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KMID : 1134820190480030390
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Journal of the Korean Society of Food Science and Nutrition
2019 Volume.48 No. 3 p.390 ~ p.393
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Assessment of the Important Factors Influencing Consistent and Accurate ABTS Assay
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Lee Ho-Hyeon
Moon Yong-Sun
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Abstract
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The ABTS (2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)) free-radical scavenging activity is the most common method for evaluating plant antioxidant activities. On the other hand, the results can be affected by a range of factors, such as ABTS adjusting solutions, sample stock solvents, sample pigments, and equations. A conventional ABTS method was assessed by these factors to obtain accurate and consistent results. Daehwang (Rheum officinale) is a medicinal herb in Asia that was used as a plant material for the experiment. Dried roots of Daehwang were pulverized using a mixer and ground with liquid nitrogen. After extracting with 70% ethanol for 24 hours at room temperature, the extracts were condensed by vacuum evaporation. Different experimental factors of the ABTS assay were analyzed. The pH was very stable when ABTS was dissolved and adjusted with water. PBS (phosphate buffered saline) also worked well unlike 100% or 70% ethanol. When the vacuum-evaporated sample was dissolved in 70% ethanol (the same extraction solvent), it gave consistent results rather than in water or dimethyl sulfoxide. As the sample concentration was increased, the ABTS result was critically affected by the yellow sample pigments and the equation. These results suggest that an ABTS+ stock solution needs to be adjusted with water or PBS buffer, and the same solvent used for extraction is required. In particular, the interference of pigments in the samples needs to be considered in the equation for accurate and consistent ABTS results of the antioxidant activity of plant samples.
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KEYWORD
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ABTS assay, antioxidant activity, Daehwang (Rheum officinale), free radical scavenging activity, in vitro evaluation
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