Synthetic fibers account for approximately 60% of the total global fiber production, and polyester (PET) and polyamide (PA) dominate. Synthetic fabrics are now widely used in clothing, upholstery, carpets and other such materials. Textiles based on these materials have the potential to release microplastics (<5 mm in size) into the environment during production and cleaning actions. These particles are released in sewage effluents, as washing machine filters and wastewater treatment plants are not specifically designed to retain them and represent an environmental pollution that continuously increases the scientific and societal concern about their effects on marine biota and ecosystems. This study was focused on the determination of the amount of microfibers release from 100% polyester fabrics, in different washing conditions (programs and temperatures), comparing the use of detergent alone vs detergent with a stain remover. Microplastics released were characterized and quantified with gravimetric analysis, different microscopic, spectroscopic and thermal techniques. Tests were carried out in replicates to assess the data reproducibility and to show statistical differences between washing conditions.