There is a strong need for certified reference materials in the quality assurance of nonionic soil contaminant bioavailability estimations through physicochemical methods. We applied desorption extraction, a method recently standardized as ISO16751, to determine the bioavailable concentration of the most commonly regulated polycyclic aromatic hydrocarbon (PAH), benzo(a)pyrene (BaP), in the reference industrial soil BCR-524 with a certified BaP total concentration of 8.60 mg kg−1. This concentration represented BaP levels found in many PAH-polluted soils. The method, based on single-point extraction of the analyte desorbed into the aqueous phase by a receiving phase (Tenax or cyclodextrin), was applied ten times. The data fulfilled highly demanding quality criteria based on recovery and repeatability. The bioavailable BaP concentration detected through Tenax extraction, 1.82 mg kg−1, was comparable to bioavailable concentrations determined in field-contaminated soils and to environmental quality standards based on previously observed total BaP concentrations. There was good agreement (Student's t-test, P ≤ 0.05) with the bioavailable BaP concentration determined by cyclodextrin extraction (1.53 mg kg−1). The methods were extended to four other certified 4- and 5-ringed PAHs for comparative purposes. We suggest ways of improving of the ISO16751 standard related to further systematic assessment of the Tenax-to-soil ratio and incorporation of mass balances. Furthermore, BCR-524 is suitable for quality-assurance protocols with these methods when used in site-specific risk assessments of PAH-polluted environments.