FREE SHIPPING over $40
OVER 9000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
Home / Equine Research Bank / Rapid Commun Mass Spectrom / Hydroxy levamisole and its phase II conjugates as potential ...
Rapid communications in mass spectrometry : RCM2022; 37(3); e9430; doi: 10.1002/rcm.9430

Hydroxy levamisole and its phase II conjugates as potential indicators of levamisole doping in thoroughbred horses.

Abstract: According to previous research, aminorex is the major metabolite of levamisole; however, in the screening of levamisole-positive racehorse urine and plasma samples, aminorex could only be detected in trace amounts or not at all. In forensic laboratories, hydroxy levamisole and its phase II conjugates make it easier to confirm levamisole misuse and to differentiate between the abuse of levamisole and aminorex. This study aimed to identify the major levamisole metabolites that can be detected along with the parent drug. Methods: The study describes levamisole and its metabolites in thoroughbred horses following oral administration and in vitro with equine liver microsomes. The plausible structures of the detected metabolites were postulated using liquid chromatography combined with high-resolution mass spectrometry. Results: Under the experimental conditions 26 metabolites (17 phase I, 2 phase II, and 7 conjugates of phase I metabolites) were detected (M1-M26). The major phase I metabolites identified were formed by hydroxylation. In phase II, the glucuronic acid conjugates of levamisole and hydroxy levamisole were detected as the major metabolites. In plasma, the parent drug and major metabolites are detectable for up to eight days, while in urine, they are detectable for up to twenty days. Levamisole levels rapidly increased at 45 min following administration, then declined gradually until detectable levels were reached approximately 8 days after administration, according to a pharmacokinetics study. Conclusions: A prolonged elimination profile and relatively high concentration of hydroxy metabolites suggest that the detection of hydroxy metabolites is imperative for investigating levamisole doping in horses.
© 2022 John Wiley & Sons Ltd.
Get Full Text
Publication Date: 2022-11-17 PubMed ID: 36382989DOI: 10.1002/rcm.9430Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
LinkedInCopy
  • Journal Article

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research investigates substances produced by the metabolism of levamisole in thoroughbred horses, which can provide evidence of its illegal use in horse racing.

Objectives of the Study

  • The research aims to identify other metabolites of levamisole, beyond aminorex, that could be used in the detection of its use. The study specifically investigates hydroxy levamisole and its phase II conjugates.
  • The study also sets out to distinguish between the misuse of levamisole and aminorex.

Methods of the Study

  • Researchers administer levamisole orally to thoroughbred horses and analyze samples of urine and plasma. They also conduct in vitro studies using equine liver microsomes.
  • They use liquid chromatography coupled with high-resolution mass spectrometry to hypothesize about the structure of the detected metabolites.

Results of the Study

  • A total of 26 metabolites were identified: 17 from phase I (including the major ones formed by hydroxylation), 2 from phase II, and 7 conjugates of phase I metabolites.
  • The main phase II metabolites were the glucuronic acid conjugates of levamisole and hydroxy levamisole.
  • In a pharmacokinetic study, it was found that levamisole levels increase rapidly 45 minutes after administration, then decrease gradually until reaching detectable levels approximately 8 days later. In plasma, the drug and its major metabolites can be detected for up to eight days, and up to twenty days in urine.

Conclusion of the Study

  • Given their prolonged elimination profile and relatively high concentration, the researchers conclude that identifying hydroxy metabolites is crucial to detecting levamisole misuse in equestrian sports.

Cite This Article

APA
Philip M, Karakka Kal AK, Subhahar MB, Karatt TK, Mathew B, Perwad Z, Graiban FM, Caveney MR, Sayed R, Kadry AM. (2022). Hydroxy levamisole and its phase II conjugates as potential indicators of levamisole doping in thoroughbred horses. Rapid Commun Mass Spectrom, 37(3), e9430. https://doi.org/10.1002/rcm.9430

Publication

Rapid communications in mass spectrometry : RCM
ISSN: 1097-0231
NlmUniqueID: 8802365
Country: England
Language: English
Volume: 37
Issue: 3
Pages: e9430

