title: The characterization of sulphated metabolites of norethindrone in human milk after oral administration of contraceptive steroids.
abstract: The excretion of ethynyl steroids in milk from a lactating woman taking a daily dose of an oral contraceptive (Conlumin) containing 1 mg of norethindrone and 50 micrograms of mestranol has been studied. Milk was diluted with aq. triethylamine sulphate and steroids were extracted on a Sep-Pak C18 cartridge at 60-64 degrees C. Groups of unconjugated steroids, glucuronides, mono- and disulphates were separated on triethylaminohydroxypropyl Sephadex LH-20. Following hydrolysis and further purification, steroids possessing an ethynyl-substituent were isolated by chromatography on sulphohydroxypropyl Sephadex LH-20 in silver form. Gas chromatographic-mass spectrometric analysis of the O-methyloxime-trimethylsilyl ether derivatives of these steroids, showed the presence of norethindrone and mestranol in the free fraction and of tetrahydro metabolites of norethindrone with 3 alpha,5 alpha, 3 alpha,5 beta and 3 beta,5 alpha configurations in the mono- and disulphate fractions. The disulphate of the 3 alpha,5 alpha isomer was the most abundant ethynyl steroid in milk after 13 days of administration. The site of conjugation of the monosulphates was established by acetylation prior to solvolysis and analysis by gas chromatography-mass spectrometry. This showed that the 3 alpha,5 alpha isomer was conjugated mainly in the 17 beta-position while the 3 alpha,5 beta isomer was conjugated at C-3.
mesh_terms: D000328:Adult; D003278:Contraceptives, Oral, Hormonal / Q000378:metabolism*; D005260:Female; D006801:Humans; D008895:Milk, Human / Q000032:analysis*; D009640:Norethindrone / Q000378:metabolism*; D013237:Stereoisomerism; D013464:Sulfuric Acids / Q000378:metabolism*
chemical_list: D003278:Contraceptives, Oral, Hormonal; D013464:Sulfuric Acids; D009640:Norethindrone