An Ultra-High Performance Liquid Chromatography combined with Time-of-Flight Mass Spectrometry (UHPLC–ToF-MS) method has been developed for determination of nine mycotoxins, namely aflatoxins (AFB 1 , AFB 2 , AFG 1 and AFG 2 ), ochratoxin A (OTA), zearalenone (ZEA), toxin T2 (T2) and fumonisins (FB 1 and FB 2 ) in maize. The method included a two-step extraction with acetonitrile 80% (v/v). After optimization, the analytical method was validated. The different concentrations tested take in account the Maximum Levels (ML) for maize (Commission Regulation EC no. 1881/2006) and good results for repeatability (%RSD r ≤ 15.4%), reproducibility (%RSD R ≤ 15.9%) and recovery (77.8–110.4%, except for AFG2 at 2 μg/kg which presented a recovery of 73.4%) were achieved. These met the performance criteria imposed by Commission Regulation (EC) no. 401/2006. The method was applied to twenty-two samples from Portuguese producers of maize. Fumonisins were the most frequently detected mycotoxins, but the levels do not exceed those imposed by European legislation.