Verapamil is an L-type calcium channel blocker widely used in the treatment of cardiovascular diseases such as hypertension, angina and arrhythmias, requiring accurate therapeutic monitoring to maintain plasma, urine and saliva concentrations within a safe range. In this context, a novel analytical approach has been proposed to determine verapamil in biological samples, using bar adsorptive microextraction coated with reversed-phase polymers followed by high-performance liquid chromatography with diode array detection. Two adsorbents have been chosen, i.e. STRATA-CN and ENVI-18 polymers, showing recoveries from 56.01 ± 2.16% to 96.82 ± 0.61% under optimized experimental conditions, such as, sample pH: 10.0 (STRATA-CN) and 8.0 (ENVI-18), 2 h of equilibrium time, stirring speed at 990 rpm, back-extraction solvent using methanol: acetonitrile (1:1 v/v), and 1 h under sonication. The analytical method showed linearity from 20 to 600 ng mL-1 (r ≥ 0.99), as well as adequate precision (with RSD% below 15%) and accuracy (with RE% within 15% of the nominal value). Finally, the analytical method was applied to plasma, urine and saliva samples and proved to be a promising alternative for the trace analysis of verapamil in biological matrices.