Studies on the In Vitro and In Vivo Metabolic Fate of the New Psychoactive Substance N-Ethyl-N-Propyltryptamine for Analytical Purposes

Abstract

Prerequisites for the reliable identification of substances in terms of forensic and clinical toxicology or doping control include knowledge about their metabolism and their excretion patterns in urine. N-Ethyl-N-propyltryptamine (N-ethyl-N-[2-(1H-indol-3-yl)ethyl]propan-1-amine, EPT) is an N,N-dialkylated tryptamine derivative, sold as new psychoactive substance, and supposed to act as a partial agonist at the 5-HT2A receptor. The aims of the presented study were to elucidate in vitro metabolites of EPT after incubations with pooled human liver S9 fraction (pS9) and in vivo metabolites excreted into rat urine. Finally, suitable analytical target compounds should be identified. Analysis of pS9 incubations using liquid chromatography-high-resolution tandem mass spectrometry revealed EPT metabolites formed after N-dealkylation as well as alkyl and aryl hydroxylation and formation of a hydroxy sulfate. Investigations using rat urine after oral dosing showed that the metabolic pathways of EPT shifted from in vitro hydroxylation of the alkyl amine group to an increased in vivo hydroxylation of the indole ring with several N-dealkyl metabolites. A glucuronic acid conjugate after hydroxylation of the indole ring was additionally found in vivo. The parent compound could not be detected in the rat urine samples. Therefore, analytical methods using mass spectrometry should include hydroxy-EPT and two hydroxy-EPT glucuronide isomers for reliable identification.