Determination of S-nitrosothiols in biological and clinical samples using electron paramagnetic resonance spectrometry with spin trapping.
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.
Peninsula Medical School, Universities of Exeter and Plymouth, St. Luke's Campus, Exeter, United Kingdom.
Paul G
PG
Winyard
Iona A
IA
Knight
Frances L
FL
Shaw
Sophie A
SA
Rocks
Claire A
CA
Davies
Paul
P
Eggleton
Richard
R
Haigh
Matthew
M
Whiteman
Nigel
N
Benjamin
D000818:Animals; D004578:Electron Spin Resonance Spectroscopy / Q000379:methods*; D006801:Humans; D026403:S-Nitrosothiols / Q000032:analysis* / Q000097:blood* / Q000737:chemistry; D019097:Spin Trapping / Q000379:methods*
D026403:S-Nitrosothiols