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Original

Formation of TEMPOL-hydroxylamine during reaction between TEMPOL and hydroxyl radical: HPLC/ECD study

, , , , , & show all
Pages 505-512 | Received 21 Feb 2008, Published online: 07 Jul 2009
 

Abstract

Nitroxyl radicals are important antioxidants that have been used to protect animal tissues from oxidative damage. Their reaction with hydroxyl radical (OH) is generally accepted to be the mechanism of antioxidant function. However, the direct interaction of nitroxyl radicals with OH does not always provide a satisfactory explanation in various pH, because the concentration of hydrogen ion may affect the generation of secondary OH-derived radicals. In the present study, it was confirmed that the reaction between 4-hydroxy-2,2,6,6-tetramethylpiperidine-N-oxyl (TEMPOL) and OH generated TEMPOL-hydroxylamine, 4-oxo-2,2,6,6-tetramethylpiperidine-N-oxyl (TEMPON) and TEMPON-hydroxylamine using HPLC coupled with electrochemical detection. In the absence of NADH, TEMPOL-H may be generated by the reaction with secondary OH-derived radicals in acidic condition. In the presence of NADH, a large proportion of the non-paramagnetic products was TEMPOL-H. Finally, it was clarified that TEMPOL-H was generated during dopamine metabolism, which is believed to be one of the OH sources in pathological processes such as Parkinson's disease.

Abbreviations
ESR=

electron spin resonance

ECD=

electrochemical detection

HPLC=

high performance liquid chromatography

MAO=

monoamine oxidase

OH=

hydroxyl radical

TEMPOL=

4-hydroxy-2,2,6,6-tetramethylpiperidine-N-oxyl

TEMPOL-H=

4-hydroxy-2,2,6,6-tetramethyl-N-hydroxypiperidine

TEMPON=

4-oxo-2,2,6,6-tetramethylpiperidine-N-oxyl

TEMPON-H=

4-oxo-2,2,6,6-tetramethyl-N-hydroxypiperidine

Abbreviations
ESR=

electron spin resonance

ECD=

electrochemical detection

HPLC=

high performance liquid chromatography

MAO=

monoamine oxidase

OH=

hydroxyl radical

TEMPOL=

4-hydroxy-2,2,6,6-tetramethylpiperidine-N-oxyl

TEMPOL-H=

4-hydroxy-2,2,6,6-tetramethyl-N-hydroxypiperidine

TEMPON=

4-oxo-2,2,6,6-tetramethylpiperidine-N-oxyl

TEMPON-H=

4-oxo-2,2,6,6-tetramethyl-N-hydroxypiperidine

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