Comparison of blood sirolimus, tacrolimus and everolimus concentrations measured by LC-MS/MS, HPLC-UV and immunoassay methods.

摘要

OBJECTIVES: An LC-MS/MS method was developed for simultaneous quantitation of tacrolimus, sirolimus and everolimus in whole blood, and compared to HPLC-UV and immunoassay methods. DESIGN AND METHODS: Blood (0.1mL) was analysed following solid-phase extraction and chromatographic resolution using a C18 column (45 degrees C) and mobile phase of methanol/40mM ammonium acetate/glacial acetic acid (83/17/0.1) at 200muL/min, with positive electrospray ionisation and multiple reaction monitoring. RESULTS: Intra- and inter-day imprecision and inaccuracy were 30% bias in FPIA everolimus concentrations measured in pooled patient samples versus spiked drug-free whole blood. CONCLUSIONS: LC-MS/MS provides significant accuracy and precision advantages compared to HPLC and immunoassays. Discrepancies in everolimus concentrations measured by the Seradyn FPIA immunoassay require further investigation.