A reverse-phase liquid chromatographic assay for flavin-containing monooxygenase activity.
A rapid, convenient assay for flavin-containing monooxygenase activity is described. The method is based on direct analysis of quenched incubation mixtures by reverse-phase liquid chromatography, and utilizes p-nitrophenyl-1,3-oxathiolane as the substrate. The synthesis of the substrate and the product are described. The usefulness of p-nitrophenyl-1,3-oxathiolane S-oxide formation as a measure of flavin-containing monooxygenase activity was demonstrated using highly purified and microsomal hog and rat liver flavin-containing monooxygenase. The assay is especially useful for determining stereoselectivity of flavin-containing monooxygenase activity in small amounts of crude tissue preparations.
Cashman
J R
JR
Department of Pharmaceutical Chemistry, School of Pharmacy, University of California, San Francisco 94143.
Proudfoot
J
J
D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006571:Heterocyclic Compounds; D008862:Microsomes, Liver / Q000201:enzymology; D009578:Nitrobenzenes; D010105:Oxygenases / Q000032:analysis*; D051381:Rats; D013552:Swine; D013876:Thiophenes
D006571:Heterocyclic Compounds; D009578:Nitrobenzenes; D013876:Thiophenes; C059208:4-nitrophenyl-1,3-oxathiolane; D010105:Oxygenases; C020066:dimethylaniline monooxygenase (N-oxide forming)