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.

Alterra P O Box 47 Wageningen 6700 DD The Netherlands

Aquatic Ecology and Water Quality Management Group Wageningen University P O Box 47 Wageningen 6700 DD The Netherlands

Biochemistry Faculty of Science and Engineering Åbo Akademi University Tykistökatu 6A 3rd Floor Turku 20520 Finland

Department of Analytical Bioanalytical Sciences and Miniaturization UMR CBI 8231 ESPCI ParisTech CNRS PSL Research University ESPCI ParisTech 75005 Paris France

Department of Environmental Science and Analytical Chemistry Stockholm University SE 10691 Stockholm Sweden

Department of Environmental Science and Technology Cyprus University of Technology 3036 Lemesos Cyprus

Department of Marine Biotechnology University of Gdansk Al Marszalka Pilsudskiego 46 Gdynia 81 378 Poland

Drinking Water Resources and Water Treatment Federal Environment Agency Schichauweg 58 12307 Berlin Germany

Faculty of Science RECETOX Masaryk University Kamenice 5 625 00 Brno Czech Republic

Institute for Pathological Physiology School of Medicine University of Belgrade 11000 Belgrade Serbia

Kinneret Limnological Laboratory Israel Oceanographic and Limnological Research P O Box 447 Migdal 14950 Israel

Laboratory of Catalytic Photocatalytic Processes and Environmental Analysis Institute of Nanoscience and Nanotechnology National Center for Scientific Research Demokritos Patriarchou Grigoriou and Neapoleos 15310 Agia Paraskevi Athens Greece

Mass Spectrometry Research Centre and PROTEOBIO Research Groups Department of Physical Sciences Cork Institute of Technology Rossa Avenue Bishopstown V92 F9WY Co Cork Ireland

NIOO KNAW Droevendaalsesteeg 10 Wageningen 6708 PB The Netherlands

Pharmacy and Life Sciences Robert Gordon University Aberdeen AB10 7GJ UK

Saint Petersburg Scientific Research Centre for Ecological Safety Russian Academy of Sciences 18 Korpusnaya street St Petersburg 197110 Russia

Water Quality Department Division of Quality Research and Development 156 Oropou str 11146 Athens Greece

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