Researcher Affiliations

Philip, Moses
  • Equine Forensic Unit, Central Veterinary Research Laboratory, Dubai, United Arab Emirates.
Karakka Kal, Abdul Khader
  • Equine Forensic Unit, Central Veterinary Research Laboratory, Dubai, United Arab Emirates.
Subhahar, Michael Benedict
  • Equine Forensic Unit, Central Veterinary Research Laboratory, Dubai, United Arab Emirates.
Karatt, Tajudheen K
  • Equine Forensic Unit, Central Veterinary Research Laboratory, Dubai, United Arab Emirates.
Mathew, Binoy
  • Equine Forensic Unit, Central Veterinary Research Laboratory, Dubai, United Arab Emirates.
Perwad, Zubair
  • Equine Forensic Unit, Central Veterinary Research Laboratory, Dubai, United Arab Emirates.
Graiban, Fatma Mohammed
  • Equine Forensic Unit, Central Veterinary Research Laboratory, Dubai, United Arab Emirates.
Caveney, Marina Rodriguez
  • Equine Forensic Unit, Central Veterinary Research Laboratory, Dubai, United Arab Emirates.
Sayed, Ramy
  • Equine Forensic Unit, Central Veterinary Research Laboratory, Dubai, United Arab Emirates.
Kadry, Ahmed Mahmoud
  • Equine Forensic Unit, Central Veterinary Research Laboratory, Dubai, United Arab Emirates.

MeSH Terms

  • Horses
  • Animals
  • Levamisole / urine
  • Aminorex / urine
  • Mass Spectrometry
  • Microsomes, Liver / metabolism
  • Administration, Oral
  • Doping in Sports

