Optimization of detection of twelve polycyclic aromatic hydrocarbons in Carpathian barbel ( <i>Barbus carpathicus</i> ) tissues: extraction methods and GC-MS parameters

A GC-MS method in selected ion monitoring (SIM) mode was optimized and validated for the determination of twelve polycyclic aromatic hydrocarbons (PAHs) in <i>Barbus carpathicus</i> tissues (muscle and hepatopancreas). Two chromatographic columns (HP-5ms and DB-EUPAH) were evaluated with multiple temperature programs, and the DB-EUPAH column proved superior for separating critical PAH isomers within a 25-minute runtime. Method validation demonstrated excellent linearity (R² ≥ 0.995), with deviation from linearity <20%, precision expressed as relative standard deviation (RSD ≤ 10%), and detection limits (LOD) ranging from 0.3 to 2.2 ng/mL and quantification limits (LOQ) from 0.95 to 6.85 ng/mL across the twelve validated analytes. Sample preparation optimization comparing ultrasonic and shaker-assisted extraction revealed that mechanical shaker extraction yielded superior results, particularly in the 12-16 min retention time window. Subsequent solid-phase extraction (SPE) purification using dichloromethane as the eluent significantly reduced matrix interference. Overall recovery rates ranged from 63.1% to 146.0%, with eight PAHs meeting acceptable criteria (70-120% recovery) for <i>B.carpathicus</i> tissue analysis. The developed method provides a reliable, validated analytical tool suitable for routine monitoring and risk assessment of PAH contamination in <i>B.carpathicus</i>, supporting environmental monitoring and food safety protocols.