This article reports on a stability assessment in the evaluation of synthetic cannabinoid metabolites in human blood in the absence of parent compounds.
Synthetic cannabinoids are a chemically diverse class of novel psychoactive substances (NPS) responsible for large analytical and interpretative challenges for forensic toxicologists. Between 2016 and 2019, the three most prevalent synthetic cannabinoids in the United States were MMB-FUBINACA (FUB-AMB), 5F-MDMB-PINACA (5F-ADB) and 5F-MDMB-PICA, based on results from seized drug and toxicology testing. In 2018, accurate determination of synthetic cannabinoid positivity was brought into question when it was determined that the metabolites of these drug species were present in the absence of parent compounds in forensically relevant blood samples. During the current study, the stability of MMB-FUBINACA, 5F-MDMB-PINACA, and 5F-MDMB-PICA was evaluated, as well as the characterization of breakdown products. A liquid–liquid extraction method was assessed for recovery of basic parent compounds and acidic metabolites and deemed fit for use in this study. Analysis was performed by liquid chromatography–quadrupole time-of-flight mass spectrometry (LC–QTOF-MS) using a SCIEX TripleTOF® 5600+. All three synthetic cannabinoids were found to be unstable when stored in blood at either room temperature or refrigerated; all analytes were considerably more stable when stored in the freezer. All three synthetic cannabinoids degraded to their respective butanoic acid metabolites: MMB-FUBINACA 3-methylbutanoic acid, 5F-MDMB-PINACA 3,3-dimethylbutanoic acid and 5F-MDMB-PICA 3,3-dimethylbutanoic acid. All three of these metabolites were studied and determined to be stable in blood at all storage conditions. Considering these results, the authors’ laboratory continued testing for synthetic cannabinoid metabolites in blood samples and found 83 positives (21 percent) for only a synthetic cannabinoid metabolite. A case report is presented in this article which indicates that 5F-MDMB-PINACA 3,3-dimethylbutanoic acid was identified in the absence of 5F-MDMB-PINACA. Forensic toxicologists should be aware of the results of this study, since they directly impact analytical consideration for test development and implementation, as well as interpretation of findings. (publisher abstract modified)
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