Journal
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
Volume 870, Issue 2, Pages 233-240Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jchromb.2008.06.026
Keywords
estrogen; HPLC; mass spectrometry
Funding
- NIA NIH HHS [AG13319, P30 AG013319] Funding Source: Medline
- NIDDK NIH HHS [R33 DK070290-03, R33 DK070290] Funding Source: Medline
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A fast and sensitive LC-ESI-MS method is described for the comparative quantification of 16 estrogen metabolites based on the derivatization of estrogens with a novel derivatizing reagent, N-methyl-nicotinic acid N-hydroxysuccinimide ester (C1-NA-NHS). The process introduces a quaternary amine to the analytes, making the analytes permanently charged regardless of the pH of the high-performance liquid chromatography (HPLC) mobile phase. This quaternization resulted in a highly efficient separation of 16 estrogen metabolites in 7 min at a detection level below 1 ng/mL. By using a deuterated derivatizing reagent (C1-d(3)-NA-NHS), a complete set of deuterated standards was utilized and used as internal standards in a comparative quantification and recovery study, demonstrating acceptable results over a wide concentration range. A pooled breast cancer serum sample was analyzed using the described method, and 15 estrogens were detected in the range of 80-530 pg/mL. (C) 2008 Elsevier B.V. All rights reserved.
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