This analytical method was developed for the determination of three stilbene residues, diethylstilbestrol (DES), dienestrol (DEN), and hexestrol (HEX), in edible tissues of finfish including catfish, salmon, trout, and tilapia. Fortified fish samples were extracted with acetonitrile and further cleaned up using silica solid phase extraction columns. Stilbene residues were separated from matrix components by reversed phase high-performance liquid chromatography on a C8 column and analyzed using a tandem mass spectrometer with negative electrospray ionization. The overall average residue recoveries using post-fortified matrix-matched calibrants were 119, 99, and 104% with %RSDs of 18, 11, and 15% for DEN, DES, and HEX, respectively. Method detection limits of DEN, DES, and HEX in each matrix were found to be at or below 0.21 ng/g, and the limit of quantification averaged 0.3 ng/g and ranged from 0.18 to 0.65 ng/g for all analytes in all matrices.