Efficient Assay for Total Antioxidant Capacity in Human Plasma Using a 96-Well Microplate
Efficient Assay for Total Antioxidant Capacity in Human Plasma Using a 96-Well Microplate
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DOI：
10.3164/jcbn.08-162
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发表时间：
2009-01-01
影响因子：
2.4
通讯作者：
Ogino, Keiki
中科院分区：
文献类型：
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作者：
Kambayashi, Yasuhiro;Binh, Nguyen Thanh;Ogino, Keiki
在本研究中，我们试图建立一个有效的测定总抗氧化能力（TAC）在人血浆中使用96孔微孔板。使用抗氧化剂对抗肌红蛋白诱导的2，2 '-连氮基-二（3-乙基苯并噻唑啉-6-磺酸（ABTS）与过氧化氢的氧化的滞后时间来评估TAC，并表示为Trolox当量。Trolox浓度范围为2.5 μ M至25 μ M（R-2 = 0.997）时，Trolox校准曲线保持线性。将该测定法应用于健康人血浆中TAC的测量。日内测定的变异系数为2.4%。日间测定无差异。男性的血浆TAC（（569 +/- 41）μ M Trolox当量; n = 6）高于女性（（430 +/- 28）μ M Trolox当量; n = 4）。饮用蔬菜汁1周后，几乎所有志愿者的血浆TAC均升高。总之，我们开发了使用96孔微孔板的血浆TAC的有效测定。
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 mu M to 25 mu M (R-2 = 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) mu M Trolox equivalent; n = 6) was higher than that of women ((430 +/- 28) mu M 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.