Atropine and scopolamine are tropane alkaloids (TAs), which are regulated for cereal-based foods for children in the EU. For ergot alkaloids (EAs) in cereals and cereal-based food harmonised legislation is not yet established. A fast and straightforward method, which employs extraction by acidified water/methanol followed by ultra-filtration prior to analysis by LC-MS/MS, was validated in bread for 20 EAs and six TAs. LOQs for individual alkaloids ranged from 0.3 to 1.2 µg kg−1, while recoveries ranged from 65% to 94% and repeatability from 3.4% to 17%. A survey was conducted in the Netherlands on 40 retail samples of bread (wheat, rye, wheat-rye, multi-grain) collected in 2014 and 2018. TAs, including atropine and scopolamine, were not detected. Eighteen different EAs were detected and total levels varied between −1. Since EAs were detected in a wide concentration range, it is recommended to monitor their occurrence in bread more regularly.