There is an increasing global demand for, and transportation of, goods classified as Hazardous and Noxious Substances (HNS), including bio-derived oils and chemicals. With the increasing variety of bio-feedstocks and the development of processing technologies, the physical and chemical properties of commercially available and transported bio-derived oils vary significantly. The objective of this work was to analyze and compare the physical and chemical characteristics, behaviour, and risks of several bio-derived oils (four vegetable oils, one biodiesel, and two pyrolysis oils) and one petrol diesel when spilled into both fresh and salt water system. The laboratory results indicated that the vegetable oils and biodiesel remained buoyant and resulted in low total organic carbon (TOC) in water when spilled, similar behaviour to petrol diesel. However, the two pyrolysis oils exhibited remarkably different behaviours. The two oils sunk directly in the water and then rapidly dissolved the organic components into the water when mixed. This resulted in a high TOC (100 times higher than other types of oil) and an altered water chemistry, leaving a large portion of water-insoluble portion (35.0 wt% of pyrolysis oils) remaining at the water bottom. Initial Microtox findings indicated that water samples contaminated with pyrolysis oils and biodiesel (similar to petrol diesel) exhibited increased aqueous toxicity towards Vibrio fischeri. Further biological tests are required to understand their ecological risks. This work provides insights to the challenges and knowledge gaps on the fate, behaviour and risk of emerging bio-derived oils in water, while supporting HNS emergency response and remediation measures.