Abstract
Adult and pediatric endocrinology and oncology often requires measuring serum estrogens and testosterone at very low concentrations. Conventional immunoassay methods often lack the required performance to meet this analytical need, and mass spectrometry techniques must be employed. Our aim was to develop a sensitive HPLC-MS/MS assay for both estradiol (E2) and testosterone (Te) in serum, utilizing commercially available calibrators and without the need for chemical derivatization. Serum samples, after the addition of an internal standard, are combined with a hexane:ethyl acetate extraction solution. The samples are vortexed, and the organic layer is decanted into a clean sample tube and evaporated to dryness under a stream of nitrogen. The samples are reconstituted in a water:methanol solution and separated chromatographically using a reversed-phase HPLC column. Subsequent mass spectrometry is performed using both positive ion mode for Te and negative ion mode for E2.
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Acknowledgments
This work was supported by the Department of Laboratory Medicine of Memorial Sloan Kettering Cancer Center and through the NIH/NCI Cancer Center Support Grant P30 CA008748.
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Schofield, R.C., Kirchoff, D., Carlow, D.C. (2022). Quantitation of Estradiol and Testosterone in Serum Using LC-MS/MS. In: Garg, U. (eds) Clinical Applications of Mass Spectrometry in Biomolecular Analysis. Methods in Molecular Biology, vol 2546. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2565-1_13
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DOI: https://doi.org/10.1007/978-1-0716-2565-1_13
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