Objective To investigate the distribution characteristics of antibiotics in source water and their health risks to human body in Yichang City, and to provide a scientific basis for the prevention and control of new types of pollution such as antibiotics in drinking water. Methods Solid phase extraction (SPE) was used for enrichment, and ultra-high performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) was used to detect and analyze the content of five types of antibiotics including sulfonamides, tetracyclines, chloramphenicol, quinolones and β-lactams in the main drinking water sources of Yichang City. The human health risk assessment of these antibiotics was conducted. Results All five types of antibiotics were detected in the source water of Yichang City, and the detection rate was 6.25% to 81.25%. The total detection rate of β-lactam antibiotics was the highest, and the highest detection concentration of norfloxacin was 213.9 ng/L. The health risk assessment of 11 antibiotics detected in the source water showed that these antibiotics would not cause obvious health harm to human body, but the risk entropy (RQ_H) tended to increase with the younger age. Conclusion The distribution level of antibiotics in source water of Yichang City will not cause obvious health harm to human body.