Advertisement

Quantitative monitoring of tamoxifen in human plasma extended to 40 metabolites using liquid-chromatography high-resolution mass spectrometry: new investigation capabilities for clinical pharmacology

Abstract

Liquid-chromatography (LC) high-resolution (HR) mass spectrometry (MS) analysis can record HR full scans, a technique of detection that shows comparable selectivity and sensitivity to ion transitions (SRM) performed with triple-quadrupole (TQ)-MS but that allows de facto determination of “all” ions including drug metabolites. This could be of potential utility in in vivo drug metabolism and pharmacovigilance studies in order to have a more comprehensive insight in drug biotransformation profile differences in patients. This simultaneous quantitative and qualitative (Quan/Qual) approach has been tested with 20 patients chronically treated with tamoxifen (TAM). The absolute quantification of TAM and three metabolites in plasma was realized using HR- and TQ-MS and compared. The same LC-HR-MS analysis allowed the identification and relative quantification of 37 additional TAM metabolites. A number of new metabolites were detected in patients’ plasma including metabolites identified as didemethyl-trihydroxy-TAM-glucoside and didemethyl-tetrahydroxy-TAM-glucoside conjugates corresponding to TAM with six and seven biotransformation steps, respectively. Multivariate analysis allowed relevant patterns of metabolites and ratios to be associated with TAM administration and CYP2D6 genotype. Two hydroxylated metabolites, α-OH-TAM and 4′-OH-TAM, were newly identified as putative CYP2D6 substrates. The relative quantification was precise (<20 %), and the semiquantitative estimation suggests that metabolite levels are non-negligible. Metabolites could play an important role in drug toxicity, but their impact on drug-related side effects has been partially neglected due to the tremendous effort needed with previous MS technologies. Using present HR-MS, this situation should evolve with the straightforward determination of drug metabolites, enlarging the possibilities in studying inter- and intra-patients drug metabolism variability and related effects.

This is a preview of subscription content, log in to check access.

Access options

Buy single article

Instant unlimited access to the full article PDF.

US$ 39.95

Price includes VAT for USA

Subscribe to journal

Immediate online access to all issues from 2019. Subscription will auto renew annually.

US$ 157

This is the net price. Taxes to be calculated in checkout.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Abbreviations

HR-MS:

High-resolution mass spectrometry

MDF:

Mass defect filtering

TDM:

Therapeutic drug monitoring

TQ-MS:

Triple-quadrupole mass spectrometer TDM

XIC:

Extracted ions chromatogram

References

  1. 1.

    Committee on Quality of Health Care in America To Err Is Human (2000) Building a Safer Health System. In: Kohn LT, Corrigan JM, Donaldson MS (eds) National Academy Press. Washington, DC

  2. 2.

    Preventable Adverse Drug Reactions: A Focus on Drug Interactions. U.S. Food and Drug Administration. Accessed 30 Oct. 2013. http://www.fda.gov/drugs/developmentapprovalprocess/developmentresources/druginteractionslabeling/ucm110632.htm

  3. 3.

    Gautier S, Bachelet H, Bordet R, Caron J (2003) The cost of adverse drug reactions. Expert Opin Pharmacother 4:319–326

  4. 4.

    U.S. Food and Drug Administration. Guidance for Industry: Safety Testing of Drug Metabolites. Accessed 30 Oct. 2013. http://www.fda.gov/downloads/Drugs/GuidanceComplianceRegulatoryInformation/Guidances/ucm079266.pdf

  5. 5.

    International Conference on Harmonisation. Guidance on nonclinical safety studies for the conduct of human clinical trials and marketing authorization for pharmaceuticals M3(R2). Accessed 30 Oct 2013. http://www.fda.gov/downloads/Drugs/GuidanceComplianceRegulatoryInformation/Guidances/ucm073246.pdf

  6. 6.

    Gao H, Obach RS (2011) Addressing MIST (Metabolites in Safety Testing): bioanalytical approaches to address metabolite exposures in humans and animals. Curr Drug Metab 12:578–586

  7. 7.

    Testa B, Krämer SD (2009) The biochemistry of drug metabolism—an introduction: part 5. Metabolism and bioactivity. Chem Biodivers 6:591–684

  8. 8.

    Stachulski AV, Baillie TA, Kevin Park B, Scott Obach R, Dalvie DK, Williams DP, Srivastava A, Regan SL, Antoine DJ, Goldring CE, Chia AJ, Kitteringham NR, Randle LE, Callan H, Castrejon JL, Farrell J, Naisbitt DJ, Lennard MS (2013) The generation, detection, and effects of reactive drug metabolites. Med Res Rev 33:985–1080

  9. 9.

