Phospholipids (PLs) are increasingly considered as nutrients with putative health benefits. Mammalian milk is a prominent source of phospholipids. In this study, PLs from different mammalian milk powders (buffalo, camel, bovine, goat, and ewe) purchased from supermarkets were enriched by solid-phase extraction cartridges, and subsequently separated and analyzed with ultra-performance liquid chromatography-electrospray ionization-quadrupole-time of flight-mass spectrometry (UPLC-ESI-Q-TOF-MS). Phospholipid classes were separated using hydrophilic interaction liquid chromatography (100×1mm, 1.7μm particle size BEH column) by gradient elution with acetonitrile/ammonium formate as a mobile phase. 22 molecular species of phosphatidylcholine (PC) were detected in the five types of milk powders, and PC (C16:0–C18:1) was the most abundant species with relative abundances of 10.93, 9.35, 9.35, 12.92, and 11.14%, respectively. While, 15 molecular species of phosphatidylethanolamine (PE) were detected, and PE (C18:0–C18:2) was the most abundant species making up 32.70, 16.45, 31.39, 29.88, and 27.24%, respectively. Furthermore, 9 molecular species of sphingomyelin (SM) were also detected, and the predominant species was SM (d18:1/16:0). This method of analysis was rapid, sensitive and precise for the identification of milk PLs. Also, it can be suggested as an application for studying specific technology markers in the dairy processing industry, and exploring novel applications of milk phospholipids.