## 1 Introduction
In general, the chemical analysis showed the elements Al, Ca, Fe, Mg, and Cr are the most frequently found in both solid samples and eluates of FA [10]. [PERSON] et al. [13] proved _Daphnia magna_ could be suitable to evaluate the ecotoxicity of leachate from concrete. The leachates of cement samples in acute toxicity test showed 100% _Daphnia magna_ immobilization upon 24 h exposure. The authors pointed out the potential negative impact of alkali leaching from concrete into the aquatic environment [13]. The yeast species _Saccharomyces cerevisiae_ is a model eukaryotic microorganism and has been used as a test organism in the assessment of the potential toxicity of pesticides and other environmental contaminants [14]. A study developed by [PERSON] et al. [14] showed that this model is suitable for the development of simple low-cost toxicity tests, which can be useful to provide quick preliminary screening of environmental samples' potential toxicity level, before advancing further to ecotoxicity testing with more complex higher eukaryotes. [PERSON] and [PERSON] [15] studied the composition of leachates obtained from concrete mixes produced with two types of aggregates and four types of Portland cement. The authors used two leachants: acetic acid solution and deionised water. The results with acetic acid showed that Hg, Cr, Pb, and Ni were leached, whereas with the deionised water, only partial leaching of Hg, Cr, and Ni occurred. [PERSON] and [PERSON] [16] investigated various leaching tests on cementitious mortars with FA. The experiment included four types of tests: glass/polyethylene column test (pH 4 water with nitric acid); DEV-S4 batch test (demineralized water); bottle test (pH 4 water with nitric acid); and tank-leaching test (pH 4 water with nitric acid). The first two tests were considered not adequate to construction materials. In the third test, maximum toxic element concentrations were estimated and the results were considered reproducible. The diffusion leaching rates were calculated according to the fourth test, which was also considered reproducible. [PERSON] et al. [17] investigated the long-term (up to 256 days) leaching of toxic metals into deionized water from cementitious mortars. The tested samples were analysed according to Directive 80/778/EEC [18] for different metals by using atomic absorption spectroscopy. Of the metals analysed, only vanadium leaching was found in poorly cured mixes [17]. [PERSON] and [PERSON] [19] studied the leachability of heavy metals from cement, sand, and coal ash into water. Total concentrations of Zn, Pb, and Mn in coal ash were lower or similar to those in the other materials, whereas total Fe and Cu were higher in coal ash. In general, the leached concentrations of Fe, Zn, Cu, Mn, and Pb from coal ash and from concrete made with it were found low and comparable to that of conventional concrete, suggesting leaching risk. The authors suggested that coal ash can be incorporated in concrete since it is not anticipated to have harmful impact on environmental and public health related to this constituent [19]. In conclusion, the literature review shows that the incorporation of alternatives (FA and RA) to replace conventional components (cement and natural aggregates (NA)) is insufficiently studied in terms of ecotoxicological potential [20]. Moreover, studies concerning concrete mixes with the simultaneous and high incorporation ratio of FA and RA to produce sustainable RA concrete are missing. In this context, this study focuses on the application of an innovative methodology developed in a previous study by the authors [21] that reveals the environmental profile and the potential ecotoxicological risk of raw materials and building materials. In the present study, the developed methodology was applied to three concrete mixes and to raw materials incorporated in them, namely RA and FA. This was intended to assess their environmental risk when considered unusable and sent to landfills.