The first report from Ankara on the presence of xylazine abuse in blood and urine samples using a validated LC-HRMS method.

Xylazine, an alpha-2 adrenergic receptor agonist approved for veterinary use, has emerged as a significant adulterant in illicit drugs, particularly synthetic opioids like fentanyl, contributing to a rise in fatal overdoses. Despite its widespread abuse in the United States and other regions, data on xylazine prevalence in Türkiye remain limited. This study aimed to develop and validate a sensitive and rapid liquid chromatography high-resolution mass spectrometry (LC-HRMS) method for detecting xylazine and its metabolites, 2,6-dimethylaniline (DMA) and 4-hydroxyxylazine, in human blood and urine samples. Solid-phase extraction (SPE) was used for sample preparation and the method demonstrated high sensitivity with limits of detection of 0.025-0.06 and 0.068 0.19 ng/mL for xylazine and its metabolites in blood and urine, respectively. Between January and December 2024, 9,123 biological samples from suspected drug users in Ankara were analyzed for xylazine and its metabolites. Xylazine was detected in 28 cases (0.3%). While 4-hydroxyxylazine was detected in 78% of positive cases, DMA was detected in 25% of positive cases. Polysubstance use was common, with pregabalin (50%) and methamphetamine (28%) being the most frequently co-detected substances. This study represents the first report of xylazine abuse in Türkiye, highlighting its emergence in the local drug supply. The findings underscore the need for routine xylazine screening in forensic toxicology to monitor its prevalence and mitigate its impact as an emerging public health threat. The developed LC-HRMS method offers a rapid, sensitive, and reliable tool for detecting xylazine and its metabolites, supporting efforts to address this growing concern.