Hydrocortisone is essential to maintain the normal physiological status of liver in tissue-engineered systems. Using microfluidic bioreactors containing human hepatocytes with and without Kupfer cells, we conducted experiments to determine hydrocortisone clearance and metabolite production at starting concentrations ranging from 100 nM down to normal physiological concentrations, i.e., 20-30 nM. Clearance rate measurements and metabolite identifications were carried out with ultra high performance liquid chromatography and high-mass-accuracy mass spectrometry. Under non-inflamed conditions, 100 nM administered hydrocortisone was depleted to approximately 20 nM in the culture medium after 48 hours, with good first-order kinetics. Cortisone, the inactive metabolite of hydrocortisone, if present, was below the detection limit. Accurate masses corresponding to five potential hydrocortisone metabolites were observed at levels estimated at 1-14 nM, but several of these have identical molecular weights, so structural confirmation awaits additional data. The majority of these metabolites were accounted for by reductase/oxidoreductase metabolism, although a small amount of a metabolite tentatively identified as 6b-hydroxycortisol suggest the involvement of P4503A4. Results of current experiments to complete the metabolite identification will be presented.

Hydrocortisone metabolite profiling in a human microphysiological liver bioreactor

246th National Meeting (2013)

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