A Novel Oral Fluid Assay (LC-QTOF-MS) for the Detection of Fentanyl and Clandestine Opioids in Oral Fluid After Reported Heroin Overdose

J Med Toxicol. 2017 Dec;13(4):287-292. doi: 10.1007/s13181-017-0632-6. Epub 2017 Oct 2.


Introduction: The adulteration of heroin with non-pharmaceutical fentanyl and other high-potency opioids is one of the factors contributing to striking increases in overdose deaths. To fully understand the magnitude of this problem, accurate detection methods for fentanyl and other novel opioid adulterant exposures are urgently required. The objective of this work was to compare the detection of fentanyl in oral fluid and urine specimens using liquid chromatography quadrupole time-of-flight mass spectrometry (LC-QTOF-MS) in a population of heroin users presenting to the Emergency Department after overdose.

Methods: This was a prospective observational study of adult Emergency Department patients who presented after a reported heroin overdose requiring naloxone administration. Participants provided paired oral fluid and urine specimens, which were prepared, extracted, and analyzed using a dual LC-QTOF-MS workflow for the identification of traditional and emerging drugs of abuse. Analytical instrumentation included SCIEX TripleTOF® 5600+ and Waters Xevo® G2-S QTOF systems.

Results: Thirty participants (N = 30) were enrolled during the study period. Twenty-nine participants had fentanyl detected in their urine, while 27 had fentanyl identified in their oral fluid (overall agreement 93.3%, positive percent agreement 93.1%). Cohen's Kappa (k) was calculated and demonstrated moderately, significant agreement (k = 0.47; p value 0.002) in fentanyl detection between oral fluid and urine using this LC-QTOF-MS methodology. Additional novel opioids and metabolites, including norfentanyl, acetylfentanyl, and U-47700, were detected during this study.

Conclusion: In this study of individuals presenting to the ED after reported heroin overdose, a strikingly high proportion had a detectable fentanyl exposure. Using LC-QTOF-MS, the agreement between paired oral fluid and urine testing for fentanyl detection indicates a role for oral fluid testing in surveillance for nonpharmaceutical fentanyl. Additionally, the use of LC-QTOF-MS allowed for the detection of other clandestine opioids (acetylfentanyl and U-47700) in oral fluid.

Keywords: Drug test; Fentanyl; Heroin; Oral fluid; QTOF.

Publication types

  • Observational Study

MeSH terms

  • Adolescent
  • Analgesics, Opioid / analysis*
  • Analgesics, Opioid / urine
  • Chromatography, Liquid*
  • Drug Contamination*
  • Drug Overdose / diagnosis*
  • Drug Overdose / drug therapy
  • Drug Overdose / metabolism
  • Drug Overdose / urine
  • Emergency Service, Hospital
  • Female
  • Fentanyl / analysis*
  • Fentanyl / urine
  • Heroin Dependence / diagnosis*
  • Heroin Dependence / drug therapy
  • Heroin Dependence / metabolism
  • Heroin Dependence / urine
  • Humans
  • Male
  • Mass Spectrometry*
  • Naloxone / administration & dosage
  • Narcotic Antagonists / administration & dosage
  • Predictive Value of Tests
  • Prospective Studies
  • Reproducibility of Results
  • Saliva / chemistry*
  • Substance Abuse Detection / methods*
  • Urinalysis
  • Young Adult


  • Analgesics, Opioid
  • Narcotic Antagonists
  • Naloxone
  • Fentanyl