Measurement of hydroxyl radical in rat blood vessel by microbore liquid chromatography and electrochemical detection: an on-line microdialysis study

Salicylic acid (0.5 mM) is used as a trapping reagent of hydroxyl radical, and the formed 2,3- and 2,5-dihydroxybenzoic acids were collected via an on -line microdialysis device from the blood vessels. This study revealed the use of a sensitive liquid chromatographic system with electrochemical detec tion for the determination of 2,3- and 2,5-dihydroxybenzoic acids. Mobile p hase consisted of 0.1 M monochloroacetic acid, 10 mM EDTA, 0.5 mM sodium oc tylsulfate, 20% acetonitrile and 5% tetrahydrofuran in 1 l (pH 3.0 adjusted with 1 M NaOH), and the flow-rate of 0.05 ml/min were found to be optimum. Isocratic separation of these adducts on a microbore column (reversed-phas e C-18, 150X1 mm I.D., 5 mu m) was achieved within 10 min. The optimal appl ied potential of dihydroxybenzoic acids was set at 750 mV based on a hydrod ynamic study. This method has the detection limits of 1.3 pmol/ml (or 0.2 n g/ml) for 2,3- and 2,5-dihydroxybenzoic acids in Ringer solution (at signal -to-noise ratio=3). (C) 1999 Elsevier Science BN. All rights reserved.