Skip to main content
Log in

Metabolism of triacetone triperoxide (TATP) by canine cytochrome P450 2B11

  • Original Article
  • Published:
Forensic Toxicology Aims and scope Submit manuscript

Abstract

Purpose

This work is performed to determine if there is a potential for the accumulation and potential toxicity of triacetone triperoxide (TATP) in canines. Additional utility of this information may suggest human toxicity and possibly detection of biomarkers, metabolites or intact molecule of those using this material for nefarious reasons.

Methods

Liquid chromatography/mass spectrometry of dog liver microsome (DLM) incubation samples of TATP was used to measure substrate depletion. Trapping of electrophilic products was performed using glutathione (GSH) and semicarbazide. Comparisons were made to free hydroperoxides found in methyl ethyl ketone peroxides (MEKP).

Results

The non-specific Km value of 2.2 μM and a Vmax of 1.1 nmol/min/mg of protein were determined. Canine recombinant cytochrome P450 (rCYP) 2B11 with human cytochrome b5 was found to catalyze the NADPH-dependent metabolism of TATP into its only phase I metabolite, hydroxy-TATP (TATP-OH). No secondary metabolite(s) or degraded products were detected or trapped from microsomal incubations. MEKP subjected to similar conditions was found to undergo significant metabolism, semicarbazide trapping and rapid oxidation of GSH to GSSG. The synthesized TATP-OH metabolite incubated in DLM progressed three times faster than TATP metabolism with no secondary metabolites found or trapped.

Conclusions

TATP does not react as MEKP suggesting that TATP does not ring-open to form hydroperoxides. TATP and TATP-OH compete for the same enzyme, with TATP dominating this competition. Failure to detect additional metabolite(s) suggests they may be too small to detect by our system or bound covalently to a protein or polymer in the incubation reaction.

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

Access this article

Subscribe and save

Springer+ Basic
EUR 32.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or Ebook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Scheme 1
Fig. 8
Fig. 9
Fig. 10
Fig. 11

Similar content being viewed by others

References

  1. Channon HJ, Mills GT, Williams RT (1944) The metabolism of 2:4:6-trinitrotoluene (a-T.N.T.). Biochem J 38:70–85

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Buedingg E, Jolliffe N (1946) Metabolism of trinitrotoluene (TNT) in vitro. J Pharmacol Exp Ther 88:300–312. https://doi.org/10.1007/bf01571053

    Article  Google Scholar 

  3. DiCarlo FJ, Crew MC, Sklow NJ, Coutinho CB, Nonkin P et al (1966) Metabolism of pentaerythritol tetranitrate by patients with coronary artery disease. J Pharmacol Exp Ther 153:254–258

    CAS  Google Scholar 

  4. Davidson IWF, Miller HS, Dicarlo FJ (1970) Absorption, excretion and metabolism of pentaerythritol tetranitrate by humans. J Pharmacol Exp Ther 175:42–50

    CAS  PubMed  Google Scholar 

  5. Don Van Natta J, Sciolino E, Grey S (2006) Details emerge in British terror case. The New York Times. https://www.nytimes.com/2006/08/28/world/europe/28plot.html. Accessed 28 Oct 2017

  6. Fikes BJ (2012) REGION: Escondido “bomb house” controlled burn a model for nation, expert says. The San Diego Union-Tribune. http://www.sandiegouniontribune.com/sdut-region-escondido-bomb-house-controlled-burn-a-2012mar26-story.html. Accessed 3 Nov 2017

  7. Morris S (2009) Terror suspect student ‘had suicide vest and explosives’. The Gaurdian. https://www.theguardian.com/uk/2009/jun/04/terror-suspect-isa-ibrahim. Accessed 03 Nov 2017

  8. Oxley JC, Smith JL, Moran J, Nelson K, Utley WE (2004) Training dogs to detect triacetone triperoxide. In: Proceedings of SPIE—The International Society for Optical Engineering, Proc. SPIE 5403, Sensors, and Command, Control, Communications, and Intelligence (C3I) Technologies for Homeland Security and Homeland Defense III (15 September 2004). https://doi.org/10.1117/12.555791

