Abstract
Abstract— A simple fluorometric method for the estimation of endogenous 3,4‐dihydroxyphenylethylene glycol sulphate (DOPEG‐SO4) in brain tissue is described. DOPEG‐SO4 was extracted from brain tissue with 0.4 n‐perchloric acid and purified by column chromatography on QAE‐Sephadex A‐25, Cl− form. The isolated DOPEG‐SO4 was hydrolysed by incubation with a purified preparation of sulphatase from Helix pomatia. The free DOPEG thus formed was further purified on a column of aluminium oxide and determined fluorometrically after conversion to a fluorescent compound by condensation with ethylenediamine.The sensitivity of this method was about 15 ng per perchloric acid extract applied on the column (3 ml) and the reproducibility was excellent, with a variation coefficient of 1.6% (n = 5). The concentration of DOPEG‐SO4 in whole brains of rats, determined by the present method, was 79.3 ± 4.7 ng/g (mean ± S.D., n = 10) in terms of free DOPEG. No appreciable DOPEG‐SO, was detected in the brains of mice and guinea‐pigs.
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