Simultaneous determination of glucocorticoid alcohols, their succinates and hydrocortisone in plasma.

A reversed-phase high-performance liquid chromatographic (HPLC) assay is described for the simultaneous determination of methylprednisolone, methylprednisolone-21-hemisuccinate and endogenous hydrocortisone in biological fluids. This assay is also applicable to the determination of prednisolone-21-hemisuccinate in the presence of prednisolone and hydrocortisone in biological fluids. Prednisolone and hydrocortisone are determined by normal-phase HPLC. The stability of hemisuccinate esters in ampoules, in saline and in plasma has been studied. Whereas the esters were stable in saline at 37 degrees C, they were considerably hydrolysed in plasma at the same temperature. Comparison of hydrocortisone levels obtained by HPLC and radioimmunoassay (RIA) showed the large influence of the cross-reactivity of methylprednisolone and hydrocortisone in the presence of high serum concentrations of methylprednisolone.