  9. Boveris A, Cadenas E (2000) Mitochondrial production of hydrogen peroxide regulation by nitric oxide and the role of ubisemiquinone. IUBMB Life 50:245–250. https://doi.org/10.1080/713803732

    Article  CAS  PubMed  Google Scholar 

  10. Hayyan M, Hashim MA, Alnashef IM (2016) Superoxide ion: generation and chemical implications. Chem Rev 116:3029–3085. https://doi.org/10.1021/acs.chemrev.5b00407

    Article  CAS  PubMed  Google Scholar 

  11. Karuzina II, Archakov AI (1994) Hydrogen peroxide-mediated inactivation of microsomal cytochrome P450 during monooxygenase reactions. Free Radic Biol Med 17:557–567. https://doi.org/10.1016/0891-5849(94)90095-7

    Article  CAS  PubMed  Google Scholar 

  12. Kundu TK, Hille R, Velayutham M, Zweier JL (2007) Characterization of superoxide production from aldehyde oxidase: an important source of oxidants in biological tissues. Arch Biochem Biophys 460:113–121. https://doi.org/10.1016/j.abb.2006.12.032

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Bayne AC, Mockett RJ, Orr WC, Sohal RS (2005) Enhanced catabolism of mitochondrial superoxide/hydrogen peroxide and aging in transgenic Drosophila. Biochem J 391:277–284. https://doi.org/10.1042/bj20041872

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Jones DP, Eklöw L, Thor H, Orrenius S (1981) Metabolism of hydrogen peroxide in isolated hepatocytes: relative contributions of catalase and glutathione peroxidase in decomposition of endogenously generated H2O2. Arch Biochem Biophys 210:505–516. https://doi.org/10.1016/0003-9861(81)90215-0

    Article  CAS  PubMed  Google Scholar 

  15. Martins D, English AM (2014) Catalase activity is stimulated by H2O2 in rich culture medium and is required for H2O2 resistance and adaptation in yeast. Redox Biol 2:308–313. https://doi.org/10.1016/j.redox.2013.12.019

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Vaz ADN, Coon MJ (1987) Hydrocarbon formation in the reductive cleavage of hydroperoxides by cytochrome P450. Proc Natl Acad Sci 84:1172–1176

    Article  CAS  PubMed  Google Scholar 

  17. Chefson A, Zhao J, Auclair K (2006) Replacement of natural cofactors by selected hydrogen peroxide donors or organic peroxides results in improved activity for CYP3A4 and CYP2D6. ChemBioChem 7:916–919. https://doi.org/10.1002/cbic.200600006

    Article  PubMed  Google Scholar 

  18. Degousee N, Triantaphylides C, Starek S, Iacazio G, Martini D et al (1995) Measurement of thermally produced volatile alkanes: an assay for the plant hydroperoxy fatty acid evaluation. Anal Biochem 224:524–531. https://doi.org/10.1006/abio.1995.1082

    Article  CAS  PubMed  Google Scholar 

  19. Litov RE, Matthews LC, Tappel AL (1981) Vitamin E protection against in vivo peroxidation initiated in rats by methyl ethyl ketone peroxide as monitored by pentane. Toxicol Appl Pharmacol 55:96–106. https://doi.org/10.1016/0041-008x(81)90456-7

    Article  Google Scholar 

  20. Hix S, Kadiiska MB, Mason RP, Augusto O (2000) In vivo metabolism of tert-butyl hydroperoxide to methyl radicals. EPR spin-trapping and DNA methylation studies. Chem Res Toxicol 13:1056–1064. https://doi.org/10.1021/tx000130l

    Article  CAS  PubMed  Google Scholar 

  21. Hix S, Augusto O (1999) DNA methylation by tert-butyl A role for the transition metal ion in the production of DNA base adducts. Chem Biol Interact 118:141–149. https://doi.org/10.1016/s0009-2797(99)00079-4

