Metabolite-targeted LC–HRMS quantification of emerging indazole-based synthetic cannabinoids in urine samples from an addiction treatment cohort

Eroglu H. I., Arslan Z., Örüm T. G. Y., Bulut C., Kazdal F. S., Civek S., ...Daha Fazla

JOURNAL OF CHROMATOGRAPHY B: ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES, cilt.1285, ss.1-15, 2026 (SCI-Expanded, Scopus)

Özet

Abstract

The continuous emergence of structurally diverse indazole-based synthetic cannabinoids (SCs) challenges routine analytical detection because parent compounds are rapidly metabolized and often absent in urine. Therefore, sensitive metabolite-targeted analytical methods are required for reliable exposure assessment.
In this study, we adapted and fully validated a targeted high-resolution LC–HRMS method for the quantitative determination of eight contemporary indazole-based SC markers in human urine and applied it to 50 authentic urine specimens from treatment-seeking individuals attending an addiction treatment clinic. The method allowed the identification of robust analytical performance, with LOD and LOQ values of 0.01–0.46 ng/mL and 0.04–1.53 ng/mL, respectively, along with acceptable recovery (81.54–111.72%), bias (−4.47–13.17%), precision (RSD 5.12–15.03%), and matrix effects (75.92–100.03%). All target analytes were stable for one week at room temperature, 4 °C, and −20 °C, and no significant carryover was observed.
In this treatment-seeking cohort, 76% of samples were positive for at least one SC marker, with eight markers identified across MDMB-PINACA, MDMB-BINACA, and ADB-BUTINACA analogs. MDMB-4en-PINACA butanoic acid was the dominant biomarker (62%), followed by MDMB-INACA 3,3-dimethylbutanoic acid (42%), for which this study provides the first validated quantitative urine data. Other markers were detected at lower frequencies, and frequent co-detection indicates widespread polydrug SC use.
The validated high-resolution LC–HRMS method enables reliable metabolite-based assessment of exposure to emerging SCs in authentic urine samples. The predominance of MDMB-type metabolites demonstrates the applicability of the method for monitoring current trends in SC exposure, while the quantitative characterization of MDMB-INACA 3,3-dimethylbutanoic acid expands the available analytical data for newly emerging SCs. This analytical platform provides a robust tool for forensic and clinical toxicology laboratories to monitor SC exposure in rapidly evolving drug markets.