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Original Article

Discriminative EPR detection of NO and HNO by encapsulated nitronyl nitroxides

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Pages 74-81 | Received 28 Aug 2012, Accepted 31 Oct 2012, Published online: 28 Nov 2012
 

Abstract

Nitric oxide, NO, is one of the most important molecules in the biochemistry of living organisms. By contrast, nitroxyl, NO, one-electron reduced analog of NO which exists at physiological conditions in its protonated form, HNO, has been relatively overlooked. Recent data show that HNO might be produced endogenously and display unique biological effects. However, there is a lack of specific and quantitative methods of detection of endogenous HNO production. Here we present a new method for discriminative NO and HNO detection by nitronyl nitroxides (NNs) using electron paramagnetic resonance (EPR). It was found that NNs react with NO and HNO with similar rate constants of about 104 M− 1s− 1 but yield different products: imino nitroxides and the hydroxylamine of imino nitroxides, correspondingly. An EPR approach for discriminative NO and HNO detection using liposome-encapsulated NNs was developed. The membrane barrier of liposomes protects NNs against reduction in biological systems while is permeable to both analytes, NO and HNO. The sensitivity of this approach for the detection of the rates of NO/HNO generation is about 1 nM/s. The application of encapsulated NNs for real-time discriminative NO/HNO detection might become a valuable tool in nitric oxide-related studies.

Declaration of interest

The authors report no declarations of interest. The authors alone are responsible for the content and writing of the paper.

This work was partly supported by NIH grant HL089036.

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