On practical problems in estimation of antioxidant activity of compounds by DPPH method (Problems in estimation of antioxidant activity)

2,2′-Diphenyl-1-picrylhydrazyl (DPPH) method is commonly applied for the estimation of antioxidant activity of single compounds and their mixtures. The method is based on the spectrophotometric measurement of DPPH concentration changes resulting from the DPPH reaction with an antioxidant. The amount of remaining DPPH in the examined system is a measure of the antioxidative activity of compounds. The study shows that the type and amount of solvent used for the antioxidant dissolution, water content, and hydrogen or metal ion concentration in the measuring system significantly influence the differences in the amount of unreacted DPPH. The presence of ethyl acetate or dioxane or metal ions in the system decreases the kinetics of the reaction between DPPH and butylohydroxytoluene in relation to its kinetics in the system containing only methanolic solutions. The opposite effect is observed for water, low concentration of chloroform or hydrogen ions. The presented relationships confirm the necessity of standardising the DPPH method and show the complexity of the problem, even in very simple DPPH/antioxidant systems.     

Total phenolic contents and antioxidant properties of commonly consumed vegetables grown in Colorado

A total of 38 commonly consumed vegetable samples, including 3 kale, 1 rhubarb, 3 spinach, 3 broccoli, 2 green bean, 5 carrot, 10 tomato, and 11 potato samples, were investigated for their total phenolic contents (TPC) and antioxidant properties. The measured antioxidant properties included free radical scavenging activities against DPPH, superoxide anion radical (O₂⁻), ABTS⁺ and peroxyl radical, and Fe²⁺ chelating capacity. The tested vegetables differed in their antioxidant properties and TPC, although all vegetables had significant antioxidant activities and contained significant levels of phenolics. The TPC and the measured antioxidant properties were correlated to each other. The Fe⁺² chelating capacity and the scavenging capacities against ABTS⁺, DPPH and O₂⁻ were reported for these commonly consumed vegetables for the first time. Results from this study suggest that kale, spinach, broccoli, and rhubarb are the better dietary sources of natural antioxidant activities and phenolic compounds. Furthermore, kale, broccoli and spinach produced in Colorado may have greater antioxidant contents than those grown at other locations.     

Structural identification of isomallotusinin and other phenolics in Phyllanthus emblica L. fruit hull

The air-dried fruit hull of Phyllanthus emblica L. was extracted with 95% ethanol, and then the extract was partitioned by diethyl ether and ethyl acetate (EA). The EA fraction was then subjected to separation and purification using silica gel and Sephadex LH-20 column chromatography repeatedly to obtain five hydrolysable tannins. They were identified as mucic acid 1,4-lactone 3-o-gallate (C1), isocorilagin (C2), chebulanin (C3), chebulagic acid (C4) and isomallotusinin (C5) using mass spectrometry and nuclear magnetic resonance (NMR) spectrometry. Isomallotusinin and chebulanin were identified from emblica dried fruit hull for the first time, and isomallotusinin was the first time identified from Phyllanthus. Furthermore, the antioxidant abilities of these hydrolysable tannins were investigated using DPPH and ABTS⁺ radical scavenging systems. All hydrolysable tannins showed strong DPPH and ABTS⁺ radical scavenging activities. Isomallotusinin and chebulagic acid exhibited the highest antioxidant activity compared to other purified compounds tested.