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Selective LC-MS/MS method for the identification of BMAA from its isomers in biological samples

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Algal blooms are well-known sources of acute toxic agents that can be lethal to aquatic organisms. However, one such toxin, β-N-methylamino-l-alanine (BMAA) is also believed to cause amyotrophic lateral sclerosis, also known as Lou Gehrig’s disease. The detection and identification of BMAA in natural samples were challenging until the recent introduction of reliable methods. However, the issue of potential interference from unknown isomers of BMAA present in samples has not yet been thoroughly investigated. Based on a systematic database search, we generated a list of all theoretical BMAA structural isomers, which was subsequently narrowed down to seven possible interfering compounds for further consideration. The seven possible candidates satisfied the requirements of chemical stability and also shared important structural domains with BMAA. Two of the candidates, 2,4-diaminobutyric acid (DAB) and N-(2-aminoethyl) glycine (AEG) have recently been studied in the context of BMAA. A further isomer, β-amino-N-methyl-alanine (BAMA), has to be considered because it can potentially yield the fragment ion, which is diagnostic for BMAA. Here, we report the synthesis and analysis of BAMA, together with AEG, DAB, and other isomers that are of interest in the separation and detection of BMAA in biological samples by using either high-performance liquid chromatography or ultra-high-performance liquid chromatography coupled with tandem mass spectrometry. We detected for the first time BAMA in blue mussel and oyster samples. This work extends the previously developed liquid chromatography–tandem mass spectrometry platform Spacil et al. (Analyst 135:127, 2010) to allow BMAA isomers to be distinguished, improving the detection and identification of this important amino acid.

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  1. Spacil Z, Eriksson J, Jonasson S et al (2010) Analytical protocol for identification of BMAA and DAB in biological samples. Analyst 135:127

    Article  CAS  Google Scholar 

  2. Cox P, Banack S, Murch S (2003) Biomagnification of cyanobacterial neurotoxins and neurodegenerative disease among the Chamorro people of Guam. Proc Natl Acad Sci USA 100:13380

    Article  CAS  Google Scholar 

  3. Cox PA, Banack SA, Murch SJ et al (2005) Diverse taxa of cyanobacteria produce beta-N-methylamino-l-alanine, a neurotoxic amino acid. Proc Natl Acad Sci USA 102:5074

    Article  CAS  Google Scholar 

  4. Banack SA, Johnson HE, Cheng R et al (2007) Production of the neurotoxin BMAA by a marine cyanobacterium. Mar Drugs 5:180

    Article  CAS  Google Scholar 

  5. Johnson HE, King SR, Banack SA et al (2008) Cyanobacteria (Nostoc commune) used as a dietary item in the Peruvian highlands produce the neurotoxic amino acid BMAA. J Ethnopharmacol 118:159

    Article  CAS  Google Scholar 

  6. Esterhuizen M, Downing T (2008) Beta-N-methylamino-l-alanine (BMAA) in novel South African cyanobacterial isolates. Ecotoxicol Environ Saf 71:309

    Article  CAS  Google Scholar 

  7. Metcalf JS, Banack SA, Lindsay J et al (2008) Co-occurrence of beta-N-methylamino-l-alanine, a neurotoxic amino acid with other cyanobacterial toxins in British waterbodies, 1990–2004. Environ Microbiol 10:702

    Article  CAS  Google Scholar 

  8. Eriksson J, Jonasson S, Papaefthimiou D et al (2009) Improving derivatization efficiency of BMAA utilizing AccQ-Tag in a complex cyanobacterial matrix. Amino Acids 36:43

    Article  CAS  Google Scholar 

  9. Bidigare RR, Christensen SJ, Wilde SB et al (2009) Cyanobacteria and BMAA: possible linkage with avian vacuolar myelinopathy (AVM) in the south-eastern United States. Amyotroph Lateral Scler 10(Suppl 2):71

    Article  Google Scholar 

  10. Faassen EJ, Gillissen F, Zweers HAJ et al (2009) Determination of the neurotoxins BMAA (beta-N-methylamino-l-alanine) and DAB (alpha-, gamma-diaminobutyric acid) by LC-MSMS in Dutch urban waters with cyanobacterial blooms. Amyotrophic Lateral Scler 10:79

    Article  CAS  Google Scholar 

  11. Rosen J, Hellenaes K (2008) Determination of the neurotoxin BMAA (β-N-methylamino-l-alanine) in cycad seed and cyanobacteria by LC-MS/MS (liquid chromatography tandem mass spectrometry). Analyst (Cambridge, U K) 133:1785

    Article  CAS  Google Scholar 

  12. Kisby GE, Roy DN, Spencer PS (1988) Determination of β-N-methylamino-l-alanine (BMAA) in plant (Cycas circinalis L.) and animal tissue by precolumn derivatization with 9-fluorenylmethyl chloroformate (FMOC) and reversed-phase high-performance liquid chromatography. J Neurosci Methods 26:45

