Identification of the Synthetic Cannabinoid 1-(4-cyanobutyl)-N-(2-phenylpropan-2-yl)-1H-indazole-3-carboxamide (CUMYL-4CN-BINACA) in Plant Material and Quantification in Post-Mortem Blood Samples

Abstract

In May 2016, a new type of synthetic indazole-3-carboxamide cannabinoid (CUMYL-4CN-BINACA) was detected in seized plant material submitted to the Istanbul Council of Forensic Medicine by the National Police Office. The major ingredient in this material was purified using preparative liquid chromatography, and its structure was identified using liquid chromatography-high-resolution mass spectrometry (LC-HR/MS), gas chromatography-electron ionization/mass spectrometry (GC-EI/MS), nuclear magnetic resonance (NMR) spectroscopy and Fourier transform-infrared spectroscopy (FT-IR). Using HR-MS, the molecular formula of the compound was determined to be C22H24N4O (MW = 360.1950). The 1H and 13C-NMR and FT-IR spectrometric data revealed that the structure of compound was 1-(4-cyanobutyl)-N-(2-phenylpropan-2-yl)-1H-indazole-3-carboxamide (CUMYL-4CN-BINACA). After identification, it was quickly added to our generic drug list, and an ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS-MS) method was developed to determine its presence in blood samples. This study reports on the identification of CUMYL-4CN-BINACA in plant material using LC-HR/MS, GC-EI/MS, NMR and FT-IR as well as a validated method for quantification of CUMYL-4CN-BINACA in post-mortem blood samples by UPLC-MS-MS analysis. The quantification method has been validated in terms of linearity (0.1-50 ng/mL), selectivity, intra- and inter-assay accuracy and precision (CV < 15%), recovery (94-99%), limit of detection (0.07 ng/mL) and limit of quantification (0.1 ng/mL). Matrix effects, stability and process efficiency were also assessed. The method has been applied to 2,350 post-mortem blood samples from the autopsy cases in the Morgue Department of the Council of Forensic Medicine (Istanbul, Turkey) between 1 July 2016 and 31 December 2016.