The salicylate hydroxylation assay to measure hydroxyl free radicals induced by local application of glutamate in vivo or induced by the Fenton reaction in vitro

P Teismann, B Ferger

Research output: Contribution to journalArticle

43 Citations (Scopus)


The direct measurement of hydroxyl radicals in vivo is extremely difficult. Therefore, the indirect determination of hydroxyl radicals using salicylate (2-hydroxybenzoate) is widely accepted. Reverse microdialysis with glutamate led to a dose-dependent production of hydroxyl free radicals indicated by the hydroxylation adduct of salicylate, namely 2,3-dihydroxybenzoic acid. The local stimulation of hydroxyl free radical formation seems to be suitable to investigate a radical-scavenging property of potential neuroprotective drugs. In vitro experiments using the Fenton reaction may be a helpful tool to assess whether or not a substance is able to act as a radical scavenger in a cell free environment, which is easy to handle and a simple screening method before in vivo experiments were performed. In the present study we present an in vivo approach using local application of glutamate into the striatum and an in vitro screening using the Fenton reaction to induce hydroxyl radical formation. The main goal is to reliable measure hydroxyl free radicals, which are the most reactive oxygen radicals in biology and medicine.
Original languageEnglish
Pages (from-to)204-210
Number of pages7
JournalBrain Research. Brain Research Protocols
Issue number2
Publication statusPublished - 1 Apr 2000



  • animals
  • dose-response relationship, drug
  • gentisates
  • glutamic acid decarboxylase
  • hydroxybenzoic acids
  • hydroxyl radical
  • hydroxylation
  • male
  • methods
  • microdialysis
  • rats
  • rats, wistar
  • salicylates
  • excitotoxicity
  • Fenton system
  • salicylate assay
  • NMDA receptor activation
  • D-aspartate antagonists
  • rat striatum
  • cautionary note
  • MPP+ neurotoxicity
  • tert-butylnitrone PBN
  • substantia-nigra
  • nitric-oxide
  • dopamine
  • neurodegeneration

Cite this