We used statistical approach by coupling redundancy analysis with linear regression analysis, which is useful to understand potential sources of antibiotics in the tide rising and ebbing of surface water in the Yongjiang Estuary, China. This study aimed to investigate the relationship between 29 antibiotics at five sites over four seasons and 13 environmental parameters during the tide rising and ebbing durations. The results found that dissolved organic carbon (DOC), salinity, temperature and chlorophyll a (Chla) were the main factors to impact antibiotics. The concentrations of macrolides were increasing with DOC, suggesting DOC may influence the adsorption capacity of antibiotics. The concentrations of tetracyclines had significant correlation with temperature and Chla during the tide rising period. This study demonstrated a method of exploring the relationship between the concentrations of antibiotics and environmental parameters, which is beneficial to future antibiotics research in estuaries.