Abstract
Mass spectrometry (MS) is a highly specific and sensitive technique that is used for the detection of many different analytes with diverse chemical characteristics. It has been adopted by clinical laboratories for the quantification of small molecules and, by extension, has been widely used for therapeutic drug monitoring. It is an attractive alternative to immunoassay methods, because it is not subject to the same interferences. A limitation of MS (relative to immunoassays) is the turnaround time. However, this can be addressed by workflow parallelization with other assays. Herein we describe a tandem LC-MS/MS method for the detection and quantification of methotrexate in human plasma with a lower limit of quantification of 0.01 μM and within-assay and between-assay coefficients of variation of less than 15%. This method lacks interference from high-abundance metabolites and utilizes kindred chromatography to improve turnaround time in the therapeutic drug monitoring laboratory.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Jannetto PJ, Fitzgerald RL (2016) Effective use of mass spectrometry in the clinical laboratory. Clin Chem 62:92–98. https://doi.org/10.1373/clinchem.2015.248146
Strathmann FG, Hoofnagle AN (2011) Current and future applications of mass spectrometry to the clinical laboratory. Am J Clin Pathol 136:609–616. https://doi.org/10.1309/AJCPW0TA8OBBNGCK
Garg U, Zhang YV (2016) Mass spectrometry in clinical laboratory: applications in therapeutic drug monitoring and toxicology. In: Clinical applications of mass spectrometry in drug analysis. Springer, New York, pp 1–10
Al-Turkmani MR, Law T, Narla A, Kellogg MD (2010) Difficulty measuring methotrexate in a patient with high-dose methotrexate-induced nephrotoxicity. Clin Chem 56:1792. https://doi.org/10.1373/clinchem.2010.144824
Albertioni F, Rask C, Eksborg S et al (1996) Evaluation of clinical assays for measuring high-dose methotrexate in plasma. Clin Chem 42(39):39–44
Bath RK, Brar NK, Forouhar FA, Wu GY (2014) A review of methotrexate-associated hepatotoxicity. J Dig Dis 15:517–524. https://doi.org/10.1111/1751-2980.12184
Widemann BC, Adamson PC (2006) Understanding and managing methotrexate nephrotoxicity. Oncologist 11:694–703. https://doi.org/10.1634/theoncologist.11-6-694
Cavone JL, Yang D, Wang A (2014) Glucarpidase intervention for delayed methotrexate clearance. Ann Pharmacother 48:897–907. https://doi.org/10.1177/1060028014526159
Green JM (2012) Glucarpidase to combat toxic levels of methotrexate in patients. Ther Clin Risk Manag 8:403–413. https://doi.org/10.2147/TCRM.S30135
Ahmed YAAR, Hasan Y (2013) Prevention and management of high dose methotrexate toxicity. J Cancer Sci Ther 5:106–112. https://doi.org/10.4172/1948-5956.1000193
Evans WE, Crom WR, Abromowitch M et al (1986) Clinical pharmacodynamics of high-dose methotrexate in acute lymphocytic leukemia. N Engl J Med 314:471–477. https://doi.org/10.1056/NEJM198602203140803
Aumente D, Buelga DS, Lukas JC et al (2006) Population pharmacokinetics of high-dose methotrexate in children with acute lymphoblastic leukaemia. Clin Pharmacokinet 45:1227–1238. https://doi.org/10.2165/00003088-200645120-00007
Przybylski M, Preiss J, Dennebaum R, Fischer J (1982) Identification and quantitation of methotrexate and methotrexate metabolites in clinical high-dose therapy by high pressure liquid chromatography and field desorption mass spectrometry. Biomed Mass Spectrom 9:22–32. https://doi.org/10.1002/bms.1200090106
Wu D, Wang Y, Sun Y et al (2015) A simple, rapid and reliable liquid chromatography-mass spectrometry method for determination of methotrexate in human plasma and its application to therapeutic drug monitoring. Biomed Chromatogr BMC 29:1197–1202. https://doi.org/10.1002/bmc.3408
Schofield RC, Ramanathan LV, Murata K et al (2015) Development and validation of a turbulent flow chromatography and tandem mass spectrometry method for the quantitation of methotrexate and its metabolites. J Chromatogr B Analyt Technol Biomed Life Sci 1002:169–175. https://doi.org/10.1016/j.jchromb.2015.08.025
Gunther V, Mueller D, von Eckardstein A, Saleh L (2016) Head to head evaluation of the analytical performance of two commercial methotrexate immunoassays and comparison with liquid chromatography-mass spectrometry and the former fluorescence polarization immunoassay. Clin Chem Lab Med 54:823–831. https://doi.org/10.1515/cclm-2015-0578
Steinborner S, Henion J (1999) Liquid-liquid extraction in the 96-well plate format with SRM LC/MS quantitative determination of methotrexate and its major metabolite in human plasma. Anal Chem 71:2340–2345
Nair H, Lawrence L, Hoofnagle AN (2012) Liquid chromatography–tandem mass spectrometry work flow for parallel quantification of methotrexate and other immunosuppressants. Clin Chem 58:943. https://doi.org/10.1373/clinchem.2011.175067
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2019 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Winston-McPherson, G.N., Schmeling, M., Hoofnagle, A.N. (2019). Quantification of Methotrexate in Human Serum and Plasma by Liquid Chromatography Tandem Mass Spectrometry. In: Langman, L., Snozek, C. (eds) LC-MS in Drug Analysis. Methods in Molecular Biology, vol 1872. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8823-5_10
Download citation
DOI: https://doi.org/10.1007/978-1-4939-8823-5_10
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-8822-8
Online ISBN: 978-1-4939-8823-5
eBook Packages: Springer Protocols