期刊
AMINO ACIDS
卷 36, 期 1, 页码 43-48出版社
SPRINGER
DOI: 10.1007/s00726-007-0023-4
关键词
beta-N-methyl-amino-L-alanine (BMAA); Cyanobacteria; Carbamate 6-aminoquinolyl-N-hydrosuccinimidyl (AQC) Derivatization; High-performance liquid chromatography (HPLC)
资金
- Swedish Research Council for Environmental, Agricultural Sciences and Spatial Planning (FORMAS)
- VWR
Two different assays have been developed and used in order to investigate the optimal conditions for derivatization and detection of acid beta-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(A (R)) chemistry and Chromolith(A (R)) 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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