Livestock milk is important for daily cooking and food production, but some lawless traders often adulterate the milk product using low-priced raw. Therefore, it is urgent to develop a simple, sensitive and accurate fingerprinting identification method to provide consumers with early warning of risks. Herein, we first systematically separated and authenticated the triacylglycerols composition of milks from holstein cattles, goats, mongolian horses, bactrian camels, yaks and buffaloes, which were less reported in domestic and international studies, using the SFC-Q-TOF-MS technique. Subsequently, the fingerprinting of triacylglycerols from different livestock milks was modelled using chemometric methods. The results showed that the proximity of different livestock milks was consistent with the species taxonomy, and the accuracy of internal as well as external validation was satisfactory. This work not only provides an innovative strategy for authentic traceability of livestock milks, but also offers an excellent demonstration for the establishment of nutritional databases.