TY - JOUR
T1 - Separation of dansylated 17β-estradiol, 17α-estradiol, and estrone on a single HPLC column for simultaneous quantitation by LC-MS/MS
AU - Szarka, Szabolcs
AU - Nguyen, Vien
AU - Prokai, Laszlo
AU - Prokai-Tatrai, Katalin
N1 - Funding Information:
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).
PY - 2013/4
Y1 - 2013/4
N2 - 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®. [Figure not available: see fulltext.]
AB - 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®. [Figure not available: see fulltext.]
KW - C-labeled internal standards
KW - Dansylation
KW - Estrogens
KW - LC-MS/MS
KW - Phenyl-hexyl stationary phase
UR - http://www.scopus.com/inward/record.url?scp=84876156157&partnerID=8YFLogxK
U2 - 10.1007/s00216-013-6710-y
DO - 10.1007/s00216-013-6710-y
M3 - Article
C2 - 23371528
AN - SCOPUS:84876156157
SN - 1618-2642
VL - 405
SP - 3399
EP - 3406
JO - Analytical and Bioanalytical Chemistry
JF - Analytical and Bioanalytical Chemistry
IS - 10
ER -