title: Beta-N-methylamino-L-alanine: LC-MS/MS optimization, screening of cyanobacterial strains and occurrence in shellfish from Thau, a French Mediterranean lagoon.
abstract: β-N-methylamino-L-alanine (BMAA) is a neurotoxic non-protein amino acid suggested to be involved in neurodegenerative diseases. It was reported to be produced by cyanobacteria, but also found in edible aquatic organisms, thus raising concern of a widespread human exposure. However, the chemical analysis of BMAA and its isomers are controversial, mainly due to the lack of selectivity of the analytical methods. Using factorial design, we have optimized the chromatographic separation of underivatized analogues by a hydrophilic interaction chromatography coupled to tandem mass spectrometry (HILIC-MS/MS) method. A combination of an effective solid phase extraction (SPE) clean-up, appropriate chromatographic resolution and the use of specific mass spectral transitions allowed for the development of a highly selective and sensitive analytical procedure to identify and quantify BMAA and its isomers (in both free and total form) in cyanobacteria and mollusk matrices (LOQ of 0.225 and 0.15 µg/g dry weight, respectively). Ten species of cyanobacteria (six are reported to be BMAA producers) were screened with this method, and neither free nor bound BMAA could be found, while both free and bound DAB were present in almost all samples. Mussels and oysters collected in 2009 in the Thau Lagoon, France, were also screened, and bound BMAA and its two isomers, DAB and AEG, were observed in all samples (from 0.6 to 14.4 µg/g DW), while only several samples contained quantifiable free BMAA.
authors.affiliation: Ifremer (French Research Institute for the Exploitation of the Seas), Phycotoxins Laboratory, rue de l'Ile d'Yeu, BP 21105, F-44311 Nantes, France. Damien.Reveillon@ifremer.fr.
authors.forename: Damien
authors.initials: D
authors.lastname: Réveillon
authors.affiliation: Ifremer (French Research Institute for the Exploitation of the Seas), Laboratoire Environnement Ressources du Languedoc Roussillon (LER-LR) F-34203 Sète, France. Eric.Abadie@ifremer.fr.
authors.forename: Eric
authors.initials: E
authors.lastname: Abadie
authors.affiliation: Ifremer (French Research Institute for the Exploitation of the Seas), Phycotoxins Laboratory, rue de l'Ile d'Yeu, BP 21105, F-44311 Nantes, France. Veronique.Sechet@ifremer.fr.
authors.forename: Véronique
authors.initials: V
authors.lastname: Séchet
authors.affiliation: UMR Eco-Bio-Université de Rennes I, F-35042 Rennes, France. Luc.Brient@univ-rennes1.fr.
authors.forename: Luc
authors.initials: L
authors.lastname: Brient
authors.affiliation: Ifremer (French Research Institute for the Exploitation of the Seas), Phycotoxins Laboratory, rue de l'Ile d'Yeu, BP 21105, F-44311 Nantes, France. Veronique.Sechet@ifremer.fr.
authors.forename: Véronique
authors.initials: V
authors.lastname: Savar
authors.affiliation: Ifremer (French Research Institute for the Exploitation of the Seas), Phycotoxins Laboratory, rue de l'Ile d'Yeu, BP 21105, F-44311 Nantes, France. Michele.Bardouil@ifremer.fr.
authors.forename: Michèle
authors.initials: M
authors.lastname: Bardouil
authors.affiliation: Ifremer (French Research Institute for the Exploitation of the Seas), Phycotoxins Laboratory, rue de l'Ile d'Yeu, BP 21105, F-44311 Nantes, France. Philipp.Hess@ifremer.fr.
authors.forename: Philipp
authors.initials: P
authors.lastname: Hess
authors.affiliation: Ifremer (French Research Institute for the Exploitation of the Seas), Phycotoxins Laboratory, rue de l'Ile d'Yeu, BP 21105, F-44311 Nantes, France. Zouher.Amzil@ifremer.fr.
authors.forename: Zouher
authors.initials: Z
authors.lastname: Amzil
mesh_terms: D000599:Amino Acids, Diamino / Q000737:chemistry* / Q000302:isolation & purification; D000818:Animals; D002853:Chromatography, Liquid / Q000379:methods*; D000458:Cyanobacteria / Q000378:metabolism*; D000087522:Cyanobacteria Toxins; D005602:France; D008522:Mediterranean Sea; D008974:Mollusca / Q000378:metabolism; D012680:Sensitivity and Specificity; D052616:Solid Phase Extraction / Q000379:methods; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D000599:Amino Acids, Diamino; D000087522:Cyanobacteria Toxins; C001824:beta-N-methylamino-L-alanine