Free radical-scavening activity of phenolics by reversed-phase TLC |
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Authors: | Teijo?Yrj?nen Li?Peiwu Jari?Summanen Anu?Hopia Email author" target="_blank">Heikki?VuorelaEmail author |
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Affiliation: | (1) Department of Applied Chemistry and Microbiology, University of Helsinki, FIN-00014, Finland;(2) Present address: Oll Crops Research Institute of Chinese Academy of Agricultural Sciences, Wuhan, 430062 Hubei, People’s Republic of China;(3) Division of Pharmacognosy, Department of Pharmacy, University of Helsinki, P.O. Box 56, FIN-00014, Finland |
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Abstract: | A method was developed to measure the radicalscavenging 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-scavening 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. |
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Keywords: | 1 1-Diphenyl-2-picrylhydrazyl free radical scavening activity phenolic compounds reversed-phase thinlayer chromatography |
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