Analytical and Bioanalytical Chemistry

, Volume 391, Issue 1, pp 251–261 | Cite as

Determination of benzimidazole- and bicyclic hydantoin-derived selective androgen receptor antagonists and agonists in human urine using LC–MS/MS

  • Mario Thevis
  • Maxie Kohler
  • Andreas Thomas
  • Joachim Maurer
  • Nils Schlörer
  • Matthias Kamber
  • Wilhelm Schänzer
Original Paper


Selective androgen receptor modulators (SARMs) represent a novel class of drugs with tissue-specific agonistic and antagonistic properties, which are prohibited in sports from January 2008 according to the World Anti-Doping Agency. Preventive approaches to restrict the use of SARMs include early implementation of target analytes into doping control screening assays. Five model SARMs were synthesized, four of which are analogs to prostate-specific androgen receptor antagonists with a 5,6-dichloro-benzimidazole nucleus. The fifth SARM is a muscle-tissue specific agonist with a bicyclic hydantoin structure (BMS-564929). Dissociation pathways after negative electrospray ionization were studied using an LTQ-Orbitrap mass analyzer, and diagnostic product ions and common fragmentation patterns were employed to establish a screening procedure that target the intact SARMs as well as putative metabolic products. Sample preparation based on solid-phase extraction and subsequent LC–MS/MS measurement allowed for detection limits of 1–20 ng/mL, intra- and interday precisions of between 2.4 and 13.2% and between 6.5 and 24.2%, respectively. Recoveries varied from 89 to 106%, and tests for ion suppression or enhancement effects were negative for all analytes.


Precursor ion scanning as a tool to screen for the common nucleus of benzimidazole-derived SARMs


Doping Sport Mass spectrometry Precursor ion scan SARMs Fragmentation Liquid chromatography 



The study was carried out with the support of the Federal Office of Sports, Magglingen, Switzerland, and the Manfred Donike Institute for Doping Analysis, Cologne, Germany.


