Objective: This study aimed to evaluate the determination uncertainty of four quinolones residues in honey by liquid chromatography-tandem mass spectrometry （LC-MS/MS）. Methods: Based on GB 31657.2—2021, a mathematical model for sources of uncertainty evaluation was established. The uncertainty of each variable which affect the result were calculated and the weight of each uncertainty component was assessed. Results: The uncertainties in the determination of enrofloxacin, sarafloxacin, ofloxacin and ciprofloxacin in honey were （4.97±0.81）, （10.13±1.58）, （7.67±1.23）, （14.97±2.25） μg/kg （P=95%，k=2） respectively. Conclusion: The main influencing factors of the determination uncertainty were preparation and dilution process of standard solution and fitting of standard curve, which should be controlled in practical work.