Journal of Medical Toxicology

, Volume 14, Issue 4, pp 295–305 | Cite as

Monitoring Dose Response of Cyanide Antidote Dimethyl Trisulfide in Rabbits Using Diffuse Optical Spectroscopy

  • Jangwoen LeeEmail author
  • Gary Rockwood
  • Brian Logue
  • Erica Manandhar
  • Ilona Petrikovics
  • Changhoon Han
  • Vik Bebarta
  • Sari B. Mahon
  • Tanya Burney
  • Matthew Brenner
Original Article



Cyanide (CN) poisoning is a serious chemical threat from accidental or intentional exposures. Current CN exposure treatments, including direct binding agents, methemoglobin donors, and sulfur donors, have several limitations. Dimethyl trisulfide (DMTS) is capable of reacting with CN to form the less toxic thiocyanate with high efficiency, even without the sulfurtransferase rhodanese. We investigated a soluble DMTS formulation with the potential to provide a continuous supply of substrate for CN detoxification which could be delivered via intramuscular (IM) injection in a mass casualty situation. We also used non-invasive technology, diffuse optical spectroscopy (DOS), to monitor physiologic changes associated with CN exposure and reversal.


Thirty-six New Zealand white rabbits were infused with a lethal dose of sodium cyanide solution (20 mg/60 ml normal saline). Animals were divided into three groups and treated with saline, low dose (20 mg), or high dose (150 mg) of DMTS intramuscularly. DOS continuously assessed changes in tissue hemoglobin concentrations and cytochrome c oxidase redox state status throughout the experiment.


IM injection of DMTS increased the survival in lethal CN poisoning. DOS demonstrated that high-dose DMTS (150 mg) reversed the effects of CN exposure on cytochrome c oxidase, while low dose (20 mg) did not fully reverse effects, even in surviving animals.


This study demonstrated potential efficacy for the novel approach of supplying substrate for non-rhodanese mediated sulfur transferase pathways for CN detoxification via intramuscular injection in a moderate size animal model and showed that DOS was useful for optimizing the DMTS treatment.


Chemical and biological weapons Cyanide toxicity reversal Optical hemodynamic monitoring Dimethyl trisulfide Lethal cyanide poisoning Diffuse optical spectroscopy 



Diffuse optical spectroscopy










Cytochrome c oxidase


Compliance with Ethical Standards

All procedures were reviewed and approved by the University of California, Irvine, Institutional Animal Care and Use Committee (IACUC).

Conflict of Interest Statement



This work was supported, in part, by the CounterACT Program, National Institutes of Health Office of the Director (NIH OD), and the National Institute of Neurological Disorders and Stroke (NINDS) grant numbers U54 NS0792, U01 NS058030, and U54 NS063718, AMRMC W81XWH-12-2-0098, CounterACT NIH No. 1U54 NS079201 and by the Air Force Office of Scientific Research award numbers FA9550-17-1-0193 and FA9550-14-1-0193, and the Robert A. Welch Foundation (x-001) at Sam Houston State University, Huntsville, TX.

Any opinions, findings, conclusions, or recommendations expressed in this material are those of the author(s) and do not necessarily reflect the views of the United States Air Force.