    Guengerich FP (2011) Mechanisms of drug toxicity and relevance to pharmaceutical development. Drug Metab Pharmacokinet 26:3–14

  10. 10.

    Krämer SD, Testa B (2008) The biochemistry of drug metabolism—an introduction: part 6. Inter-individual factors affecting drug metabolism. Chem Biodivers 5:2465–2578

  11. 11.

    Krämer SD, Testa B (2009) The biochemistry of drug metabolism—an introduction: part 7. Intra-individual factors affecting drug metabolism. Chem Biodivers 6:1477–1660

  12. 12.

    The importance of pharmacovigilance. Safety Monitoring of medicinal products. World Health Organization 2002 http://apps.who.int/medicinedocs/pdf/s4893e/s4893e.pdf

  13. 13.

    Johnson MD, Zuo H, Lee KH, Trebley JP, Rae JM, Weatherman RV, Desta Z, Flockhart DA, Skaar TC (2004) Pharmacological characterization of 4-hydroxy-N-desmethyl tamoxifen, a novel active metabolite of tamoxifen. Breast Cancer Res Treat 85:151–159

  14. 14.

    Goetz MP, Rae JM, Suman VJ, Safgren SL, Ames MM, Visscher DW, Reynolds C, Couch FJ, Lingle WL, Flockhart DA, Desta Z, Perez EA, Ingle JN (2005) Pharmacogenetics of tamoxifen biotransformation is associated with clinical outcomes of efficacy and hot flashes. J Clin Oncol 23:9312–9318

  15. 15.

    Rochat B (2005) Role of cytochrome P450 activity in the fate of anticancer agents and in drug resistance: focus on tamoxifen, paclitaxel and imatinib metabolism. Clin Pharmacokinet 44:349–366

  16. 16.

    Teunissen SF, Rosing H, Seoane MD, Brunsveld L, Schellens JH, Schinkel AH, Beijnen JH (2011) Investigational study of tamoxifen phase I metabolites using chromatographic and spectroscopic analytical techniques. J Pharm Biomed Anal 55:518–526

  17. 17.

    Testa B, Pedretti A, Vistoli G (2012) Reactions and enzymes in the metabolism of drugs and other xenobiotics—a meta-analysis of the primary literature. Drug Discov Today 17:549–560

  18. 18.

    Teunissen SF, Rosing H, Schinkel AH, Schellens JH, Beijnen JH (2010) Bioanalytical methods for determination of tamoxifen and its phase I metabolites: a review. Anal Chim Acta 683:21–37

  19. 19.

    Kaufmann A, Butcher P, Maden K, Walker S, Widmer M (2011) Quantitative and confirmative performance of liquid chromatography coupled to high-resolution mass spectrometry compared to tandem mass spectrometry. Rapid Commun Mass Spectrom 25:979–992

  20. 20.

    Henry H, Sobhi HR, Scheibner O, Bromirski M, Nimkar SB, Rochat B (2012) Comparison between a high-resolution single-stage Orbitrap and a triple quadrupole mass spectrometer for quantitative analyses of drugs. Rapid Commun Mass Spectrom 26:499–509

  21. 21.

    Ramanathan R, Korfmacher W (2012) The emergence of high-resolution MS as the premier analytical tool in the pharmaceutical bioanalysis arena. Bioanalysis 4:467–469

  22. 22.

    Rochat B, Kottelat E, McMullen J (2012) The future key role of LC-high-resolution-MS analyses in clinical laboratories: a focus on quantification. Bioanalysis 4:2939–2958

  23. 23.

    Kaufmann A, Butcher P, Maden K, Walker S, Widmer M (2010) Comprehensive comparison of liquid chromatography selectivity as provided by two types of liquid chromatography detectors (high resolution mass spectrometry and tandem mass spectrometry) “where is the crossover point?”. Anal Chim Acta 673:60–72

  24. 24.

    Rochat B (2012) Quantitative/qualitative analysis using LC-HRMS: the fundamental step forward for clinical laboratories and clinical practice. Bioanalysis 4:1709–1711

  25. 25.

    FDA Adverse Event Reporting System (FAERS) and European database of suspected adverse reaction reports. Human Drug Information. Division of Drug Information (CDER). Accessed 30 Oct. 2013. http://www.fda.gov/Drugs/GuidanceComplianceRegulatoryInformation/Surveillance/AdverseDrugEffects/default.htm

  26. 26.

    European Medicines Agency. Accessed 30 Oct. 2013. http://www.ema.europa.eu/docs/en_GB/document_library/Press_release/2012/05/WC500127957.pdf http://www.adrreports.eu

  27. 27.

