We use cookies to improve your experience. By continuing to browse this site, you accept our cookie policy.×

A simple and robust LC–MS/MS method for measuring sirolimus and everolimus in whole blood

    Chao Yuan

    Department of Clinical Pathology, Cleveland Clinic, Cleveland, OH 44195, USA

    ,
    Drew Payto

    Department of Clinical Pathology, Cleveland Clinic, Cleveland, OH 44195, USA

    ,
    Jessica Gabler

    Thermo Fisher Scientific, West Palm Beach, FL 33407, USA

    &
    Sihe Wang

    *Author for correspondence:

    E-mail Address: wangs2@ccf.org

    Department of Clinical Pathology, Cleveland Clinic, Cleveland, OH 44195, USA

    Published Online:https://doi.org/10.4155/bio.14.43

    Therapeutic drug monitoring of immunosuppressants sirolimus and everolimus is mandatory and liquid chromatography tandem mass spectrometry (LC–MS/MS) is the preferred technology for the measurement. Due to the high hydrophobicity these analytes bind to reverse-phase columns tightly and need column heating to elute. Column heating not only requires extra instrument preparation but also causes permanent column damage if the heater is left on while elution pumps stop by the end of the run. The primary improvement in the current method was to elute the analytes at room temperature using special buffers. This new LC–MS/MS method has been validated for clinical use and offers improved simplicity and robustness by eliminating column heating yet with high sensitivity, precision and accuracy.

    Papers of special note have been highlighted as: • of interest; •• of considerable interest

