LC-MS/MS-based method for the multiplex detection of 24 fentanyl analogues and metabolites in whole blood at Sub ng mL−1 concentrations

Kraig E. Strayer, Heather M. Antonides, Matthew P. Juhascik, Raminta Daniulaityte, Ioana E. Sizemore

Research output: Contribution to journalArticlepeer-review

83 Scopus citations


The United States and numerous other countries worldwide are currently experiencing a public health crisis due to the abuse of illicitly manufactured fentanyl (IMF) and its analogues. This manuscript describes the development of a liquid chromatography-tandem mass spectrometry-based method for the multiplex detection of N = 24 IMF analogues and metabolites in whole blood at concentrations as low as 0.1−0.5 ng mL−1. These available IMFs were fentanyl, norfentanyl, furanyl norfentanyl, remifentanil acid, butyryl norfentanyl, remifentanil, acetyl fentanyl, alfentanil, AH-7921, U-47700, acetyl fentanyl 4-methylphenethyl, acrylfentanyl, para-methox-yfentanyl, despropionyl fentanyl (4-ANPP), furanyl fentanyl, despropionyl para-fluorofentanyl, carfentanil, (±)-cis-3-methyl fentanyl, butyryl fentanyl, isobutyryl fentanyl, sufentanil, valeryl fentanyl, para-fluorobutyryl fentanyl and para-fluoroisobutyryl fentanyl. Most IMF analogues (N = 22) could be easily distinguished from one another; the isomeric forms butyryl/isobutyryl fentanyl and para-fluorobutyryl/para-fluoroisobutyryl fentanyl could not be differentiated. N = 13 of these IMF analogues were quantified for illustrative purposes, and their forensic quality control standards were also validated for limit of detection (0.017−0.056 ng mL−1), limit of quantitation (0.100−0.500 ng mL−1), selectivity/sensitivity, ionization suppression/enhancement (87−118%), process efficiency (60−95%), recovery (64−97%), bias (<20%), and precision (>80%). This flexible, time- and cost-efficient method was successfully implemented at the Montgomery County Coroner’s Office/Miami Valley Regional Crime Laboratory in Dayton, Ohio, where it aided in the analysis of N = 725 postmortem blood samples collected from February 2015 to November 2016.

Original languageEnglish (US)
Pages (from-to)514-523
Number of pages10
JournalACS Omega
Issue number1
StatePublished - Jan 31 2018
Externally publishedYes

ASJC Scopus subject areas

  • General Chemistry
  • General Chemical Engineering


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