title: Marketplace Analysis of Conjugated Estrogens: Determining the Consistently Present Steroidal Content with LC-MS.
abstract: Conjugated estrogens purified from pregnant mares urine has been used as estrogen hormone replacement therapy since 1942. Previously, methods were proposed to identify and quantify the components of this complex mixture but ultimately were withdrawn due to incomplete characterization of the product and difficulties in transferring the method between laboratories. The aim of the current study is to develop a LC method that can reliably detect multiple steroidal components in conjugated estrogen tablets and measure their relative amount. The method developed was optimized for UHPLC columns, and the elution profile was analyzed using high-resolution mass spectrometry. A total of 60 steroidal components were identified using their exact m/z, product ion spectra of known, and predicted conjugated estrogen structures. These components were consistently present in 23 lots of Premarin tablets spanning two production years. The ten conjugated estrogens identified in the USP monograph and other additional estrogens reported elsewhere are among the 60 steroidal components reported here. The LC-MS method was tested in different laboratories using multiple samples, and the obtained results were reproducible among laboratories.
authors.affiliation: Division of Pharmaceutical Analysis, Office of Testing and Research, Center for Drug Evaluation and Research, U.S. Food and Drug Administration, 645 S. Newstead Avenue, St. Louis, Missouri, 63110, USA.
authors.forename: Michaella J
authors.initials: MJ
authors.lastname: Levy
authors.affiliation: Division of Pharmaceutical Analysis, Office of Testing and Research, Center for Drug Evaluation and Research, U.S. Food and Drug Administration, 645 S. Newstead Avenue, St. Louis, Missouri, 63110, USA.|Biotechlogic, Inc., Glenview, Illinois, USA.
authors.forename: Michael T
authors.initials: MT
authors.lastname: Boyne
authors.affiliation: Division of Pharmaceutical Analysis, Office of Testing and Research, Center for Drug Evaluation and Research, U.S. Food and Drug Administration, 645 S. Newstead Avenue, St. Louis, Missouri, 63110, USA.
authors.forename: Sarah
authors.initials: S
authors.lastname: Rogstad
authors.affiliation: Office of New Drug Products, Office of Pharmaceutical Quality, Center for Drug Evaluation and Research, U.S. Food and Drug Administration, 10903 New Hampshire Ave, Silver Spring, Maryland, 20993, USA.
authors.forename: David J
authors.initials: DJ
authors.lastname: Skanchy
authors.affiliation: Office of Research and Standards, Office of Generic Drugs, Center for Drug Evaluation and Research, U.S. Food and Drug Administration, 10903 New Hampshire Ave, Silver Spring, Maryland, 20993, USA.
authors.forename: Xiaohui
authors.initials: X
authors.lastname: Jiang
authors.affiliation: Division of Pharmaceutical Analysis, Office of Testing and Research, Center for Drug Evaluation and Research, U.S. Food and Drug Administration, 645 S. Newstead Avenue, St. Louis, Missouri, 63110, USA. ilan.geerlof-vidavsky@fda.hhs.gov.
authors.forename: Ilan
authors.initials: I
authors.lastname: Geerlof-Vidavsky
mesh_terms: D000818:Animals; D002853:Chromatography, Liquid / Q000379:methods; D004340:Drug Contamination; D004966:Estrogens, Conjugated (USP) / Q000032:analysis* / Q000737:chemistry*; D005260:Female; D006736:Horses; D013058:Mass Spectrometry / Q000379:methods*; D011247:Pregnancy
chemical_list: D004966:Estrogens, Conjugated (USP)
keywords: conjugated estrogens; mass spectrometry; steroidal content