Development and in-house validation of a method for quantification of BMAA in mussels using dansyl chloride derivatization and ultra performance liquid chromatography tandem mass spectrometry

• A new approach for the detection and quantification of beta-N-methylamino-L-alanine (BMAA) in mussels using chemical derivatization with dansyl chloride and ultra performance liquid chromatography-electrospray ionization-tandem mass spectrometry (UPLC-ESI-MS/MS) is presented. The method, using dansyl chloride as the reagent, is simple, robust and cost efficient. Comparing the fragmentation patterns for derivatized BMAA and its isomer L-2,4-diaminobutyric acid derivatized a selective fragment for the derivatized BMAA was formed m/z 585 > m/z 71. To ensure an actual detection of BMAA, the ion ratio of the daughter ions m/z 71 and m/z 277 was calculated and compared with the calculated mean ion ratio. The method development resulted in a simplified sample preparation excluding solid phase extraction and, instead, performing filtration and dilution of the samples before the derivatization. Validation was performed and the limit of quantification was determined to be 0.15 mu g g(-1) wet mussel homogenate (33 fmol per injection) and the limit of detection was estimated to be 16 ng g(-1) (4 fmol per injection). The intra-and inter-day precisions were within the accepted criteria and the recovery was about 83%. The stability of BMAA in the stock solution was at least 3 months and the stability of derivatized extracts in vials for the quantification was good for 27 days. This is the first quantification method for BMAA in mussels with extensive validation data published and the method was also applied on real mussel samples collected on the west coast of Sweden. The concentrations of BMAA in the mussel samples were determined to be between 0.27 and 1.6 mu g g(-1) wet mussel homogenates.

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