Objective To investigate and analyze the contamination of 12 mycotoxins in unprocessed wheat grain samples from four provinces of China in 2019. Methods A total of 289 wheat grain samples harvested in 2019 from He'nan, Hebei, Anhui and Shandong provinces of China were collected and analyzed for 12 kinds of mycotoxins, including 8 Fusarium toxins and 4 aflatoxins by high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). Results Deoxynivalenol (DON) was the predominant mycotoxin in wheat samples, and the positive rate and the average level of DON were 95.5% (276/289) and 135.9 μg/kg. While the positive rates and the average levels of 15-acetyl-deoxynivalenol (15-Ac-DON) and 3-acetyl-deoxynivalenol (3-Ac-DON) were 37.4% (108/289) and 4.4 μg/kg, 36.7% (106/289) and 5.2 g/kg, respectively. The positive rates and the average levels of the rest of 8 Fusarium toxins and 4 aflatoxins were relatively low. There was no significant difference in mycotoxins contamination levels among the four provinces. Among 289 wheat samples, 59.9% (173/289) samples were simultaneously contaminated with more than 2 mycotoxins with DON and its derivatives 15-Ac-DON and/or 3-Ac-DON being the frequent combination. Conclusion DON is the predominant mycotoxin in wheat samples in China and the incidence of co-occurrence of DON and its derivatives, 3-Ac-DON and/or 15-Ac-DON are high. The positive rate and contamination levels of aflatoxins and other Fusarium toxins are relatively low. These results suggest that there should be a continuous monitoring for the contamination of fusarium mycotoxins, especially DON and its derivatives, in wheat in China.