Quantifying steroid hormones in amniotic fluid by ultra-performance liquid chromatography and tandem mass spectrometry

Noëllie Rivet,Gunther Meinlschmidt,Nathalie Reix,Carole Jamey,Peter Kozlowski,Marion Tegethoff,Judith Lawrenz,Lisa M Körner,Jean-Sébastien Raul,Kathrin Erdmann,Pascal Kintz,Martin Heil,Susanne Fröhlich

doi:10.12688/f1000research.15848.1

Abstract

Background: Simultaneous assessment of steroid hormone concentrations in amniotic fluid is of importance for elucidating long-term consequences of intrauterine processes, and of broad scientific and clinical relevance. The objective of the study was to develop sensitive and specific analytical ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) methods for determination of steroid hormone concentrations in human amniotic fluid, and to provide proof-of-concept of their applicability. Methods: Methods were validated according to linearity, limit-of-detection, limit-of-quantification, recovery, intra- and inter-assay precision, and applied to 275 amniotic fluid samples. Results: Limits-of-quantification (S/N=10:1) were 0.05 ng/mL for cortisol, dehydroepiandrosterone sulfate (DHEAS), estradiol, estriol, and testosterone, and 0.01 ng/mL and 1.0 ng/mL for cortisone and dehydroepiandrosterone (DHEA), respectively. Good inter- and intra-assay precision were observed. Cortisol, cortisone, DHEAS, estradiol and estriol concentration were quantified in all samples. By lack of sensitivity of the analytical method, DHEA was quantified in 11 samples only. Testosterone was quantified in 119 of 275 samples analyzed. 116 of the quantified testosterone samples were from male offspring (out of 138 male offspring). Conclusion: These specific and sensitive methods offer a simple and non-invasive way to measure cortisol, cortisone, DHEAS, estradiol, estriol, and testosterone concentrations in human amniotic fluid.

Highlights

Amniotic fluid, with its nutritive and protective functions, is essential for fetal well-being and development[1]
Validation Cortisol, cortisone, DHEA, dehydroepiandrosterone sulfate (DHEAS), testosterone assessment LOD, LOQ and linearity The LOD and LOQ, as well as the coefficient of variation (CV) are presented in Table 2, for both cortisol, cortisone, DHEA, DHEAS, testosterone
We developed an analytical method to simultaneously measure cortisol, cortisone, DHEAS, and testosterone, and an analytical method to simultaneously measure estradiol and estriol in human amniotic fluid, using an LC-MS/MS assay, with good linearity, sensitivity, specificity and accuracy without interferences between the molecules and a low LOQ

Summary

Introduction

With its nutritive and protective functions, is essential for fetal well-being and development[1]. It is contained by the amniotic cavity enclosed by the amnion and chorion[2]. The objective of the study was to develop sensitive and specific analytical ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) methods for determination of steroid hormone concentrations in human amniotic fluid, and to provide proof-of-concept of their applicability. Results: Limits-of-quantification (S/N=10:1) were 0.05 ng/mL for cortisol, dehydroepiandrosterone sulfate (DHEAS), estradiol, estriol, and testosterone, and 0.01 ng/mL and 1.0 ng/mL for cortisone and dehydroepiandrosterone (DHEA), respectively. Cortisone, DHEAS, estradiol and estriol concentration were quantified in all samples. By lack of sensitivity of the analytical method, DHEA was quantified in 11 samples only.

Objectives

Methods

Results

Conclusion