Banerjee Dibyajyoti, Madhusoodanan U K, Sharanabasappa M, Ghosh Sandip, Jacob Jose
Department of Biochemistry, Kasturba Medical College, Manipal 576119, Karnataka State, India.
Clin Chim Acta. 2003 Nov;337(1-2):147-52. doi: 10.1016/j.cccn.2003.08.004.
Hydroperoxides are well-recognized reactive oxygen species which are associated with oxidative stress, a phenomenon of current clinical interest as oxidative stress is associated with a number of disease condition. Ferrous ion oxidation xylenol orange (FOX) methods of hydroperoxide estimation has outdated other methods available for hydroperoxide estimation. Two versions FOX assays are described in the literature, FOX-1 and FOX-2, in which FOX-1 is more sensitive.
we increased the sensitivity of FOX-1 assay by stabilizing the reagent pH 1.7-1.8. Analogous to FOX-2 assay, we have modified FOX-1 assay by using it in conjunction with triphenylphosphine and butylated hydroxytoluene, thus increasing the specificity of FOX-1 assay for hydroperoxide. By modified FOX-1 method, we estimated plasma hydroperoxide concentration of normal human subjects and of diabetic patients and compared with FOX-2 method.
The FOX-1 method showed a significant high value of plasma hydroperoxide concentration compared to FOX-2 method both in normal subjects and diabetic patients with a significant correlation. By modified FOX-1 method, the recovery percentage of cumene hydroperoxide was better in biological samples when compared to FOX-2 method.
The modified FOX-1 method is equally specific for hydroperoxide determination when compared to FOX-2 but is more sensitive.
氢过氧化物是公认的活性氧物质,与氧化应激相关,氧化应激是当前临床关注的一种现象,因为它与多种疾病状况有关。亚铁离子氧化二甲酚橙(FOX)法用于氢过氧化物的测定,已使其他可用的氢过氧化物测定方法过时。文献中描述了两种版本的FOX测定法,即FOX-1和FOX-2,其中FOX-1更灵敏。
我们通过将试剂pH稳定在1.7 - 1.8来提高FOX-1测定法的灵敏度。与FOX-2测定法类似,我们通过将FOX-1测定法与三苯基膦和丁基化羟基甲苯结合使用来对其进行改进,从而提高FOX-1测定法对氢过氧化物的特异性。通过改进的FOX-1方法,我们测定了正常人和糖尿病患者的血浆氢过氧化物浓度,并与FOX-2方法进行了比较。
在正常受试者和糖尿病患者中,FOX-1方法显示的血浆氢过氧化物浓度值均显著高于FOX-2方法,且具有显著相关性。与FOX-2方法相比,通过改进的FOX-1方法,生物样品中氢过氧化异丙苯的回收率更高。
与FOX-2相比,改进的FOX-1方法在氢过氧化物测定方面同样具有特异性,但更灵敏。