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

Glen E. Kisby, Dwijendra N. Roy, Peter Spencer

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58 Citations (Scopus)

Abstract

A high-performance liquid chromatography (HPLC) method is described for determining subpicomole concentrations of β-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 γ-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 μM 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.

Original languageEnglish (US)
Pages (from-to)45-54
Number of pages10
JournalJournal of Neuroscience Methods
Volume26
Issue number1
DOIs
StatePublished - 1988
Externally publishedYes

Fingerprint

Cycas
Reverse-Phase Chromatography
Alanine
High Pressure Liquid Chromatography
Amino Acids
Seeds
Aminobutyrates
Amyotrophic Lateral Sclerosis
Parkinsonian Disorders
Glutamine
Aspartic Acid
Glycine
Haplorhini
Cerebrospinal Fluid
Dementia
Glutamic Acid
1-(9-fluorenyl)methyl chloroformate
Brain
Serum

Keywords

  • Amyotrophic lateral sclerosis
  • Brain tissue
  • Cerebrospinal fluid
  • Cycad
  • Serum
  • β-N-Methylamino-l-alanine (BMAA)

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

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title = "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",
abstract = "A high-performance liquid chromatography (HPLC) method is described for determining subpicomole concentrations of β-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 γ-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 μM 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.",
keywords = "Amyotrophic lateral sclerosis, Brain tissue, Cerebrospinal fluid, Cycad, Serum, β-N-Methylamino-l-alanine (BMAA)",
author = "Kisby, {Glen E.} and Roy, {Dwijendra N.} and Peter Spencer",
year = "1988",
doi = "10.1016/0165-0270(88)90128-8",
language = "English (US)",
volume = "26",
pages = "45--54",
journal = "Journal of Neuroscience Methods",
issn = "0165-0270",
publisher = "Elsevier",
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TY - JOUR

T1 - 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

AU - Kisby, Glen E.

AU - Roy, Dwijendra N.

AU - Spencer, Peter

PY - 1988

Y1 - 1988

N2 - A high-performance liquid chromatography (HPLC) method is described for determining subpicomole concentrations of β-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 γ-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 μM 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.

AB - A high-performance liquid chromatography (HPLC) method is described for determining subpicomole concentrations of β-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 γ-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 μM 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.

KW - Amyotrophic lateral sclerosis

KW - Brain tissue

KW - Cerebrospinal fluid

KW - Cycad

KW - Serum

KW - β-N-Methylamino-l-alanine (BMAA)

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