Analysis of S-nitrosothiols via copper cysteine (2C) and copper cysteine-carbon monoxide (3C) methods

Methods. 2013 Aug 1;62(2):123-9. doi: 10.1016/j.ymeth.2012.10.008. Epub 2012 Oct 29.

Abstract

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.

Keywords: Analysis; Carbon monoxide; Copper cysteine; Nitric oxide; S-nitrosothiol.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Calibration
  • Carbon Monoxide / chemistry*
  • Copper / chemistry*
  • Cysteine / chemistry*
  • Erythrocytes / chemistry
  • Gases / chemistry
  • Humans
  • Luminescent Measurements / standards
  • Reference Standards
  • S-Nitrosothiols / chemistry*

Substances

  • Gases
  • S-Nitrosothiols
  • Copper
  • Carbon Monoxide
  • Cysteine