Abstract: A method for continuous preparation of highly pure monoglyceride comprises the steps of esterifying fatty acid dissolved in a nonpolar solvent at the interface between the nonpolar solvent phase and a polar solvent phase containing glycerol, water and dispersed lipase, the polar phase being located below the nonpolar phase, continuously taking out the nonpolar solvent phase, cooling the taken out phase to isolate the resulting monoglyceride by precipitation, and bringing back the remaining solution to the esterification system.By this method, monoglycerides having high purity can be produced continuously and at low cost, and further monoglycerides of straight-chained fatty acids can preferentially obtained even though the fatty acids used contain branched fatty acids.