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Quantitation of Teriflunomide in Human Serum/Plasma Across a 40,000-Fold Concentration Range by LC/MS/MS

  • Geoffrey S. RuleEmail author
  • Alan L. Rockwood
  • Kamisha L. Johnson-Davis
Part of the Methods in Molecular Biology book series (MIMB, volume 1383)

Abstract

Leflunomide is a prodrug used primarily for treatment of rheumatoid arthritis. The active metabolite, teriflunomide (A77 1726), inhibits the enzyme dihydroorotate dehydrogenase and thereby reduces the synthesis of pyrimidine ribonucleotides. Teriflunomide is also administered directly and finds use in treating multiple sclerosis. Therapeutic concentrations are generally in the tens of μg/mL serum or plasma and, due to adverse effects and the time required to reach steady state, therapeutic drug monitoring is beneficial. The drug is also a potential teratogen.

A method was developed and validated to quantify the drug teriflunomide over a 40,000-fold concentration range of 5 ng/mL to 200 μg/mL in serum or plasma. This is accomplished by dividing the quantitative range into two separate but overlapping regions; a high curve and a low curve range. Samples are evaluated first against the high curve after a 100-fold dilution of the sample extract. Samples falling below the upper curve region are evaluated again without dilution and quantified, if possible, against the low curve calibration standards. Appropriate choice of a concentration for the deuterated internal standard (D4-teriflunomide) allows for a single, identical, extraction procedure to be performed for both curve regions but with the dilution performed for high curve samples. The method is rugged and reliable with good accuracy and precision statistics.

Key words

Leflunomide Teriflunomide Dynamic range LC/MS/MS Arthritis 

Notes

Acknowledgements

The authors would like to thank the ARUP Institute for Clinical and Experimental Pathology for making this work possible.

References

  1. 1.
    Schattenkirchner M (2000) The use of leflunomide in the treatment of rheumatoid arthritis: an experimental and clinical review. Immunopharmacology 47(2–3):291–298CrossRefPubMedGoogle Scholar
  2. 2.
    Oh J, O’Connor PW (2013) Teriflunomide for the treatment of multiple sclerosis. Semin Neurol 33(1):45–55CrossRefPubMedGoogle Scholar
  3. 3.
    Chacko B, John GT (2012) Leflunomide for cytomegalovirus: bench to bedside. Transpl Infect Dis 14(2):111–120CrossRefPubMedGoogle Scholar
  4. 4.
    Teschner S, Gerke P, Geyer M, Wilpert J, Krumme B, Benzing T et al (2009) Leflunomide therapy for polyomavirus-induced allograft nephropathy: efficient BK virus elimination without increased risk of rejection. Transplant Proc 41(6):2533–2538CrossRefPubMedGoogle Scholar
  5. 5.
    Babic-Naglic D, Anic B, Novak S, Grazio S, Martinavic Kaliterna D (2010) Treatment of rheumatoid and psoriatic arthritis—review of leflunomide. Reumatizam 57(2):161–162PubMedGoogle Scholar
  6. 6.
    Schmidt A, Schwind B, Gillich M, Brune K, Hinz B (2003) Simultaneous determination of leflunomide and its active metabolite, A77 1726, in human plasma by high-performance liquid chromatography. Biomed Chromatogr 17(4):276–281CrossRefPubMedGoogle Scholar
  7. 7.
    Brent RL (2001) Teratogen update: reproductive risks of leflunomide (Arava); a pyrimidine synthesis inhibitor: counseling women taking leflunomide before or during pregnancy and men taking leflunomide who are contemplating fathering a child. Teratology 63(2):106–112CrossRefPubMedGoogle Scholar
  8. 8.
    Chan V, Charles BG, Tett SE (2004) Rapid determination of the active leflunomide metabolite A77 1726 in human plasma by high-performance liquid chromatography. J Chromatogr B Analyt Technol Biomed Life Sci 803(2):331–335CrossRefPubMedGoogle Scholar
  9. 9.
    van Roon EN, Yska JP, Raemaekers J, Jansen TL, van Wanrooy M, Brouwers JR (2004) A rapid and simple determination of A77 1726 in human serum by high-performance liquid chromatography and its application for optimization of leflunomide therapy. J Pharm Biomed Anal 36(1):17–22CrossRefPubMedGoogle Scholar
  10. 10.
    Parekh JM, Vaghela RN, Sutariya DK, Sanyal M, Yadav M, Shrivastav PS (2010) Chromatographic separation and sensitive determination of teriflunomide, an active metabolite of leflunomide in human plasma by liquid chromatography-tandem mass spectrometry. J Chromatogr B Analyt Technol Biomed Life Sci 878(24):2217–2225CrossRefPubMedGoogle Scholar
  11. 11.
    Rule GS, Clark ZD, Yue B, Rockwood AL (2013) Correction for isotopic interferences between analyte and internal standard in quantitative mass spectrometry by a nonlinear calibration function. Anal Chem 85(8):3879–3885CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Geoffrey S. Rule
    • 1
    Email author
  • Alan L. Rockwood
    • 2
  • Kamisha L. Johnson-Davis
    • 3
  1. 1.Institute for Clinical and Experimental PathologyARUP LaboratoriesSalt Lake CityUSA
  2. 2.Department of PathologyUniversity of Utah School of MedicineSalt Lake CityUSA
  3. 3.Department of PathologyUniversity of Utah School of Medicine, ARUP Institute for Clinical and Experimental PathologySalt Lake CityUSA

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