Assessment of QuEChERS extraction protocol for the LC-MS/MS detection of emerging fentanyl analogs in biological specimens for clinical and forensic purposes

Abstract

Drug abuse within the United States is a dynamic and pervasive issue that affects the quality and life expectancy of our society. One of the major drug classes that are constantly utilized is opioids. Opioids are prescribed for pain management in patients suffering from cancer and other medical illnesses. Long-term use of opioids could lead to tolerance in humans, which poses a risk of reaching fatal overdose levels. Unfortunately, opioids are abused recreationally, and in some instances, individuals are unaware of the presence of dangerous synthetic drugs like fentanyl, hidden in materials such as heroin, cocaine, oxycodone, and methamphetamine. Recent drug reports highlight the increase in seized opioids, which is also reflected by the number of associated deaths, escalating to epidemic levels. According to the National Institute of Drug Abuse (NIDA), opioid overdose deaths amount to 68,630 in 2020 (1). Therefore, the incorporation of novel approaches that offer superior sample preparation schemes for specimens of toxicological interest is greatly needed. To this end, this study aims to evaluate the presence and prevalence of a suite of synthetic opioids and other drugs in several postmortem biological specimens obtained from a cohort of individuals. Specifically, a sample preparation procedure called QuEChERs is used to assess its effectiveness when dealing with complex matrices like blood, liver, and brain. The comprehensive drug panel covers several drug entities, including fentanyl and other synthetic opioids, cathinones, D9-tetrahydrocanbinol (THC), amphetamine, PCP, and cocaine, to a maximum of thirty-five analytes and metabolites using a targeted LC-QQQ-MS system to quantify results. This study has two objectives: [1] to determine the drug distribution in multiple matrices from the same source individuals, and [2] to evaluate the utility of oral fluid in outpatient clinical settings for patients receiving addiction mediating medication. This research thesis explores the use of QuEChERs over a non-traditional toxicological matrix called oral cavity fluid as an initial effort to understand the advantages and limitations of this specimen in postmortem toxicology casework. Another goal is to correlate the drug or metabolites concentration across matched matrices from the same individual. Oral fluid was identified as an alternative matrix of interest in toxicology by the Organization for Scientific Area Committee (OSAC), and in response to this need, this project provides data that assist in filling this gap of knowledge. Additionally, a separate study will be conducted to evaluate the utility of oral fluid in outpatient clinical settings for patients receiving