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A collaborative evaluation of LC-MS/MS based methods for BMAA analysis: soluble bound BMAA found to be an important fraction.

Faassen, Elisabeth J.; Antoniou, Maria G.; Beekman-Lukassen, Wendy; Blahova, Lucie; Chernova, Ekaterina; Christophoridis, Christophoros; Combes, Audrey; Edwards, Christine; Fastner, Jutta; Harmsen, Joop; Hiskia, Anastasia; Ilag, Leopold L.; Kaloudis, Triantafyllos; Lopicic, Srdjan; Lürling , Miquel; Mazur-Marzec, Hanna; Meriluoto, Jussi; Porojan, Cristina; Viner-Mozzini, Yehudit; Zguna, Nadezda


Elisabeth J. Faassen

Maria G. Antoniou

Wendy Beekman-Lukassen

Lucie Blahova

Ekaterina Chernova

Christophoros Christophoridis

Audrey Combes

Jutta Fastner

Joop Harmsen

Anastasia Hiskia

Leopold L. Ilag

Triantafyllos Kaloudis

Srdjan Lopicic

Miquel Lürling

Hanna Mazur-Marzec

Jussi Meriluoto

Cristina Porojan

Yehudit Viner-Mozzini

Nadezda Zguna


Exposure to β-Ν-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.


FAASSEN, E.J., ANTONIOU, M.G., BEEKMAN-LUKASSEN, W., BLAHOVA, L., CHERNOVA, E., CHRISTOPHORIDIS, C., COMBES, A., EDWARDS, C., FASTNER, J., HARMSEN, J., HISKIA, A., ILAG, L.L., KALOUDIS, T., LOPICIC, S., LURLING, M., MAZUR-MARZEC, H., MERILUOTO, J., POROJAN, C., VINER-MOZZINI, Y. and ZGUNA, N. 2016. A collaborative evaluation of LC-MS/MS based methods for BMAA analysis: soluble bound BMAA found to be an important fraction. Marine drugs [online], 14(3), article 45. Available from:

Journal Article Type Article
Acceptance Date Feb 6, 2016
Online Publication Date Feb 29, 2016
Publication Date Mar 31, 2016
Deposit Date May 23, 2016
Publicly Available Date May 23, 2016
Journal Marine drugs
Electronic ISSN 1660-3397
Publisher MDPI
Peer Reviewed Peer Reviewed
Volume 14
Issue 3
Article Number 45
Keywords β-Ν-methylamino-L-alanine; 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate; Alpha,gamma-diaminobutyric acid; Cycad; Daphnia magna; Hydrophilic interaction liquid chromatography; Liquid chromatography-tandem mass spectrometry; N-(2-aminoethyl)glycine; Ph
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