Abstract
Over the last decade the amino acid beta-N-methylamino-L-alanine (BMAA) has come under intense scrutiny. International laboratory and epidemiological research continues to support the hypothesis that environmental exposure to BMAA (e.g., through dietary practices, water supply) can promote the risk of various neurodegenerative diseases. A wide variety of cyanobacteria spp. have previously been reported to produce BMAA, with production levels dependent upon species, strain and environmental conditions. Since spirulina (Arthrospira spp.) is a member of the cyanobacteria phylum frequently consumed via dietary supplements, the presence of BMAA in such products may have public health implications. In the current work, we have analyzed ten spirulina-containing samples for the presence of BMAA; six pure spirulina samples from two separate raw materials suppliers, and four commercially-available multi-ingredient products containing 1.45 g of spirulina per 8.5 g serving. Because of controversy surrounding the measurement of BMAA, we have used two complementary liquid chromatography-tandem mass spectrometry (LC-MS/MS) methods: one based on reversed phase LC (RPLC) with derivatization and the other based on hydrophilic interaction LC (HILIC). Potential matrix effects were corrected for by internal standardization using a stable isotope labeled BMAA standard. BMAA was not detected at low limits of detection (80 ng/g dry weight) in any of these product samples. Although these results are reassuring, BMAA analyses should be conducted on a wider sample selection and, perhaps, as part of ongoing spirulina production quality control testing and specifications.
Highlights
Over the last decade, an increasingly robust body of bench and epidemiological research has suggested that β-N-methylamino-L-alanine (BMAA) may play a role in various neurodegenerative diseases [1,2]
2,4-diaminobutyric acid (DAB) and N-(2-aminoethyl)-glycine (AEG), were monitored since they are isomeric with BMAA and have been reported in cyanobacterial samples [19,20]
A stable-isotope labeled internal standard, d3-BMAA, was used as internal standard in both methods to correct for extraction recovery and matrix effects, the latter being commonly observed in electrospray LC-MS/MS
Summary
An increasingly robust body of bench and epidemiological research has suggested that β-N-methylamino-L-alanine (BMAA) may play a role in various neurodegenerative diseases [1,2]. BMAA is a non-proteinogenic amino acid first identified in the seeds of Cycas micronesia in 1967 [3]. It was noted in the 1980s [4] and revisited again in 2002 [5], that cycad pulp and seed flour (as well as animals that fed upon cycad seeds) have been a significant part of the traditional dietary and medicinal agents among specific Western Pacific-dwelling communities (e.g. Guam). BMAA was not found in controls that had died from causes other than neurodegenerative This suggested that BMAA may not merely be a local concern for those in Micronesia. Since the strains of cyanobacteria used in the study were drawn from diverse taxa, and given the ubiquity of the phylum cyanobacteria itself, the implications to public health could be significant
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