MSACL 2019 EU Abstract

Self-Classified Topic Area(s): Small Molecules / Tox / TDM

Analysis of Fentanyl and Metabolite in Clinical and Post-Mortem Samples

Karen S. Scott1, Rudra Maharajh2, Frank Kero2; Jamie Foss2, Joseph Jones3, Sabra Botch-Jones4
(1)Arcadia University, Glenside, PA, USA (2)PerkinElmer, Waltham, MA, USA (3)PinPoint Testing, LLC, Little Rock, AR (4)Biomedical Forensic Sciences, Boston University School of Medicine, Boston, MA, USA

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Karen Scott (Presenter)
Arcadia University

Presenter Bio: I am an academic Forensic Toxicologist. I have been involved with both clinical and forensic casework since I started my PhD at the University of Glasgow in 1994. After completing my post-doc at the Institute of Health Science in Tokyo, Japan, I started my first academic position in Cambridge, UK, before returning to Glasgow University where I developed a MSc in Forensic Toxicology in additional to working as a Forensic Toxicologist for 5 years. I moved to the USA in 2012 to take up the role of Director and Associate Professor of Forensic Science. In addition to teaching, research and administration, I remain casework active, mainly in the area of DUID.

Relevant Financial Disclosures (within past 24 months)

Committee/Board/Advisory Board NSC ADID executive committee ; COFSE president

Salary Arcadia University

Discussing Products? NO, presenter will NOT present educational content related to products/services of the ineligible company(ies) listed above.

Conflict mitigation NOT REQUIRED.

Abstract

Introduction
Fentanyl is prescribed for the treatment of chronic pain, and is used as an adjunct drug during procedures requiring anesthesia. However, in 2016, fentanyl and its analogues were the most common cause of overdose deaths in the United States, with most of these overdose deaths due to illegal fentanyl. As part of the so called opioid epidemic, analysis of fentanyls places a huge strain in both clinical and forensic settings.
Objectives
The aim of this project was to develop and implement a fast and efficient method for the determination of fentanyl and its metabolite, norfentanyl, in clinical blood and urine samples as well as in post-mortem blood samples. A UPLC-MS, method utilizing a phospholipid depletion clean up method was developed, to ensure minimal down time which can result from analysis of complex clinical and forensic matrices.
Methods
Aliquots of blood or urine were pre-treated using Biotage Phospholipid Depletion Cartridges (PLD). Calibrators and controls were spiked using certified reference materials with calibrators ranging from 0.5-500 ng/mL and controls at 1, 75 and 300 ng/mL. To 100 L of blood or urine, 300 L of acetonitrile was added along with deuterated internal standards (200 ng/mL), then applied directly to the PLD cartridges. Once eluted, samples were evaporated under nitrogen and reconstituted in 90:10 water:methanol. Analysis was carried out on a LX-50 UHPLC, using a Restek Raptor biphenyl 2.7 micron 100 x 3.0 mm column with a QSight® 220 CR MS/MS in positive ion mode. A binary gradient of 5 mM ammonium formate in water and methanol, both containing 0.1% formic acid was used for a total run time of 7 minutes. Two ions were monitored for each analyte transition and one for the deuterated standards. Human clinical and post-mortem samples were analyzed as well as samples as well as rat femoral and heart blood samples obtained from a post-mortem redistribution study.
Results
The method was validated following parameters set by ASB Standard 036-Standard Practices for Method Validation in Forensic Toxicology. The method was successfully applied to human clinical and forensic blood samples, human clinical urine samples as well as rat femoral and heart blood samples taken at various stages of decomposition.
Conclusions
A sensitive, clean and accurate method has been developed and validated for the determination of fentanyl and its primary metabolite norfentanyl in a range of different type of blood samples as well as from human urine. The use of PDL cartridges allows for quick and efficient sample clean up and increased instrument up-time.