ENZYME-BASED HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY STATIONARY PHASESAS METABOLIC REACTORS - IMMOBILIZATION OF NON-SOLUBILIZED RAT-LIVER MICROSOMES ON AN IMMOBILIZED ARTIFICIAL MEMBRANE HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY SUPPORT

Non-solubilized rat liver microsomes have been non-covalently immobili zed on an immobilized artificial membrane (IAM) HPLC stationary phase. The microsomes were immobilized on the IAM interphase of both loose I AM support and in a 1 cm x 3 mm I.D. HPLC column filled with IAM stati onary phase. The activity of the immobilized microsomes was assessed b y following the formation of 7-hydroxycoumarin (7-OHC) from the O-deet hylation of 7-ethoxycoumarin. The 7-OHC was quantified by either HPLC analysis of an aliquot from an incubation mixture containing the loose microsome-IAM support or directly using the chromatogram from the mic rosome-IAM HPLC column. Both forms of the IAM-immobilized microsomes w ere active. The result of this study indicate that the IAM stationary phase can be used to produce immobilized microsomal reactors which can be used in HPLC systems for direct on-line determination of biosynthe tic and metabolic processes.