title: Multicenter evaluation of the thermo scientific prelude for measurement of immunosuppressant drugs using sample preparation liquid chromatography-tandem mass spectrometry.
abstract: Immunosuppressant drugs (ISDs) are commonly prescribed to solid organ transplant patients. Their narrow therapeutic index and potential for toxicity necessitates careful monitoring of blood concentrations. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) methods are increasingly used for ISD measurement. However, there remain many challenges with this methodology, particularly regarding interassay variability. The Thermo Scientific Prelude is an online extraction/liquid chromatography platform that uses turbulent flow technology coupled with MS/MS. A multicenter evaluation of the Prelude for the measurement of cyclosporine A, tacrolimus, and sirolimus is described. ISDs were measured at each site using standardized protocols. Sample preparation liquid chromatography-MS/MS was performed using the Prelude coupled to a TSQ Vantage. Chromatography was achieved with a Cyclone-P TurboFlow/Accucore C8 column combination using a multisolvent loading and eluting pump system. Mass spectrometry acquisitions were performed in selective reaction monitoring mode and data processed using TraceFinder (version 3.1). Multisite mean imprecision for cyclosporine A ranged from 8.8% (54 mcg/L) to 9.8% (450 mcg/L); for tacrolimus, 4.7% (15.5 mcg/L) to 12.6% (2.5 mcg/L); for sirolimus, 7.4% (19.9 mcg/L) to 16.5% (2.6 mcg/L). Approximately 110 specimens were used for method comparison. For cyclosporine A, mean bias against the multisite mean ranged from -18% to 1%; for tacrolimus, values ranged from -7% to 4%; for sirolimus, values ranged from -4% to 2%. Comparisons of multisite mean Prelude results with routine ISD method results was also performed for cyclosporine A (slope = 0.7878, intercept = 24.16, r = 0.98), tacrolimus (slope = 0.9391, intercept = 0.1017, r = 98), and sirolimus (slope = 0.9618, intercept = 0.1483, r = 0.97). The Prelude ISD method offers acceptable and comparable multisite performance. This study has also highlighted the importance of adopting standardized protocols and LC-MS/MS methods for better comparability between ISD assays.
authors: *Department of Laboratory Medicine, Boston Children's Hospital, MA; †Thermo Scientific, Franklin, MA; ‡Department of Clinical Pathology, Cleveland Clinic LL Lab, OH; and §Department of Pathology, Johns Hopkins University Hospital, Baltimore, MD.
authors: Roy W A
authors: RW
authors: Peake
authors: Christina R
authors: CR
authors: Hartigan
authors: Christopher L
authors: CL
authors: Esposito
authors: Mark D
authors: MD
authors: Kellogg
authors: Jessica
authors: J
authors: Gabler
authors: Sihe
authors: S
authors: Wang
authors: Autumn
authors: A
authors: Breaud
authors: Joseph
authors: J
authors: Di Bussolo
authors: Joshua
authors: J
authors: Moskowitz
authors: William
authors: W
authors: Clarke
mesh_terms: D002853:Chromatography, Liquid / Q000379:methods*; D016572:Cyclosporine / Q000097:blood; D016903:Drug Monitoring / Q000379:methods*; D006801:Humans; D007166:Immunosuppressive Agents / Q000097:blood*; D020123:Sirolimus / Q000097:blood; D016559:Tacrolimus / Q000097:blood; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D007166:Immunosuppressive Agents; D016572:Cyclosporine; D020123:Sirolimus; D016559:Tacrolimus