Detection of S-nitrosothiols and other nitric oxide derivatives by photolysis-chemiluminescence spectrometry

Abstract

We have developed a method for the detection of biologically relevant derivatives of NO· that couples photolysis to chemiluminescence. S- Nitrosothiols, N-nitroso-L-tryptophan, and iron-nitrosyl derivatives can all be detected by this methodology, and these derivatives can be distinguished from free NO· by making measurements with and without photolysis. The limits of detection of this system are in the 10 nM range, with an intraassay variability of less than 2% and an interassay variability of less than 1.5%. The use of cutoff filters and alterations in pH and flow rate through the photolysis cell provide additional useful parameters with which to characterize specific NO· derivatives based on their intrinsic stability. This methodology is a useful addition to the growing armamentarium of techniques used to characterize the biochemistry and metabolism of NO·.