Determination of 19 Psychoactive Substances in Premortem and Postmortem Whole Blood Samples Using Ultra-High-Performance Liquid Chromatography–Tandem Mass Spectrometry

Sevasti Karampela,Irene Panderi,Jessica Smith

doi:10.3390/separations8060078

Abstract

An ever-increasing need exists within the forensic laboratories to develop analytical processes for the qualitative and quantitative determination of a broad spectrum of new psychoactive substances. Phenylethylamine derivatives are among the major classes of psychoactive substances available on the global market and include both amphetamine analogues and synthetic cathinones. In this work, an ultra-high-performance liquid chromatography-positive ion electrospray ionization tandem mass spectrometric method (UHPLC-ESI-MS/MS) has been developed and fully validated for the determination of 19 psychoactive substances, including nine amphetamine-type stimulants and 10 synthetic cathinone derivatives, in premortem and postmortem whole blood. The assay was based on the use of 1 mL premortem or postmortem whole blood, following solid phase extraction prior to the analysis. The separation was achieved on a Poroshell 120 EC-C18 analytical column with a gradient mobile phase of 0.1% formic acid in acetonitrile and 0.1% formic acid in water in 9 min. The dynamic multiple reaction monitoring used in this work allowed for limit of detection (LOD) and lower limit of quantitation (LOQ) values of 0.5 and 2 ng mL−1, respectively, for all analytes both in premortem and postmortem whole blood samples. A quadratic calibration model was used for the 12 quantitative analytes over the concentration range of 20–2000 ng mL−1, and the method was shown to be precise and accurate both in premortem and postmortem whole blood. The method was applied to the analysis of real cases and proved to be a valuable tool in forensic and clinical toxicology.

Highlights

Over the past decade, the trend towards substance abuse has changed, and many new psychoactive substances have emerged as “legal high” alternatives to traditional illicit drugs [1]
A quantitation method was developed for the 12 psychoactive substances, including the nine amphetamine-type stimulants and three synthetic cathinone derivatives MPHD, bupropion, which is used for both therapeutic and recreational purposes, and its metabolite
The maximum permitted tolerance for the relative fragment ions’ intensity was set at ± 20% for a positive result of all the analytes in accordance with the standard for mass spectral data acceptance in forensic toxicology originally conceived by the Scientific Working Group of Forensic Toxicology (SWGTOX) and further developed by the toxicology subcommittee of the Organizational Scientific Area Committee (OSAC) [51]

Summary

Introduction

The trend towards substance abuse has changed, and many new psychoactive substances have emerged as “legal high” alternatives to traditional illicit drugs [1] This phenomenon was accompanied by increased incidences of people in intensive care units of hospitals after substance use with unknown pharmacological and/or toxicological action [2]. Phenylethylamine derivatives are serotoninergic receptor agonists leading to psychedelic effects and in some cases inhibiting monoamine reuptake [5]. These compounds are among the major classes of psychoactive substances available on the global market and include both amphetamine and β-keto amphetamine analogues (synthetic cathinones). The Centers for Disease Control and Prevention (CDC) warns that overdose deaths in the USA from psychostimulants, including methamphetamine, have steadily increased from 1999 to 2019 [12]

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