Efficient Assay for Total Antioxidant Capacity in Human Plasma Using a 96-Well Microplate

• Published in: Journal of Clinical Biochemistry and Nutrition Vol. 44; no. 1; pp. 46 - 51
• Main Authors: Kambayashi, Yasuhiro, Binh, Nguyen Thanh, Asakura, Hiroki W., Hibino, Yuri, Hitomi, Yoshiaki, Nakamura, Hiroyuki, Ogino, Keiki
• Format: Journal Article
• Language: English
• Published: Japan SOCIETY FOR FREE RADICAL RESEARCH JAPAN 01.01.2009; Japan Science and Technology Agency; the Society for Free Radical Research Japan
• Subjects: ABTS; human plasma; lag time; microplate; Original; total antioxidant capacity; microplate; lag time; total antioxidant capacity; ABTS; human plasma
• ISSN: 0912-0009; 1880-5086
• DOI: 10.3164/jcbn.08-162

In the present study, we tried to establish an efficient assay for total antioxidant capacity (TAC) in human plasma using a 96-well microplate. TAC was assessed using lag time by antioxidants against the myoglobin-induced oxidation of 2,2'-azino-di(3-ethylbenzthiazoline-6-sulfonic acid (ABTS) with hydrogen peroxide, and expressed as Trolox equivalent. The linearity of the calibration curve with Trolox was maintained with the Trolox concentration range from 2.5 μM to 25 μM (R2 = 0.997). The assay was applied to the measurement of TAC in healthy human plasma. Coefficient of variation in intraday assay was 2.4%. Difference was not observed in interday assay. Plasma TAC of men ((569 ± 41) μM Trolox equivalent; n = 6) was higher than that of women ((430 ± 28) μM Trolox equivalent; n = 4). After the vegetable juice was drunk for 1 week, the increase in plasma TAC was observed in almost all the volunteers. In summary, we developed the efficient assay for plasma TAC using a 96-well microplate.