A radioenzymatic technique for the measurement of free and conjugated 3,4-dihydroxyphenylethylene-glycol in brain tissue and biological fluids.
A simple, sensitive and specific radioenzymatic assay for the measurement of 3,4-dihydroxyphenylethyleneglycol (DOPEG) was developed. The assay is based on the conversion of the compound to its O-methylated derivative in the presence of catechol-O-methyltransferase and [3H]S-adenosyl-methionine. The tritiated 3-methoxy-4-hydroxyphenylethyleneglycol formed is selectively extracted in organic solvents and isolated by thin layer chromatography. After oxidation to vanillin the O-methylated compound is extracted and measured by liquid scintillation spectrophotometry. This assay has been applied to the measurement of free and conjugated DOPEG in a variety of biological tissues and fluids. Both free and conjugated DOPEG were readily detected in discrete rat brain areas. Substantial amounts of free and conjugated DOPEG were also measured in ventricular perfusates from freely moving rats. Finally, the presence of DOPEG was also demonstrated in human cerebrospinal fluid, plasma and urine. Only the free form of DOPEG was found in cerebrospinal fluid, whereas both unconjugated and conjugated forms were present in plasma and urine.
D000818:Animals; D001923:Brain Chemistry; D002250:Carbon Radioisotopes; D002394:Catechol O-Methyltransferase; D006018:Glycols / Q000032:analysis*; D008297:Male; D008734:Methoxyhydroxyphenylglycol / Q000031:analogs & derivatives / Q000032:analysis*; D011865:Radioisotope Dilution Technique; D051381:Rats; D011919:Rats, Inbred Strains; D012436:S-Adenosylmethionine; D014316:Tritium
D002250:Carbon Radioisotopes; D006018:Glycols; D014316:Tritium; D008734:Methoxyhydroxyphenylglycol; D012436:S-Adenosylmethionine; D002394:Catechol O-Methyltransferase; C010117:3,4-dihydroxyphenylglycol