To facilitate the study of potential harmful compounds sorbed to microplastics, an effect-directed analysis using the DR CALUX® assay as screening tool for Aryl hydrocarbon receptor (AhR)-active compounds in extracts of marine deployed microplastics and chemical analysis of hydrophobic organic compounds (HOCs) was conducted. Pellets of three plastic polymers (low-density polyethylene (LDPE), high-density polyethylene (HDPE) and high-impact polystyrene (HIPS)) were deployed at Heron Island in the Great Barrier Reef, Australia, for up to 8 months. Detected AhR-mediated potencies (bio-TEQs) of extracted plastic pellets ranged from 15 pg/g to 100 pg/g. Contributions of target HOCs to the overall bioactivities were negligible. To identify the major contributors, remaining plastic pellets were used for fractionation with a gas chromatography (GC) fractionation platform featuring parallel mass spectrometric (MS) detection. The bioassay analysis showed two bioactive fractions of each polymer with bio-TEQs ranging from 5.7 pg/g to 14 pg/g. High resolution MS was used in order to identify bioactive compounds in the fractions. No AhR agonists could be identified in fractions of HDPE or LDPE. Via a multivariate statistical approach the polystyrene (PS) trimer 1e- Phenyl-4e-(1- phenylethyl)-tetralin was identified in fractions of HIPS and in fractions of the blank polymer of HIPS.