Hazardous and Noxious Substances (HNS) are defined as substances that are likely to create a significant impact on human health and marine ecosystem when they are released into the marine environment. Recently, as the volume of HNS transported by ships increases, the rate of leakage accidents also increases. Therefore, research should be conducted to control and monitor sunken materials from the viewpoint of technology development for hazardous material leakage accident response. In this paper, acoustic detection experiments were carried out using HNS substitute materials in order to confirm the possibility of acoustic detection of sunken HNS on the sediment. The castor oil, which has a similar acoustic impedance with chloroform, is used as a substitute. 200 kHz high frequency signals were used to discriminate the reflected signals and measure reflection loss from the interface between water and castor oil. The reflection loss measured is in good agreement with the modeling results, showing a possibility of acoustic detection for sunken HNS.