Monitoring in Situ Liver Metabolism in Rats Using Microdialysis. Comparison of Microdialysis Mass-Transport Model Predictions to Experimental Metabolite Generation Data

Abstract

□ The generation of metabolites from two model compounds, phenacetin and acetaminophen, included in the perfusion fluid of a microdialysis probe implanted into rat liver was studied. When 60μM phenacetin was included in the perfusion fluid using a flow rate of 1.0μL/min, acetaminophen and acetaminophen sulfate were recovered at concentrations that ranged between 0.4 and 1.6μM. Acetaminophen sulfate ([AS]gain) diffused back into the microdialysis probe on a micromolar percentage basis of 8.9±2.4% (n=3) when acetaminophen was passed through the probe at a concentration between 11 and 12μM. When 220–240μM acetaminophen was passed through the probe, the percentage of acetaminophen sulfate recovered was 4.8±1.4% (n=3) (P<0.1 compared to the 11μM group). No acetaminophen glucuronide was detected in the dialysate samples. A mathematical model that describes mass transport in microdialysis sampling was used to predict the concentration of metabolite that could be recovered into the dialysate after the loss of a substrate compound that undergoes metabolism. The model predicts a metabolite recovery of 23.6% using estimates for phenacetin metabolism and 21.5% using estimates for acetaminophen metabolism. The results presented here indicate that microdialysis has potential to be used to study local in situ metabolism and with further refinements of the microdialysis mass-transport model may be used to estimate in vivo metabolic formation rates.