The relative proportions of sirolimus metabolites in blood using HPLC with mass-spectrometric detection

Measurement of sirolimus in blood as a guide to dose adjustment is an accepted practice. To date, most data have resulted from the use of a chromatographic technique. With the imminent introduction of an immunoassay into this field, there is a need to know whether metabolites that could interfere with the performance of this assay, causing a bias compared with measurements made by a chromatographic assay, vary over a period of time or with changes in concomitant immunosuppressive therapy. This preliminary study measured several sirolimus metabolites in blood samples from a variety of clinical settings, using high-performance liquid chromatography with tandem mass-spectrometric detection. Two metabolites known to cross-react in one immunoassay system, single hydroxylation products and 41-O-demethyl rapamycin, were found to constitute the bulk of the metabolic products. They were also found to form a remarkably stable proportion of all metabolites measured, both with respect to the time since transplantation and the concomitant use of cyclosporine or tacrolimus. It is concluded that the analytical bias due to cross-reactivity with metabolites, inherent in this immunoassay, should be consistent across a wide spectrum of patients receiving the drug.