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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References
Brand-Williams, W., M.-E. Cuvelier, and C. Berset, Use of a Free Radical Method to Evaluate Antioxidant Activity, Food Sci. Technol. 28:25–30 (1995).
Hopia, A.I., S.-W. Huang, K. Schwarz, J.B. German, and E.N. Frankel, Effect of Different Lipid Systems on Antioxidant Activity of Rosemary Constituents Carnosol and Carnosol and Carnosic Acid With and Without α-Tocopherol, J. Agric. Food Chem. 44:2030–2036 (1996).
Heinonen, I.M., P.J. Lehtonen, and A.I. Hopia, Antioxidant Activity of Berry and Fruit Wines and Liquors, 46:25–31 (1998).
Blois, M.S., Antioxidant Determinations by the Use of a Stable Free Radical, Nature 181:1199–1200 (1958).
Yordanov, N.D., and A.G. Christova, Quantitative Spectrophotometric and EPR-Determination of 1,1-Diphenyl-2-picryl-hydrazyl (DPPH), Fresenius J. Anal Chem. 358:610–613 (1997).
Cotelle, N., J.-L. Bernier, J.-P. Catteau, J. Pommery, J.-C. Wallet, and E.M. Gaydou, Antioxidant Properties of Hydroxyflavones, Free Radic. Biol. Med. 20:35–43 (1996).
Duh, P.-D., Antioxidant Activity of Burdock (Arctium lappa Linne): Its Scavenging Effect on Free-Radical and Active Oxygen, J. Am. Oil Chem. Soc. 75:455–461 (1998).
Gamez, E.J.C., L. Luyengi, S.K. Lee, L.-F. Zhu, B.-N. Zhou, H.H.S. Fong, J.M. Pezzuto, and A.D. Kinghorn, Antioxidant Flavonoid Glycosides from Daphniphyllum calycinum, J. Nat. Prod. 61:706–708 (1998).
Goupy, P., M. Hugues, P. Boivin, and M.J. Amiot, Antioxidant Composition and Activity of Barley (Hordeum vulgare) and Malt Extracts and of Isolated Phenolic Compounds, J. Sci. Food Agric. 79:1625–1634 (1999).
Amarowicz, R., M. Naczk, and F. Shahidi, Antioxidant Activity of Crude Tannins of Canola and Rapeseed Hulls, J. Am. Oil Chem. Soc. 77:957–961 (2000).
Glavind, J., and G. Holmer, Thin-Layer Chromatographic Determination of Antioxidants by the Stable Free Radical α,α′-Diphenyl-β-picrylhydrazyl, 44:539–542 (1967).
Takao, T., F. Kitatani, N. Watanbe, A. Yagi, and K. Sakata, A Simple Screening Method for Antioxidants and Isolation of Several Antioxidants Produced by Marine Bacteria from Fish and Shellfish, Biosci. Biotechnol. Biochem. 58:1780–1783 (1994).
Cuendet, M., K. Hostettmann, O. Potterat, and W. Dyatmiko, Iridoid glucosides with Free Radical Scavenging Properties from Fagraea blumei, Helv. Chim. Acta 80:1144–1152 (1997).
Cavin, A., K. Hostettmann, W. Dyatmyko, and O. Potterat, Antioxidant and Lipophilic Constituents of Tinospora crispa, Planta Med. 64:393–396 (1998).
Nyiredy, S., K. Dallenbach-Tölke, and O. Sticher, The “PRISMA” Optimization System in Planar Chromatography, J. Planar Chromatogr. 1:336–342 (1988).
Porter, W.L., E.D. Black, and A.M. Drolet, Use of Polyamide Oxidative Fluorescence Test on Lipid Emulsions: Contrast in Relative Effectiveness of Antioxidants in Bulk versus Dispersed Systems, J. Agric. Food Chem. 37:615–623 (1989).
Yoshida, T., K. Mori, T. Hatano, T. Okumura, I. Uehara, K. Komagoe, Y. Fujita, and T. Okuda, Studies on Inhibition Mechanism of Autoxidation by Tannins and Flavonoids. V. Radical-Scavenging Effects of Tannins and Related Polyphenols on 1,1-Diphenyl-2-picrylhydrazyl Radical, Chem. Pharm. Bull. 37:1919–1921 (1989).
Cuvelier, M.-E., H. Richard, and C. Berset, Comparison of the Antioxidative Activity of Some Acid-Phenols: Structure-Activity Relationship, Biosci. Biotechnol. Biochem. 56:324–325 (1992).
Vovk, I., A. Golc-Wondra, and M. Prosek, Validation of an HPTLC Method for Determination of Caffeine, J. Planar Chromatogr. 10:416–419 (1997).
Summanen, J., T. Yrjönen, R. Hiltunen, and H. Vuorela, Influence of Densitometer and Video-Documentation Settings in the Detection of Plant Phenolics by TLC, 11:421–427 (1998).
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Yrjönen, T., Peiwu, L., Summanen, J. et al. Free radical-scavening activity of phenolics by reversed-phase TLC. J Amer Oil Chem Soc 80, 9–14 (2003). https://doi.org/10.1007/s11746-003-0642-z
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DOI: https://doi.org/10.1007/s11746-003-0642-z