    Article  CAS  PubMed  Google Scholar 

  22. Mlochowski J, Brzaszcz M, Giurg M, Palus J, Wojtowicz H (2003) Selenium-promoted oxidation of organic compounds: reactions and mechanisms. Eur J Org Chem 2003:4329–4339. https://doi.org/10.1002/ejoc.200300230

    Article  CAS  Google Scholar 

  23. Tapiero H, Townsend DM, Tew KD (2003) The antioxidant role of selenium and seleno-compounds. Biomed Pharmacother 57:134–144. https://doi.org/10.1016/s0753-3322(03)00035-0

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Tinggi U (2008) Selenium: its role as antioxidant in human health. Environ Health Prev Med 13:102–108. https://doi.org/10.1007/s12199-007-0019-4

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Bozdemir MN, Yildiz M, Seyhanli ES, Gurbuz S, Kilicaslan I et al (2011) Narrowing of airway caused by ingestion of methyl ethyl ketone peroxide. Hum Exp Toxicol 30:2002–2006. https://doi.org/10.1177/0960327111407230

    Article  CAS  PubMed  Google Scholar 

  26. Muraleedharan KM, Avery MA (2009) Progress in the development of peroxide-based anti-parasitic agents. Drug Discov Today 14:793–803. https://doi.org/10.1016/j.drudis.2009.05.008

    Article  CAS  PubMed  Google Scholar 

  27. Jefford CW (2007) New developments in synthetic peroxidic drugs as artemisinin mimics. Drug Discov Today 12:487–495. https://doi.org/10.1016/j.drudis.2007.04.009

    Article  CAS  PubMed  Google Scholar 

  28. Opsenica DM, Solaja BA (2009) Antimalarial peroxides. J Serbian Chem Soc 74:1155–1193. https://doi.org/10.2298/jsc0911155

    Article  CAS  Google Scholar 

  29. Antoine T, Fisher N, Amewu R, O’Neil PM, Ward S et al (2014) Rapid kill of malaria parasites by artemisinin and semi-synthetic endoperoxides involves ROS-dependent depolarization of the membrane potential. J Antimicrob Chemother 69:1005–1016. https://doi.org/10.1093/jac/dkt486

    Article  CAS  PubMed  Google Scholar 

  30. Haynes RK, Cheu KW, Chan HW, Wong HN, Li KY et al (2012) Interactions between artemisinins and other antimalarial drugs in relation to the cofactor model—A unifying proposal for drug action. ChemMedChem 7:2204–2226. https://doi.org/10.1002/cmdc.201200383

    Article  CAS  PubMed  Google Scholar 

  31. Ramasarma T (1982) Generation of H2O2 in biomembranes. Biochim Biophys Acta Rev Biomembr 694:69–93. https://doi.org/10.1016/0304-4157(82)90014-4

    Article  CAS  Google Scholar 

  32. Oxley JC, Smith JL, Luo W, Brady J (2009) Determining the vapor pressures of diacetone diperoxide (DADP) and hexamethylene triperoxide diamine (HMTD). Propellants Explos Pyrotech 34:539–543. https://doi.org/10.1002/prep.200800073

    Article  CAS  Google Scholar 

  33. Oxley JC, Smith JL, Porter M, McLennan L, Colizza K et al (2016) Synthesis and degradation of hexamethylene triperoxide diamine (HMTD). Propellants Explos Pyrotech 41:334–350. https://doi.org/10.1002/prep.201500151

    Article  CAS  Google Scholar 

  34. Oxley JC, Smith JL, Shinde K, Moran J (2005) Determination of the vapor density of triacetone triperoxide (TATP) using a gas chromatography headspace technique. Propellants Explos Pyrotech 30:127–130. https://doi.org/10.1002/prep.200400094

    Article  CAS  Google Scholar 

  35. Colizza K, Yevdokimov A, Mclennan L, Smith JL, Oxley JC (2018) Reactions of organic peroxides with alcohols in atmospheric pressure chemical ionization—the pitfalls of quantifying triacetone triperoxide (TATP). J Am Soc Mass Spectrom 29:393–404. https://doi.org/10.1007/s13361-017-1836-3

