Covalent organic framework (COF) nanosheets (NSs) are a new member in the family of two-dimensional (2D) nanomaterials that received increasing attention. The ability to prepare COF NSs with rapid acquisition is of great importance to explore their distinctive properties and potential applications. Herein, we elaborate design a new COF hybrid material EB-TFP:Eu(BTA)4 as a sensing platform. In the process of ratiometric fluorescence detection of 5-fluorouracil (5FU), an anticancer drug, we realize the preparation of COF NSs. Interaction occurs between 5FU and COF hybrid material, where the interlayer π-π stacking of COF was weakened, benefiting the exfoliation of bulk COF to acquire 2D COF NSs. This strategy provides not only a sensitive and selective 5FU sensor but also a significant inspiration for engineering 2D COF NSs.