Background: The prevention and treatment of dairy cow mastitis during the dry period are crucial for the dairy industry. In this work, 200 dairy cows without mastitis and normal entering dry period were selected and randomly divided into 4 groups, for the first time, to explore the prevention effects using the different formulations of Cefquinome Sulfate Breast Injection with varied excipients. Methods: Three formulations with various excipients were further evaluated based on the drug residue analysis, somatic cell reduction in the milk and clinical symptoms of mastitis. In the control group, cloxacillin benzathine was utilized for comparisons. Result: All three Cefquinome test groups showed a better effect on prevention than the benzathine cloxacillin group in terms of drug release time, post-partum drug residue, reducing somatic cell count and preventing mastitis in the dry period. Various excipients showed slightly different impacts on the drug residues, the changes in somatic cell counts of dairy cows before and after medication. The prevention effects on mastitis from different excipients were similar after 30 d.