In the industries across the petroleum chain and those involved in energy generation, the use of petroderivatives as fuel oils is common. To clean parts, equipment and environments contaminated by hydrocarbons, they use expensive, toxic products, bringing risks to the environment as well as to workers’ health. Thus, the aim of this study was to check the stability of a biodetergent prepared using atoxic substances for large-scale production and industrial energy sector application. The relationship between volume (4 to 10 L) and stirring time (5 to 10 min) of the formulation at 3200 rpm and 80 °C was evaluated. The hydrophilic lipophilic balance (HLB), long-term stability (365 days), toxicity and efficiency of low-sulfur, viscous fuel oil removal from metal pieces and floors were investigated. The interaction among operating conditions was shown to influence the features of the product, which achieved approximately 100% stability after a stirring time of 7 min. The emulsion HBL index varied between 4.3 and 11.0. The biodetergent maintained its physicochemical properties during its 365 days of storage and showed high efficiency, removing 100% of the OCB1 impregnated on the metallic surfaces and floors tested. The formulation showed reliability in scale up when submitted to the study of physicochemical factors in the productive process, and safe application, by reducing risks for workers’ health and environment.