Rapid identification of MDMB-CHMINACA metabolites using Zebrafish and Human Liver microsomes as the Biotransformation system by LC-QE-HF-MS

Abstract

MDMB-CHMINACA is a newly synthetic cannabinoid which scoped in NMS Lab, USA. Since there are currently no published data on MDMB-CHMINACA metabolism, we aimed to identify its biotransformation pathways and major metabolites. Liquid chromatography Q-Extractive HF Hybrid Quadrupole-Orbitrap mass spectrometry (LC-QE-HF-MS) using full scan positive ion mode and targeted MS/MS (ddms2) techniques with accurate mass measurement were employed to analyze the metabolic sites and pathways. An in vivo metabolic animal model of zebrafish was established to verify the metabolic pathways of MDMB-CHMINACA obtained from human liver microsomal experiment in vitro. The results showed that 29 metabolites were generated in the zebrafish animal model and human liver microsomes model. Biotransformations mainly occurred at the cyclohexylmethyl tail of the compound, minor reactions also occurred at the tert-butyl chain, and no reaction was analysised at the indazole ring. We recommend M1 group (MDMB-CHMINACA ester hydroxylation), and M2 group (MDMB-CHMINACA monohydroxylation) as the potential poisoning markers to document MDMB-CHMINACA intake in clinical and forensic cases. Additionally, this study provides preliminary information regarding the metabolism of MDMB-CHMINACA that will guide analytical standard manufacturers to better provide suitable references for further studies on newly encountered designer drugs.

Metabolic stability studies of lead compounds supported by separation techniques and chemometrics analysis

“…Most popular workflow assumes incubations of studied compounds with enzymatic systems (such as Human Liver Microsomes, or hepatocytes) [8] followed by quantitative (for metabolic stability estimation) analysis. There are other approaches and enzymatic systems, including microsomes from animals [9–19], however human liver microsomes were used in the vast majority of cited research. Such analysis enables to estimate how the drug concentration in the incubation mix changes over time.…”

Metabolism of ADB‐FUBIATA in zebrafish and pooled human liver microsomes investigated by liquid chromatography–high‐resolution mass spectrometry

“…ADB-FUBIATA, AD-18, FUB-ACADB), a structural analog of ADB-FUBINACA and ADB-FUBICA, which were known as the eighth generation of SCs with an indazole/indole amide moiety. 13,14 In 2021, this substance appeared in Russia, United States, and China. [15][16][17] The chemical structure consists of three parts: an indole or an indazole core, a dimethylbutanamide side chain, and 4-fluoropentyl tail.…”

“…11 Researchers have studied the metabolism of 4F-MDMB-BICA, AMB-FUBINACA, and MDMB-CHMINACA in the zebrafish model, which provides a more authentic characterization of metabolites. 13,37,40 Zebrafish resemble mammals in their physiology, morphology, and histology, and they show 70% genetic homology with humans. [41][42][43][44] The small size, easy maintenance, low cost, and high reproductive rate are considerable advantages for whole-organism studies with ethical permission in biology, toxicology, drug discovery, and neurobiology.…”

Metabolism of 4F‐MDMB‐BICA in zebrafish by liquid chromatography–high resolution mass spectrometry

“…Studies on the metabolism of MDMB-CHMINACA and AMB-FUBINACA by zebrafish were recently published. 32,33 The specific goals of this work are to study in vivo metabolism of 4F-MDMB-BICA by zebrafish model.…”