Abstract
The laboratory analysis of new psychoactive substances and related drugs is crucial for accurate clinical and forensic diagnosis of poisonings. Given this, a new LC-MS/MS method for analyzing hallucinogens, synthetic cathinones, and synthetic cannabinoids in urine was developed. Urine samples were extracted using a liquid-liquid extraction protocol optimized via a multivariate experimental design. An aliquot of 400 μL of urine was enzymatically hydrolyzed with β-glucuronidase and extracted with 700 μL of ethyl acetate. The resulting extracts were analyzed using LC-MS/MS, with a total chromatographic run time of 8 min. The method was validated according to the ANSI/ASB Standard 036 guideline and was applied to 24 samples from suspected poisoning cases. The lower limits of quantification ranged from 0.1 to 1 ng/mL. Within-run and between-run precision (CV) were < 16%, and bias ranged from -12.8% to 19.8%. Nine of the 20 analytes investigated showed significant ionization suppression or enhancement (> 25%). Only two analytes (2C-E and 2-oxo-3-OH-LSD) had a recovery rate lower than 70%. Among the 24 analyzed urine samples, one tested positive for 25B-NBOH, one for LSD, and two for 2-oxo-3-OH-LSD. The developed method enables the simultaneous quantification of 20 illicit drugs, serving as an efficient diagnostic tool for clinical and forensic laboratories.