Tryptamines constitute a class of organic compounds structurally derived from tryptamine, a natural alkaloid found in plants, fungi, and the human body. This group encompasses biologically significant molecules such as serotonin and melatonin, as well as psychoactive compounds like DMT and psilocybin. Tryptamines are typically classified into simple tryptamines and ergolines, with further subclassifications based on structural modifications on the indole ring system. Their primary mechanism of action involves modulation of serotonergic receptors, especially the 5-HT2A, often resulting in psychedelic effects. While some tryptamines are naturally occurring, others are synthetically produced and increasingly encountered as new psychoactive substances. The study presents a rapid and robust LC-MS/MS method for the simultaneous detection and quantification of 37 tryptamines in human urine and plasma. The method employs a ThermoFisher UHPLC-LC-MS/MS system using a Hypersil GOLD C18 column and gradient elution, with MS^n acquisition in positive ESI mode. Analytical validation demonstrated high selectivity, sensitivity, and reproducibility. Sample preparation was performed through a combination of liquid-liquid and protein precipitation extraction procedures. The method successfully identified tryptamines using product ion spectra compared to a reference spectral library, with detection limits, recovery, and matrix effects systematically evaluated. This analytical approach provides a reliable tool for forensic and clinical toxicology, particularly in the context of the increasing interest in tryptamines for both recreational and therapeutic purposes.
