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The water contamination by hormones is a major cause of endocrine disorders observed in animals. Environmental impacts caused by hormones are still poorly reported. The biosorption process is used for removal contaminants and it can be combined to the fixed bed technique. Wastes from Shiitake mushroom production were submitted to the pyrolysis process at three different temperatures to increase the surface area and develop a biochar for greater 17α-ethynyl estradiol removal. The quantification of 17α-ethynyl estradiol before and after the use of biochar was performed through the analytical analysis of High Performance Liquid Chromatography. The biochar obtained from the mushroom substrate at temperatures below 500ºC, showed the best hormone removal in the biosorption test. In conclusion the substrate biochar after the pyrolysis process showed results that demonstrate the viability of this method for the removal of 17α-ethynyl estradiol
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