Exposure to endogenous estrogens and their metabolites is associated with the development and growth of breast cancer. The mechanisms of the carcinogenic effects of estrogen have not been firmly established. This is due in part to insufficient sensitivity of the assays employed for quantification. We developed a new derivatization procedure for estrogens and their metabolites by forming analytes as pre-ionized N-methyl pyrdine sulfonyl derivatives. Here, we focus on quantification of estradiol (E2) and its metabolites in human serum. We are able to determine 2- hydroxyl estradiol (2-OHE2) and 4- hydroxyl estradiol (4-OHE2) which are thought to play key roles in estrogen-induced breast cancer. Our analysis was performed on a Thermo Scientific Vantage triple stage quadrupole mass spectrometer equipped with a CaptiveSpray� ion source in positive ion mode. This method required only 0.1 mL of human serum, yet was capable of quantifying simultaneously estradiol and its metabolites within 20 minutes. The lower limit of quantitation for E2, E3, 2-MeOE2 and 4-MeOE2 was 0.312 fg on column, and was 3.12 fg on column for 2-OHE2 and 4-OHE2. All analytes demonstrated a linear response from 0.156 to 100 pg/mL (2-OHE2 and 4-OHE2, from 1.56-1000 pg/mL). The enhanced sensitivity offered by this method will allow for a more complete analysis of estrogens and their metabolites, shedding light on previously unknown mechanisms that might be implicated in the development of breast cancer.

Development of an ultra-sensitive stable isotope dilution liquid chromatography-tandem mass spectrometry method for quantification of estradiol and its metabolites in human serum

248th National Meeting (2014)

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.