Quantitation of Fentanyl and Metabolites from Blow Fly Tissue and Development Effects of Fentanyl on Lucilia sericata

Abstract

Death investigations often involve collaborative work between different forensic disciplines, especially with regards to pathology, toxicology, and entomology. This dissertation investigates the effects of designer opioids on the development of blow fly (Lucilia sericata) larvae and the ability to detect and quantify the drugs and/or metabolites present in each stage of the fly’s life cycle. Therefore, we evaluated human tissue fortified with fentanyl at four treatment concentrations, and blow fly larvae were placed to feed on the drug infused tissue. A quick, easy, cheap, effective, rugged, and safe (QuEChERS) extraction method was developed, validated, and applied to extract fentanyl and metabolites from the larva, pupa, pupa casings, adults of Lucilia sericata. Additionally, since liver tissue was utilized as feeding media, a complete validation extraction was also developed, followed by the quantitative analysis of target drugs and metabolites via liquid-chromatography-tandem mass spectrometry (LC-MS/MS). This research represents an advancement in the combat of the opioid epidemic by offering a fully validated protocol that complements the analytical toolkit in forensic toxicology laboratories and medical examiners offices for complex biological matrices by 1) providing a protocol to extract and quantify fentanyl and metabolites from complex matrices using QuEChERS to aid in forensic investigations, 2) an evaluation any growth effects fentanyl has on the development of the insects and the potential implications on the impact of minPMI estimations, 3) conclusions on the ability to correlate the concentration of fentanyl and metabolites from the insect tissue to the initial concentration of the feeding media.