Determination of methylprednisolone in rat tissue by high-performance liquid chromatography

Methylprednisolone was determined in various types of rat tissue following an intravenous injection of methylprednisolone sodium succinate. Two modes of tissue work-up were investigated: digestion with subtilisin Carlsberg, a proteolytic enzyme, and homogenization with methanol. The final determination was by reversed-phase high-performance liquid chromatography with dexamethasone as internal standard. The extraction yields of methylprednisolone and dexamethasone from tissue homogenate and the extraction yield of methylprednisolone after incubation with viable tissue were determined. The experiments show that methylprednisolone and the internal standard are extracted in similar yields from tissue homogenates and that methylprednisolone can be recovered in a good yield after incubation with viable tissue, provided that the tissue does not have a high metabolic activity. There was a good agreement between the analytical results from the two different types of tissue work-up. The method of analysis proved feasible for pharmacokinetic work.