Separation of dansylated 17β-estradiol, 17α-estradiol, and estrone on a single HPLC column for simultaneous quantitation by LC-MS/MS.

Publisher: Springer-Verlag Country of Publication: Germany NLM ID: 101134327 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1618-2650 (Electronic) Linking ISSN: 16182642 NLM ISO Abbreviation: Anal Bioanal Chem Subsets: MEDLINE

Original Publication: Heidelberg : Springer-Verlag, 2002-

Chromatography, High Pressure Liquid/*methods ; Estradiol/*chemistry ; Estrogens/*chemistry ; Estrone/*chemistry ; Tandem Mass Spectrometry/*methods; Estradiol/administration & dosage ; Estradiol/isolation & purification ; Estrogens/administration & dosage ; Estrogens/isolation & purification ; Estrone/administration & dosage ; Estrone/isolation & purification ; Animals ; Humans ; Isomerism ; Male ; Rats ; Rats, Sprague-Dawley

We show here that baseline separation of dansylated estrone, 17β-estradiol, and 17α-estradiol can be done, contrary to previous reports, within a short run time on a single RP-LC analytical column packed with particles bonded with phenyl-hexyl stationary phase. The chromatographic method coupled with isotope dilution tandem MS offers a simple assay enabling the simultaneous analysis of these analytes. The method employs (13)C-labeled estrogens as internal standards to eliminate potential matrix effects arising from the use of deuterated estrogens. The assay also offers adequate accuracy and sensitivity to be useful for biological samples. The practical applicability of the validated method is demonstrated by the quantitative analyses of in vivo samples obtained from rats treated with Premarin®.

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R01 AG031535 United States AG NIA NIH HHS; R21 AG031421 United States AG NIA NIH HHS; AG031535 United States AG NIA NIH HHS; AG031421 United States AG NIA NIH HHS

0 (Estrogens)
2DI9HA706A (Estrone)
4TI98Z838E (Estradiol)

Date Created: 20130202 Date Completed: 20130828 Latest Revision: 20211021

20260130

PMC3600063

10.1007/s00216-013-6710-y

23371528