Lateral flow immunoassay and enzyme linked immunosorbent assay as effective immunomethods for the detection of synthetic cannabinoid JWH-200 based on the newly synthesized hapten

Lucie Fojtíková; Anna Šuláková; Martina Blažková; Barbora Holubová; Martin Kuchař; Petra Mikšátková; Oldřich Lapčík; Ladislav Fukal

doi:10.1016/j.toxrep.2017.12.004

Lateral flow immunoassay and enzyme linked immunosorbent assay as effective immunomethods for the detection of synthetic cannabinoid JWH-200 based on the newly synthesized hapten

Toxicol Rep. 2017 Dec 6:5:65-75. doi: 10.1016/j.toxrep.2017.12.004. eCollection 2018.

Authors

Lucie Fojtíková¹, Anna Šuláková², Martina Blažková¹, Barbora Holubová¹, Martin Kuchař², Petra Mikšátková², Oldřich Lapčík², Ladislav Fukal¹

Affiliations

¹ Department of Biochemistry and Microbiology, Faculty of Food and Biochemical Technology, University of Chemistry and Technology Prague, Technická 3, 166 28 Prague, Czech Republic.
² Department of Chemistry of Natural Compounds, Faculty of Food and Biochemical Technology, University of Chemistry and Technology Prague, Technická 3, 166 28 Prague, Czech Republic.

Abstract

In recent years, the use of synthetic cannabinoids (SCs) as drugs of abuse has greatly increased. SCs are associated with a risk of severe poisoning or even death. Therefore, more rapid, cost effective and reliable methods are needed, especially for the screening of drivers after traffic accidents and for detailed toxicological analysis in forensic laboratories. In this study, we developed a lateral flow immunoassay (LFIA) and an enzyme linked immunosorbent assay (ELISA) for the detection of JWH-200 in oral fluids. For this purpose a new hapten was prepared using a ten-step synthetic route. The developed immuno methods are based on antibodies obtained from rabbit immunized with synthesized hapten conjugated to carrier protein. The proposed methods are highly sensitive (LOD_LFIA = 0.08 ± 0.04 ng mL^-1; LOD_ELISA = 0.04 ± 0.02 ng mL^-1). They were applied to the quantification of JHW-200 in spiked oral fluids. The recoveries ranged from 82 to 134% for both methods. The results correlated excellently with results obtained using UHPLC-MS/MS (R²_LFIA = 0.99; R²_ELISA = 0.99). Our developed methods could be an important tool for analyses of JWH-200 in human oral fluids. The one-step LFIA is particularly suitable for roadside and on-site monitoring due to the rapid qualitative results it delivers, while the ELISA is especially useful for laboratory quantitative analyses of positive samples captured by LFIA.

Keywords: BSA, bovine serum albumin; DCC, N,N’-dicyclohexylcarbodiimide; DIBAH, diisobutylaluminium hydride; DMF, N,N-dimethylformamide; ELISA; ELISA, enzyme-linked immunosorbent assay; GAR, goat anti-rabbit antibody; GAR-Po, peroxidase labelled goat anti-rabbit antibody; Hapten synthesis; Immunomethods; JWH-200; LFIA; LFIA, lateral flow immunoassay; LOD, limit of detection; NBS, N-bromosuccinimide; NHS, N-hydroxysuccinimide; NPS, new psychoactive substances; PEG, polyethylene glycol; RSA, rabbit serum albumin; RSD, relative standard deviation; SCs, synthetic cannabinoids; Synthetic cannabinoid; THC, thin layer chromatography.