False Immunosuppressant Measurement by LC-MS/MS Method Due to Radiopaque Agents

Author(s): Ataman Gönel*, Ismail Koyuncu .

Journal Name: Combinatorial Chemistry & High Throughput Screening

Volume 22 , Issue 2 , 2019

Become EABM
Become Reviewer


Background: Although liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) is preferred as a reliable method, some molecules in the blood matrix may lead to false positive or false negative results. False positive or negative results show the direction of the deviation rate from the target value.

Aim: The aim of this study was to investigate the effect of seven different radiopaque agents on four immunosuppressant drugs.

Methods: Every agent coded with RM1 to RM7 was added to control materials containing tacrolimus, everolimus, sirolimus, and cyclosporine A drugs. Measurements were performed using an LC-MS/MS instrument. Bias values were calculated to detect the deviation rates.

Results: All RMs led to false negative results in the tacrolimus and cyclosporine A levels at a rate of -19.77% (95% CI, -27.16 to 12.52) to -44.45% (95% CI, -49.20 to -39.69). The smallest deviations were seen in the everolimus levels with the administration of RM6 (gadodiamide) and in the sirolimus levels with RM1 (gadobutrol) at the rates of 4.04% (95% CI, -11.36 to -3.17) and 2.11% (95% CI, -7.18 to 7.11), respectively. The most affected drug by RM4 (gadopentetate dimeglumine salt) was sirolimus at the rate of 114.01% (95% CI, 97.31 - 130.76). RM5 (gadodiamide) interfered cyclosporine A at the most. The highest deviations were observed with the administration of RM3 (iohexol) in the everolimus and sirolimus levels at the rates of 153.72% (95% CI, 142.44 to 164.78) and 171.41% (95% CI, 157.91 to 184.97), respectively.

Conclusion: Radiopaque agents interfered the measurement of immunosuppressant drugs. Especially, everolimus and sirolimus levels were affected due to using iohexol. The choice of gadodiamide or ioversol is important to reduce the risk of interference for everolimus measurement. The blood samples should be obtained for measurement of drug levels before contrast-enhanced imaging.

Keywords: Immunosuppressant, radiopaque agents, interference, drug levels, blood, LC-MS/MS method.

