DETERMINATION OF THE NEUROLATHYROGEN BETA-N-OXALYL-L-ALPHA,BETA-DIAMINOPROPIONIC ACID USING HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY WITH FLUOROMETRIC DETECTION

This paper describes a sensitive spectrofluorometric HPLC method suitable for determining picogram levels of beta-N-oxalyl-L-alpha,beta-diaminopropionic acid (ODAP), a neurotoxin. Aqueous extracts of powdered Lathyrus sativus seeds were treated with 9-fluorenylmethyl chloroformate (FMOC) and the ODAP-FMOC derivative analyzed by reversed-phase chromatography using a mu Bondapak C18 column. The excitation and emission wavelengths were 254 and 315 nm, respectively. The mobile phase was sodium acetate buffer (0.05 M, pH 6.35)-acetonitrile (72:28, v/v) at a flow-rate of 1 ml/min. This method represents a major advance over the standard spectrophotometric assays used currently.