On the application of 4-hydroxybenzoic acid as a trapping agent to study hydroxyl radical generation during cerebral ischemia and reperfusion

  • Liu Miao
  • L. Steven Peterson
  • Miyake Minoru
  • Liu Miao
  • Liu KeJian
Part of the Developments in Molecular and Cellular Biochemistry book series (DMCB, volume 37)


Aromatic hydroxylation from the reaction between hydroxyl radical and salicylate or its related compounds has been often utilized as a marker for the generation of hydroxyl radicals. We have investigated several technical aspects of applying this method to study hydroxyl radical production during cerebral ischemia and reperfusion using the hydroxylation of 4-hydroxybenzoic acid (4-HBA) to form 3,4-dihydroxybenzoic acid (3,4-DHBA). 4-HBA was administered to rats either through intravenous infusion, or by way of an in vivo microdialysis probe implanted in the brain. Dialysate containing 3,4-DHBA was collected and analyzed by HPLC with electrochemical detection. An endogenous compound was found to co-elute with 3,4 -DHBA but could be separated by varying the chromatographic conditions. Because of interrupted blood flow during cerebral ischemia and reperfusion, delivery of 4-HBA through the microdialysis probe is a preferred method to systemic administration such as intravenous infusion. It is concluded that the oxidation of 4-HBA to 3,4-DHBA can be a reliable and accurate indicator for the formation of hydroxyl radical in vivo if the experiments are well designed to avoid potential pitfalls associated with technical difficulties of the method. (Mol Cell Biochem 234/235: 379-385, 2002)

Key words

hydroxyl radical 4-DHBA 4-HBA microdialysis HPLC cerebral ischemia and reperfusion 


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Copyright information

© Springer Science+Business Media New York 2002

Authors and Affiliations

  • Liu Miao
    • 1
  • L. Steven Peterson
    • 1
  • Miyake Minoru
    • 1
  • Liu Miao
    • 1
    • 2
  • Liu KeJian
    • 1
  1. 1.1College of PharmacyUniversity of New MexicoAlbuquerqueUSA
  2. 2.2Kagawa Medical UniversityKagawaJapan

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