The invention discloses a method for 5'-O-ethylene adipyl uridine online synthesis through lipase catalysis. The method comprises the steps that dimethyl sulfoxide and tert-amyl alcohol serve as reactive solvents, uridine and adipic acid divinyl ester serve as raw materials, and 0.5-1.0 g of a lipase, namely Lipozyme TLIM, serves as a catalyst, wherein the volume ratio of the dimethyl sulfoxide to the tert-amyl alcohol is 1:(8-16), and the molar ratio of the uridine to the adipic acid divinyl ester is 1:(5-13). The raw materials and the reactive solvents are placed into an injector, and a reaction passage of a microfluidic passage reactor is uniformly filled with the lipase, namely the Lipozyme TLIM. Under pushing of an injection pump, the raw materials and the reactive solvents are continuously pumped into the reaction passage for acylation reaction. The inner diameter of the reaction passage of the microfluidic passage reactor is 0.8-2.4 mm, the length of the reaction passage is 0.5-1.0 m, the acylation reaction temperature is controlled to be 15-50 DEG C, and the acylation reaction time is 20-35 min. Through a product collector, reactive solution online collection is conducted, and after a reactive solution is subjected to conventional after-treatment, 5'-O-ethylene adipyl uridine is obtained. The method for 5'-O-ethylene adipyl uridine online synthesis through lipase catalysis has the advantages that the reaction time is short, the selectivity is high, and the productive rate is high.