Ethylglucoside lactate, a novel Alpha-Hydroxy Acids Derivative, was synthesized by transesterification in non-aqueous phase using immobilized lipase as biocatalyst. Based on the studies of the factors effecting initial rate and conversion under atmospheric pressure (solvent, acyl donor, different immobilized lipase, substrate concentration, enzyme concentration and temperature), the results show that solvent-free medium using butyllactate as acyl donor is suitable to the ester synthesis. The reaction conditions have been optimized as the following: the amount of enzyme = 75 g/L, the ethylglucoside concentration = 0.4 mol/L, 70 degrees C, 200 r/min, 50 h, which the conversion was 71%. A 90% conversion and a 60.7 mmol.L-1.h-1 initial rate can be obtained under reduced pressure, which the conditions are enzyme 75 g/L, ethylglucoside 0.35 mol/L, 65 degrees C, 200 r/min and 40 h. The product purified by extraction and SIO2 chromatography was identified by infrared spectroscopy and 1H NMR.