Improving derivatization efficiency of BMAA utilizing AccQ-Tag® in a complex cyanobacterial matrix

  • Eriksson, Johan
  • Jonasson, Sara
  • Papaefthimiou, Dimitra
  • Rasmussen, Ulla
  • Bergman, Birgitta

  • 1Department of Botany, Stockholm University, 10691, Stockholm, Sweden

e-mail: [email protected]

Received: 16 October 2007/Accepted: 19 December 2007/Published online: 9 January 2008

Johan Eriksson and Sara Jonasson contributed equally to this work.

Amino Acids 36(1):p 43-48, January 2009. | DOI: 10.1007/s00726-007-0023-4

Abstract

Two different assays have been developed and used in order to investigate the optimal conditions for derivatization and detection of acid β- N-methyl-amino-L-alanine (BMAA) in a cyanobacterial sample. BMAA was extracted from cyanobacterial cultures both from the cytosolic (“free”) fraction and in the precipitated (“protein”) fraction using a newly developed extraction scheme and the sample matrix was standardized according to protein concentration to ensure the highest possible derivative yield. A rapid and sensitive HPLC method for fluorescence detection of the non-protein amino acid BMAA in cyanobacteria, utilizing the Waters AccQ-Tag® chemistry and Chromolith® Performance RP-18e columns was developed. Using this new method and utilizing a different buffer system and column than that recommended by Waters, we decreased the time between injections by 75%. The limit of quantification was determined to be 12 nmol and limit of detection as 120 fmol. The linear range was in the range of 8.5 nmol-84 pmol. Accuracy and precision were well within FDA guidelines for bioanalysis.

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