Advertisement

Analytical and Bioanalytical Chemistry

, Volume 405, Issue 12, pp 4077–4088 | Cite as

Metabolism of levamisole and kinetics of levamisole and aminorex in urine by means of LC-QTOF-HRMS and LC-QqQ-MS

  • Cornelius HessEmail author
  • Natalie Ritke
  • Sebastian Broecker
  • Burkhard Madea
  • Frank Musshoff
Research Paper

Abstract

The antihelminthic drug Levamisole can enhance cocaine effects by conversion into the amphetamine-like drug aminorex. We describe an LC-MS method for the determination of levamisole and its metabolite aminorex in human urine. Selectivity is given, calibration curves were linear within the calibration range 2.5–250 ng/mL; limits of the method were LoD 0.51 ng/mL, LoQ 1.02 ng/mL for levamisole and LoD 0.65 ng/mL, LoQ 0.76 ng/mL for aminorex. Precision data was in accordance with the guidelines (intraday precision for aminorex ranged between 5.75 and 11.0 % for levamisole between 8.36 and 10.9 %; interday precision for levamisole 10.9–16.9 % and for aminorex 7.64–12.7 %; accuracy data for levamisole −1.96 to –14.3 % and for aminorex−11.9 to–18.5 %). The validated method was successfully applied to study the urinary excretion of levamisole after the administration of 100 mg of levamisole orally. Levamisole and aminorex could be detected in post-administration urine samples. Levamisole could be detected up to 39 h after ingestion, while aminorex was detectable up to 54 h. Maximum aminorex concentrations were 45 ng/mL urine. Further metabolites of levamisole after oral ingestion by means of liquid chromatography hybrid quadrupole time-of-flight high-resolution mass spectrometry (LC-QTOF-HRMS) were identified. Only 0.5 % of the ingested drug was quantified as unchanged levamisole in urine. Besides aminorex, five isomers of aminorex and 4 hydroxy-metabolites of aminorex or its isomers were found. Furthermore, levamisole is also hydroxylated and eliminated free or conjugated with sulfate or glucuronide into urine.

