The measurement of nitric oxide and its metabolites in biological samples by ozone-based chemiluminescence

Methods Mol Biol. 2008;476:11-28. doi: 10.1007/978-1-59745-129-1_2.


A plethora of publications on techniques and methodologies for measuring nitric oxide (NO) or reaction products of NO (NO metabolites) has served in recent years to complicate and confuse the majority of researchers interested in this field. Here, we provide a practical approach and summarize the key issues and corresponding solutions regarding quantification with the use of ozone-based chemiluminescence, which is the most accurate, sensitive, and widely used NO detection method. We have drawn on the vast experience of leaders in the field to produce this consensus, but the views and implications presented herein represent our own, and we limit our advice to those techniques with which we have direct experience. Hopefully, this guide will allow authors to make more informed decisions regarding NO metabolite measurement methodology, without the need for each subsequent group to rediscover previously observed advantages and pitfalls.

MeSH terms

  • Body Fluids / chemistry*
  • Carbon Monoxide / chemistry
  • Cell Extracts
  • Erythrocytes / chemistry
  • Freezing
  • Heme / chemistry
  • Hydrogen-Ion Concentration
  • Indicators and Reagents
  • Luminescent Measurements / methods*
  • Nitric Oxide / analysis*
  • Nitric Oxide / metabolism*
  • Nitrites / blood
  • Ozone / chemistry*
  • Photolysis
  • Reference Standards
  • Time Factors


  • Cell Extracts
  • Indicators and Reagents
  • Nitrites
  • Nitric Oxide
  • Heme
  • Ozone
  • Carbon Monoxide