The invention relates to a traditional Chinese medicine extract with adenosine deaminase inhibition activity and an application of the traditional Chinese medicine extract. A preparation method of thetraditional Chinese medicine extract comprises the steps of taking traditional Chinese medicinal material powder, adding methanol, performing ultrasound extraction under a sealed condition, after theultrasound extraction, complementing the lost weight with methanol, performing centrifugation, taking a supernate, performing nitrogen blow-drying to form an extract, namely the traditional Chinese medicine extract, wherein traditional Chinese medicines are radix bupleuri, ligusticum wallichii, Chinese angelica, radix sophorae flavescentis, artemisia capillaris, bighead atractylodes rhizome, folium isatidis, a glossy privet fruit, schisandra chinensis, gardenia, rheum palmatum, sinensis, rhizoma cimicifugae, pollen pini, polygonum cuspidatum or sedum sarmentosum. After determination by established liquid chromatography-tandem mass spectrometry, the traditional Chinese medicine extract has a stronger inhibiting effect on adenosine deaminase, and cell experiments and animal experiments further confirm that the traditional Chinese medicine extract has a significant treatment effect on leukemia and the like.