Abstract
Abstract Diethylstilbestrol (DES) and its monoglucuronide conjugate (DES-G) are extracted from homogenized beef liver by blending with acetonitrile containing triethylamine sulfate. An alkaline solution saturated with sodium chloride extracts DES and DES-G from the acetonitrile. The alkaline extract is acidified with sulfuric acid, triethylamine is added, and DES and DESG are extracted with 1-heptanol. DES is cleaved from DES-G by refluxing the 1-heptanol extract after adding ammonium chloride and monobasic ammonium phosphate. Solvent partitioning is used to isolate the DES, which is irradiated in a buffer solution with ultraviolet light to form the phenanthrenedione derivative. The dione is converted to the phenanthrenediol derivative in an acidified bisulfite solution. The diol derivative is isolated by solvent partitioning and determined fluorometrically in methanol. Thin layer chromatography is used for confirmation of DES. Recovery on 5 samples spiked with DES-G equivalent to 1, 2, and 4 ppb DES ranged from 82.5 to 104%.
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