title: Simple Detection Methods for Antinutritive Factor β-ODAP Present in Lathyrus sativus L. by High Pressure Liquid Chromatography and Thin Layer Chromatography.
abstract: Lathyrus sativus L. (Grass pea) is the source for cheap and nutritious food choice in drought and famine susceptible zones in greater part of North India and Africa. The non-protein amino acid β-N-oxalyl-L-α,β-diaminopropionic acid (β-ODAP) has been known for decades for its potent neurotoxic effect, causing irreversible neurodegenerative disease "neurolathyrism", present in both seed and leaf of Lathyrus sativus L. and other species in varying proportions. It is crucial to establish a rapid as well as reliable detection methodology for β-ODAP content in various Lathyrus plants. Currently available HPLC based methods involve multi-step derivatization of the sample. To overcome this, we have developed β-ODAP analysis method by HPLC without any prior derivatization. This method is statistically significant in the range of 2 to 100μg/ml and exhibited linear response with r2 > 0.99. Limit of detection and quantitation of the later method was determined to be 5.56 μg/ml and 16.86 μg/ml, respectively. In addition to this, a TLC based method has also been developed. The limit of detection of β-ODAP is 0.6μg and for its substrate, L-1,2-diaminopropionic acid is 5μg. Both HPLC and TLC methods were validated by conducting in-vitro bioconversion test to detect the presence of biocatalyst in plant extract. This method is economical, rapid and simple.
authors.affiliation: Protein Technology Core, Centre for Cellular and Molecular Platforms, NCBS-TIFR, Bangalore, Karnataka, India.
authors.forename: Bidisha
authors.initials: B
authors.lastname: Ghosh
authors.affiliation: Advanced Laboratory for Plant Genetic Engineering, Indian Institute of Technology Kharagpur, Kharagpur, West Bengal, India.
authors.forename: Joy
authors.initials: J
authors.lastname: Mitra
authors.affiliation: Advanced Laboratory for Plant Genetic Engineering, Indian Institute of Technology Kharagpur, Kharagpur, West Bengal, India.
authors.forename: Saikat
authors.initials: S
authors.lastname: Chakraborty
authors.affiliation: Advanced Laboratory for Plant Genetic Engineering, Indian Institute of Technology Kharagpur, Kharagpur, West Bengal, India.
authors.forename: Jagannath
authors.initials: J
authors.lastname: Bhattacharyya
authors.affiliation: Advanced Laboratory for Plant Genetic Engineering, Indian Institute of Technology Kharagpur, Kharagpur, West Bengal, India.
authors.forename: Anirban
authors.initials: A
authors.lastname: Chakraborty
authors.affiliation: Advanced Laboratory for Plant Genetic Engineering, Indian Institute of Technology Kharagpur, Kharagpur, West Bengal, India.
authors.forename: Soumitra Kumar
authors.initials: SK
authors.lastname: Sen
authors.affiliation: Protein Technology Core, Centre for Cellular and Molecular Platforms, NCBS-TIFR, Bangalore, Karnataka, India.
authors.forename: Muniasamy
authors.initials: M
authors.lastname: Neerathilingam
mesh_terms: D000349:Africa; D000599:Amino Acids, Diamino / Q000302:isolation & purification*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D002855:Chromatography, Thin Layer / Q000379:methods*; D007194:India; D027807:Lathyrus / Q000737:chemistry*; D009498:Neurotoxins / Q000302:isolation & purification; D010936:Plant Extracts / Q000032:analysis; D018515:Plant Leaves / Q000737:chemistry; D012639:Seeds / Q000737:chemistry
chemical_list: D000599:Amino Acids, Diamino; D009498:Neurotoxins; D010936:Plant Extracts; C100126:oxalyldiaminopropionic acid