title: A new method for analysis of underivatized free β-methylamino-alanine: Validation and method comparison.
abstract: A new method was developed for analysis of free β-Methylamino-alanine (BMAA) in biological matrices. The method is based on direct analysis of the underivatized molecule, using an amide column for separation by Hydrophilic Interaction Liquid Chromatography (HILIC) and detection by tandem mass spectrometry (MS/MS) using a deuterium labeled internal standard. The use of Ultra-High Performance Liquid Chromatography (UHPLC) combined with MS/MS detection allowed for high chromatographic resolution and a low limit of detection (0.025 μg/g wet weight (ww) in mussels). The method was validated by analyzing spiked blank mussels from the Baltic Sea (0.15-4.4 μg/g (ww), trueness 99%-105%, RSD 2%-8%). An inter-laboratory comparative analysis of extracts of mussel was performed. The mussels were extracted according to an established protocol for analysis of free BMAA, and the extracts were then analyzed in parallel by the new method and a validated procedure based on detection of BMAA derivatized with dansyl chloride. Both methods detected BMAA in similar concentrations. Thus, derivatization with dansyl chloride did not influence the results compared to direct detection. The new method presents an alternative to the commonly applied derivatization step, and is proved through validation and method comparison to reliably identify and quantify free BMAA at low concentration levels.
authors: National Food Agency, Box 622, SE-751 26, Uppsala, Sweden. Electronic address: johan.rosen@slv.se.
authors: Johan
authors: J
authors: Rosén
authors: National Food Agency, Box 622, SE-751 26, Uppsala, Sweden.
authors: Erik
authors: E
authors: Westerberg
authors: National Food Agency, Box 622, SE-751 26, Uppsala, Sweden.
authors: Karl-Erik
authors: KE
authors: Hellenäs
authors: Section of Chemical Analysis, Department of Chemistry, Environment and Feed Hygiene, National Veterinary Institute (SVA), SE-751 89 Uppsala, Sweden; Division of Analytical Pharmaceutical Chemistry, Uppsala University, BMC, SE-751 23 Uppsala, Sweden.
authors: Matilda L
authors: ML
authors: Salomonsson
mesh_terms: D000599:Amino Acids, Diamino / Q000032:analysis*; D000818:Animals; D049872:Bivalvia; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D000087522:Cyanobacteria Toxins; D057230:Limit of Detection; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*
chemical_list: D000599:Amino Acids, Diamino; D000087522:Cyanobacteria Toxins; C001824:beta-N-methylamino-L-alanine
keywords: Analysis; BMAA; HILIC-UHPLC-MS/MS; Interlaboratory comparison; Mussels