[Objective] We studied the inhibition of Cronobacter sakazakii biofilm by baicalin. [Methods] XTT reduction assay was used to evaluate the effects of baicalin on the early adhesion of C. sakazakii planktonic cells and the activity of sessile cells in biofilms. Quantitative real-time PCR (qRT-PCR) was used to determine the expression level of the biofilm-associated genes, including glpQ, nlpD, gsiB, deoB, luxS, and sdiA. [Results] The inhibition of C. sakazakii biofilms by baicalin was dose dependent. MIC80 of baicalin against C. sakazakii was 1 024 mg/L, with 83.7% inhibition of C. sakazakii ATCC BAA-894 and 53.2% of C. sakazakii IQCC10423. In addition, 2 048 mg/L of baicalin inhibited the adherence of C. sakazakii at its earlier development of new biofilms. qRT-PCR results demonstrated that baicalin could down-regulate the expression of biofilm-associated genes in C. sakazakii and further inhibited its biofilm formation. [Conclusion] Baicalin could be potentially used as an antimicrobial to inhibit C. sakazakii biofilms.