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Simultaneous analysis of fourteen endogenous steroid hormones by liquid chromatography tandem mass spectrometry with atmospheric pressure photoionization
Citation:
Blackwell, B., S. Poole, G. Ankley, AND Dan Villeneuve. Simultaneous analysis of fourteen endogenous steroid hormones by liquid chromatography tandem mass spectrometry with atmospheric pressure photoionization. SETAC North America, Minneapolis, MN, November 12 - 16, 2017.
Impact/Purpose:
Exposure to endocrine active chemicals can lead to perturbations of the hypothalamic-pituitary-gonadal (HPG) axis, ultimately leading to adverse reproductive or developmental effects. Steroid hormone production is a key biomarker of HPG disruption. Current methods target only one or two steroid hormones in a single sample, limiting the information that can be obtained. A new method was developed to screen fourteen steroid hormones simultaneously, including androgen, estrogen, glucocorticoid, and progestogen classes of steroids. Application of this method to aquatic vertebrate toxicity testing will lead to better understanding of specific mechanisms of HPG axis disruption and inform development of adverse outcome pathways (AOPs).
Description:
Product Description: To understand how some chemicals affect the endocrine system, controlled lab experiments often monitor how chemicals impact natural steroid hormones in fish. Current methods can target only one or two hormones in a single sample, limiting the information that can be obtained. A new method was developed to identify fourteen steroid hormones in a single sample. Using this method in toxicity testing will aid our understanding of how chemicals induce toxic effects along the endocrine axis of vertebrates. Abstract: Exposure to endocrine active chemicals can lead to perturbations of the hypothalamic-pituitary-gonadal (HPG) axis, ultimately leading to adverse reproductive or developmental effects. To evaluate potential effects, studies with possible HPG-active chemicals often rely on radioimmunoassay (RIA) for determination of biologically-active steroids and their precursors in biological matrices. While RIAs provide high sensitivity, there is potential for cross reactivity of antibodies, and assays are limited to a single steroid. To address these limitations, in the present study analytical methods were developed for the simultaneous analysis of fourteen steroid hormones including androgens, estrogens, progestogens, and glucocorticoids by liquid chromatography tandem mass spectrometry (LC-MS/MS). Atmospheric pressure photoionization (APPI) with a toluene dopant was utilized to allow ionization of all compounds in positive ionization mode without the need for derivatization. This method was applied to the analysis of fathead minnow (Pimephales promelas) plasma and exposure tank water from zebrafish (Danio rerio) experiments. Application to tank water analysis during flow-through chemical exposures provides a possible non-invasive endpoint for time-course experiments. The method demonstrated high sensitivity in both matrices with detection limits of most steroids at low µg/L in plasma and sub ng/L in water. Application of this method to aquatic vertebrate toxicity testing will lead to better understanding of specific mechanisms of HPG axis disruption and inform development of adverse outcome pathways (AOPs). The contents of this presentation neither constitute nor necessarily reflect US EPA policy.