A general procedure to study the biodegradation of endosulfan under laboratory conditions by white rot fungi isolated from native sources growing in YNB (yeast nitrogen base) media with 1% of glucose is presented. The evaluation of endosulfan biodegradation as well as endosulfan sulfate, endosulfan ether and endosulfan alcohol production throughout the whole bioremedation process was performed using an original and straightforward validated analytical procedure with recoveries between 78 and 112% at all concentration levels studied except for endosulfan sulfate at 50 mg L−1 that yielded 128% and RSDs < 20%. Under the developed conditions, the basidiomycete Bjerkandera adusta was able to degrade 83% of (alpha + beta) endosulfan after 27 days, 6 mg kg−1 of endosulfan diol were determined; endosulfan ether and endosulfan sulfate were produced below 1 mg kg−1 (LOQ, limit of quantitation).