Barbiturate N-demethylase activity in isolated rat hepatocytes.
The isolated hepatocyte preparation has become an increasingly popular system for studying xenobiotic metabolism, and there is a need for methods of determining metabolic activity. Described here is a simple and sensitive radiometric assay for barbiturate N-demethylase activity in the isolated hepatocyte suspension. The demethylation of 1,3-dimethylphenobarbital is determined by measuring the formation of its only significant metabolite, N-methylphenobarbital. The reaction is a typical P-450-mediated dealkylation.
D000818:Animals; D001463:Barbiturates / Q000378:metabolism*; D002845:Chromatography / Q000379:methods; D066298:In Vitro Techniques; D008099:Liver / Q000201:enzymology*; D008297:Male; D010089:Oxidoreductases, N-Demethylating / Q000302:isolation & purification*; D010634:Phenobarbital / Q000031:analogs & derivatives / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains
D001463:Barbiturates; C026540:dimethylphenobarbital; D010089:Oxidoreductases, N-Demethylating; D010634:Phenobarbital