The invention discloses a method for detecting the content of 2'-fucosyllactose in milk powder, and belongs to the technical field of food detection. The method comprises the following steps: (1) performing sample pretreatment: dissolving a milk powder sample by adopting a precipitant, and performing centrifuging, diluting and membrane passing-through; (2) quantitatively detecting the content of 2'-fucosyllactose in the pretreated sample by adopting high performance anion exchange chromatography, wherein the precipitant is one of methanol, ethanol, acetonitrile and a buffer solution. The adding standard recovery rate of the 2'-fucosyllactose detected by the method is 96.6%-103.6%, RSD is less than 3%, the detection result is accurate, the sensitivity is high, interference is avoided, and the method has a good application value for industrial batch detection.