The oxygen and strontium isotopic composition of 125 teeth of the hybodont sharks Lissodus and Hybodus from various late Triassic (Rhaetian) bone bed localities in the Central European Basin are used as environmental and ecological tracers for the shallow epicontinental Rhaetian Sea. The preservation of tooth enameloid was ascertained by cathodoluminescence microscopy. Mean δ18OP values differ regionally across the Rhaetian Sea with values of 18.6 ± 0.6‰ near the western gate, 16.1 ± 0.8‰ in the east, 15.4 ± 0.6‰ in the southeast, and 17.7 ± 0.9‰ in the south. The δ18OP values in the eastern Rhaetian Sea are 4–5‰ lower relative to Tethyan seawater, while teeth from the southern and western parts close to the marine gates are less fractionated. The strontium isotopic composition of teeth from most localities is more radiogenic in comparison to late Triassic seawater. The observed spatial δ18OP trend in the Rhaetian Sea indicates a shift from marine to brackish conditions in the south and west towards extensively brackish conditions with salinities of less than 16‰ in the east, thus confirming the existence of two marine gates. Brackish conditions persisted throughout early Rhaetian times, and were maintained by increased fluvial discharge from the Vindelician–Bohemian and Fennoscandian Highs as a result of enhanced atmospheric moisture availability in the course of the Rhaetian transgression. The δ18OP values of Lissodus and Hybodus indicate euryhaline behaviour of Rhaetian hybodonts by exceeding modern shark intraspecific variability. The lack of accordance in δ18OP values between different isotopically conspicuous localities in the Rhaetian Sea contradicts a basin-wide migration pattern. Differences between co-site taxa might indicate some degree of niche partitioning.