Author ORCID Identifier
https://orcid.org/0000-0002-0463-6263
Defense Date
2023
Document Type
Directed Research Project
First Advisor
Emanuele Alves
Second Advisor
D. Joshua Cohen
Third Advisor
Justin Poklis
Abstract
Introduction: The growing opioid epidemic in North America presents a call to action to develop new techniques for the identification and quantitation of novel synthetic opioids in uncommon matrices. In instances of extreme decomposition and/or significant scavenging, bone may be the only matrix available. In order to develop an extraction method for synthetic opioids in bone, bone samples were obtained from animal models. The animal model of choice is the rabbit due to their exhibition of spontaneous cortical bone remodeling, a process that is exhibited in humans but not in rats and mice.
Methods: An extraction method was developed to extract the following analytes: fentanyl, norfentanyl, AP-237, and AP-238. Extraction optimization was performed by adding 1000 ng/g of target analytes to the surface of the bone. Different extraction techniques were tested, including liquid-liquid extraction and solid phase extraction, while using the Bead Ruptor Elite to homogenize the bone quickly and efficiently inside an extraction solvent. Extraction solvents tested included methanol, 3% NH4OH in 20:80 isopropanol:ethyl acetate, and 4:1:0.5 butyl chloride:ethyl ether:NH4OH. Finalized extraction method underwent method validation following guidelines set by ANSI/ASB Standard 036, First Edition 2019.
Results: Analytes were able to be extracted, identified, and quantitated using gas chromatography-mass spectrometry (GC-MS). Extraction method yields for fentanyl, norfentanyl, AP-237, and AP-238 were 80%, 30%, 98%, and 84% respectively. Method validation was successful for fentanyl, AP-237, and AP-238.
Conclusion: The finalized extraction method was capable of extracting target analytes while being quicker than traditional extraction methods for bone. Norfentanyl was observed to have a lower yield than the acceptable range (80-120%). Further work will be done to increase optimization for norfentanyl. The applicability for this extraction method will be tested on samples obtained from animal model trials, and this extraction method will be validated using liquid chromatography-tandem mass spectrometry (LC-MS/MS).
Rights
© The Author(s)
Is Part Of
VCU Master of Science in Forensic Science Directed Research Projects
Date of Submission
5-3-2023