A1 Refereed original research article in a scientific journal
Free radical-scavenging activity of phenolics by reversed-phase TLC
Authors: Yrjonen T, Li PW, Summanen J, Hopia A, Vuorela H
Publisher: AMER OIL CHEMISTS SOC A O C S PRESS
Publishing place: USA
Publication year: 2003
Journal: Journal of the American Oil Chemists' Society
Journal name in source: JOURNAL OF THE AMERICAN OIL CHEMISTS SOCIETY
Journal acronym: J AM OIL CHEM SOC
Volume: 80
Issue: 1
First page : 9
Last page: 14
Number of pages: 6
ISSN: 0003-021X
DOI: https://doi.org/10.1007/s11746-003-0642-z
Abstract
A method was developed to measure the radical-scavenging activity of compounds separated by reversed-phase TLC (RP-TLC) using phenolic acids as model analytes. TLC separation was followed by dipping the plate in a 0.04% (wt/vol) solution of 1,1-diphenyl-2-picrylhydrazyl (DPPH) in methanol. The compounds possessing radical-scavenging activity were detected as bright yellow bands against a purple background. A video documentation system based on a CCD video camera was used for the detection and quantification of the activity. The developed RP-TLC-DPPH method was compared to the widely used spectrophotometric DPPH assay. The results obtained by the two methods correlated well, apart from syringic acid, ascorbic acid, and n-propyl gallate, which proved to be outliers in the regression analyses. The correlation coefficient, after excluding outliers, was r(2) = 0.923. The RP-TLC-DPPH method was applied for the measurement of free radical-scavenging activity of rapeseed meal fractions. A total of 10 separated zones with free radical-scavenging activity were detected, with R-f values ranging from 0.04 to 0.85. The results show that the method can be used for the effective fractionation and analysis of potential antioxidative compounds in natural extracts.
A method was developed to measure the radical-scavenging activity of compounds separated by reversed-phase TLC (RP-TLC) using phenolic acids as model analytes. TLC separation was followed by dipping the plate in a 0.04% (wt/vol) solution of 1,1-diphenyl-2-picrylhydrazyl (DPPH) in methanol. The compounds possessing radical-scavenging activity were detected as bright yellow bands against a purple background. A video documentation system based on a CCD video camera was used for the detection and quantification of the activity. The developed RP-TLC-DPPH method was compared to the widely used spectrophotometric DPPH assay. The results obtained by the two methods correlated well, apart from syringic acid, ascorbic acid, and n-propyl gallate, which proved to be outliers in the regression analyses. The correlation coefficient, after excluding outliers, was r(2) = 0.923. The RP-TLC-DPPH method was applied for the measurement of free radical-scavenging activity of rapeseed meal fractions. A total of 10 separated zones with free radical-scavenging activity were detected, with R-f values ranging from 0.04 to 0.85. The results show that the method can be used for the effective fractionation and analysis of potential antioxidative compounds in natural extracts.
Keywords:
1,1-diphenyl-2-picrylhydrazyl, free radical scavenging activity, phenolic compounds, reversed-phase thinlayer chromatography
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