Chemical fractions from 4 shale oils including one produced in an above-ground retort (Paraho), 2 produced below ground by in situ method (Occidental and Geokinetics), and 1 produced by a simulated modified in situ process (Livermore), were tested for mutagenicity against Salmonella typhimurium TA98 in the Ames assay system. Their mutagenic activities were compared with 2 other complex materials: a high-boiling coal distillate (The Solvent Refined Coal Process I (SRC II) heavy distillate), and a crude petroleum (Prudhoe Bay). Each material was fractionated by Sephadex LH-20 partition chromatography and by acid-base solvent extraction to provide chemical fractions for bioassay. Mutagenic activity was detected in the coal liquid (and their fractions), but not in the natural crude oil nor in any of its fractions. The basic and tar fractions derived by solvent extraction, and the LH-20 methanol fractions contained most of the mutagenic activity recovered from the unfractionated material. The heavy distillate was approx. 10 times more active in the Ames assay than the most mutagenically active shale oil. The most mutagenically active of the shale oils was about 10 times more active than the least active shale oil.