Forensic Toxicology

, Volume 37, Issue 1, pp 231–237 | Cite as

Immunoassay-based detection of fentanyl analogs in forensic toxicology

  • Madison Schackmuth
  • Sarah KerriganEmail author
Short Communication



The abuse of fentanyl and its analogs (fentalogs) is of growing concern globally. Forensic toxicology laboratories must detect these emerging drugs in biological evidence, and immunoassay is the most widely used screening technique. In this study, the cross-reactivity of 13 fentalogs were investigated using five commercially available kits.


Dose–response curves were generated using six N-acyl-substituted fentalogs (4-ANPP, acetylfentanyl, butyrylfentanyl, furanylfentanyl, isobutyrylfentanyl, valerylfentanyl), one desphenethyl fentalog (norfentanyl), two piperidine-modified [(+)-cis-3-methylfentanyl, carfentanil], and four phenethyl and piperidine substituted fentalogs (alfentanil, norcarfentanil, remifentanil, sufentanil). Cross-reactivities were estimated for each assay to determine its overall effectiveness for fentalog screening in toxicological samples.

Results and conclusions

Several commercial assays were able to detect either N-acyl or piperidine-modified fentalogs, but none was capable of detecting both. Although this is an inherent disadvantage of the immunoassay approach, it arises from the diverse structural nature of the fentanyl analogs themselves.


Fentanyl Fentalogs ELISA Immunoassay Urine 


Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.


