Objective To explore the effect of different intensities of irritant chemotherapy agents infused at varied sequences on venous injury, and to provide references for standardization of clinical medication. Methods The vesicant daunorubicin hydrochloride, and non-vesicant highly irritant cisplatin and non- vesicant weakly irritant cyclophosphamide were selected for venous infusion. A total of 24 ordinary healthy New Zealand rabbits were randomly divided into 8 groups,with 3 rabbits in each group: group A were given daunorubicin first and then cisplatin infusion, group B cisplatin and then daunorubicin, group C a single dose of daunorubicin, group D a single dose of cisplatin, group E cisplatin and then cyclophosphamide, group F cyclophosphamide and then cisplatin, group G single cyclophosphamide, and group H normal saline alone（the control group）.The incidence rates and pathological changes of phlebitis in the ear-vein the rabbits were compared after 48 h. Results Varied degrees of phlebitis occurred in all the chemotherapy agents infusion groups, and had significant differences with the control group （P＜0.05 for all）. No significant differences in phlebitis degree were found among the chemotherapy agents infusion groups （P＞0.05）. There were different degrees of pathological damage in the ear-vein of rabbits among the chemotherapy agents infusion groups, though no significant differences were found among the groups （P＞0.05）. Conclusion The irritant property of chemotherapy drugs is an important risk factor of chemotherapeutic phlebitis, and the infusion sequence of two or more chemotherapy drugs had minimal effect on venous injury.