An Improved HILIC HPLC-MS/MS Method for the Determination of ß-ODAP and Its α Isomer in Lathyrus sativus.

ABSTRACT

ß-N-Oxalyl-l-α,ß-diaminopropionic acid (ß-ODAP) is a non-protein amino acid present in Lathyrus sativus (grass pea) and other Lathyrus species, in parallel with its nontoxic isomer, α-ODAP. When consuming grass pea for several months as staple food, ß-ODAP may cause neurolathyrism, a motor neuron degeneration syndrome. Therefore, the independent quantification of both ODAP isomers instead of only the total amount in grass pea allows the identification of less toxic varieties and the development of tools to support breeding for improving grass pea quality. In this work, a simple and fast HPLC-MS/MS method was developed without sample derivatization, using a hydrophilic interaction chromatography (HILIC) column and an isocratic gradient of eluents for 18 min, which allowed the determination of both α- and ß-ODAP. The proposed method was fully validated and applied to the determination of α- and ß-ODAP contents in a diverse collection of 107 grass pea accessions representative of the main grass pea-growing geographical regions in the world, with the prompt identification of contrasting accessions. ß-ODAP content in the analyzed grass pea samples ranged from 0.45 ± 0.02 to 6.04 ± 0.45 mg g-1. The moderate correlation found between α- and ß-ODAP contents (0.65) in this collection reinforces the importance of the independent quantification of both ODAP isomers.