Different schemes in the production of structured triglycerides based on solvent-free esterification of oleic acid and palmitic acid with glycerol were evaluated in terms of reactant conversion, product composition, and yield. Two-step processes, with or without glycerol dosing, were compared with onestep versions carried out at temperature conditions within 40
C–60
C to a final oleic acid/palmitic acid/glycerol ratio of 2:1:1, the stoichiometric ratio in 1,3-dioleoyl-2-palmitoyl glycerol (OPO). Substantial amounts of C52 triglycerides, including OPO, were produced in all schemes at 34.98–39.55 wt.% of the total triglycerides and triglyceride yields of 50.10–56.56 wt.% based on the raw materials. The two-step process with glycerol dosing at 60
C showed the highest palmitic acid incorporation at the sn-2 position of 46.46%, but the same process carried out with a step increase in temperature from 40
C to 60
C gave the highest OPO yield at 19.30 wt.%. These results demonstrate the possibility of manipulating regioselectivity and triglyceride specificity by varying reactant feeding schemes without the use of solvents and sn-1,3-specific lipases common in conventional acidolysis-based processes.