MSACL 2024 Abstract

Copper State Opioid Crisis: Developing a Fentanyl and Fentanyl Analog in Blood Method for Fatal and Non-Fatal Overdoses

Dawn R Parker, Josette Madonia, Albert J Kind, Lucas C Erickson, Austin Echelmeier
Toxicology Program, Arizona State Public Health Laboratory (ASPHL), Phoenix, AZ

Austin Echelmeier, PhD (Presenter) Arizona Department of Health Services

Presenter Bio: Austin analyzes clinical samples with the Arizona State Public Health Laboratory (ASPHL) in the Chemical Emergency Response program and the Toxicology program.

Relevant Financial Disclosures (within past 24 months, reported on Jan 16, 2024) Grant/Research Support Center for Disease Control and Prevention Conflict mitigation NOT REQUIRED.

INTRODUCTION: According to the Centers for Disease Control and Prevention (CDC), nearly 300 lives are lost daily in the United States due to drug overdoses. To reduce overdoses and health disparities, the CDC launched the Overdose Data to Action (OD2A) program in 2019. ASPHL was awarded the OD2A grant in 2019 to assist with fatal overdose testing for county Medical Examiners. ASPHL has continued this work and is currently working to expand to non-fatal overdose testing. As part of this OD2A grant work, ASPHL continues to improve toxicology testing. One recent improvement was to develop and validate a method for fentanyl and fentanyl analogs in blood.

OBJECTIVES: The primary objective was to develop and validate a quantitative method on liquid chromatography tandem mass spectrometry (LC-MS/MS) to identify and quantify fentanyl and fentanyl analogs in human blood at clinically relevant concentrations.

METHODS: Human whole blood was prepared by agitation, centrifugation, supported liquid extraction (SLE) on a Biotage Extrahera liquid handler, dry down on a Biotage Turbovap, and reconstitution on a Biotage Extrahera. The samples were analyzed on an Agilent 1260 Infinity II LC Stack with a Sciex 7500 MS. The LC method used an Agilent Poroshell 120 EC-C18, 2.1 x 100 mm, 1.9 µm column with a 16-minute gradient elution of Mobile Phases A (10 mM Ammonium Formate with 0.05% Formic Acid in Water) and B (0.05% Formic Acid in Acetonitrile). The MS method used Multiple Reaction Monitor (MRM) transitions and retention times for a total of 26 fentanyl and fentanyl analog analytes. Data was processed and evaluated using Sciex OS software. The validation data was evaluated by the ANSI/ASB Standard 036, First Edition 2019, Standard Practices for Method Validation in Forensic Toxicology.

RESULTS: The method was evaluated for the following criteria: reportable range, calibration model, carryover, precision, accuracy, ionization suppression or enhancement, interference, stability, and uncertainty.

CONCLUSION: ASPHL developed and validated a quantitative LC-MS/MS method to detect fentanyl and fentanyl analogs in blood. The method will be used for fatal overdose patient samples that fall under the OD2A Strategy 3 grant work and non-fatal overdose patient samples that fall under the OD2A Strategy 4 grant work.