Abstract
A method based on double isotope derivative principle and with sensitivity of about 0.005μg pure steroid was developed for determination of estradiol-17β and estriol. This method includes esterification of estradiol-17βor estriol with p-iodobenzenesulfonyl chloride (pipsyl chloride), separation of ester by thin layer chromatography, acetylation of ester with 3H-labeled acetic anhydride, addition of known amount of pipsylated estrogen acetate-14C, purification of doubly esterified estrogen by 3 different systems of thin layer chromatography, and counting of 3H and 14C. The data on specificity of pipsylation, mobility of pipsylated estrogen acetate in thin layer chromatography, ratio of acetic anhydride to pyridine in acetylation of pipsylated estrogen, and recovery experiment are recorded.
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