Direct Measurement of Free Estradiol in Human Serum and Plasma by Equilibrium Dialysis-Liquid Chromatography-Tandem Mass Spectrometry

Julie A Ray; Mark M Kushnir; Alan L Rockwood; A Wayne Meikle

doi:10.1007/978-1-4939-3182-8_12

Direct Measurement of Free Estradiol in Human Serum and Plasma by Equilibrium Dialysis-Liquid Chromatography-Tandem Mass Spectrometry

Methods Mol Biol. 2016:1378:99-108. doi: 10.1007/978-1-4939-3182-8_12.

Authors

Julie A Ray¹, Mark M Kushnir², Alan L Rockwood^{3

4}, A Wayne Meikle^{5

6

7}

Affiliations

¹ ARUP Institute for Clinical and Experimental Pathology, 500 Chipeta Way, Salt Lake City, UT, 84108, USA. [email protected].
² ARUP Institute for Clinical and Experimental Pathology, 500 Chipeta Way, Salt Lake City, UT, 84108, USA. [email protected].
³ ARUP Institute for Clinical and Experimental Pathology, 500 Chipeta Way, Salt Lake City, UT, 84108, USA. [email protected].
⁴ Department of Pathology, University of Utah School of Medicine, Salt Lake City, UT, USA. [email protected].
⁵ ARUP Institute for Clinical and Experimental Pathology, 500 Chipeta Way, Salt Lake City, UT, 84108, USA.
⁶ Department of Pathology, University of Utah, Salt Lake City, UT, USA.
⁷ Department of Medicine, University of Utah, Salt Lake City, UT, USA.

PMID: 26602122
DOI: 10.1007/978-1-4939-3182-8_12

Abstract

We describe a direct method of measurement of free estradiol using equilibrium dialysis followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Serum aliquots and internal standards are extracted by liquid-liquid extraction using methyl-tert-butyl ether (MTBE) followed by derivatization with dansyl chloride. An API 5500 mass spectrometer operated in positive electrospray mode is used for detection.

Keywords: Derivatization; Equilibrium dialysis; Free estradiol; LC-MS/MS; SHBG.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Blood Chemical Analysis / methods*
Chromatography, Liquid / methods*
Estradiol / blood*
Estradiol / chemistry
Estradiol / isolation & purification
Humans
Liquid-Liquid Extraction
Methyl Ethers / chemistry
Tandem Mass Spectrometry / methods*

Substances

Methyl Ethers
methyl tert-butyl ether
Estradiol