Chemical Determination of Diethylstilbestrol Residues in the Tissues of Treated Chickens

Abstract

Abstract Bioassays based on the oral mouse uterine weight method have demonstrated the presence of estrogenic residues due to diethylstilbestrol in the edible tissues of chickens implanted with the drug. Chemical methods have now been developed which demonstrate conclusively that the estrogenic residues are diethylstilbestrol. Alcoholic extracts of the tissues are hydrolyzed with acid and carried through several extraction procedures. The residues are irradiated with ultraviolet light in a buffered alcohol solution, giving a yellow product which is measured colorimetrically. The colored product can be further converted to a colorless phenanthrene derivative that can be measured fluorometrically. The fluorophor gives characteristic activation and fluorescence spectra which can be used for identification. The diethylstilbestrol can also be identified by paper chromatography. Hydrolysis of the extracts with betaglucuronidase frees only a portion of the diethylstilbestrol in liver tissue.