Citation:

Blackwell, B., D. Martinovic-Weigelt, G. Ankley, AND Dan Villeneuve. Simultaneous analysis of thirteen endogenous steroid hormones by liquid chromatography tandem mass spectrometry with atmospheric pressure photoionization. SETAC North America, Salt Lake City, UT, November 01 - 05, 2015.

Impact/Purpose:

not applicable

Description:

Exposure to endocrine active chemicals can lead to perturbations of the hypothalamic-pituitary-gonadal (HPG) axis, ultimately leading to adverse reproductive effects. To evaluate potential reproductive effects, many aquatic toxicity assessments still rely on radioimmunoassay (RIA) for determination of specific steroids in biological matrices. While RIAs do provide high sensitivity, there is potential for cross reactivity of antibodies, and assays are limited to a single analyte. To address these limitations, analytical methods were developed for the simultaneous analysis of thirteen 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 biological matrices including fathead minnow (Pimephales promelas) plasma and gonad ex vivo steroidogenesis media. Additionally, tank water from fathead minnow exposures was collected and concentrated by solid phase extraction for analysis of excreted steroids. The method demonstrated high sensitivity in all three matrices with detection limits at sub ng/mL for plasma and ex vivo and low ng/L in tank water. Method performance for each sample matrix will be presented and discussed. Application of this method in aquatic vertebrate toxicity testing will lead to better understanding of specific mechanisms of HPG axis disruption and inform development of adverse outcome pathways (AOPs).

Record Details: