T. Alebickolbah et Iw. Wainer, 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, Journal of chromatography, 646(2), 1993, pp. 289-295
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.