Oil spills upset the delicate balance of the environment. Remediation methods are employed to recover areas affected by contaminants. Green surfactants have considerable potential with regards to enhancing environmental remediation methods. The aim of the present study was to formulate a green natural detergent containing the biosurfactant produced by Starmerella bombicola ATCC 22214 grown in mineral medium containing 1.2% canola oil, 10% sucrose, and 0.5% corn steep liquor. The biosurfactant reduced surface tension to 31.84 mN/m and was produced at a yield of 22 g/L. Twelve formulations were initially proposed using cottonseed oil as the natural solvent and three different stabilisers (vegetable wax, hydroxyethyl cellulose, and sodium alginate). The natural detergent was assessed with regards to emulsifying capacity and stability over a 10-day period. Ecotoxicity of the formulated detergent was investigated using the marine recruitment test on metal plates covered with paint into which the biosurfactant was incorporated as well as tests with the microcrustacean Artemia salina and vegetable seeds. Dispersing capacity and the potential for removing oil from rock surfaces were also investigated. The formulation designated H, which was composed of 1% biosurfactant, 40% cottonseed oil and 2% hydroxyethyl cellulose, achieved the best results. The formulation exhibited both stability and emulsifying capacity (100% of petroleum). The ecotoxicity tests revealed no significant toxicity, demonstrating the safety of the natural detergent. The detergent achieved satisfactory oil dispersion and solubilised 98% of oil impregnated on rock. The results indicate the potential application of the natural detergent formulated in this study for the efficient remediation of environmental compartments contaminated with oil and petroleum derivatives.