Simultaneous detection of ubiquinol and ubiquinone in human plasma as a marker of oxidative stress
Simultaneous detection of ubiquinol and ubiquinone in human plasma as a marker of oxidative stress
复制标题
DOI：
10.1006/abio.1997.2187
复制
发表时间：
1997-07-15
影响因子：
2.9
通讯作者：
Yamamoto, Y
中科院分区：
文献类型：
--
作者：
Yamashita, S;Yamamoto, Y
描述了一种同时检测人血浆中泛醇-10 和泛醌-10 的方法。在此过程中，将肝素化人血浆与 5 体积甲醇和 10 体积己烷混合。剧烈摇动和离心后，立即将己烷相的等分试样（5μl）直接注射到反相HPLC上，以尽量减少泛醇到泛醌的氧化。分离后的在线还原柱将泛醌转化为泛醇，并通过电化学检测进行定量。血浆 ubiquinol-10 和 ubiquinone-10 的检测限约为 4 nM，具有良好的重现性，该方法还可以检测生育酚、番茄红素和 β-胡萝卜素。此外，游离胆固醇和胆固醇酯可以通过其在 210 nm 处的吸收来定量。使用这种方法，我们确定健康捐赠者的人血浆中泛醇与泛醌的比例约为 95/5。我们认为这种方法将会很有用，因为泛醇与泛醌的比例已被认为是氧化应激的良好标志。 (C) 1997 年学术出版社。
A method is described for the simultaneous detection of ubiquinol-10 and ubiquinone-10 in human plasma. In this procedure, heparinized human plasma was mixed with 5 vol of methanol and 10 vol of hexane. After vigorous shaking and centrifugation, an aliquot of the hexane phase (5 mu l) was injected immediately and directly onto a reversed-phase HPLC to minimize the oxidation of ubiquinol to ubiquinone. A post-separation, on-line reduction column converts ubiquinone to ubiquinol which is quantified by electrochemical detection. The detection limit of plasma ubiquinol-10 and ubiquinone-10 is about 4 nM With excellent reproducibilities, Tocopherols, lycopene, and beta-carotene are also detectable in this method. In addition, free cholesterol, and cholesteryl esters can be quantified by their absorption at 210 nm. Using this method we have determined the ratio of ubiquinol to ubiquinone is about 95/5 in human plasma from healthy donors. We suggest that this method will be useful since the ratio of ubiquinol to ubiquinone has been suggested as a good marker of oxidative stress. (C) 1997 Academic Press.