In recent years, the presence of acrylamide (AA) in food has been a cause of concern, since it has been classified by the IARC as probably carcinogenic to humans (Group 2A). Within the EU, benchmark values were established in 2017, and in 2019, the importance of monitoring this contaminant was further reinforced by a recommendation from the European Commission. The new legislation was the result of a recognition that data on AA levels were still insufficient—not only for foods covered by the 2017 regulation but also for other food categories not covered by the regulation that may nevertheless contain significant levels of AA and/or contribute significantly to overall dietary exposure. This chapter presents a sensitive, precise, accurate, and straightforward in-house analytical protocol for the determination of AA in cereals and cereal-based products using UPLC–MS/MS. The analytical workflow encompasses critical pre-treatment steps, including sample defatting, aqueous extraction, SPE for sample cleanup. Another critical step is the AA quantification by UPLC-MS/MS, where the extract is then analyzed using a triple quadrupole mass spectrometer operating in ESI+, with AA-d3 as an IS. The analyte and IS are separated on an HSS column by gradient elution using water and methanol as the mobile phase. The method achieves a limit of quantification of 19 μg/kg, aligning with regulatory monitoring requirements for AA. Additionally, the main analytical challenges and critical control points throughout the procedure are discussed, with particular emphasis on sample preparation and instrumental parameters that influence quantification accuracy.