ASSAY OF NEUROTOXIN β-ODAP AND NON-PROTEIN AMINO ACIDS IN LATHYRUS SATIVUS BY HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY WITH DANSYLATION

Abstract

A new HPLC method was developed for simultaneous quantitative determination of the neurotoxin, β-N-L-oxalyl-α, β-diaminopropionic acid (β-ODAP) and its nontoxic α-isomer and other nonprotein amino acids in Lathyrus sativus based on precolumn derivatization with 5-dimethylaminenaphthalene-1-sulfonylchlorid (DnsCl). These Dns derivatives were separated by reversed-phase chromatography with a phosphate buffer-acetonitrile gradient and by UV absorption at 254 nm. The method showed high sensitivity and precision. The detection limits for both isomers were 2.3 ng and the average recovery was 98% and the correlation coefficients r > 0.999.

Keywords:

• β-N-L-oxalyl-α
• β-Diaminopropionic acid
• HPLC
• Lathyrus sativus
• 5-Dimethylaminenaphthalene-1-sulfonylchlorid
• Assay

ACKNOWLEDGMENT

This study was financially supported by NKBRSF Project G 2000018603 and YS001-A25-004.