References

This article includes 25 references
  1. Tanphaichitr P, Tanphaichitr D, Sureeratanan J, Chatasingh S. Treatment of nephrotic syndrome with levamisole. J Pediatr 1980;96(3):490-493.
    doi: 10.1016/s0022-3476(80)80707-4google scholar: lookup
  2. Taylor MA, Coop RL, Wall RL. Veterinary Parasitology. John Wiley & Sons; 2015:313-335.
  3. Keiser J, Utzinger J. Efficacy of current drugs against soil-transmitted helminth infections: Systematic review and meta-analysis. JAMA 2008;299(16):1937-1948.
  4. Dillman RO. Cancer immunotherapy. Cancer Biother Radiopharm 2011;26(1):1-64.
    doi: 10.1089/cbr.2010.0902google scholar: lookup
  5. Schneider S, Meys F. Analysis of illicit cocaine and heroin samples seized in Luxembourg from 2005-2010. Forensic Sci Int 2011;212(1-3):242-246.
    doi: 10.1016/j.forsciint.2011.06.027google scholar: lookup
  6. Larocque A, Hoffman RS. Levamisole in cocaine: Unexpected news from an old acquaintance. Clin Toxicol (Phila) 2012;50(4):231-241.
    doi: 10.3109/15563650.2012.665455google scholar: lookup
  7. Wolford A, McDonald TS, Eng H. Immune-mediated agranulocytosis caused by the cocaine adulterant levamisole: A case for reactive metabolite(s) involvement. Drug Metab Dispos 2012;40(6):1067-1075.
    doi: 10.1124/dmd.112.045021google scholar: lookup
  8. Zhu NY, Legatt DF, Turner AR. Agranulocytosis after consumption of cocaine adulterated with levamisole. Ann Intern Med 2009;150(4):287-289.
    doi: 10.7326/0003-4819-150-4-200902170-00102google scholar: lookup
  9. Buchanan JA, Lavonas EJ. Agranulocytosis and other consequences due to use of illicit cocaine contaminated with levamisole. Curr Opin Hematol 2012;19(1):27-23.
    doi: 10.1097/moh.0b013e32834da9efgoogle scholar: lookup
  10. Farhat EK, Muirhead TT, Chaffins ML, Douglass MC. Levamisole-induced cutaneous necrosis mimicking coagulopathy. Arch Dermatol 2010;146(11):1320-1321.
    doi: 10.1001/archdermatol.2010.335google scholar: lookup
  11. Walsh NM, Green PJ, Burlingame RW, Pasternak S, Hanly JG. Cocaine-related retiform purpura: Evidence to incriminate the adulterant, levamisole. J Cutan Pathol 2010;37(12):1212-1219.
    doi: 10.1111/j.1600-0560.2010.01613.xgoogle scholar: lookup
  12. Hess C, Ritke N, Broecker S, Madea B, Musshoff F. Metabolism of levamisole and kinetics of levamisole and aminorex in urine by means of LC-QTOF-HRMS and LC-QqQ-MS. Anal Bioanal Chem 2013;405(12):4077-4088.
    doi: 10.1007/s00216-013-6829-xgoogle scholar: lookup
  13. Bertol E, Mari F, Milia MG, Politi L, Furlanetto S, Karch SB. Determination of aminorex in human urine samples by GCMS after use of levamisole. J Pharm Biomed Anal 2011;55(5):1186-1189.
    doi: 10.1016/j.jpba.2011.03.039google scholar: lookup
  14. Ho EN, Leung DK, Leung GN. Aminorex and rexamino as metabolites of levamisole in the horse. Anal Chim Acta 2009;638(1):58-68.
    doi: 10.1016/j.aca.2009.02.033google scholar: lookup
  15. Lynch KL, Dominy SS, Graf J, Kral AH. Detection of levamisole exposure in cocaine users by liquid chromatography-tandem mass spectrometry. J Anal Toxicol 2011;35(3):176-178.
    doi: 10.1093/anatox/35.3.176google scholar: lookup
  16. Handley SA, Belsey SL, Couchman L, Flanagan RJ. Plasma and urine levamisole in clinical samples containing benzoylecgonine: Absence of aminorex. J Anal Toxicol 2019;43(4):299-306.
    doi: 10.1093/jat/bky102google scholar: lookup
  17. Shea JL. Bioanalytical methods for quantitation of levamisole, a widespread cocaine adulterant. Clin Chem Lab Med 2013;51(1):205-212.
    doi: 10.1515/cclm-2012-0519google scholar: lookup
  18. Karakka Kal AK, Karatt TK, Philip M, Meissir S, Nalakath J. Separation and determination of the enantiomeric levamisole and dexamisole in equine plasma samples using chiral polysaccharide column/LC-MS/MS. Curr Anal Chem 2020;16(6):761-767.
    doi: 10.2174/1573411015666190808103143google scholar: lookup
  19. Mathew B, Philip M, Perwad Z. Identification of hypoxia-inducible factor (HIF) stabilizer roxadustat and its possible metabolites in thoroughbred horses for doping control. Drug Test Anal 2021;13(6):1203-1215.
    doi: 10.1002/dta.3014google scholar: lookup
  20. Subhahar MB, Singh J, Albert PH, Kadry AM. Pharmacokinetics, metabolism, and excretion of celecoxib, a selective cyclooxygenase-2 inhibitor in horses. J Vet Pharmacol Therap 2019;42(5):1-7.
    doi: 10.1111/jvp.12757google scholar: lookup
  21. Philip M, Karakka Kal AK, Subhahar MB, Karatt TK, Mathew B, Koshy SA. Characterization of growth hormone secretagogue small molecule ibutamoren (MK-0677) and its possible metabolites in thoroughbred horses for doping control. Rapid Commun Mass Spectrom 2022;36(18):e9337.
    doi: 10.1002/rcm.9337google scholar: lookup
  22. Philip M, Karakka Kal AK, Subhahar MB, Karatt TK, Mathew B, Perwad Z. In vitro studies of hypoxia-inducible factor-prolyl hydroxylase inhibitors daprodustat, desidustat, and vadadustat for equine doping control. Drug Test Anal 2022;14(2):317-348.
    doi: 10.1002/dta.3188google scholar: lookup
  23. Bogan JA, Marriner SE, Galbraith EA. Pharmacokinetics of levamisole in sheep. Res Vet Sci 1982;32(1):124-126.
    doi: 10.1016/s0034-5288(18)32451-2google scholar: lookup
  24. Watson ADJ, van Gogh H, Sangster NC, Church DB. Levamisole pharmacokinetics and bioavailability in dogs. Res Vet Sci 1988;45(3):411-413.
    doi: 10.1016/s0034-5288(18)30975-5google scholar: lookup
  25. Kouassi E, Caillé G, Léry L, Larivière L, Vézina M. Novel assay and pharmacokinetics of levamisole and p-hydroxy levamisole in human plasma and urine. Biopharm Drug Dispos 1986;7(1):71-89.
    doi: 10.1002/bdd.2510070110google scholar: lookup

Citations

This article has been cited 0 times.
Subscribe to our newsletter
Join 99,344 horse owners like you who receive our email updates on horse health and nutrition.