  1. 1.
    Martin CO, Adams HP Jr. Neurological aspects of biological and chemical terrorism: a review for neurologists. Arch Neurol. 2003;60(1):21–5.CrossRefGoogle Scholar
  2. 2.
    Gracia R, Shepherd G. Cyanide poisoning and its treatment. Pharmacotherapy. 2004;24(10):1358–65.CrossRefGoogle Scholar
  3. 3.
    Eckstein M. Cyanide as a chemical terrorism weapon. JEMS. 2004;29(8):suppl 22–31.PubMedGoogle Scholar
  4. 4.
    Baskin SI, Brewer TG. Medical aspects of chemical and biological warfare. Chapter 10, Cyanide Poisoning. In: Sidell FR, Takafuji ET, Franz DR, Borden Institute (U.S.), editors. Textbook of military medicine. Part I, Warfare, weaponry, and the casualty. Washington, D.C.: Borden Institute, Walter Reed Army Medical Center; Office of the Surgeon General, U.S. Army; U.S. Army Medical Dept. Center and School; U.S. Army Medical Research and Material Command; Uniformed Services University of the Health Sciences; 1997. p. 272–86.Google Scholar
  5. 5.
    Hydrogen cyanide, potassium cyanide and sodium cyanide [MAK Value Documentation, 2003]. In: Tschickardt M, editor. The MAK-collection for occupational health and safety. Wiley Online Library; 2012. pp. 193–95.
  6. 6.
    Wong-Chong GM, Nakles DV, Luthy RG. Manufacture and the use of cyanide. In: Dzombak DA, Ghosh RS, Wong-Chong GM, editors. Cyanide in water and soil: chemistry, risk, and management. Boca Raton: CRC Press; 2006. pp. 41–57.Google Scholar
  7. 7.
    Suskind R, editor. The one percent doctrine: deep inside America’s pursuit of its enemies since 9/11. New York: Simon & Schuster; 2006.Google Scholar
  8. 8.
    Cummings TF. The treatment of cyanide poisoning. Occup Med (Lond). 2004;54(2):82–5.CrossRefGoogle Scholar
  9. 9.
    Kovacs K, Jayanna PK, Duke A, Winner B, Negrito M, Angalakurthi S, et al. A lipid base formulation for intramuscular administration of a novel sulfur donor for cyanide antagonism. Curr Drug Deliv. 2016;13(8):1351–7. doi:CDD-EPUB-74480 [pii]CrossRefGoogle Scholar
  10. 10.
    Rockwood GA, Thompson DE, Petrikovics I. Dimethyl trisulfide: a novel cyanide countermeasure. Toxicol Ind Health. 2016;32(12):2009–16. doi:0748233715622713 [pii]Google Scholar
  11. 11.
    DeLeon SM, Downey JD, Hildenberger DM, Rhoomes MO, Booker L, Rockwood GA, et al. DMTS is an effective treatment in both inhalation and injection models for cyanide poisoning using unanesthetized mice. Clin Toxicol (Phila). 2018;56(5):332–41. Scholar
  12. 12.
    Rockwood GA, Petrikovics I, Baskin SI, inventors; Sam Houston State University, US Secretary of Army, assignee. Dimethyl trisulfide as a cyanide antidote. US Patent US9375407B2, 28 June 2016.Google Scholar
  13. 13.
    Petrikovics I, Kovacs KI, inventors; Sam Houston State University, assignee. Formulations of dimethyl trisulfide for use as a cyanide antidote. US Patent US9456996B2, 04 Oct 2016.Google Scholar
  14. 14.
    Lee J, Cerussi AE, Saltzman D, Waddington T, Tromberg BJ, Brenner M. Hemoglobin measurement patterns during noninvasive diffuse optical spectroscopy monitoring of hypovolemic shock and fluid replacement. J Biomed Opt. 2007;12(2):024001.CrossRefGoogle Scholar
  15. 15.
    Lee J, Kim JG, Mahon S, Tromberg BJ, Mukai D, Kreuter K, et al. Broadband diffuse optical spectroscopy assessment of hemorrhage- and hemoglobin-based blood substitute resuscitation. J Biomed Opt. 2009;14(4):044027. Scholar
  16. 16.
    Lee J, El-Abaddi N, Duke A, Cerussi AE, Brenner M, Tromberg BJ. Noninvasive in vivo monitoring of methemoglobin formation and reduction with broadband diffuse optical spectroscopy. J Appl Physiol. 2006;100(2):615–22.CrossRefGoogle Scholar
  17. 17.
    Lee J, Armstrong J, Kreuter K, Tromberg BJ, Brenner M. Non-invasive in vivo diffuse optical spectroscopy monitoring of cyanide poisoning in a rabbit model. Physiol Meas. 2007;28(9):1057–66. doi:S0967-3334(07)47812-1 [pii]CrossRefGoogle Scholar
  18. 18.