    Article  CAS  PubMed  Google Scholar 

  36. Minh VD, Dolan GF, Brach BB, Moser KM (1978) Functional residual capacity and body position in the dog. J Appl Physiol 44:291–296. https://doi.org/10.1152/jappl.1978.44.2.291

    Article  CAS  PubMed  Google Scholar 

  37. Barrett K, Brooks H, Boitano S, Barman S (2010) Ganong’s review of medical physiology, 23rd edn. McGraw Hill, New York, pp 261–272

    Google Scholar 

  38. Colizza K, Yevdokimov A, McLennan L, Smith JL, Oxley JC (2018) Using gas phase reactions of hexamethylene triperoxide diamine (HMTD) to improve detection in mass spectrometry. J Am Soc Mass Spectrom 29:675–684. https://doi.org/10.1007/s13361-017-1879-5

    Article  CAS  PubMed  Google Scholar 

  39. Colizza K, Mahoney KE, Yevdokimov AV, Smith JL, Oxley JC (2016) Acetonitrile ion suppression in atmospheric pressure ionization mass spectrometry. J Am Soc Mass Spectrom. https://doi.org/10.1007/s13361-016-1466-1

    Article  PubMed  Google Scholar 

  40. Oxley JC, Smith JL, Bowden PR, Rettinger RC (2013) Factors influencing triacetone triperoxide (TATP) and diacetone diperoxide (DADP) formation: part 1. Propellants Explos Pyrotech 38:244–254. https://doi.org/10.1002/prep.201200116

    Article  CAS  Google Scholar 

  41. Smith ME, Wall C, Fitzgerald M (2012) Characterisation of the major synthetic products of the reactions between butanone and hydrogen peroxide. Propellants Explos Pyrotech 37:282–287. https://doi.org/10.1002/prep.201100091

    Article  CAS  Google Scholar 

  42. Milas NA, Golubovic A (1959) Studies in organic peroxides. XXV. Preparation, separation and identification of peroxides derived from methyl ethyl ketone and hydrogen peroxide. J Am Chem Soc 81:5824–5826. https://doi.org/10.1021/ja01530a068

    Article  CAS  Google Scholar 

  43. Court MH (2014) Canine cytochrome P450 (CYP) pharmacogenetics. NIH Public Access 43:1027–1038. https://doi.org/10.1016/j.cvsm.2013.05.001

    Article  Google Scholar 

  44. Linder CD, Renaud NA, Hutzler JM (2009) Is 1-Aminobenzotriazole an appropriate in vitro tool as a nonspecific cytochrome P450 inactivator? Drug Metab Dispos 37:10–13. https://doi.org/10.1124/dmd.108.024075

    Article  CAS  PubMed  Google Scholar 

  45. Bondoc FY, Bao Z, Hu WY, Gonzalez FJ, Wang Y et al (1999) Acetone catabolism by cytochrome P450 2E1: studies with CYP2E1-null mice. Biochem Pharmacol 58:461–463. https://doi.org/10.1016/s0006-2952(99)00111-2

    Article  CAS  PubMed  Google Scholar 

  46. Koop DR, Casazza JP (1985) Identification of ethanol-inducible P-450 isozyme 3a as the acetone and acetol monooxygenase of rabbit microsomes. J Biol Chem 260:13607–13612

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This material is based upon work supported by U.S. Department of Homeland Security (DHS), Science & Technology Directorate, Office of University Programs, under Grant 2013-ST-061-ED0001. Views and conclusions are those of the authors and should not be interpreted as necessarily representing the official policies, either expressed or implied, of DHS.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Jimmie C. Oxley.

Ethics declarations

Conflict of interest

The authors have no conflict of interest to declare.

Ethical approval

Samples produced using human cytochrome b5 or human reductase bactosomes are not recovered from any human patients and are not subject to human biological sample management required by GxP.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (PDF 531 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Colizza, K., Gonsalves, M., McLennan, L. et al. Metabolism of triacetone triperoxide (TATP) by canine cytochrome P450 2B11. Forensic Toxicol 37, 174–185 (2019). https://doi.org/10.1007/s11419-018-0450-9

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11419-018-0450-9

Keywords

Navigation