    References

    • 1 Vezina C, Kudelski A, Sehgal S. Rapamycin (AY-22,989), a new antifungal antibiotic. I. Taxonomy of the producing streptomycete and isolation of the active principle. J. Antibiot. (Tokyo) 28(10), 721–726 (1975).
    • 2 Kirchner G, Meier-Wiedenbach I, Manns M. Clinical pharmacokinetics of everolimus. Clin. Pharmacokinet. 43(2), 83–95 (2004).
    • 3 Monchaud C, Marquet P. Pharmacokinetic optimization of immunosuppressive therapy in thoracic transplantation: part I. Clin. Pharmacokinet. 48(7), 419–462 (2009).
    • 4 Faivre S, Kroemer G, Raymond E. Current development of mTOR inhibitors as anticancer agents. Nat. Rev. Drug Discov. 5(8), 671–688 (2006).
    • 5 Sedrani R, Cottens S, Kallen J, Schuler W. Chemical modification of rapamycin: the discovery of SDZ RAD. Transplant. Proc. 30(5), 2192–2194 (1998).
    • 6 Neumayer H, Paradis K, Korn A et al. Entry-into-human study with the novel immunosuppressant SDZ RAD in stable renal transplant recipients. Br. J. Clin. Pharmacol. 48(5), 694–703 (1999).
    • 7 Strom T, Haschke M, Zhang Y et al. Identification of everolimus metabolite patterns in trough blood samples of kidney transplant patients. Ther. Drug Monit. 29(5), 592–599 (2007).
    • 8 Gallant-Haidner H, Trepanier D, Freitag D, Yatscoff R. Pharmacokinetics and metabolism of sirolimus. Ther. Drug Monit. 22(1), 31–35 (2000).
    • 9 Kovarik J, Tedesco H, Pascual J et al. Everolimus therapeutic concentration range defined from a prospective trial with reduced-exposure cyclosporine in de novo kidney transplantation. Ther. Drug Monit. 26(5), 499–505 (2004).
    • 10 Holt D, Denny K, Lee T, Johnston A. Therapeutic monitoring of sirolimus: its contribution to optimal prescription. Transplant. Proc. 35(Suppl. 3), S157–S161 (2003).
    • 11 Kahan B, Napoli K, Kelly P et al. Therapeutic drug monitoring of sirolimus: correlations with efficacy and toxicity. Clin. Transplant. 14(2), 97–109 (2000).
    • 12 Kelly P, Kahan B. Review: metabolism of immunosuppressant drugs. Curr. Drug Metab. 3(3), 275–287 (2002).
    • 13 Johnson-Davis K, De S, Jimenez E, McMillin Ga, De B. Evaluation of the Abbott ARCHITECT i2000 sirolimus assay and comparison with the Abbott IMx sirolimus assay and an established liquid chromatography-tandem mass spectrometry method. Ther. Drug Monit. 33(4), 453–459 (2011). •• Two commercial immunoassays showed positive bias in sirolimus measurement relative to an LC–MS/MS method.
    • 14 Dasgupta A, Davis B, Chow L. Evaluation of QMS everolimus assay using Hitachi 917 Analyzer: comparison with liquid chromatography/mass spectrometry. Ther. Drug Monit. 33(2), 149–154 (2011).
    • 15 Yang Z, Wang S. Recent development in application of high performance liquid chromatography-tandem mass spectrometry in therapeutic drug monitoring of immunosuppressants. J. Immunol. Methods 336(2), 98–103 (2008). •• Recent developments in LC–MS/MS analysis of immunosuppressants are reviewed.
    • 16 Bogusz M, Enazi E, Hassan H, Abdel-Jawaad J, Ruwaily , J, Tufail M. Simultaneous LC–MS-MS determination of cyclosporine A, tacrolimus, and sirolimus in whole blood as well as mycophenolic acid in plasma using common pretreatment procedure. J. Chromatogr. B Anal. Technol. Biomed. Life Sci. 850(1–2), 471–480 (2007).
    • 17 Heideloff C, Payto D, Wang S. Comparison of a stable isotope-labeled and an analog internal standard for the quantification of everolimus by a liquid chromatography-tandem mass spectrometry method. Ther. Drug Monit. 35(2), 246–250 (2013).
    • 18 Korecka M, Solari S, Shaw L. Sensitive, high throughput HPLC–MS/MS method with on-line sample clean-up for everolimus measurement. Ther. Drug Monit. 28(4), 484–490 (2006).
    • 19 Korecka M, Patel R, Shaw L. Evaluation of performance of new, isotopically labeled internal standard ([13c2d4]RAD001) for everolimus using a novel high-performance liquid chromatography tandem mass spectrometry method. Ther. Drug Monit. 33(4), 460–463 (2011).
    • 20 Mueller D, Rentsch K. Sensitive quantification of sirolimus and everolimus by LC–MS/MS with online sample cleanup. J. Chromatogr. B Anal. Technol. Biomed. Life Sci. 878(13–14), 1007–1012 (2010).
    • 21 Meinitzer A, Gartner G, Pilz S, Stettin M. Ultra fast liquid chromatography-tandem mass spectrometry routine method for simultaneous determination of cyclosporin A, tacrolimus, sirolimus, and everolimus in whole blood using deuterated internal standards for cyclosporin A and everolimus. Ther. Drug Monit. 32(1), 61–66 (2010).
    • 22 Taylor P, Franklin M, Graham K, Pillans P. A HPLC-mass spectrometric method suitable for the therapeutic drug monitoring of everolimus. J. Chromatogr. B Anal. Technol. Biomed. Life Sci. 848(2), 208–214 (2007).
    • 23 Mano N, Sato M, Nozawa M et al. An accurate quantitative LC/ESI-MS/MS method for sirolimus in human whole blood. J. Chromatogr. B Anal. Technol. Biomed. Life Sci. 879(13–14), 987–992 (2011).
    • 24 Wang S, Miller A. A rapid liquid chromatography-tandem mass spectrometry analysis of whole blood sirolimus using turbulent flow technology for online extraction. Clin. Chem. Lab. Med. 46(11), 1631–1634 (2008). • This is our original LC–MS/MS method for measuring sirolimus with the use of a column heater.
    • 25 Yuan C, Heideloff C, Kozak M, Wang S. Simultaneous quantification of 19 drugs/metabolites in urine important for pain management by liquid chromatography-tandem mass spectrometry. Clin. Chem. Lab. Med. 50(1), 95–103 (2012).
    • 26 Bonfiglio R, King R, Olah T, Merkle K. The effects of sample preparation methods on the variability of the electrospray ionization response for model drug compounds. Rapid Commun. Mass Spectrom. 13(12), 1175–1185 (1999).
    • 27 Yuan C, Kosewick J, He X, Kozak M, Wang S. Sensitive measurement of serum 1alpha,25-dihydroxyvitamin D by liquid chromatography/tandem mass spectrometry after removing interference with immunoaffinity extraction. Rapid Commun. Mass Spectrom. 25(9), 1241–1249 (2011).
    • 28 Capone D, Gentile A, Polichetti G et al. Stability of sirolimus and everolimus measured by immunoassay techniques in whole blood samples from kidney transplant patients. Int. J. Immunopathol. Pharmacol. 21(2), 297–307 (2008). • Stability of sirolimus and everolimus was investigated at different temperatures.
    • 29 Salm P, Tresillian M, Taylor P, Pillans P. Stability of sirolimus (rapamycin) in whole blood. Ther. Drug Monit. 22(4), 423–426 (2000).
    • 30 Mano N, Nozawa M, Sato M et al. Identification and elimination of ion suppression in the quantitative analysis of sirolimus in human blood by LC/ESI-MS/MS. J. Chromatogr. B Anal. Technol. Biomed. Life Sci. 879(13–14), 968–974 (2011). • Matrix effect (ion supression) in the LC–MS/MS analysis of sirolimus was systematically investigated, and a way of reducing matrix effect was demonstrated.
    • 31 Ivanova M, Artusi C, Polo G, Zaninotto M, Plebani M. High-throughput LC–MS/MS method for monitoring sirolimus and everolimus in the routine clinical laboratory. Clin. Chem. Lab. Med. 49(7), 1151–1158 (2011). • A very fast (3.5 min) and sensitive (lower limit of quantification: 0.2 ng/ml) LC–MS/MS method is presented for measuring sirolimus and everolimus.