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Microalgae as the third generation of biomass sources for biofuel production has a promising future, but its economic challenges have not been resolved yet. One of the major challenges in order to practical implementation of biofuel production is separation of microalgae from culture medium, since this step devotes a significant part of the energy and time consumption. Although various methods have been reported for harvesting, electro-coagulation–flotation (EC) method was preferred due to the simplicity and capability of process scale up. In this study, the effect of three parameters, i.e., initial pH, electric current density and duration on microalgae separation efficiency and operating costs, as the response variables, was investigated by applying “Response Surface Methodology” (RSM) technique. It was revealed that duration was the most effective parameter on the microalgae separation efficiency, as well as the …