Separate determination of human urinary conjugated and unconjugated 3-methoxy-4-hydroxyphenylethyleneglycol

A new determination procedure for human urinary 3-methoxy-4-hydroxyphenylethyleneglycol (MHPG) was established. In addition to the previously established solid-phase extraction method for unconjugated MHPG, another solid-phase extraction method for conjugated MHPG was developed. Unconjugated MHPG was adsorbed on a Sep-Pak Diol cartridge and selectively recovered by elution with ethyl acetate. The eluate was evaporated and the residue was redissolved and analyzed by reversed-phase high-performance liquid chromatography with fluorimetric detection. Conjugated (sulfate plus glucuronide) MHPG was adsorbed on a Sep-Pak Accell QMA cartridge and quantitatively eluted with 0.2 M NaCl. After enzymatic hydrolysis, deconjugated MHPG was extracted using a Sep-Pak Diol cartridge and analyzed in the same manner as unconjugated MHPG. The new method is simple and rapid and can quantitate conjugated and unconjugated MHPG discriminatively.