Estrogen and its oxidative metabolites have been implicated as main drivers of hormone-responsive breast cancers. Inhibition of aromatase, the enzyme responsible for aromatization of androgen precursors into estrogens, has proven effective in both the prevention and treatment of breast cancer. However, resistance frequently arises and there has been a lack of sensitive methodology for reliably tracking therapeutic response to aromatase inhibitors. Here we present a sensitive and specific stable isotope dilution nano-flow liquid chromatography tandem mass spectrometry assay capable of quantifying low picomolar levels of estrogens and androgens from human serum. Estrogen and androgen analysis was performed on a Vantage triple stage quadrupole mass spectrometer interfaced with a Waters NanoAcquity Ultra-Performance Liquid Chromatograpy (UPLC) system with gradient elution. Sensitivity of detection was greatly enhanced with use of pre-ionized derivatives. Assay validation has been performed in 0.5 mL double charcoal stripped human serum.  This method allows estrogen and androgen metabolites to be quantified with high sensitivity and specificity from human serum.

Sensitive quantitation of estrogens and androgens from human serum with stable isotope dilution nanoflow liquid chromatography tandem mass spectrometry

246th National Meeting (2013)

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