Separation of dihydroxybenzoates, indicators of in-vivo hydroxyl free radical formation, in the presence of transmitter amines and some metabolites in rodent brain, using high-performance liquid chromatography with electrochemical detection.
Based on a detailed study of retention parameters, reversed-phase ion-pair chromatographic methods were developed for the simultaneous determination of dihydroxybenzoates, indicators of in-vivo hydroxyl free radical formation, transmitter amines and some metabolites to facilitate neurochemical investigations in rodent brain. Coupling of the separation methods with electrochemical detection and the use of short-chain perfluorinated carboxylic acids for ion-pairing, allowed for a fast and sensitive determination of salicylate-derived 2,3- and 2,5-dihydroxybenzoic acids and the major electroactive, hydroxylated aromatic compounds present in brain samples. Detection limits for the dihydroxybenzoates (signal-to-noise ratio = 2) were 18-22 fmol injected on the column. Basal levels of 2,3-dihydroxybenzoate and 2,5-dihydroxybenzoate in the the striatum of mice treated with salicylate were 72 +/- 13 and 94 +/- 11 ng/g wet tissue, respectively.
Patthy
M
M
Institute for Drug Research, Budapest, Hungary.
Király
I
I
Sziráki
I
I
D000818:Animals; D001679:Biogenic Amines / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003342:Corpus Striatum / Q000378:metabolism*; D004563:Electrochemistry; D005841:Gentisates; D062385:Hydroxybenzoates / Q000302:isolation & purification*; D017665:Hydroxyl Radical; D051379:Mice; D008810:Mice, Inbred C57BL
D001679:Biogenic Amines; D005841:Gentisates; D062385:Hydroxybenzoates; D017665:Hydroxyl Radical; C009135:2,3-dihydroxybenzoic acid; C010925:2,5-dihydroxybenzoic acid