    Article  CAS  Google Scholar 

  13. Snyder LR, Cruz-Aguado R, Sadilek M et al (2009) Parkinson-dementia complex and development of a new stable isotope dilution assay for BMAA detection in tissue. Toxicol Appl Pharmacol 240:180

    Article  CAS  Google Scholar 

  14. Snyder LR, Hoggard JC, Montine TJ et al (2010) Development and application of a comprehensive two-dimensional gas chromatography with time-of-flight mass spectrometry method for the analysis of l-beta-methylamino-alanine in human tissue. J Chromatogr A 1217:4639

    Article  CAS  Google Scholar 

  15. Marler TE, Snyder LR, Shaw CA (2010) Cycas micronesica (Cycadales) plants devoid of endophytic cyanobacteria increase in β-methylamino-l-alanine. Toxicon 56:563

    Article  CAS  Google Scholar 

  16. Caller TA, Doolin JW, Haney JF et al (2009) A cluster of amyotrophic lateral sclerosis in New Hampshire: a possible role for toxic cyanobacteria blooms. Amyotrophic Lateral Scler 10:101

    Article  CAS  Google Scholar 

  17. Cox PA, Richer R, Metcalf JS et al (2009) Cyanobacteria and BMAA exposure from desert dust: a possible link to sporadic ALS among Gulf War veterans. Amyotrophic Lateral Scler 10:109

    Article  CAS  Google Scholar 

  18. Craighead D, Metcalf JS, Banack SA et al (2009) Presence of the neurotoxic amino acids beta-N-methylamino-l-alanine (BMAA) and 2,4-diamino-butyric acid (DAB) in shallow springs from the Gobi Desert. Amyotrophic Lateral Scler 10:96

    Article  CAS  Google Scholar 

  19. Roney BR, Li R, Banack SA et al (2009) Consumption of fa cai Nostoc soup: a potential for BMAA exposure from Nostoc cyanobacteria in China? Amyotrophic Lateral Scler 10:44

    Article  CAS  Google Scholar 

  20. Jonasson S, Eriksson J, Berntzon L et al (2010) Transfer of a cyanobacterial neurotoxin within a temperate aquatic ecosystem suggests pathways for human exposure. Proc Natl Acad Sci USA 107:9252

    Article  CAS  Google Scholar 

  21. Kruger T, Monch B, Oppenhauser S et al (2010) LC-MS/MS determination of the isomeric neurotoxins BMAA (beta-N-methylamino-l-alanine) and DAB (2,4-diaminobutyric acid) in cyanobacteria and seeds of Cycas revoluta and Lathyrus latifolius. Toxicon 55:547

    Article  Google Scholar 

  22. Moad G, Benkovic SJ (1978) On the mechanism of decomposition of geminal diamines. J Am Chem Soc 100:5495

    Article  CAS  Google Scholar 

  23. Banack S, Downing T, Spacil Z et al (2010) Distinguishing the cyanobacterial neurotoxin beta-N-methylamino-l-alanine (BMAA) from its structural isomer 2,4-diaminobutyric acid (2,4-DAB). Toxicon 56:868

    Article  CAS  Google Scholar 

  24. Banack SA, Metcalf JS, Spacil Z et al (2011) Distinguishing the cyanobacterial neurotoxin β-N-methylamino-l-alanine (BMAA) from other diamino acids. Toxicon 57:730

    Article  CAS  Google Scholar 

  25. Dietrich RF, Sakurai T, Kenyon GL (1979) Substituted beta.-alanines. Preparation of.alpha.-substituted.beta.-alanine derivatives from 5-substituted uracils and dihydrouracils. J Org Chem 44:1894

    Article  CAS  Google Scholar 

  26. Apffel A, Fischer S, Goldberg G et al (1995) Enhanced sensitivity for peptide mapping with electrospray liquid chromatography–mass spectrometry in the presence of signal suppression due to trifluoroacetic acid-containing mobile phases. J Chromatogr A 712:177

    Article  Google Scholar 

  27. Aruda WO, Aruda KO (2009) Review of volatile perfluorocarboxylic acids as ion-pair reagents in LC, part II. LCGC North Am 27:916–928

    CAS  Google Scholar 

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We thank Ulla Rasmussen (Department of Botany, Stockholm University) for kindly providing the biological samples in this study. We also thank Frank Turecek (University of Washington, Seattle, WA 98195, USA) for his suggestions concerning the gas-phase chemistry and Peter B. Wyatt (School of Biological and Chemical Sciences at Queen Mary University of London) for helpful discussions.

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Correspondence to Zdenek Spacil or Leopold L. Ilag.

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Jiang, L., Aigret, B., De Borggraeve, W.M. et al. Selective LC-MS/MS method for the identification of BMAA from its isomers in biological samples. Anal Bioanal Chem 403, 1719–1730 (2012).

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