Chemical Confirmation of Diethylstilbestrol Residues in Beef Livers

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Abstract
A simple chemical technique was developed to confirm the presence of diethylstilbestrol (DES) residues in beef tissues at 2 ppb. The DES-trifluoroacetate (TFA) from the previously published method is hydrolyzed in water-methanol at 55°C for 20 min to free DES which is then extracted into benzene. Trimethylamine and a small amount of heptafluorobutyric (HFB) anhydride are added to effect the smooth conversion of DES to its di-HFB at either room temperature or 60°C. DES-HFB is determined by gas-liquid chromatography with electron capture detection on a 6′ × 4 mm id column of 3% OV-210 at 190°C. This procedure yields 2 peaks for DES-HFB (retention times 2.7 and 4.8 min for the cis and trans, respectively) which are resolved from the 2 DES-TFA peaks (retention times 2.3 and 3.3 min for cis and trans, respectively). The sensitivity of detection of DES-HFB is increased by a factor of 4–5 over that for DES-TFA. This chemical technique was successfully applied to purified extracts of beef liver fortified with DES at 2–10 ppb with recoveries from 60 to 110%.

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