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

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Abstract

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 13C-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®.

Quantification of estrogens from rat samples by LC–MS/MS

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Acknowledgments

We thank Ms. Shastazia White for her excellent technical assistance. This work was supported in part by the National Institutes of Health (grant number AG031535 to LP and AG031421 to KPT) and the Robert A. Welch Foundation (endowment BK-0031 to LP).

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Authors and Affiliations

  1. Department of Molecular Biology and Immunology, University of North Texas Health Science Center, 3500 Camp Bowie Boulevard, Fort Worth, TX, 76107-2699, USA

    Szabolcs Szarka, Vien Nguyen & Laszlo Prokai

  2. Department of Pharmacology and Neuroscience, University of North Texas Health Science Center, 3500 Camp Bowie Boulevard, Fort Worth, TX, 76107-2699, USA

    Katalin Prokai-Tatrai

  3. Department of Pharmaceutical Sciences, University of North Texas System College of Pharmacy, 3500 Camp Bowie Boulevard, Fort Worth, TX, 76107-2699, USA

    Katalin Prokai-Tatrai

Authors
  1. Szabolcs Szarka
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  2. Vien Nguyen
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  3. Laszlo Prokai
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  4. Katalin Prokai-Tatrai
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Correspondence to Katalin Prokai-Tatrai.

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Szarka, S., Nguyen, V., Prokai, L. et al. Separation of dansylated 17β-estradiol, 17α-estradiol, and estrone on a single HPLC column for simultaneous quantitation by LC–MS/MS. Anal Bioanal Chem 405, 3399–3406 (2013). https://doi.org/10.1007/s00216-013-6710-y

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  • DOI: https://doi.org/10.1007/s00216-013-6710-y

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