A post-column enzyme reactor for detection of oxidized cholesterols in h.p.l.c. separations

An enzyme reactor is used as a post-column detector after the h.p.l.e. separation of a mixture of cholesterol and some auto-oxidation products. The enzyme reactor contains cholesterol oxidase immobilized on controlled-pore glass. A method for purification of the enzyme is described. The properties and the solvent dependence of the reactor are discussed. A post-column system for dilution of the eluent with aqueous buffer is described.