INTRODUCTION: The use of new psychoactive substances as drugs of abuse has dramatically increased over the last years. Hallucinogenic phenethylamines gained particular popularity as they have both stimulating and psychedelic effects. Although generally perceived as safe, these illicit drugs pose a serious health risk; they have been linked to cases of severe poisoning or even deaths. Therefore, simple, cost-effective and reliable methods are needed for rapid determination of abused hallucinogens. METHODS: For this purpose, two haptens derived from 2C-H were designed, synthesized and subsequently attached to a carrier protein. Polyclonal antibodies obtained from a rabbit immunized with one of the prepared immunogens were used for the development of two immunoassays. RESULTS: In this study, a lateral flow immunoassay (LFIA) and an enzyme linked immunosorbent assay (ELISA) for the detection of 2C-B and related hallucinogenic phenethylamines in urine were developed. The presented LFIA is primarily suitable for on-site monitoring as it is simple and can provide a visual evidence of 2C-B presence within a few minutes. Its reasonable sensitivity (LODLFIA = 15 ± 7 ng mL-1) allows detection of the drug presence in urine after acute exposure. For greater accuracy, highly sensitive ELISA (LODELISA = 6 ± 3 pg mL-1) is proposed for toxicological quantitative analyses of positive samples captured by the LFIA. DISCUSSION: The comparison of the ELISA with the well-established UHPLC-MS-MS method shows excellent agreement of results, which confirms good potential of the ELISA to be used for routine analyses of 2C-B and related hallucinogenic phenethylamines of both main sub-families.

Department of Biochemistry and Microbiology University of Chemistry and Technology Prague Technická 3 166 28 Praha 6 Dejvice Czechia

Department of Chemistry of Natural Compounds University of Chemistry and Technology Prague Technická 5 166 28 Praha 6 Dejvice Czechia

Department of Chemistry of Natural Compounds University of Chemistry and Technology Prague Technická 5 166 28 Praha 6 Dejvice Czechia; Department of Biochemistry and Microbiology University of Chemistry and Technology Prague Technická 3 166 28 Praha 6 Dejvice Czechia

Department of Chemistry of Natural Compounds University of Chemistry and Technology Prague Technická 5 166 28 Praha 6 Dejvice Czechia; Forensic Laboratory of Biologically Active Substances University of Chemistry and Technology Prague Technická 3 166 28 Praha 6 Dejvice Czechia

Department of Chemistry of Natural Compounds University of Chemistry and Technology Prague Technická 5 166 28 Praha 6 Dejvice Czechia; Forensic Laboratory of Biologically Active Substances University of Chemistry and Technology Prague Technická 3 166 28 Praha 6 Dejvice Czechia; Department of Experimental Neurobiology National Institute of Mental Health Topolová 748 250 67 Klecany Czechia

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