussi
authors.initials: J
authors.lastname: Meriluoto
authors.affiliation: Mass Spectrometry Research Centre (MSRC) and PROTEOBIO Research Groups, Department of Physical Sciences, Cork Institute of Technology, Rossa Avenue, Bishopstown, V92 F9WY, Co. Cork, Ireland. cristina.porojan@mycit.ie.
authors.forename: Cristina
authors.initials: C
authors.lastname: Porojan
authors.affiliation: Kinneret Limnological Laboratory, Israel Oceanographic &amp; Limnological Research, P.O. Box 447, Migdal 14950, Israel. diti@ocean.org.il.
authors.forename: Yehudit
authors.initials: Y
authors.lastname: Viner-Mozzini
authors.affiliation: Department of Environmental Science and Analytical Chemistry, Stockholm University, SE-10691 Stockholm, Sweden. nadezda.kiselova@aces.su.se.
authors.forename: Nadezda
authors.initials: N
authors.lastname: Zguna
mesh_terms: D000599:Amino Acids, Diamino / Q000032:analysis* / Q000378:metabolism; D000818:Animals; D001921:Brain / Q000378:metabolism; D002853:Chromatography, Liquid / Q000379:methods*; D000458:Cyanobacteria / Q000378:metabolism; D000087522:Cyanobacteria Toxins; D003621:Daphnia; D009498:Neurotoxins / Q000032:analysis* / Q000378:metabolism; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods*; D014238:Trichloroacetic Acid / Q000737:chemistry
chemical_list: D000599:Amino Acids, Diamino; D000087522:Cyanobacteria Toxins; D009498:Neurotoxins; C001824:beta-N-methylamino-L-alanine; D014238:Trichloroacetic Acid
keywords: ">l-alanine (BMAA); 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC); Daphnia magna; Internal standard; Liquid chromatography-tandem mass spectrometry (LC-MS/MS); N-(2-aminoethyl) glycine (AEG); cycad; hydrophilic interaction liquid chromatography (HILIC); phytoplankton; seafood; α,γ-diaminobutyric acid (DAB); β-N-methylamino-