Determination of beta-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.
A high-performance liquid chromatography (HPLC) method is described for determining subpicomole concentrations of beta-N-methylamino-L-alanine (BMAA) in plant and animal tissue. BMAA and other amino acids were reacted with 9-fluorenylmethyl chloroformate (FMOC) for 10 min under alkaline conditions to form highly fluorescent and stable derivatives. All amino acids, including BMAA, eluted from the column within 22 min. BMAA (tr = 18.02 +/- 0.07 min) was detected in Cycas circinalis L. seed and in serum, cerebrospinal fluid and brain tissue from BMAA-treated monkeys and rats. The primary amino acids glutamine, glutamic acid, aspartic acid, alanine, glycine and gamma-aminobutyric acid (GABA) could also be detected since they were well resolved from BMAA. These amino acids and BMAA were linear over the concentration range of 0.15-7.5 microM with a relative standard deviation ranging from 2.1-6.7%. This method should prove useful in studies to determine the role of BMAA in the Western Pacific amyotrophic lateral sclerosis/Parkinsonism-dementia complex for which cycad seed is the principal etiological candidate.
Kisby
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GE
Institute of Neurotoxicology, Albert Einstein College of Medicine, Bronx, NY 10461.
Roy
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Spencer
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D000599:Amino Acids, Diamino / Q000032:analysis* / Q000097:blood / Q000493:pharmacokinetics; D000818:Animals; D002540:Cerebral Cortex / Q000187:drug effects / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D000087522:Cyanobacteria Toxins; D005449:Fluorenes; D008252:Macaca fascicularis; D010944:Plants / Q000032:analysis*; D051381:Rats; D011919:Rats, Inbred Strains
D000599:Amino Acids, Diamino; D000087522:Cyanobacteria Toxins; D005449:Fluorenes; C001824:beta-N-methylamino-L-alanine; C054007:1-(9-fluorenyl)methyl chloroformate