LC-MS/MS Analysis of 30+ Fentanyl Analogs in Human Liver to Support Medicolegal Death Investigations

Kylea Mathison*, Joseph Cox, Colby Ott and Luis Arroyo, Department of Forensic and Investigative Science, West Virginia University, Morgantown, WV 26506

Field (Broad Category): Forensics (Physical Sciences & Engineering) 

Student’s Major: Forensic Chemistry 

As opioid abuse continues to increase across the United States; illicit modification to the fentanyl core structure results in novel analog molecules that have shown higher toxicity to humans. This continuous change in the synthetic opioid landscape represents an issue during the drug toxicological screening testing. Samples submitted in drug overdose death investigations include blood, urine, liver, and other biological tissues to determine if any identifiable drugs played a role in the death of an individual. From the medical examiner viewpoint, the interpretation of the postmortem redistribution of therapeutic and illicit drugs is a complicated task. To this end, the liver constitutes the ideal matrix as it is less susceptible to postmortem redistribution. The effective extraction of target drugs from this crucial biological endpoint represents a continuous battle among practitioners. This research presents a simple, clean, and efficient extraction of 34 fentanyl analogs from postmortem human liver specimens. These specimens were extracted using a modified quick, easy, cheap, effective, rugged, and safe (QuEChERS) method and analyzed through liquid chromatography-tandem mass spectrometry (LC-MS/MS). This in-house modified QuEChERS extraction allows for the analysis of the minimal sample (0.1g liver tissue) and prevents crosscontamination. The resulting average bias was -1.48% and -0.58%, with an average precision of 9.91% and 7.60% at low and high concentrations, respectively. The matrix effects, recovery, process efficiency, and application to authentic postmortem liver samples were evaluated for the QuEChERS extraction. 

Funding: 

Program/mechanism supporting research/creative efforts: WVU 497-level course