A method for the simultaneous determination of β-ODAP, α-ODAP, homoarginine and polyamines in Lathyrus sativus by liquid chromatography using a new extraction procedure

The paper proposes anew method to extract beta-N-oxalyl-(L)-alpha,beta-diaminopropionic acid (beta-ODAP), alpha-N-oxalyl-(L)-alpha,beta-diaminopropionic acid (alpha-ODAP), homoarginine and polyamines using 0.2 M HClO4 for only 1 h at 0-4 degrees C. Based on the proposed extraction, an assay is presented for simultaneously determining polyamines, the neurotoxin P-ODAP, its non-neurotoxic isomer alpha-ODAP and homoarginine in Lathyrus sativus extracts using a liquid chromatography (LC) system after derivatization with para-nitrobenzyloxycarbonyl chloride (PNZ-Cl). The method exhibits some prominent advantages, which include: shorter extraction-time, simultaneous extraction of non-protein amino acids and polyamines and inhibition of the isomerization of beta-ODAP to alpha-ODAP. The assay proved satisfactory for the quantitative determination of L. sativus extracts of seeds and seedlings. (c) 2004 Elsevier B.V. All rights reserved.