Department of Pediatrics, Division of Critical Care Medicine, Washington University School of Medicine, 1 Children’s Place, St. Louis, MO 63110, United States.
Methods. 2013 Aug 1;62(2):123-9. doi: 10.1016/j.ymeth.2012.10.008. Epub 2012 Oct 29.
This chapter summarizes the principles of RSNO measurement in the gas phase, utilizing ozone-based chemiluminescence and the copper cysteine (2C)±carbon monoxide (3C) reagent. Although an indirect method for quantifying RSNOs, this assay represents one of the most robust methodologies available. It exploits the NO detection sensitivity of ozone based chemiluminescence, which is within the range required to detect physiological concentrations of RSNO metabolites. Additionally, the specificity of the copper cysteine (2C and 3C) reagent for RSNOs negates the need for sample pretreatment, thereby minimizing the likelihood of sample contamination (false positive results), or the loss of certain highly labile RSNO species. Herein, we outline the principles of this methodology, summarizing key issues, potential pitfalls and corresponding solutions.
本章总结了利用臭氧化学发光和铜半胱氨酸（2C）±一氧化碳（3C）试剂测量气相中 RSNO 的原则。虽然这是一种间接定量 RSNO 的方法，但该测定方法代表了目前可用的最可靠方法之一。它利用了臭氧化学发光对 NO 的检测灵敏度，该灵敏度在检测生理浓度的 RSNO 代谢物所需的范围内。此外，铜半胱氨酸（2C 和 3C）试剂对 RSNO 的特异性消除了对样品预处理的需求，从而最大程度地减少了样品污染（假阳性结果）或某些高度不稳定的 RSNO 物种损失的可能性。在此，我们概述了该方法的原理，总结了关键问题、潜在的陷阱和相应的解决方案。