Grinyó, J.M. Why is organ transplantation clinically important? Cold Spring Harb. Perspect. Med., 2013, 3(6)a014985
Humar, A.; Ramcharan, T.; Denny, R.; Gillingham, K.J.; Payne, W.D.; Matas, A.J. Are wound complications after a kidney transplant more common with modern immunosuppression? Transplantation, 2001, 72(12), 1920-1923.
Duncan, M.D.; Wilkes, D.S. Transplant-related immune suppression: a review of immunosuppression and pulmonary infections. Proc. Am. Thorac. Soc., 2005, 2(5), 449-455.
Krasowski, M.D.; Drees, D.; Morris, C.S.; Maakestad, J.; Blau, J.L.; Ekins, S. Cross-reactivity of steroid hormone immunoassays: Clinical significance and two-dimensional molecular similarity prediction. BMC Clin. Pathol., 2014, 14(1), 33.
Simpson, J.; Zhang, Q.; Ozaeta, P.; Aboleneen, H. A specific method for the measurement of cyclosporin A in human whole blood by liquid chromatography-tandem mass spectrometry. Ther. Drug Monit., 1998, 20(3), 294-300.
Kim, H-J.; Kang, J-S. Matrix effects: Hurdle for development and validation of bioanalytical LC–MS methods in biological samples analyses. Biodesign, 2016, 4(2), 46-58.
Matuszewski, B.; Constanzer, M.; Chavez-Eng, C. Strategies for the assessment of matrix effect in quantitative bioanalytical methods based on HPLC− MS/MS. Anal. Chem., 2003, 75(13), 3019-3030.
Park, Y.J.; Rim, J.H.; Yim, J.; Lee, S-G.; Kim, J-H. Effects of two types of medical contrast media on routine chemistry results by three automated chemistry analyzers. Clin. Biochem., 2017, 50(12), 719-725.
Dams, R.; Huestis, M.A.; Lambert, W.E.; Murphy, C.M. Matrix effect in bio-analysis of illicit drugs with LC-MS/MS: Influence of ionization type, sample preparation, and biofluid. J. Am. Soc. Mass Spectrom., 2003, 14(11), 1290-1294.
Matuszewski, B.; Constanzer, M.; Chavez-Eng, C. Matrix effect in quantitative LC/MS/MS analyses of biological fluids: a method for determination of finasteride in human plasma at picogram per milliliter concentrations. Anal. Chem., 1998, 70(5), 882-889.
Tang, L.; Kebarle, P. Dependence of ion intensity in electrospray mass spectrometry on the concentration of the analytes in the electrosprayed solution. Anal. Chem., 1993, 65(24), 3654-3668.
Buchwald, A.; Winkler, K.; Epting, T. Validation of an LC-MS/MS method to determine five immunosuppressants with deuterated internal standards including MPA. BMC Clin. Pharmacol., 2012, 12(1), 2.
Srinivas, T.R.; Meier-Kriesche, H-U. Minimizing immune-suppression, an alternative approach to reducing side effects: objectives and interim result. Clin. J. Am. Soc. Nephrol., 2008, 3(Suppl. 2), S101-S16.
Christians, U.; Klawitter, J.; Klawitter, J.; Brunner, N.; Schmitz, V. Biomarkers of immunosuppressant organ toxicity after transplantation: status, concepts and misconceptions. Expert Opin. Drug Metab. Toxicol., 2011, 7(2), 175-200.
McShane, A.J.; Bunch, D.R.; Wang, S. Therapeutic drug moni-toring of immunosuppressants by liquid chromatography–mass spectrometry. Clin. Chim. Acta, 2016, 454, 1-5.
Thomas, B.; Weir, M.R. Rejection of the Kidney Transplant; Nephrology Secrets: First South Asia Edition-E-Book Elsevier:. , 2018, p. 410.
Freudenberger, K.; Hilbig, U.; Gauglitz, G. Recent advances in therapeutic drug monitoring of immunosuppressive drugs. TrAC. Trends Analyt. Chem., 2016, 79, 257-268.
de Jonge, H.; Geerts, I.; Declercq, P.; de Loor, H.; Claes, K.; Desmet, K.; Kuypers, D.R. Apparent elevation of cyclosporine whole blood concentrations in a renal allograft recipient. Ther. Drug Monit., 2010, 32(5), 529-531.
Morris, R.G.; Salm, P.; Taylor, P.J.; Wicks, F.A.; Theodossi, A. Comparison of the reintroduced MEIA assay with HPLC-MS/MS for the determination of whole-blood sirolimus from transplant recipients. Ther. Drug Monit., 2006, 28(2), 164-168.
Schmid, R.W.; Lotz, J.; Schweigert, R.; Lackner, K.; Aimo, G.; Friese, J.; Rosiere, T.; Dickson, D.; Kenney, D.; Maine, G.T. Multi-site analytical evaluation of a chemiluminescent magnetic microparticle immunoassay (CMIA) for sirolimus on the Abbott ARCHITECT analyzer. Clin. Biochem., 2009, 42(15), 1543-1548.
Dasgupta, A.; Davis, B.; Chow, L. Evaluation of QMS everolimus assay using Hitachi 917 Analyzer: Comparison with liquid chromatography/mass spectrometry. Ther. Drug Monit., 2011, 33(2), 149-154.
Hoffer, E.; Kurnik, D.; Efrati, E.; Scherb, I.; Karasik, M.; Ring, G.; Bentur, Y. Comparison of everolimus QMS immunoassay on architect ci4100 and liquid chromatography/mass spectrometry: Lack of agreement in organ-transplanted patients. Ther. Drug Monit., 2015, 37(2), 214-219.
Sallustio, B.C.; Noll, B.D.; Morris, R.G. Comparison of blood sirolimus, tacrolimus and everolimus concentrations measured by LC-MS/MS, HPLC-UV and immunoassay methods. Clin. Biochem., 2011, 44(2), 231-236.
Armendariz, Y.; Garcia, S.; Lopez, R.M.; Pou, L. Hematocrit influences immunoassay performance for the measurement of tacrolimus in whole blood. Ther. Drug Monit., 2005, 27(6), 766-769.
Westley, I.S.; Taylor, P.J.; Salm, P.; Morris, R.G. Cloned enzyme donor immunoassay tacrolimus assay compared with high-performance liquid chromatography-tandem mass spectrometry and microparticle enzyme immunoassay in liver and renal transplant recipients. Ther. Drug Monit., 2007, 29(5), 584-591.
Bazin, C.; Guinedor, A.; Barau, C.; Gozalo, C.; Grimbert, P.; Duvoux, C.; Furlan, V.; Massias, L.; Hulin, A. Evaluation of the Architect tacrolimus assay in kidney, liver, and heart transplant recipients. J. Pharm. Biomed. Anal., 2010, 53(4), 997-1002.
Altinier, S.; Varagnolo, M.; Zaninotto, M.; Boccagni, P.; Plebani, M. Heterophilic antibody interference in a non-endogenous molecule assay: an apparent elevation in the tacrolimus concentration. Clin. Chim. Acta, 2009, 402(1-2), 193-195.
Taylor, P.J. Matrix effects: The Achilles heel of quantitative high-performance liquid chromatography–electrospray–tandem mass spectrometry. Clin. Biochem., 2005, 38(4), 328-334.
George, R.; Haywood, A.; Khan, S.; Radovanovic, M.; Simmonds, J.; Norris, R. Enhancement and suppression of ionization in drug analysis using HPLC-MS/MS in support of therapeutic drug monitoring: A review of current knowledge of its minimization and assessment. Ther. Drug Monit., 2018, 40(1), 1-8.
Bonfiglio, R.; King, R.C.; Olah, T.V.; Merkle, K. The effects of sample preparation methods on the variability of the electrospray ionization response for model drug compounds. Rapid Commun. Mass Spectrom., 1999, 13(12), 1175-1185.
King, R.; Bonfiglio, R.; Fernandez-Metzler, C.; Miller-Stein, C.; Olah, T. Mechanistic investigation of ionization suppression in electrospray ionization. J. Am. Soc. Mass Spectrom., 2000, 11(11), 942-950.
Tong, X.S.; Wang, J.; Zheng, S.; Pivnichny, J.V.; Griffin, P.R.; Shen, X.; Donnelly, M.; Vakerich, K.; Nunes, C.; Fenyk-Melody, J. Effect of signal interference from dosing excipients on pharmacokinetic screening of drug candidates by liquid chromatography/mass spectrometry. Anal. Chem., 2002, 74(24), 6305-6313.

Rights & PermissionsPrintExport Cite as

Article Details

Year: 2019
Page: [129 - 134]
Pages: 6
DOI: 10.2174/1386207322666190418125307
Price: $58

Article Metrics

PDF: 23
PRC: 1