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High Sensitivity LC-MS Methods for Quantitation of Hydroxy- and Keto-androgens

Overview
Journal Methods Enzymol
Specialty Biochemistry
Date 2023 Oct 6
PMID 37802578
Authors
Andrea J Detlefsen
Ryan D Paulukinas
Clementina Mesaros
Affiliations
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Abstract

The quantitation of androgens is necessary to diagnose and monitor the development of diseases such as prostate cancer and polycystic ovary syndrome. Androgen measurements also support the laboratory-based study of androgen metabolism in cellular and animal models. The methods described in this chapter combine chemical derivatization of hydroxy- and keto-androgens with stable isotope dilution liquid chromatography mass spectrometry (SID-LC-MS). Chemical derivatization of hydroxy-androgens by picolinic acid and keto-androgens by Girard P enhances the ionization and detection sensitivity of androgens, while chromatographic separation and [C]-labeled internal standards add specificity that allow for simultaneous quantitation of multiple androgens. This chapter describes the materials and protocols necessary for chemical derivatization, enzymatic synthesis of internal standards, and LC-MS detection of keto- and hydroxy-androgens.

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