  1. 1.
    United Nations Office on Drugs and Crime (2017) World Drug Report 2017. Available at. Accessed 10 Apr 2018
  2. 2.
    United Nations Office on Drugs and Crime (2016) Global smart update 17. Available at. Accessed 29 Oct 2017
  3. 3.
    P.A. Janssen (1962) A review of the chemical features associated with strong morphine-like activity. Br J Anaesth 34:260–8. Available at. Accessed 23 Aug 2018
  4. 4.
    Lotsch J, Walter C, Parnham MJ, Oertel BG, Geisslinger G (2013) Pharmacokinetics of non-intravenous formulations of fentanyl. Clin Pharmacokinet 52(1):23–36. CrossRefGoogle Scholar
  5. 5.
    U.S. Drug Enforcement Administration Diversion Control Division (2017) NFLIS brief: fentanyl and fentayl-related substances reported in NFLIS, 2015–2016. Available at. Accessed 24 Oct 2017
  6. 6.
    Mather LE (1983) Clinical pharmacokinetics of fentanyl and its newer derivatives. Clin Pharmacokinet 8(5):422–446. CrossRefGoogle Scholar
  7. 7.
    Patel PM, Patel HH, Rot DM (2011) General anesthetics and therapeutic gases. In: Brunton LL (ed) Goodman & Gilman’s the pharmacological basis of therapeutics. McGraw-Hill, New York, pp 527–564Google Scholar
  8. 8.
    EMCDDA (2017) European Drug Report 2017: Trends and developments. Available at. Accessed 23 Aug 2017
  9. 9.
    Lozier MJ, Boyd M, Stanley C, Ogilvie L, King E, Martin C, Lewis L (2015) Acetyl fentanyl, a novel fentanyl analog, causes 14 overdose deaths in Rhode Island, March–May 2013. J Med Toxicol 11(2):208–217. CrossRefGoogle Scholar
  10. 10.
    MC Mercado-Crespo, SA Sumner, MB Spelke, DE Sugerman, Stanley (2014) Notes from the field: increase in fentanyl-related overdose deaths—Rhode Island, November 2013—March 2014. MMWR: Morb Mortal Wkly Rep 63(24):531–531. Available at. Accessed 15 Oct 2017
  11. 11.
    Riches JR, Read RW, Black RM, Cooper NJ, Timperley CM (2012) Analysis of clothing and urine from Moscow theatre siege casualties reveals carfentanil and remifentanil use. J Anal Toxicol 36(9):647–656. CrossRefGoogle Scholar
  12. 12.
    Breindahl T, Kimergard A, Andreasen MF, Pedersen DS (2017) Identification of a new psychoactive substance in seized material: the synthetic opioid N-phenyl-N-[1-(2-phenethyl)piperidin-4-yl]prop-2-enamide (Acrylfentanyl). Drug Test Anal 9(3):415–422. CrossRefGoogle Scholar
  13. 13.
    Armenian P, Olson A, Anaya A, Kurtz A, Ruegner R, Gerona RR (2017) Fentanyl and a novel synthetic opioid U-47700 masquerading as street “Norco” in central california: a case report. Ann Emerg Med 69(1):87–90. CrossRefGoogle Scholar
  14. 14.
    Vo KT, van Wijk XMR, Lynch KL, Wu AHB, Smollin CG (2016) Counterfeit Norco poisoning outbreak—San Francisco Bay Area, California, March 25–April 5, 2016. Morb Mortal Wkly Rep 65(16):420–423. CrossRefGoogle Scholar
  15. 15.
    Swanson DM, Hair LS, Strauch Rivers SR, Smyth BC, Brogan SC, Ventoso AD, Vaccaro SL, Pearson JM (2017) Fatalities Involving carfentanil and furanyl fentanyl: two case reports. J Anal Toxicol 41(6):498–502. CrossRefGoogle Scholar
  16. 16.
    Klar SA, Brodkin E, Gibson E, Padhi S, Predy C, Green C, Lee V (2016) Furanyl-fentanyl overdose events caused by smoking contaminated crack cocaine—British Columbia, Canada, July 15–18, 2016. Mal Chron Blessures Can 36(9):200–201. Google Scholar
  17. 17.
    Stogner JM (2014) The potential threat of acetyl fentanyl: legal issues, contaminated heroin, and acetyl fentanyl “disguised” as other opioids. Ann Emerg Med 64(6):637–639. CrossRefGoogle Scholar
  18. 18.
    United States Department of Justice, Drug Enforcement Administration (2018) Schedules of controlled substances: temporary placement of fentanyl-related substance in schedule I. 21 CFR Part 1308:5188–5192. Available at. Accessed 27 Feb 2018
  19. 19.
    Smith ML (2013) Immunoassay. In: Levine B (ed) Principles of forensic toxicology. AACC Press, Washington, pp 149–170Google Scholar
  20. 20.
    Logan BK, D’Orazio AL, Mohr ALA, Limoges JF, Miles AK, Scarneo CE, Kerrigan S, Liddicoat LJ, Scott KS, Huestis MA (2018) Recommendations for toxicological investigation of drug-impaired driving and motor vehicle fatalities-2017 Update. J Anal Toxicol 42(2):63–68. CrossRefGoogle Scholar
  21. 21.
    Valdez CA, Leif RN, Mayer BP (2014) An efficient, optimized synthesis of fentanyl and related analogs. PLoS One 9(9):e108250–e108250. CrossRefGoogle Scholar
  22. 22.
    Package insert, Fentanyl ELISA Kit (2016) Immunalysis, Ponoma, CAGoogle Scholar
  23. 23.
    Package insert, Fentanyl ELISA Kit (2016) Randox Laboratories Ltd, Crumlin, UKGoogle Scholar
  24. 24.
    Package insert, Carfentanil/Remifentanil ELISA Kit (2017) Randox Laboratories Ltd, Crumlin, UKGoogle Scholar
  25. 25.
    Package insert, Fentanyl ELISA Kit (2017) Neogen Corporation, Lansing, MIGoogle Scholar
  26. 26.
    Package insert, Fentanil ELISA Kit (2017) Neogen Corporation, Lansing, MIGoogle Scholar
  27. 27.
    Coopman V, Cordonnier J, Pien K, Van Varenbergh D (2007) LC-MS/MS analysis of fentanyl and norfentanyl in a fatality due to application of multiple Duragesic transdermal therapeutic system. Forensic Sci Int 169(2–3):223–227CrossRefGoogle Scholar
  28. 28.
    Cummings OT, Enders J, McIntire GL (2017) Fentanyl-norfentanyl concentrations during transdermal patch application: lC-MS-MS urine analysis. J Anal Toxicol 41(2):165–166. Google Scholar
  29. 29.
    Van Rooy H, Vermeulen M, Bovill J (1981) The assay of fentanyl and its metabolites in plasma of patients using gas chromatography with alkali flame ionisation detection and gas chromatography-mass spectrometry. J Chromatogr B 223(1):85–93. CrossRefGoogle Scholar

Copyright information

© Japanese Association of Forensic Toxicology and Springer Japan KK, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Forensic ScienceSam Houston State UniversityHuntsvilleUSA

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