Ogasawara Yuki, Mukai Yoshio, Togawa Tadayasu, Suzuki Toshihiro, Tanabe Shinzo, Ishii Kazuyuki
Departments of Environmental Biology, Meiji Pharmaceutical University, 2-522-1 Noshio, Kiyose, Tokyo 204-8588, Japan.
J Chromatogr B Analyt Technol Biomed Life Sci. 2007 Jan 1;845(1):157-63. doi: 10.1016/j.jchromb.2006.08.006. Epub 2006 Sep 8.
We examined the influence of oxidative stress on the relative amounts of various albumin-bound thiols in human plasma. To determine the ratio of thiols existing as mixed disulfides following oxidation, we developed a method combining fast purification of albumin using affinity columns and high-performance liquid chromatography (HPLC) with fluorescence detection for low molecular weight thiols which were labeled after reduction. When the effect of exposure of plasma to radical oxygen species on binding of thiols to albumin was determined by the present method, significant increases in the ratio of cysteine bound to albumin (Alb-Cys) to total cysteine were clearly demonstrated.
我们研究了氧化应激对人血浆中各种与白蛋白结合的硫醇相对含量的影响。为了确定氧化后以混合二硫键形式存在的硫醇比例，我们开发了一种方法，该方法将使用亲和柱快速纯化白蛋白与高效液相色谱（HPLC）相结合，并对还原后标记的低分子量硫醇进行荧光检测。当用本方法测定血浆暴露于活性氧对硫醇与白蛋白结合的影响时，与白蛋白结合的半胱氨酸（Alb-Cys）与总半胱氨酸的比例显著增加得到了明确证实。
