Antibiotics are emerging contaminants and widely used in human healthcare, livestock, and aquaculture. The toxicity posed by antibiotics and their mixtures in sediments depends on their bioavailability. Now, the bioavailability of organic materials can be determined accurately by the diffusive gradients in thin films (DGT) technique. This technique was used for the first time ever in this study to evaluate in detail the integral toxicity of antibiotics in sediments to aquatic biota. Zhelin Bay was selected as a case study, because it is the largest mariculture area in eastern Guangdong, South China. Two antibiotics, chlortetracycline (CTC) (A) and sulfachlorpyridazine (SCP), were detected at average concentrations of 2.83 and 1.14 ng/ml, respectively. The other fifteen antibiotics were undetectable. The single risk assessment based on the risk quotient (RQ) of CTC and SCP shows that a relatively low risk has occurred. After this careful assessment of probabilistic ecotoxicological risks, the combined toxicity of antibiotic mixtures (CTC and SCP) clearly indicates that the toxicity probability of surface sediments to aquatic organisms was relatively low (0.23%).