Metabolite markers for three synthetic tryptamines N‐ethyl‐N‐propyltryptamine, 4‐hydroxy‐N‐ethyl‐N‐propyltryptamine, and 5‐methoxy‐N‐ethyl‐N‐propyltryptamine

N-Ethyl-N-propyltryptamine (EPT), 4-hydroxy-N-ethyl-N-propyltryptamine (4-OH-EPT), and 5-methoxy-N-ethyl-N-propyltryptamine (5-MeO-EPT) are tryptamine-class new psychoactive substances sold online. Their metabolism was previously undescribed. Incubating these compounds with pooled human liver microsomes for up to 4 hours and analyzing with high-performance liquid chromatography and mass spectrometry revealed major metabolic pathways. EPT was primarily metabolized by hydroxylation, N-dealkylation, and carbonylation. 4-OH-EPT metabolism involved double bond formation, N-dealkylation, hydroxylation, and carbonylation. 5-MeO-EPT underwent O-demethylation, hydroxylation, and N-dealkylation. Unique metabolites for 4-OH-EPT were identified in a human postmortem blood sample from a suspected EPT or 4-OH-EPT intoxication, demonstrating the markers' forensic utility.

Study at a glance

Abstract

N-Ethyl-N-propyltryptamine (EPT), 4-hydroxy-N-ethyl-N-propyltryptamine (4-OH-EPT), and 5-methoxy-N-ethyl-N-propyltryptamine (5-MeO-EPT) are new psychoactive substances classified as tryptamines, sold online. Many tryptamines metabolize rapidly, and identifying the appropriate metabolites to reveal intake is essential. While the metabolism of 4-OH-EPT and 5-MeO-EPT are not previously described, EPT is known to form metabolites by indole ring hydroxylation among others. Based on general knowledge of metabolic patterns, 5-MeO-EPT is also expected to form ring hydroxylated EPT (5-OH-EPT). In the present study, the aim was to characterize the major metabolites of EPT, 4-OH-EPT, and 5-MeO-EPT, to provide markers for substance identification in forensic casework. The tryptamines were incubated with pooled human liver microsomes at 37°C for up to 4 h. The generated metabolites were separated and detected by ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry analysis. The major in vitro EPT metabolites were formed by hydroxylation, N-dealkylation, and carbonylation. In comparison, 4-OH-EPT metabolism was dominated by double bond formation, N-dealkylation, hydroxylation, and carbonylation in vitro and hydroxylation or carbonylation combined with double bond loss, carbonylation, N-dealkylation, and hydroxylation in vivo. 5-MeO-EPT was metabolized by O-demethylation, hydroxylation, and N-dealkylation in vitro. The usefulness of the characterized metabolites in forensic casework was demonstrated by identification of unique metabolites for 4-OH-EPT in a human postmortem blood sample with suspected EPT or 4-OH-EPT intoxication.