Molecular Networking: A Useful Tool for the Identification of New Psychoactive Substances in Seizures by LC–HRMS

Abstract

New Psychoactive Substances (NPS) are a global concern since they are spreading at an unprecedented rate. Despite their commerce still being limited compared to traditional illicit drugs, the identification of NPS in seizures may represent a challenge because of the variety of possible structures. In this study we report the successful application of molecular networking (MN) to identify unexpected fentanyl analogs in two seizures. The samples were extracted with 1 mL of methanol and analyzed with an untargeted data-dependent acquisition approach by LC–HRMS. The obtained data were examined using the MN workflow within the Global Natural Product Search (GNPS). A job was submitted to GNPS by including both seizures and standard mixtures containing synthetic cannabinoids and fentanyls raw files; spectra obtained from standards were used to establish representative networks for both molecular classes. All synthetic cannabinoids in the mixture were linked together resulting in a molecular network despite their different fragmentation spectra. Looking at fentanyls, all the molecules with the typical 188.143 and 105.070 fragments were combined in a representative network. By exploiting the standard networks two unexpected fentanyls were found in the analyzed seizures and were putatively annotated as para-fluorofuranylfentanyl and (iso)butyrylfentanyl. The identity of these two fentanyl analogs was confirmed by NMR analysis. Other m/z ratios in the seizures were compatible with fentanyl derivatives; however, they appeared to be minor constituents, probably impurities or synthetic byproducts. The latter might be of interest for investigations of common fingerprints among different seizures.

Simultaneous Quantification of 25 Fentanyl Derivatives and Metabolites in Oral Fluid by Means of Microextraction on Packed Sorbent and LC–HRMS/MS Analysis

“…To obtain a complete fragmentation pattern of the analytes, different runs were performed in full-scan data-dependent acquisition mode, each one with three different collision energies (CE) as described in a previously published method [ 19 ].…”

New psychoactive substance cocktail in an intensive care intoxication case elucidated by molecular networking

“…As molecular 3 networking method is based on the same principle with a mass range restricted to the only molecules of interest and their known metabolites, we here detected masses between 150 and 450 m/z using only positive ionization. Molecular networking has already proven to be a powerful tool for identifying NPS from drug seizures [3] and biological samples as exemplified with a 3-MeO-PCP intoxication [2]. The comparison of molecular network data obtained from patient's urine and from the seven powders contained in the bags found next to the patient allowed us to confirm the consumption of five molecules: 2F-DCK (2-fluorodeschloroketamine), CMC (chloromethcathinone), Nethylhexedrone, 3-MeO-PCP and 3-OHPCP (Figure 1).…”

Recent Advances in the Heterologous Biosynthesis of Natural Products from Streptomyces

“…The putative structure of molecules is closely linked to the structure of compounds found in the GNPS. The GNPS is a powerful tool for the discovery of new drugs and metabolites [52].…”

“…The advantage of molecular networking is the possibility of detecting analog compounds and identifying the biosynthetic pathways. Molecular networking permits finding unknown compounds and connecting "standard networks" for the annotation of new compounds [52]. For example, a saponin glycoside from the British Bluebell, a map of matlystatin congeners in various media, and the proposal of bottromycin pathway were explored via molecular networking [53][54][55].…”