A simple method to evaluate liver microsomal enzyme activity for phenol with a high-performance liquid chromatograph.
A method to evaluate liver microsomal cytochrome P-450-related enzyme activity for phenol is described. Phenol of a known quantity q was incubated with enzyme and cofactor for a period of time t. At the end of the incubation, phenol remaining unchanged in the reaction mixture was analyzed with a high-performance liquid chromatograph (HPLC). No direct determination of phenol concentration was necessary: only the ratio of chromatographic peak area a1 obtained from a sample vessel (cofactor solution contained NADP) to the area a from a reference vessel (cofactor solution devoid of NADP) was needed to calculate the rate of enzymatic reaction V, i.e., V = q(1 - a'/a)/t. The time-activity and enzyme-activity studies proved the method to be useful for the evaluation of enzyme activity. This method is expected to have wide application in the field of drug metabolism study.
D000818:Animals; D002851:Chromatography, High Pressure Liquid; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D007700:Kinetics; D008297:Male; D008862:Microsomes, Liver / Q000201:enzymology*; D019800:Phenol; D010636:Phenols / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains; D013379:Substrate Specificity
D010636:Phenols; D019800:Phenol; D003577:Cytochrome P-450 Enzyme System