Residence time and time of production were investigated during the enzymatic production of a specific structured lipid/human milk fat substitute (SL-HMFS), on a kg scale, made from lard and soybean oil fatty acids, using a packed-bed reactor and short path distillation. There were no effects of residence time or time of production on C18:2 and C18:3 incorporation or on acyl migration in the sn-2 position. In addition, the SL-HMFS was compared to commercial human milk fat substitute (HMFS) regarding fatty acid composition, content of antioxidants and oxidative stability. Fats were stored at 60 °C for four days and the oxidative stability evaluated by analysis of peroxide value (PV) and volatile secondary oxidation products. SL-HMFS had a lower oxidative stability than did commercial HMFS products or lard, probably due to a lower level of tocopherol in SL-HMFS.