Keywords

Levamisole Aminorex LC-MS LC-QTOF-HRMS Metabolism 

References

  1. 1.
    Dillman RO (2011) Cancer immunotherapy. Cancer Biother Radiopharm 26:1–64CrossRefGoogle Scholar
  2. 2.
    Schneider S, Meys F (2011) Analysis of illicit cocaine and heroin samples seized in Luxembourg from 2005–2010. Forensic Sci Int 212:242–246CrossRefGoogle Scholar
  3. 3.
    Larocque A, Hoffman RS (2012) Levamisole in cocaine: unexpected news from an old acquaintance. Clin Toxicol (Phila) 50:231–241CrossRefGoogle Scholar
  4. 4.
    Bertol E, Mari F, Milia MG, Politi L, Furlanetto S, Karch SB (2011) Determination of aminorex in human urine samples by GC-MS after use of levamisole. J Pharm Biomed Anal 55:1186–1189CrossRefGoogle Scholar
  5. 5.
    Ho EN, Leung DK, Leung GN et al (2009) Aminorex and rexamino as metabolites of levamisole in the horse. Anal Chim Acta 638:58–68CrossRefGoogle Scholar
  6. 6.
    Karch SB, Mari F, Bartolini V, Bertol E (2012) Aminorex poisoning in cocaine abusers. Int J Cardiol 158:344–346CrossRefGoogle Scholar
  7. 7.
    Zhu NY, Legatt DF, Turner AR (2009) Agranulocytosis after consumption of cocaine adulterated with levamisole. Ann Intern Med 150:287–289Google Scholar
  8. 8.
    Wolford A, McDonald TS, Eng H et al (2012) Immune-mediated agranulocytosis caused by the cocaine adulterant levamisole: a case for reactive metabolite(s) involvement. Drug Metab Dispos 40:1067–1075CrossRefGoogle Scholar
  9. 9.
    Buchanan JA, Lavonas EJ (2012) Agranulocytosis and other consequences due to use of illicit cocaine contaminated with levamisole. Curr Opin Hematol 19:27–31CrossRefGoogle Scholar
  10. 10.
    Walsh NM, Green PJ, Burlingame RW, Pasternak S, Hanly JG (2010) Cocaine-related retiform purpura: evidence to incriminate the adulterant, levamisole. J Cutan Pathol 37:1212–1219CrossRefGoogle Scholar
  11. 11.
    Farhat EK, Muirhead TT, Chaffins ML, Douglass MC (2010) Levamisole-induced cutaneous necrosis mimicking coagulopathy. Arch Dermatol 146:1320–1321CrossRefGoogle Scholar
  12. 12.
    Wolford A, McDonald TS, Eng H et al (2012) Immune-mediated agranulocytosis caused by the cocaine adulterant levamisole: a case for reactive metabolite(s) involvement. Drug Metab Dispos 40:1067–1075CrossRefGoogle Scholar
  13. 13.
    Heykants JMW&VPA. Levamisole pharmacokinetics in healthy volunteers and in cancer patients. A review of the data available until January 1990. Unpublished Report Serial No. R12564/117 from Janssen Research Foundation. Submitted to WHO by Janssen Pharmaceutica, Beerse, Belgium. 1990Google Scholar
  14. 14.
    Ho EN, Leung DK, Leung GN et al (2009) Aminorex and rexamino as metabolites of levamisole in the horse. Anal Chim Acta 638:58–68CrossRefGoogle Scholar
  15. 15.
    Lynch KL, Dominy SS, Graf J, Kral AH (2011) Detection of levamisole exposure in cocaine users by liquid chromatography-tandem mass spectrometry. J Anal Toxicol 35:176–178CrossRefGoogle Scholar
  16. 16.
    Bertol E, Mari F, Milia MG, Politi L, Furlanetto S, Karch SB (2011) Determination of aminorex in human urine samples by GC-MS after use of levamisole. J Pharm Biomed Anal 55:1186–1189CrossRefGoogle Scholar
  17. 17.
    Ho EN, Leung DK, Leung GN et al (2009) Aminorex and rexamino as metabolites of levamisole in the horse. Anal Chim Acta 638:58–68CrossRefGoogle Scholar
  18. 18.
    Trehy ML, Brown DJ, Woodruff JT et al (2011) Determination of levamisole in urine by gas chromatography–mass spectrometry. J Anal Toxicol 35:545–550CrossRefGoogle Scholar
  19. 19.
    Woestenborghs R, Michielsen L, Heykants J (1981) Determination of levamisole in plasma and animal tissues by gas chromatography with thermionic specific detection. J Chromatogr 224:25–32CrossRefGoogle Scholar
  20. 20.
    Kouassi E, Caille G, Lery L, Lariviere L, Vezina M (1986) Novel assay and pharmacokinetics of levamisole and p-hydroxylevamisole in human plasma and urine. Biopharm Drug Dispos 7:71–89CrossRefGoogle Scholar
  21. 21.
    Shea JL (2012) Bioanalytical methods for quantitation of levamisole, a widespread cocaine adulterant. Clin Chem Lab Med 1–8Google Scholar
  22. 22.
    Ho EN, Leung DK, Leung GN et al (2009) Aminorex and rexamino as metabolites of levamisole in the horse. Anal Chim Acta 638:58–68CrossRefGoogle Scholar
  23. 23.
    Peters FT, Drummer OH, Musshoff F (2007) Validation of new methods. Forensic Sci Int 165:216–224CrossRefGoogle Scholar
  24. 24.
    Broecker S, Herre S, Wust B, Zweigenbaum J, Pragst F (2011) Development and practical application of a library of CID accurate mass spectra of more than 2,500 toxic compounds for systematic toxicological analysis by LC-QTOF-MS with data-dependent acquisition. Anal Bioanal Chem 400:101–117CrossRefGoogle Scholar
  25. 25.
    Broecker S, Koch M, Pragst F (2011) Detection of metabolites in the general unknown screening of hair by LC-QTOF-MS. Ann Tox Anal 23:S1–S12CrossRefGoogle Scholar
  26. 26.
    Peters F, Drummer O, Musshoff F. Anhang B zur Richtlinie der GTFCh zur Qualitätssicherung bei forensich-toxikologischen Untersuchungen: Anforderung an die Validierung von Analysensystemen. https://www.gtfch.org/cms/files/GTFCh_Richtlinie_Anhang%20B_Validierung_Version%201.pdf, accessed 12.12.2011: 2011
  27. 27.
    Matuszewski BK, Constanzer ML, Chavez-Eng CM (1998) 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 70:882–889CrossRefGoogle Scholar
  28. 28.
    DIN EN ISO/IEC 32645. 1994Google Scholar
  29. 29.
    Lynch KL, Dominy SS, Graf J, Kral AH (2011) Detection of levamisole exposure in cocaine users by liquid chromatography-tandem mass spectrometry. J Anal Toxicol 35:176–178CrossRefGoogle Scholar
  30. 30.
    Bertol E, Mari F, Milia MG, Politi L, Furlanetto S, Karch SB (2011) Determination of aminorex in human urine samples by GC-MS after use of levamisole. J Pharm Biomed Anal 55:1186–1189CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • Cornelius Hess
    • 1
    Email author
  • Natalie Ritke
    • 1
  • Sebastian Broecker
    • 2
  • Burkhard Madea
    • 1
  • Frank Musshoff
    • 1
  1. 1.Institute of Forensic MedicineUniversity of BonnBonnGermany
  2. 2.Institute of Legal MedicineUniversity Hospital CharitéBerlinGermany

Personalised recommendations