This study was performed to investigate the effects of different cooking methods (boiling, roasting, stir-frying, and deep-frying) on folate retention in 6 kinds of mushrooms (Beech-, button-, Juda’s ear-, oak-, oyster-, and winter-mushrooms) frequently consumed in Korea. In order to assure reliability of analytical data, trienzyme extraction-L casei method was verified and analytical quality control was also evaluated. Folate contents of mushrooms varied by 6.04-64.82 g/100 g depending on the type of mushrooms. and were significantly affected by cooking methods. Depending on cooking methods, folate contents of mushrooms decreased by 22-48%, 2-31%, and 17-56% for Juda’s ear-, oak- and oyster-mushrooms, respectively, while 17-90% of folate was increased in Beech mushroom. Overall, the largest weight loss was found in boiled mushrooms, but the lowest one in deep-fried samples. True folate retention rates considering processing factor were less than 100% for all cooked mushrooms except for Beech samples. Overall, folate loss was the largest by boiling with water but the smallest by deep-frying. Both accuracy and precision of trienzyme extraction-L-casei method were excellent based on a recovery close to 100% and coefficient variations less than 3%. Quality control chart of folate analysis (n=26) obtained during the entire study and an international proficiency test (z-score=-0.5) showed that trienzyme extraction-L casei method is reliable enough for production of national folate database.