Detection of hydroxyl radicals in the post-ischemic reperfused heart using salicylate as a trapping agent

Tomoya Onodera, Muhammad Ashraf

Research output: Contribution to journalArticle

70 Citations (Scopus)

Abstract

The formation of hydroxyl radical in the post-ischemic reperfused heart was measured with high performance liquid chromatography and ultraviolet detection using salicylic acid. Hydroxyl radicals react with salicylic acid yielding 2,3- and 2,5-dihydroxybenzoic acid, which can be separated by the liquid chromatography. Isolated rat hearts were perfused with 1 mm salicylic acid and were subjected to 30 mins of global ischemia followed by aerobic or anaerobic reperfusion at 37°C. The effluent from the hearts was collected at various intervals, extracted with ether, and injected into the high performance liquid chromatography unit. 2,5-dihydroxybenzoic acid was present only after aerobic reperfusion and was not detected before ischemia. The liquid chromatography peak of 2,3-dihydroxybenzoic acid was too small for quantitation. The concentration of 2,5-dihydroxybenzoic acid was the highest within 300 s of reperfusion. 2,5-dihydroxybenzoic acid was not detected in the ischemic hearts during anaerobic reperfusion. In ischemic hearts perfused with mannitol, the amount of 2,5-dihydroxybenzoic acid after reperfusion was reduced. These data suggest that hydroxyl radicals are produced in the post-ischemic reperfused heart and that the present method is useful and reliable for the measurement of hydroxyl radicals in the heart.

Original languageEnglish (US)
Pages (from-to)365-370
Number of pages6
JournalJournal of molecular and cellular cardiology
Volume23
Issue number3
DOIs
StatePublished - Jan 1 1991
Externally publishedYes

Fingerprint

Salicylates
Hydroxyl Radical
Reperfusion
Salicylic Acid
Liquid Chromatography
Ischemia
High Pressure Liquid Chromatography
Mannitol
Ether
2,5-dihydroxybenzoic acid

Keywords

  • Hydroxyl radical
  • Oxygen-free radicals
  • Radical trapping agent
  • Salicylic acid
  • liquid chromatography
  • post-ischemic reperfusion

ASJC Scopus subject areas

  • Molecular Biology
  • Cardiology and Cardiovascular Medicine

Cite this

Detection of hydroxyl radicals in the post-ischemic reperfused heart using salicylate as a trapping agent. / Onodera, Tomoya; Ashraf, Muhammad.

In: Journal of molecular and cellular cardiology, Vol. 23, No. 3, 01.01.1991, p. 365-370.

Research output: Contribution to journalArticle

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AB - The formation of hydroxyl radical in the post-ischemic reperfused heart was measured with high performance liquid chromatography and ultraviolet detection using salicylic acid. Hydroxyl radicals react with salicylic acid yielding 2,3- and 2,5-dihydroxybenzoic acid, which can be separated by the liquid chromatography. Isolated rat hearts were perfused with 1 mm salicylic acid and were subjected to 30 mins of global ischemia followed by aerobic or anaerobic reperfusion at 37°C. The effluent from the hearts was collected at various intervals, extracted with ether, and injected into the high performance liquid chromatography unit. 2,5-dihydroxybenzoic acid was present only after aerobic reperfusion and was not detected before ischemia. The liquid chromatography peak of 2,3-dihydroxybenzoic acid was too small for quantitation. The concentration of 2,5-dihydroxybenzoic acid was the highest within 300 s of reperfusion. 2,5-dihydroxybenzoic acid was not detected in the ischemic hearts during anaerobic reperfusion. In ischemic hearts perfused with mannitol, the amount of 2,5-dihydroxybenzoic acid after reperfusion was reduced. These data suggest that hydroxyl radicals are produced in the post-ischemic reperfused heart and that the present method is useful and reliable for the measurement of hydroxyl radicals in the heart.

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