Akademik Veri Yönetim Sistemi
MICROCHEMICAL JOURNAL, cilt.218, sa.115793, ss.115793, 2025 (SCI-Expanded, Scopus)
Xylazine, a substance abused in doping, drug-facilitated crimes, and accidental or intentional poisonings, has emerged as a significant global public health concern amid its rapid rise as an adulterant in illicit drugs over the past decade. This study aimed to develop a UHPLC-Q Exactive Focus Orbitrap tandem MS method for the determination of substances (fentanyl, ketamine, and cocaine) commonly adulterated with xylazine and their metabolites in urine samples following enzymatic hydrolysis. The method was validated in terms of interference studies, carryover, detection and quantification limits, linearity, bias, precision, extraction efficiency, and stability. It showed no significant carryover or matrix interference and yielded high recovery (93–112 %), acceptable bias (−9.00 % to 15.48 %), strong linearity (R2 ≥ 0.9925), and precise results (RSD ≤ 11.35 %) across all analytes. Application to an authentic urine sample confirmed, for the first time, the simultaneous presence of all target analytes and their key metabolites at concentrations ranging from 1.9 to 62.6 ng/mL. Additionally, 4-hydroxy-xylazine, a xylazine metabolite, was qualitatively monitored. In this context, the toxicity and alpha-2 adrenergic receptor binding potentials of 4-hydroxy-xylazine and xylazine were evaluated for the first time in silico, contributing novel insight to the field. This study highlights the increasing threat of xylazine adulteration in polydrug exposures, contributes to forensic and clinical analysis, and provides a valuable reference for future studies. As the xylazine crisis continues at an alarming rate in the world, necessary measures should be taken in our country and internationally before it becomes a widespread public health emergency.