Currently, active chlorine is considered the most effective treatment for preventing biofouling of structures in contact with seawater. This compound falls under the scope of the EU Biocidal Products Regulation, which includes since 2018 a requirement to assess all active ingredients for their potential endocrine-disrupting properties on humans and non-target organisms. Therefore, this study examines the endocrine-disrupting (ED) potential of active chlorine based on the European Chemicals Agency and European Food Safety Authority guidance (ED TGD). It includes two approaches: (i) a systematic literature review using appropriate search terms and (ii) an in silico assessment, both supported by expert judgement. Finally, the feasibility and relevance of in vitro tests were examined by considering the stability of chlorine and the applicability domain of the recommended in vitro assays.