Assay for dopamine beta-hydroxylase in rat serum and adrenal medulla by high-performance liquid chromatography with fluorescence detection.
A highly sensitive method for the assay of dopamine beta-hydroxylase in rat serum and in sample solution prepared from rat adrenal medulla is described which employs high-performance liquid chromatography with fluorescence detection. Octopamine, formed enzymatically from the substrate tyramine, is separated by Dowex 50/-X4 column chromatography and oxidized with periodate to p-hydroxybenzaldehyde, which is then converted into a fluorescent compound with 2,2'-dithiobis(1-aminonaphthalene). The derivative, after extraction with n-hexane--chloroform, is separated by normal-phase chromatography on Alox T. The limit of detection for octopamine formed enzymatically is 10 pmol. This method requires as little as 5 microliter of rat serum.
D000313:Adrenal Medulla / Q000201:enzymology*; D000367:Age Factors; D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004299:Dopamine beta-Hydroxylase / Q000097:blood*; D008297:Male; D051381:Rats; D011919:Rats, Inbred Strains; D012016:Reference Values
D004299:Dopamine beta-Hydroxylase