The growing presence of New Psychoactive Substances, especially synthetic opioids, represents a critical challenge for public health authorities. These compounds, including fentanyl analogues and the more recent class of nitazenes, are known for their extreme potency and high risk of fatal overdoses. Additionally, classical psychoactive agents such as hallucinogens are regaining relevance, both in the context of intoxication and through emerging clinical applications. Considering these developments, this study aimed to establish a straightforward and robust method for the simultaneous analysis of synthetic opioids and hallucinogens in forensic whole blood samples.
A quantitative method was designed for the analysis of selected synthetic opioids and hallucinogens, including fentanyl, isotonitazene, LSD, and their analogues. The analytical workflow involved protein precipitation from small volumes of whole blood, using a Full Factorial Design of Experiments to fine-tune the preparation parameters. The resulting extract was directly introduced into an UHPLC system coupled with tandem mass spectrometry (UHPLC-MS/MS). The total run time was 5 minutes, and compounds were monitored in multiple-reaction monitoring mode with two specific transitions per analyte. Method validation was conducted following the ANSI/ASB Standard 036 guidelines.
The method showed excellent linearity between 0.1 and 20 ng/mL. The detection limits ranged from 0.05 and 1 ng/mL. No significant matrix interference or carryover was detected. Both the intra- and inter-day precision and accuracy met the acceptance criteria of <20% and ±20%, respectively. Selectivity was demonstrated through an analysis of spiked samples containing common therapeutic drugs and illicit substances.
This newly developed approach offers forensic toxicology laboratories an efficient and reliable tool for monitoring both synthetic opioids and hallucinogens in whole blood samples. Its operational simplicity, high sensitivity, and effectiveness make it well suited to routine toxicological investigations in both postmortem and clinical scenarios.