    Dahmane E, Mercier T, Zanolari B, Cruchon S, Guignard N, Buclin T, Leyvraz S, Zaman K, Csajka C, Decosterd LA (2010) An ultra performance liquid chromatography-tandem MS assay for tamoxifen metabolites profiling in plasma: first evidence of 4′-hydroxylated metabolites in breast cancer patients. J Chromatogr B Anal Technol Biomed Life Sci 878:3402–3414

  28. 28.

    Anari MR, Sanchez RI, Bakhtiar R, Franklin RB, Baillie TA (2004) Integration of knowledge-based metabolic predictions with liquid chromatography data-dependent tandem mass spectrometry for drug metabolism studies: application to studies on the biotransformation of indinavir. Anal Chem 76:823–832

  29. 29.

    Chen G, Yin S, Maiti S, Shao X (2002) 4-hydroxytamoxifen sulfation metabolism. J Biochem Mol Toxicol 16:279–285

  30. 30.

    Mürdter TE, Schroth W, Bacchus-Gerybadze L, Winter S, Heinkele G, Simon W, Fasching PA, Fehm T, German Tamoxifen and AI Clinicians Group, Eichelbaum M, Schwab M, Brauch H (2011) Activity levels of tamoxifen metabolites at the estrogen receptor and the impact of genetic polymorphisms of phase I and II enzymes on their concentration levels in plasma. Clin Pharmacol Ther 89:708–717

  31. 31.

    Testa B, Krämer SD (2007) The biochemistry of drug metabolism—an introduction. Part 2: redox reactions and their enzymes. Chem Biodivers 4:257–405

  32. 32.

    Tang BK (1990) Drug glucosidation. Pharmacol Ther 46:53–56

  33. 33.

    Testa B, Krämer SD (2008) The biochemistry of drug metabolism—an introduction. Part 4: reactions of conjugation and their enzymes. Chem Biodivers 5:2171–2336

  34. 34.

    Bock KW (2003) Vertebrate UDP-glucuronosyltransferases: functional and evolutionary aspects. Biochem Pharmacol 66:691–696

  35. 35.

    Tang C, Hochman JH, Ma B, Subramanian R, Vyas KP (2003) Acyl glucuronidation and glucosidation of a new and selective endothelin ETA receptor antagonist in human liver microsomes. Drug Metab Dispos 31:37–45

  36. 36.

    Bowersox SS, Lightning LK, Rao S, Palme M, Ellis D, Coleman R, Davies AM, Kumaraswamy P, Druzgala P (2011) Metabolism and pharmacokinetics of naronapride (ATI-7505), a serotonin 5-HT4 receptor agonist for gastrointestinal motility disorders. Drug Metab Dispos 39:1170–1180

  37. 37.

    Rochat B, Fayet F, Widmer N, Lahrichi S, Pesse B, Décosterd L, Biollaz J (2008) Imatinib metabolite profiling in parallel to imatinib quantification in plasma of treated patients using Liquid chromatography–mass spectrometry. J Mass Spectrom 43:736–752

  38. 38.

    Schulz M, Iwersen-Bergmann S, Andresen H, Schmoldt A (2012) Therapeutic and toxic blood concentrations of nearly 1,000 drugs and other xenobiotics. Crit Care 16:R136–R140

  39. 39.

    The International Association of Forensic Toxicologists. Accessed 30 Oct. 2013. http://www.gtfch.org/cms/images/stories/Updated_TIAFT_list_202005.pdf

Download references

Acknowledgments

The authors are thankful to Drs. Anton Kaufmann and Olaf Scheibner for fruitful discussions on the Exactive Plus-HR-MS instrument to Prof. Laurent Décosterd for laboratory issues and Dr. Hugues Henry for his kind help with laboratory and software issues.

Conflict of interest statement

The authors have no conflicting financial interests.

Author information

Correspondence to Bertrand Rochat.

Additional information

Parts of this work were presented at the 62nd ASMS Conference on Mass Spectrometry and Allied Topics in Baltimore Maryland, 2013.

Electronic supplementary material

Below is the link to the electronic supplementary material.

ESM 1

(PDF 1.23 MB)

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Dahmane, E., Boccard, J., Csajka, C. et al. Quantitative monitoring of tamoxifen in human plasma extended to 40 metabolites using liquid-chromatography high-resolution mass spectrometry: new investigation capabilities for clinical pharmacology. Anal Bioanal Chem 406, 2627–2640 (2014). https://doi.org/10.1007/s00216-014-7682-2

Download citation

Keywords

  • Drug metabolites
  • High-resolution mass spectrometry
  • Plasma
  • Quantification
  • Therapeutic drug monitoring
  • Triple-quadrupole