Tacrolimus (FK 506) measurement by immunoassays in clinical samples of organ transplant patients often lacks a specific reference method. A method combining liquid chromatography (LC) with tandem mass spectrometry (MS/MS) was developed to quantify tacrolimus in whole blood. Liquid-liquid extraction was performed on 1 ml of sample before narrow-bore LC/MS/MS analysis. Ascomycin was used as an internal standard. The standard curve was composed of seven points ranging from 1 to 50 micrograms/l (average r2 = 0.9999). Limits of detection and quantitation were 0.25 and 0.75 microgram/l, respectively. Imprecision was < 5% across the therapeutic range. Tacrolimus recovery averaged 62%. The most abundant metabolites detected in clinical samples were 13-O- and 15-O-demethyl tacrolimus. This method was used in a comparison study with a microparticle enzyme immunoassay (MEIA): MEIA = 1.03 LC/MS/MS -0.084 (microgram/l), (Sy/x = 1.43), r2 = 0.933. With its high sensitivity and specificity, this LC/MS/MS method presents a good reference method for immunoassay evaluation as well as a valuable tool for metabolism studies.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.