  1. 1.
    Bhasin S, Calof OM, Storer TW, Lee ML, Mazer NA, Jasuja R, Montori VM, Gao W, Dalton JT (2006) Nat Clin Pract Endocrinol Metab 2:146–159CrossRefGoogle Scholar
  2. 2.
    Hengge UR (2003) AIDS Read 13:S15–S21Google Scholar
  3. 3.
    Moretti C, Frajese GV, Guccione L, Wannenes F, De Martino MU, Fabbri A, Frajese G (2005) J Endocrinol Invest 28:56–64Google Scholar
  4. 4.
    Gooren LJ, Bunck MC (2004) Drugs 64:1861–1891CrossRefGoogle Scholar
  5. 5.
    Dalton JT, Mukherjee A, Zhu Z, Kirkovsky L, Miller DD (1998) Biochem Biophys Res Commun 244:1–4CrossRefGoogle Scholar
  6. 6.
    Mohler ML, Nair VA, Hwang DJ, Rakov IM, Patil R, Miller DD (2005) Exp Opin Ther Patents 15:1565–1585CrossRefGoogle Scholar
  7. 7.
    Chen J, Kim J, Dalton JT (2005) Mol Interv 5:173–188CrossRefGoogle Scholar
  8. 8.
    Negro-Vilar A (1999) J Clin Endocrinol Metab 84:3459–3462CrossRefGoogle Scholar
  9. 9.
    Gao W, Dalton JT (2007) Drug Discov Today 12:241–248CrossRefGoogle Scholar
  10. 10.
    Thevis M, Schänzer W (2007) Mini-Rev Med Chem 7:533–539CrossRefGoogle Scholar
  11. 11.
    Omwancha J, Brown TR (2006) Curr Opin Investig Drugs 7:873–881Google Scholar
  12. 12.
    World Anti-Doping Agency (2008) The 2008 Prohibited List. Accessed 28 Nov 2007
  13. 13.
    Thevis M, Kohler M, Maurer J, Schlörer N, Kamber M, Schänzer W (2007) Rapid Commun Mass Spectom 21:3477–3486CrossRefGoogle Scholar
  14. 14.
    Thevis M, Kamber M, Schänzer W (2006) Rapid Commun Mass Spectrom 20:870–876CrossRefGoogle Scholar
  15. 15.
    Thevis M, Kohler M, Schänzer W (2007) Mass spectrometry of new growth promoting drugs: hydantoin-derived selective androgen receptor modulators and growth hormone secretagogues. In: Schänzer W, Geyer H, Gotzmann A, Mareck U (eds) Recent advances in doping analysis. Sport und Buch Strauß, Cologne (in press)Google Scholar
  16. 16.
    Ng RA, Guan J, Alford VC, Jr., Lanter JC, Allan GF, Sbriscia T, Lundeen SG, Sui Z (2007) Bioorg Med Chem Lett 17:955–958CrossRefGoogle Scholar
  17. 17.
    Ng RA, Guan J, Alford VC Jr, Lanter JC, Allan GF, Sbriscia T, Linton O, Lundeen SG, Sui Z (2007) Bioorg Med Chem Lett 17:784–788Google Scholar
  18. 18.
    Ostrowski J, Kuhns JE, Lupisella JA, Manfredi MC, Beehler BC, Krystek SR, Jr., Bi Y, Sun C, Seethala R, Golla R, Sleph PG, Fura A, An Y, Kish KF, Sack JS, Mookhtiar KA, Grover GJ, Hamann LG (2006) Endocrinology 48:4–12CrossRefGoogle Scholar
  19. 19.
    Sun C, Robl JA, Wang TC, Huang Y, Kuhns JE, Lupisella JA, Beehler BC, Golla R, Sleph PG, Seethala R, Fura A, Krystek SR, Jr., An Y, Malley MF, Sack JS, Salvati ME, Grover GJ, Ostrowski J, Hamann LG (2006) J Med Chem 49:7596–7599CrossRefGoogle Scholar
  20. 20.
    Ng RA, Lanter JC, Alford VC, Allan GF, Sbriscia T, Lundeen SG, Sui Z (2007) Bioorg Med Chem Lett 17:1784–1787CrossRefGoogle Scholar
  21. 21.
    Handelsman DJ (2004) Sci STKE 244:pe41CrossRefGoogle Scholar
  22. 22.
    Catlin DH, Ahrens BD, Kucherova Y (2002) Rapid Commun Mass Spectrom 16:1273–1275CrossRefGoogle Scholar
  23. 23.
    Catlin DH, Sekera MH, Ahrens BD, Starcevic B, Chang Y-C, Hatton CK (2004) Rapid Commun Mass Spectrom 18:1245–1249CrossRefGoogle Scholar
  24. 24.
    Sekera MH, Ahrens BD, Chang YC, Starcevic B, Georgakopoulos C, Catlin DH (2005) Rapid Commun Mass Spectrom 19:781–784CrossRefGoogle Scholar
  25. 25.
    Thevis M, Schänzer W (2005) J Chromatogr Sci 43:22–31Google Scholar
  26. 26.
    ICH (2005) Validation of analytical procedures: text and methodology. Accessed 28 Jan 2008
  27. 27.
    Kromidas A (1999) Validierung in der Analytik. Wiley-VCH, WeinheimGoogle Scholar
  28. 28.
    Dams R, Huestis MA, Lambert WE, Murphy CM (2003) J Am Soc Mass Spectrom 14:1290–1294CrossRefGoogle Scholar
  29. 29.
    Matuszewski BK, Constanzer ML, Chavez-Eng CM (2003) Anal Chem 75:3019–3030CrossRefGoogle Scholar
  30. 30.
    Thevis M, Geyer H, Mareck U, Schänzer W (2005) J Mass Spectrom 40:955–962CrossRefGoogle Scholar
  31. 31.
    Mazzarino M, Botre F (2006) Rapid Comm Mass Spectrom 20:3465–3476CrossRefGoogle Scholar
  32. 32.
    Kolmonen M, Leinonen A, Pelander A, Ojanpera I (2007) Anal Chim Acta 585:94–102CrossRefGoogle Scholar
  33. 33.
    Georgakopoulos CG, Vonaparti A, Stamou M, Kiousi P, Lyris E, Angelis YS, Tsoupras G, Wuest B, Nielen MW, Panderi I, Koupparis M (2007) Rapid Commun Mass Spectrom 21:2439–2446CrossRefGoogle Scholar
  34. 34.
    Nielen MW, Bovee TF, van Engelen MC, Rutgers P, Hamers AR, van Rhijn JH, Hoogenboom LR (2006) Anal Chem 78:424–431CrossRefGoogle Scholar
  35. 35.
    Jemal M, Ouyang Z, Zhao W, Zhu M, Wu WW (2003) Rapid Commun Mass Spectrom 17:2732–2740CrossRefGoogle Scholar
  36. 36.
    Hakala KS, Kostiainen R, Ketola RA (2006) Rapid Commun Mass Spectrom 20:2081–2090CrossRefGoogle Scholar
  37. 37.
    Li AC, Gohdes MA, Shou WZ (2007) Rapid Commun Mass Spectrom 21:1421–1430CrossRefGoogle Scholar
  38. 38.
    Xia YQ, Miller JD, Bakhtiar R, Franklin RB, Liu DQ (2003) Rapid Commun Mass Spectrom 17:1137–1145CrossRefGoogle Scholar
  39. 39.
    Thevis M, Kohler M, Schlörer N, Kamber M, Kuhn A, Linscheid MW, Schänzer W (2007) J Mass Spectrom. DOI  10.1002/jms.1364

Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Mario Thevis
    • 1
  • Maxie Kohler
    • 1
  • Andreas Thomas
    • 1
  • Joachim Maurer
    • 1
  • Nils Schlörer
    • 2
  • Matthias Kamber
    • 3
  • Wilhelm Schänzer
    • 1
  1. 1.Center for Preventive Doping Research, Institute of BiochemistryGerman Sport University CologneCologneGermany
  2. 2.Institute of Organic ChemistryUniversity of CologneCologneGermany
  3. 3.Department of Doping PreventionFederal Office of SportsMagglingenSwitzerland

Personalised recommendations