Abstract
Cyclosporine A, everolimus, sirolimus, and tacrolimus are the most commonly used immunosuppressant drugs in organ transplant and auto-immune patients. The narrow therapeutic window of these immunosuppressant drugs requires close monitoring of drug blood levels to ensure proper therapeutic response. A quick, robust high-throughput liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was introduced for monitoring whole blood levels of these immunosuppressant drugs with the use of the MassTrak™ Immunosuppressant kit. The assay was carried out in 96-well plate format and requires a simple precipitation step; after which, the supernatant is subjected to liquid chromatography separation (2 min total run time) using a C18 Cartridge column. Identification and quantitation of cyclosporine A, everolimus, sirolimus, and tacrolimus was achieved by employing multiple reaction monitoring (MRM) in positive mode electrospray ionization (ESI). The method exhibits a linear measuring range from 10 to 1500 ng/mL (Cyclosporine A), 1.0–30.0 ng/mL (Everolimus), 1.0–26.0 ng/mL (Sirolimus), and 1.0–30.0 ng/mL (Tacrolimus) and has a within-run and between-run imprecision of <10%.
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References
Morris PJ (2004) Transplantation--a medical miracle of the 20th century. N Engl J Med 351(26):2678–2680
Duncan MD, Wilkes DS (2005) Transplant-related immunosuppression. Proc Am Thorac Soc 2(5):449–455
Claeys E, Vermeire K (2019) Immunosuppressive drugs in organ transplantation to prevent allograft rejection: mode of action and side effects. J Immunol Sci 3(4):14–21
Koster RA et al (2009) Robust, high-throughput LC-MS/MS method for therapeutic drug monitoring of cyclosporine, tacrolimus, everolimus, and sirolimus in whole blood. Ther Drug Monit 1(1):116–125
Bjergum MW, Jannetto PJ, Langman LJ (2019) Simultaneous determination of tacrolimus and cyclosporine A in whole blood by ultrafast LC-MS/MS. In: Langman L, Snozek C (eds) LC-MS in drug analysis, Methods in molecular biology, vol 1872. Humana Press, New York
Hober S et al (2020) Evaluation of the first immunosuppressive drug assay available on a fully automated LC-MS/MS-based clinical analyzer suggests a new era in laboratory medicine. Clin Chem Lab Med 59(5):913–920
Seger C, Salzmann L (2020) After another decade: LC-MS/MS became routine in clinical diagnostics. Clin Biochem 82:2–11
Upadhyay V et al (2012) Rapid and sensitive UPLC-MS-MS determination of tacrolimus in Wistar rats and human blood. J Chromatogr Sci 52(1):59–67
Buchwald A et al (2012) Validation of an LC-MS/MS method to determine five immunosuppressants with deuterated internal standards including MPA. BMC Clin Pharmacol 12(2):2–11
Waters Literature (2023) Waters Inc. https://www.waters.com/waters/library.htm?locale=en_US&lid=134946076. Accessed 27 Feb 2023
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Zhang, A., Conklin, S.E. (2024). Multiplex Immunosuppressant (ISD) Method for the Measurement of Cyclosporine A, Tacrolimus, Sirolimus/Rapamycin, and Everolimus in Whole Blood Using MassTrak™ Kit. In: Garg, U. (eds) Clinical Applications of Mass Spectrometry in Drug Analysis. Methods in Molecular Biology, vol 2737. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3541-4_28
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DOI: https://doi.org/10.1007/978-1-0716-3541-4_28
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Publisher Name: Humana, New York, NY
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