Limits...
Simple Detection Methods for Antinutritive Factor β-ODAP Present in Lathyrus sativus L. by High Pressure Liquid Chromatography and Thin Layer Chromatography.

Ghosh B, Mitra J, Chakraborty S, Bhattacharyya J, Chakraborty A, Sen SK, Neerathilingam M - PLoS ONE (2015)

Bottom Line: 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.It is crucial to establish a rapid as well as reliable detection methodology for β-ODAP content in various Lathyrus plants.The limit of detection of β-ODAP is 0.6μg and for its substrate, L-1,2-diaminopropionic acid is 5μg.

View Article: PubMed Central - PubMed

Affiliation: Protein Technology Core, Centre for Cellular and Molecular Platforms, NCBS-TIFR, Bangalore, Karnataka, India.

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.

No MeSH data available.


Related in: MedlinePlus

Results for β-ODAP detection by TLC.A) Visualization of standard β-ODAP on TLC plate. B) Calibration curve for β-ODAP C) Determination of Limit of detection for β-ODAP. The lowest concentration 0.4μg visible is circled.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4629898&req=5

pone.0140649.g004: Results for β-ODAP detection by TLC.A) Visualization of standard β-ODAP on TLC plate. B) Calibration curve for β-ODAP C) Determination of Limit of detection for β-ODAP. The lowest concentration 0.4μg visible is circled.

Mentions: The relative mobility of β-ODAP was determined to be 0.37 (Fig 4A). Calibration plot showed linear relationship between the concentrations of β-ODAP applied to the spot area obtained. A seven point calibration graph was plotted with r2 value obtained was greater than 0.97 (Fig 4B). In order to determine the lowest detection range, 0.2, 0.4 and 0.6μg of β-ODAP were applied. As seen in Fig 4C, this method enabled visualization of compound as low as 0.6μg making it a sensitive method. β-ODAP develops as circular shape and L-1,2-DAPAas oval shape which was seen after ninhydrin staining.


Simple Detection Methods for Antinutritive Factor β-ODAP Present in Lathyrus sativus L. by High Pressure Liquid Chromatography and Thin Layer Chromatography.

Ghosh B, Mitra J, Chakraborty S, Bhattacharyya J, Chakraborty A, Sen SK, Neerathilingam M - PLoS ONE (2015)

Results for β-ODAP detection by TLC.A) Visualization of standard β-ODAP on TLC plate. B) Calibration curve for β-ODAP C) Determination of Limit of detection for β-ODAP. The lowest concentration 0.4μg visible is circled.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4629898&req=5

pone.0140649.g004: Results for β-ODAP detection by TLC.A) Visualization of standard β-ODAP on TLC plate. B) Calibration curve for β-ODAP C) Determination of Limit of detection for β-ODAP. The lowest concentration 0.4μg visible is circled.
Mentions: The relative mobility of β-ODAP was determined to be 0.37 (Fig 4A). Calibration plot showed linear relationship between the concentrations of β-ODAP applied to the spot area obtained. A seven point calibration graph was plotted with r2 value obtained was greater than 0.97 (Fig 4B). In order to determine the lowest detection range, 0.2, 0.4 and 0.6μg of β-ODAP were applied. As seen in Fig 4C, this method enabled visualization of compound as low as 0.6μg making it a sensitive method. β-ODAP develops as circular shape and L-1,2-DAPAas oval shape which was seen after ninhydrin staining.

Bottom Line: 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.It is crucial to establish a rapid as well as reliable detection methodology for β-ODAP content in various Lathyrus plants.The limit of detection of β-ODAP is 0.6μg and for its substrate, L-1,2-diaminopropionic acid is 5μg.

View Article: PubMed Central - PubMed

Affiliation: Protein Technology Core, Centre for Cellular and Molecular Platforms, NCBS-TIFR, Bangalore, Karnataka, India.

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.

No MeSH data available.


Related in: MedlinePlus