Objective: This study aimed to establish a method using ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) to determine 10-hydroxy-2-decenoic acid in honey. Methods: The samples were extracted with methanol-water and cleaned up by a PLS solid-phase extraction column. The separation was performed on an Agilent ZOXBAR SB-C18 column (2.1 mm×50 mm, 1.8 μm) and methanol -5 mmol/L ammonium acetate solution was used as the mobile phase for gradient elution. Mass spectrometry was carried out under the multiple reaction monitoring (MRM) mode with negative electrospray ionization (ESI-) and standard external method for quantitative analysis. Results: Good linearity of the developed analytical method was obtained in the range of 0.05 ~ 0.80 μg/mL with the correlation coefficient of 0.995. The limit of detection (LOD) and the limit of quantitation (LOQ) was 0.04 mg/kg and 0.10 mg/kg, respectively. The recoveries at 0.10, 0.20, 1.00 mg/kg spiked levels ranged from 96.9% to 108.3% and the relative standard deviations (RSDs) ranged from 4.5% to 10.7%. Intra-day RSDs ranged from 5.9% to 9.2%, and inter-day RSDs ranged from 3.5% to 14.2%. Conclusion: The established method is rapid, sensitive, and accurate, and suitable for determining 10-hydroxy-2-decenoic acid in honey. It was found that 10-hydroxy - 2-decanoic acid was ubiquitous in honey as an endogenous component.