Environmental exposures causing ingestions of toxic chemicals, such as the polycyclic aromatic hydrocarbon 2-aminoanthracene (2-AA), may increase the risk of developing cancer and other diseases such as diabetes. To understand the mode of action of 2-AA as it relates to diabetogenic processes and pancreatic cancer, 2-AA binding to soluble protein mixtures was investigated using a novel technique called dynamic isoelectric anisotropy ligand binding assay (DIABLA). Twenty four post-weaning 3-4 week old Fischer-344 (F-344) male rats were fed 0 mg/kg (control), 50 mg/kg (low dose), 75 mg/kg (medium dose) and 100 mg/kg (high dose) 2-AA diet for 14 and 28 days. Total proteins extracted from the pancreas and liver were evaluated for their binding potential using DIABLA. This technique utilizes capillary isoelectric focusing and fluorescence anisotropy to separate proteins in their active form as well as evaluate the chemical interactions. Isoelectric point (pI) values for protein binding as well as experimental mass spectra data were determined. Investigation of 2-AA binding through screening a complex mixture of proteins is a step towards understanding the mode of action and the biological activities.