The ash reduction effects and fuel property changes of spent mushroom substrates by acid solution extraction, and the production possibilities of wood pellet fuel using them were studied. The ash weight of spent Pleurotus eryngii substrates was reduced from 8.81%(w/w) to 3.33%(w/w), and calorific value was increased from 3,958.3 kcal/kg to 4219.2 kcal/kg when extracted with a mixture of Acetic acid 2%(w/w) and Anhydrous citric acid 1%(w/w) in condition of liquid ratio 1:8, extraction temperature 55 ℃ and extraction time 180 min. The ash weight of spent Flammulina velutipes substrates was reduced from 14.91%(w/w) to 4.07%(w/w), and calorific value was increased from 4,190.3 kcal/kg to 4,219.2 kcal/kg when extracted with a mixture of Acetic acid 3%(w/w) and Anhydrous citric acid 1%(w/w) in condition of liquid ratio 1:8, extraction temperature 65 ℃ and extraction time 180 min. The ash weight of spent Pleurotus osteratus substrates was reduced from 3.31%(w/w) to 0.59%(w/w), and the smallest reduction was in calorific value from 4,558.6 kcal/kg to 4,216.2 kcal/kg when extracted with a mixture of Acetic acid 1%(w/w) and Anhydrous citric acid 1%(w/w) in condition of liquid ratio 1:8, extraction temperature 65 ℃ and extraction time 180 min.