Chemical and food industries are focusing on reducing the economic and environmental impact of cleaning operations. This study aims to analyse the use of alkaline ozonated formulations to remove starch adhering to stainless steel surfaces, improving disinfection of mature biofilms with Listeria and Pseudomonas, and reducing the environmental impact of wastewater. Three key cleaning parameters have been analysed in a lab-simulated Clean-In Place system (CIP): temperature (20-60 degrees C), time (20-120 min), and cleaning formulations containing ozone and surfactant (linear alkylbenzene sulfonate, alkylpolyglucoside, fatty ethoxylated alcohol, lauramine oxide) were assayed. Higher temperature, time, and ozone concentration improved starch removal and disinfection of the stainless-steel surfaces, as well as the environmental impact of cleaning wastewater. Therefore, ozone-based CIP protocols could provide new opportunities to achieve cleaner, greener, and safer industries by intensifying cleaning, disinfection, and wastewater treatment in one step, saving operational costs compared to conventional CIP techniques. (C) 2021 Institution of Chemical Engineers. Published by Elsevier B.V. All rights reserved.