Work

Development and validation of a simple and fast method for routine analysis of new synthetic opioids and hallucinogens in whole blood using protein precipitation and UHPLC-MS/MS

Year: 2025

Type: article

Abstract: In forensic toxicology, the rapid and reliable detection of emerging synthetic opioids and hallucinogens is crucial for case investigations and public health monitoring. This work describes the development, optimization and validation of a simple, fast and sensitive methodology for the simultaneous analysis of 6 new synthetic opioids (carfentanil, fentanyl, isotonitazene, metonitazene, norfentanyl, and sufentanil) and 2 hallucinogens (lysergide [LSD] and mescaline), together with the main LSD metabolite 2-oxo-3-hydroxy-lysergide [LSD-OH], in whole blood samples by liquid chromatography coupled to tandem mass spectrometry. Under optimized experimental conditions, linearity was verified between 0.1 and 20 ng/mL for all analytes except mescaline (2.5 to 500 ng/mL), with r 2 > 0.99 for 1/x weighting, and no significant carryover or matrix effects were observed. Good precision (% RSD < 13%) and trueness (% Bias within ± 20%) values were achieved. The estimated limit of quantification (LOQ) was 0.1 ng/mL for all compounds except mescaline (2.5 ng/mL). Authentic forensic samples were also analyzed, and positive samples for fentanyl, norfentanyl, and sufentanil were identified. The proposed methodology allows the simultaneous analysis of compounds from different families of psychoactive substances, in both postmortem and in vivo samples, using only 50 µL of whole blood. The demonstrated speed, simplicity, and effectiveness make it particularly advantageous for routine implementation in forensic toxicology laboratories. • First known UHPLC-MS/MS method for nitazenes and hallucinogens analysis; • Validation of fast method for in vivo and postmortem whole blood samples analysis; • Low sample volume required is advantageous for forensic toxicology laboratories; • Sensitive and extensive linear range for a broad spectrum of concentrations; • Applied to suspected intoxication cases and feasible to routine forensic work.

Source: Forensic Science International

DOI: 10.1016/j.forsciint.2025.112713

Authors: Joana R.P. Pereira, Mónica Antunes, Nuno R. Neng, Carla Mustra, João Franco, Suzana Fonseca