Quantitation of Estradiol and Testosterone in Serum Using LC-MS/MS

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 de...

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Bibliographic Details
Published inMethods in molecular biology (Clifton, N.J.) Vol. 2546; p. 141
Main Authors Schofield, Ryan C, Kirchoff, Daniel, Carlow, Dean C
Format Journal Article
LanguageEnglish
Published United States 2022
Subjects
Adult
Child
Chromatography, Liquid - methods
Estradiol
Estrogens
Hexanes
Humans
Methanol
Nitrogen
Reference Standards
Tandem Mass Spectrometry - methods
Testosterone
Water
Commercial calibrators
Testosterone
Serum
Liquid chromatography
Estradiol
Mass spectrometry
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Summary: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.
ISSN:1940-6029
DOI:10.1007/978-1-0716-2565-1_13