The invention discloses a ratio fluorescence probe for identification of nilotinib, and a preparation and identification method thereof. With a molecular formula of C114H118O20N42, the ratio fluorescence probe is prepared from cucurbit[10]uril and a guest B with a molecular formula of C27H29N, during identification, the probe is dissolved in water to prepare a reagent, a to-be-identified sample is dripped into the reagent to obtain a sample solution, the sample solution is subjected to fluorescence excitation, and the fluorescence excited fluorescence wavelength is detected. The ratio fluorescence probe provided by the invention has the characteristics of low detection and analysis cost, simple operation, simple preparation process, visible test result and more direct analysis.