    Lee J, Keuter KA, Kim J, Tran A, Uppal A, Mukai D, et al. Noninvasive in vivo monitoring of cyanide toxicity and treatment using diffuse optical spectroscopy in a rabbit model. Mil Med. 2009;174(6):615–21.CrossRefGoogle Scholar
  19. 19.
    Brenner M, Mahon SB, Lee J, Kim J, Mukai D, Goodman S, et al. Comparison of cobinamide to hydroxocobalamin in reversing cyanide physiologic effects in rabbits using diffuse optical spectroscopy monitoring. J Biomed Opt. 2010;15(1):017001. Scholar
  20. 20.
    Brenner M, Kim JG, Mahon SB, Lee J, Kreuter KA, Blackledge W, et al. Intramuscular cobinamide sulfite in a rabbit model of sublethal cyanide toxicity. Ann Emerg Med. 2010;55(4):352–63. Scholar
  21. 21.
    Kovacs K, Duke AC, Shifflet M, Winner B, Lee SA, Rockwood GA, et al. Parenteral dosage form development and testing of dimethyl trisulfide, as an antidote candidate to combat cyanide intoxication. Pharm Dev Technol. 2016;22:1–6. Scholar
  22. 22.
    Bartling CM, Andre JC, Howland CA, Hester ME, Cafmeyer JT, Kerr A, et al. Stability characterization of a polysorbate 80-dimethyl trisulfide formulation, a cyanide antidote candidate. Drugs R D. 2016;16(1):109–27. Scholar
  23. 23.
    Lee J, Kim JG, Mahon SB, Mukai D, Yoon D, Boss GR, et al. Noninvasive optical cytochrome c oxidase redox state measurements using diffuse optical spectroscopy. J Biomed Opt. 2014;19(5):055001.CrossRefGoogle Scholar
  24. 24.
    Zijlstra WG, Buursma A, Assendelft OW. Visible and near-infrared absorption spectra of human and animal hemoglobin. AH Zeist: VSP BV; 2000.Google Scholar
  25. 25.
    Kou LH, Labrie D, Chylek P. Refractive indices of water and ice in the 0.65 to 2.5 mm spectral range. Appl Opt. 1993;32:3531–40.CrossRefGoogle Scholar
  26. 26.
    Eker C. Optical characterization of tissue for medical diagnostics [Ph.D. dissertation]. Lund: Lund University; 1999.Google Scholar
  27. 27.
    Wray S, Cope M, Delpy DT, Wyatt JS, Reynolds EO. Characterization of the near infrared absorption spectra of cytochrome aa3 and haemoglobin for the non-invasive monitoring of cerebral oxygenation. Biochim Biophys Acta. 1988;933(1):184–92.CrossRefGoogle Scholar
  28. 28.
    Brenner M, Kim JG, Lee J, Mahon SB, Lemor D, Ahdout R, et al. Sulfanegen sodium treatment in a rabbit model of sub-lethal cyanide toxicity. Toxicol Appl Pharmacol. 2010;248(3):269–76.CrossRefGoogle Scholar
  29. 29.
    Kim JG, Lee J, Mahon SB, Mukai D, Patterson SE, Boss GR, et al. Noninvasive monitoring of treatment response in a rabbit cyanide toxicity model reveals differences in brain and muscle metabolism. J Biomed Opt. 2012;17(10):105005.CrossRefGoogle Scholar
  30. 30.
    Villringer A, Chance B. Non-invasive optical spectroscopy and imaging of human brain function. Trends Neurosci. 1997;20(10):435–42.CrossRefGoogle Scholar
  31. 31.
    Cooper CE, Cope M, Springett R, Amess PN, Penrice J, Tyszczuk L, et al. Use of mitochondrial inhibitors to demonstrate that cytochrome oxidase near-infrared spectroscopy can measure mitochondrial dysfunction noninvasively in the brain. J Cereb Blood Flow Metab. 1999;19(1):27–38.CrossRefGoogle Scholar

Copyright information

© American College of Medical Toxicology 2018

Authors and Affiliations

  1. 1.Beckman Laser InstituteUniversity of CaliforniaIrvineUSA
  2. 2.Analytical Toxicology DivisionUS Army Medical Research Institute of Chemical DefenseAberdeenUSA
  3. 3.Department of Chemistry and BiochemistrySouth Dakota UniversityBrookingsUSA
  4. 4.Department of ChemistrySam Houston State UniversityHuntsvilleUSA
  5. 5.Department of Internal MedicineNational Health Insurance Service Ilsan HospitalGoyang-siSouth Korea
  6. 6.Department of Emergency MedicineUniversity of Colorado School of MedicineAuroraUSA
  7. 7.Division of Pulmonary and Critical Care Medicine, Department of MedicineUniversity of CaliforniaIrvineUSA

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