Pancreatic lipase activity and bile salt concentration are very low in infants. Therefore, infant formula (IF) preparation with structured lipids (SLs) containing essential fatty acids and medium-chain fatty acids (MCFAs) with easy absorption features has attracted great attention. In this work, the lipid composition and physiochemical properties of novel SLs enriched with medium- and long-chain triacylglycerols (MLCTs) and their incorporation into a powdered IF compared to commercial IFs were evaluated. The obtained results showed that the SL contains arachidonic (ARA), caprylic, and capric as major fatty acids (FAs). Further, the evaluation of commercial IFs oil showed that oleic, linoleic, palmitic, and lauric were the most abundant FAs. A novel SL possessed highest levels of MCFAs (36.33 ± 1.05%) at sn-1,3 positions and LCUFAs (65.91 ± 1.32%) at the sn-2 position especially ARA (42.82 ± 0.89%). A total of 45, 38, and 25 triacylglycerols molecular species were revealed in a novel SL and commercial IFs available in China and the USA, respectively. Thermoprofle revealed that all IFs oil samples melted below the body temperature. The formula produced with microencapsulated SL and the dry-mixing process showed better optical properties and oxidative stability having lower moisture content. The study has potential applications in the development of IF with improved properties.