The main analytical targets of synthetic cannabinoids are often metabolites. With the high number of new psychoactive substances entering the market, suitable workflows are needed for analytical target identification in biological samples. The aims of this study were to identify the main metabolites of the synthetic cannabinoids, AMB-CHMICA and 5C-AKB48, using an in silico-assisted workflow with analytical data acquired using ultra-high-performance liquid chromatography–(ion mobility spectroscopy)–high resolution–mass spectrometry in data-independent acquisition mode (UHPLC-(IMS)-HR-MS^E). The metabolites were identified after incubation with rat and pooled human hepatocytes using UHPLC-HR-MS^E, followed by UHPLC-IMS-HR-MS^E. Metabolites of AMB-CHMICA and 5C-AKB48 were predicted with Meteor (Lhasa Ltd) and imported to the UNIFI software (Waters). The predicted metabolites were assigned to analytical components supported by the UNIFI in silico fragmentation tool. The main metabolic pathway of AMB-CHMICA was O-demethylation and hydroxylation of the methylhexyl moiety. For 5C-AKB48, the main metabolic pathways were hydroxylation(s) of the adamantyl moiety and oxidative dechlorination with subsequent oxidation to the ω-COOH. The matrix components in the metabolite spectra were reduced with IMS, which improved the accuracy of the spectral interpretation; however, this left fewer fragment ions for assigning sites of metabolism. Meteor was able to predict the majority of the metabolites, with the most notable exception being the oxidative dechlorination and, consequently, all metabolites that underwent that transformation pathway. Oxidative dechlorination of ω-chloroalkanes in humans has not been previously reported in the literature. The postulated metabolites can be used for screening of biological samples, with four-dimensional identification based on retention time, collision cross section, precursor ion, and fragment ions.

合成大麻素的主要分析目标通常是代谢产物。随着大量新型精神活性物质进入市场，需要合适的工作流程来鉴定生物样品中的分析目标。本研究的目的是确定的合成大麻素，AMB-CHMICA和5C-AKB48的主要代谢物，使用在硅片-assisted工作流程与使用超高性能液相色谱（离子迁移光谱法）所获取的分析数据-与数据无关的采集模式下的高分辨率质谱（UHPLC-（IMS）-HR-MS ^E）。在使用UHPLC-HR-MS ^E与大鼠和合并的人类肝细胞温育后，鉴定出代谢物，然后使用UHPLC-IMS-HR-MS ^E。用Meteor（Lhasa Ltd）预测AMB-CHMICA和5C-AKB48的代谢物，并将其导入UNIFI软件（Waters）。将预测的代谢物分配给由UNIFI in silico破碎工具支持的分析成分。AMB-CHMICA的主要代谢途径是O-甲基己基部分的-脱甲基化和羟基化。对于5C-AKB48，主要的代谢途径是金刚烷基部分的羟基化和氧化脱氯，随后氧化成ω-COOH。IMS减少了代谢物光谱中的基质成分，提高了光谱解释的准确性；但是，这留下了更少的碎片离子来分配新陈代谢的位点。流星能够预测大多数代谢产物，最显着的例外是氧化脱氯，因此，所有经历该转化途径的代谢产物。先前尚未在文献中报道人类中ω-氯烷烃的氧化脱氯。假定的代谢物可用于筛选生物样品，