Winyard Paul G, Knight Iona A, Shaw Frances L, Rocks Sophie A, Davies Claire A, Eggleton Paul, Haigh Richard, Whiteman Matthew, Benjamin Nigel
Peninsula Medical School, Universities of Exeter and Plymouth, St. Luke's Campus, Exeter, United Kingdom.
Methods Enzymol. 2008;441:151-60. doi: 10.1016/S0076-6879(08)01208-1.
S-Nitroso moieties, such as the S-nitroso group within S-nitrosated albumin, constitute a potential endogenous reservoir of nitric oxide (NO.) in human tissues and other biological systems. Moreover, S-nitroso compounds are under investigation as therapeutic agents in humans. Therefore, it is important to be able to detect S-nitrosothiols (RSNOs) in human extracellular fluids, such as plasma and synovial fluid, as well as other biological samples. This chapter describes a method for the determination of S-nitrosothiols in biofluids. The method is based on electron paramagnetic resonance (EPR) spectrometry, in combination with spin trapping using a ferrous ion complex of the iron chelator N-methyl-d-glucamine dithiocarbamate under alkaline conditions. This iron complex mediates the decomposition of RSNO to NO., as well as spin trapping the generated NO.. The resulting spin adduct has a unique EPR signal that can be quantified.
S-亚硝基部分，如S-亚硝基化白蛋白中的S-亚硝基基团，是人体组织和其他生物系统中一氧化氮（NO·）的潜在内源性储存库。此外，S-亚硝基化合物正在作为人类治疗剂进行研究。因此，能够检测人体细胞外液（如血浆和滑液）以及其他生物样品中的S-亚硝基硫醇（RSNOs）非常重要。本章介绍了一种测定生物流体中S-亚硝基硫醇的方法。该方法基于电子顺磁共振（EPR）光谱法，并结合在碱性条件下使用铁螯合剂N-甲基-d-葡糖胺二硫代氨基甲酸盐的亚铁离子络合物进行自旋捕获。这种铁络合物介导RSNO分解为NO·，同时自旋捕获生成的NO·。产生的自旋加合物具有独特的EPR信号，可以进行定量分析。
