Dried Blood Spot analysis as a sustainable and innovative approach for the detection of psychoactive substances and markers of alcohol abuse

The PhD project entitled “Dried Blood Spot analysis as a sustainable and innovative approach for the detection of psychoactive substances and markers of alcohol abuse”, had as its main purpose the development and validation of innovative, sustainable and effective analytical methods for the detection of common drugs of abuse (e.g. cocaine, THC, MDMA), new psychoactive substances (NPS) (e.g. fentanyl, nitazenes) and alcohol biomarkers (Phosphatidyl ethanol, known as PEth). The results of this work should be applicable in the daily routine of forensic toxicology laboratories and will introduce the non-invasive micro sampling dried blood spots (DBS) technique that can be performed by individuals outside a clinical context. Apart from the main goals of this study, other methods (ELISA), monitoring tools (hair analysis), matrices (whole blood, urine, toads) and other alarming substances (EtG-EtS, bufotenine, PFAS) were explored and included. In detail, one analytical method for the detection of PEth was developed and validated in Dried Blood Spots using HPLC-HRMS-Q-TOF using only 30 μL. Moreover, one analytical method for the detection of Nitazenes, a new group of potent synthetic opioids with a 2-benzyl-benzimidazole structural core, was developed and validated in volumetric DBS devises, the Capitainer®B, that use only 10 μL of sample. The quantification was performed by UHPLC-MS/MS. The 10 μL Capitainer®B devices were also used for the detection of 18 drugs of abuse and metabolites, including THC by UHPLC-MS/MS. The results of all the three studies resulted in participation in conferences (oral and poster) (PEthNET 2022, TIAFT 2022, 2023, 2024) and in scientific publications. The PEth method is currently applied as a routine method During the research stay at the Laboratory of Toxicology, Ghent University, Belgium the work was focused on three research projects that are related to the determination of alcohol biomarkers, mainly Phosphatidyl ethanol (PEth), in biological matrices but the research frame was broadened to the detection of PEth with ELISA, the detection of Ethyl glucuronide (EtG) and Ethyl sulfate (EtS) in urine samples and the simultaneous detection of PEth, EtG, EtS in dried blood spots and in dried urine spots. Lastly, an analytical Reference Measurement Procedure for the detection of PEth in whole blood was examined and developed which also resulted in a poster submission (TIAFT 2023) and in a best presentation award (PEthNET 2024). Moreover, the development of an analytical method for the detection of of 5-OH-DMT (Bufotenine) in the Asian toad Duttyphrynus melanostictus in Madagascar was achieved as bufotenine, which is a tryptamine derivative related to the neurotransmitter serotonin, is proven to have psychoactive effects when found in large amounts. The results of this work were presented as poster submission (EUROTOX 2024) and a publication to scientific journal is prepared.