Analytical and Bioanalytical Chemistry

, Volume 405, Issue 10, pp 3399–3406

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

Authors

  • Szabolcs Szarka
    • Department of Molecular Biology and ImmunologyUniversity of North Texas Health Science Center
  • Vien Nguyen
    • Department of Molecular Biology and ImmunologyUniversity of North Texas Health Science Center
  • Laszlo Prokai
    • Department of Molecular Biology and ImmunologyUniversity of North Texas Health Science Center
    • Department of Pharmacology and NeuroscienceUniversity of North Texas Health Science Center
    • Department of Pharmaceutical SciencesUniversity of North Texas System College of Pharmacy
Original Paper

DOI: 10.1007/s00216-013-6710-y

Cite this article as:
Szarka, S., Nguyen, V., Prokai, L. et al. Anal Bioanal Chem (2013) 405: 3399. doi:10.1007/s00216-013-6710-y

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®.

https://static-content.springer.com/image/art%3A10.1007%2Fs00216-013-6710-y/MediaObjects/216_2013_6710_Figa_HTML.gif
Figure

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

Keywords

EstrogensPhenyl-hexyl stationary phaseLC–MS/MSDansylation13C-labeled internal standards

Supplementary material

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Copyright information

© Springer-Verlag Berlin Heidelberg 2013