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Development of a Comprehensive Steroid Hormone Screening Method using Liquid Chromatography/Tandem Mass Spectrometry (LC-MS/MS)

Published

Author(s)

Ashley S. Boggs-Russell, John Bowden, Louis J. Guillette, John R. Kucklick

Abstract

The ability to monitor endocrine pathways is often limited by techniques that use indirect measurement or single-hormone/class analysis. As our knowledge of the interconnections of the endocrine system grows, there is a need to develop highly specific methods for the simultaneous measurement of biologically relevant concentrations of multiple classes of hormones (estrogens, androgens, progestogens, and corticosteroids) from a single analysis. The objectives of this study were to develop a liquid chromatography tandem mass spectrometry (LC-MS/MS) method using biologically relevant concentrations of multiple classes of steroid hormones, then test detection of steroid hormones in a biological matrix by spiking stripped fetal bovine serum (FBS) extract. All steroids were measured on an AB Sciex API4000 QTRAP hybrid triple quadrupole/linear ion trap mass spectrometer in positive mode using electrospray ionization. Standards of 17 hormones were examined in methanol (MeOH) on a 10 point calibration curve from 60 pg to 500 ng with seven internal standards. The 10 point calibration curve was repeated, spiked into FBS extract. Additional sets of MeOH and FBS calibration curves were derivatized with dansyl chloride (DC) to investigate improvement of estrogen detection. Biologically relevant limits of detection (LODs) were calculated for 15 of the 17 hormones analyzed. DC derivatization improved response curves of estrogens two to eight fold, and provided better chromatographic separation than without derivatization. FBS extract increased background and elevated LODs compared to the MeOH standards, but LODs were still within biologically relevant ranges. This method quantified biologically relevant concentrations of multiple classes of hormones in a single sample which could vastly increase the data collection capabilities and thereby expound the knowledge of endocrinology.
Citation
Journal of Chromatography A

Keywords

androgens, corticosteroids, estrogens, hormones, liquid chromatography, progestogens, steroids, tandem mass spectrometry

Citation

Boggs-Russell, A. , Bowden, J. , Guillette, L. and Kucklick, J. (2016), Development of a Comprehensive Steroid Hormone Screening Method using Liquid Chromatography/Tandem Mass Spectrometry (LC-MS/MS), Journal of Chromatography A, [online], https://doi.org/10.1007/s00216-016-9512-1 (Accessed April 24, 2024)
Created April 2, 2016, Updated January 27, 2020