Abstract: Objective: To establish an evaluation method for the uncertainty of high-performance liquid chromatography in the determination of sucralose in soy sauce. Methods: Testing was conducted in accordance with the national food safety standard GB 22255-2014. The content of sucralose in soy sauce was analyzed in accordance with the national metrological technical specification JJF 1059.1-2012. An uncertainty evaluation mathematical model was established to analyze the uncertainty factors during the measurement process. The uncertainty of the test results was evaluated and calculated through repeated sample measurements, standard solution purity, standard curve, glassware, weighing process, recovery rate, and testing instruments. Results: According to the confidence interval of 95%, the content of sucralose in soy sauce was (0.048±0.007) g/kg, k=2. Conclusion: The evaluation method for the uncertainty of sucralose in soy sauce by high performance liquid chromatography was established. The sources of the uncertainty of the determination results of sucralose in soy sauce were finally determined as follows: The fitting of the standard curve established by sucralose standard solution, and the recovery rate. And the proportion of various uncertainty evaluation factors in the process of determining sucralose content in soy sauce was cleared.