BMAA extraction of cyanobacteria samples: which method to choose?
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β-N-Methylamino-l-alanine (BMAA), a neurotoxin reportedly produced by cyanobacteria, diatoms and dinoflagellates, is proposed to be linked to the development of neurological diseases. BMAA has been found in aquatic and terrestrial ecosystems worldwide, both in its phytoplankton producers and in several invertebrate and vertebrate organisms that bioaccumulate it. LC-MS/MS is the most frequently used analytical technique in BMAA research due to its high selectivity, though consensus is lacking as to the best extraction method to apply. This study accordingly surveys the efficiency of three extraction methods regularly used in BMAA research to extract BMAA from cyanobacteria samples. The results obtained provide insights into possible reasons for the BMAA concentration discrepancies in previous publications. In addition and according to the method validation guidelines for analysing cyanotoxins, the TCA protein precipitation method, followed by AQC derivatization and LC-MS/MS analysis, is now validated for extracting protein-bound (after protein hydrolysis) and free BMAA from cyanobacteria matrix. BMAA biological variability was also tested through the extraction of diatom and cyanobacteria species, revealing a high variance in BMAA levels (0.0080–2.5797 μg g−1 DW).
Keywordsβ-N-Methylamino-l-alanine Extraction Validation Cyanobacteria Diatoms
We wish to thank the Swedish Research Council Formas for its financial support and the Baltic 2020 and Olle Engkvist Byggmästare foundations for funding the Acquity UPLC coupled to the Xevo-TQ-MS system (Waters) via the MiMeBS grant awarded to Birgitta Bergman. We are grateful to Nicola Wannicke of the Leibniz Institute for Baltic Sea Research, Germany and to Katja Pasdzierny of GEOMAR, Germany for isolating the diatom species from the Baltic Sea and North Sea samples.
Conflict of Interest
The authors declare no conflict of interest.
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