Lipid rafts may affect the coalescence of milk fat globules through phase transition after thermal treatment

•Size of fat globules increased while its ζ-potential decreased.•GPL and Cho of fat globule membrane decreased while SM increased.•Cer is potential marker during thermal treatment of lipid rafts.•Sphingomyelins and ceramides increased in rafts while sterol lipids decreased.•Acyl chain unsaturation manifests decreased during thermal treatment. This study aimed to investigate changes of fat globules and their membranes, and further analyze evolution of lipid profile of lipid rafts in membranes during heat processing of cow milk. Size of fat globules increased from 3.16 μm to 3.70 μm and ζ-potential decreased from −0.53 mV to −0.38 mV after thermal treatment, suggesting that fat globule membrane was destroyed and fat globule occurred coalescence. Glycerophospholipids and cholesterol in fat globule membrane decreased while sphingomyelin increased after thermal treatment. Results of lipidomics show that total of 38 species of 5 lipids molecule showed ability to differentiate the samples. At high temperatures, highly unsaturated glycerophospholipids and sterol lipids were lost from rafts, meanwhile, sphingomyelin and ceramide increased in this region. Significant change of lipid profile in the raft region during thermal treatment suggested a potential relationship between lipid rafts and fat globule coalescence behavior.