Abstract: To promote the application of Leucosceptrum canum honey in food processing and improve its potential development and utilization value, in this study, Leucosceptrum canum honey yogurt was prepared with Leucosceptrum canum honey and cow milk as raw materials. The preparation process of Leucosceptrum canum honey yogurt was optimized based on a single-factor experiment and response surface test. Gas chromatography-mass spectrometer (GC-MS) and headspace-gas chromatography-ion mobility spectrometry (HS-GC-IMS) were used to analyze the free fatty acids and volatile flavor components of Leucosceptrum canum honey yogurt. The results showed that the best formula for Leucosceptrum canum honey yogurt was 1.2% Leucosceptrum canum honey, 7% sucrose, and 9 h fermentation time. The sensory score was 87.5±1.3, and the related physical and chemical health indicators of the product met the standard requirements. At the same time, compared with the yogurt without Leucosceptrum canum honey, the content of unsaturated fatty acids in Leucosceptrum canum honey yogurt increased. There were 74 volatile flavor components in Leucosceptrum canum honey yogurt, among which the contents of hydrocarbons and esters were higher. And the relative contents of phenols, esters, and ketones were increased by 4.06%, 1.35%, and 1.35%, respectively, compared with the blank yogurt. The overall flavor quality was improved. The yogurt prepared with Leucosceptrum canum honey and cow milk, had delicate tissue, good stability, and good flavor. The yogurt product was health with certain characteristics.