Abstract:The changes in volatile compounds during the preparation of double-layer steamed milk custard (DLSMC) were investigated in this study. Volatile substances at four key stages of the preparation process, namely the raw material stage (group A), boiling stage (group B), skin separation stage (group C), and steaming stage (group D), were analyzed using an electronic nose and headspace-gas chromatography-ion mobility spectrometry (HS-GC-IMS). The analysis identified methyl compounds, alcohols, ketones, and aldehydes as volatile components with highly significant differences across the stages (P<0.01). Meanwhile, HS-GC-IMS identified 38 compounds, including esters, alcohols, ketones, aldehydes, ethers, acids, and pyrazine derivatives. Principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA) indicated that the changes in volatile compounds were primarily attributed to the boiling and steaming stages. During the boiling stage (B group), the unpleasant flavor of 1-pentanol in raw buffalo milk was significantly ameliorated (P<0.05), while the levels of 2-butanone, 2-butanol, and dimethyl sulfide were significantly increased (P<0.05). In the steaming stage (group D), the levels of 2-pentanone, 2-heptanone, acetone, and diallyl disulfide were significantly increased (P<0.05). Ultimately, flavor characteristics, such as grassy, oily, fruity, milky, sweet, buttery, fruity, and roasted-nut aromas, were formed in the DLSMC. The present study revealed changes in volatile compounds during the DLSMC preparation process and provided a theoretical basis for elucidating the mechanisms underlying its flavor characteristics.