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.

Therapeutic drug monitoring of immunosuppressants by liquid chromatography–mass spectrometry

“…The largest mean difference from the reference measurement (exact matching-isotope dilution MS) was 4.4% [52]. A unified method by Thermo Fisher's Prelude online extraction and LC system coupled to a TSQ Vantage MS platform was evaluated for measuring cyclosporine A, tacrolimus, and sirolimus by 3 hospital labs [53]. The site bias for each immunosuppressant was calculated against the multisite mean.…”

“…Lastly, a fourth group found that differences in hematocrit (20% to 45%), spotted blood volume (25,35,50,75, and 100 μL), and punch location did not affect immunosuppressant quantitation [39]. [46,47] Online solid phase extraction (analytical run time of 1 min) [48] No LC separation (analytical run time of 13 s) [49] Liquid handler [50] Commercialization/ Standardization 7-Lab tacrolimus unification [52] 3-Lab cyclosporine A, tacrolimus, and sirolimus unification [53] Much of these DBS validation studies involve the deposition of venous blood from a collection tube onto the paper. Two recent studies evaluated capillary blood from a finger stick.…”

Recent advances in analytical methods for the therapeutic drug monitoring of immunosuppressive drugs

“…Evaluations of LC–MS/MS for commercially available immunosuppressant assays have also been reported . Peake et al reported a multicenter evaluation of Thermo Fisher's Prelude for the measurement of CsA, TAC, and SIR. A unified method by the Prelude online extraction and LC system coupled to a TSQ Vantage MS platform was evaluated in 3 hospital laboratories.…”

“…55,56 Levine et al 55 Evaluations of LC-MS/MS for commercially available immunosuppressant assays have also been reported. [57][58][59]…”

Antineoplastic drugs and their analysis: a state of the art review

“…589 The sensitivity and selectivity provided by MS helped to ease sample preparation in clinical chemistry laboratories. Column-switching systems coupling the extraction support to the chromatographic column before MS detection were also used for the analysis of sirolimus [598][599][600][601][602][603][604][605][606][607][608][609][610][611] and everolimus 598,599,602,603,[605][606][607]609,[612][613][614] in whole blood. LLE 615 or SPE 616 on 96-well plates constitutes another approach for sample preparation automatisation.…”