The time course study of high monoester mixtures from soybean oil (HMMS) synthesis, as healthier alternatives totransfood products, in a supercritical CO2(SCCO2) medium with and without enzyme, was investigated. Phosphorous nuclear magnetic resonance (P-31-NMR) was used to quantify the absolute amount of partially esterified acylglycerols (PEGs). Carbon NMR was utilized to determine the type and position of the fatty acids (FAs) of HMMS. Enzyme and time significantly influenced the synthesis of 1-monoglycerides (1-MGs), 2-MGs, and 1,2-diglycerides (1,2-DGs) in this alcoholysis of soybean oil with 1,2-propanediol, based on high catalytic activity and operational stability of Novozym 435 in SCCO(2)during short reaction time. Results suggest that 4 h is a suitable reaction time for this lipase-catalyzed interesterification (LIE) system for the synthesis of 2-MGs with a yield of 20%. The highest polyunsaturated fatty acid (PUFA) (65%) in the triglyceride (TG) of HMMS was produced after 4 h of reaction. After 6 h of reaction, a high level (20%) of saturated fatty acids (SFAs) was found in the TGs of HMMS, which were distributed between thesn-2 (5%) andsn-1, 3 (15%) positions.