Target metabolomics for forensic analysis: Biomarker identification of synthetic opioids using LC-MS/MS

The rapid global spread of new psychoactive substances (NPS) in the illicit drug market, accompanied by a steady increase in the number and diversity of these substances, poses an increasingly complex challenge for forensic scientists, doctors and regulatory authorities.[1] It is therefore, essential to develop innovative analytical approaches to contrast the new drugs phenomenon. In recent years, several studies conducted on NPS have shown that metabolomics can be a valuable solution for identifying biomarkers useful for detecting NPS consumption. [2] The purpose of this work is to develop a target metabolomics method to investigate systemic effects induced by opioids belonging to different classes, regardless of their chemical structure. Specifically, we investigated the impact of four opioids – morphine, brorphine, etonitazene, and fentanyl – on the urinary metabolic profile of CD-1 mice. The analysis was conducted using liquid chromatography coupled to mass spectrometry LC-MS, using QTRAP mass spectrometer, working in Multiple Reaction Monitoring (MRM) mode. Twenty-four metabolites have been included in the study, mostly of them belong to the amino acid, catecholamines and tricarboxylic acids classes, that were shown to be significantly altered following opioid intake in previous untargeted metabolomics studies.