Quantitative determination of conjugated estrogens in formulations by capillary GLC

A rapid capillary GLC method for the analysis of conjugated estrogen tablets and injectable formulations is described. The method involves the hydrolytic cleavage of the sodium sulfate ester conjugates by sulfatase enzyme. The free phenolic steroids are reacted sequentially with hydroxylamine hydrochloride and N,O-bis(trimethylsilyl)trifluoroacetamide. The resulting dual derivatives are analyzed on a 15-m glass capillary column wall coated with a cyanopropylmethyl silicone phase.