Separation of dansylated 17β-estradiol, 17α-estradiol, and estrone on a single HPLC column for simultaneous quantitation by LC-MS/MS.
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®.
Department of Molecular Biology and Immunology, University of North Texas Health Science Center, Fort Worth, TX 76107-2699, USA.
Szabolcs
S
Szarka
Vien
V
Nguyen
Laszlo
L
Prokai
Katalin
K
Prokai-Tatrai
D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004958:Estradiol / Q000008:administration & dosage / Q000737:chemistry* / Q000302:isolation & purification; D004967:Estrogens / Q000008:administration & dosage / Q000737:chemistry* / Q000302:isolation & purification; D004970:Estrone / Q000008:administration & dosage / Q000737:chemistry* / Q000302:isolation & purification; D006801:Humans; D007536:Isomerism; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley; D053719:Tandem Mass Spectrometry / Q000379:methods*
D004967:Estrogens; D004970:Estrone; D004958:Estradiol