Department of Nutrition and Food Science, XaRTA-INSA, University of Barcelona, 08028 Barcelona, Spain.
Anal Biochem. 2013 Mar 1;434(1):172-7. doi: 10.1016/j.ab.2012.11.021. Epub 2012 Dec 7.
Plasma hydroperoxides (HPs) are widely accepted to be good indicators of oxidative stress. By means of the method proposed here, which uses diphenyl-1-pyrenylphosphine (DPPP) as a fluorescent probe, all types of plasma HP were determined. The limits of detection and quantification of the method were 0.08 and 0.25 nmol of cumene hydroperoxide (CHP) equivalents in 40 μl of plasma, respectively. The method is satisfactory in terms of precision (5.3% for 14.5 μM CHP eq., n=8), and the recoveries were 91% and 92% after standard additions of 26 and 52 μM CHP, respectively. The selectivity of the proposed method is higher than 96%. Moreover, optimization of the reaction conditions and the addition of ethylenediaminetetraacetic acid (EDTA) disodium salt and 2,6-di-tert-butyl-4-methylphenol (BHT) prevented the formation of HP artifacts during the analysis. Therefore, the proposed method is useful for simple and quantitative determination of total plasma HPs.
血浆过氧化物（HPs）被广泛认为是氧化应激的良好指标。通过使用二苯基-1-芘基膦（DPPP）作为荧光探针的方法，可以测定所有类型的血浆 HP。该方法的检测限和定量限分别为 40μl 血浆中 0.08 和 0.25nmol 过氧化枯烯（CHP）当量。该方法在精度方面令人满意（14.5μM CHP eq. 的 5.3%，n=8），标准添加 26 和 52μM CHP 后的回收率分别为 91%和 92%。所提出方法的选择性高于 96%。此外，通过优化反应条件并添加乙二胺四乙酸二钠（EDTA）和 2,6-二叔丁基-4-甲基苯酚（BHT），可以防止在分析过程中形成 HP 伪影。因此，该方法可用于简单定量测定总血浆 HPs。
