title: Steroidomic Footprinting Based on Ultra-High Performance Liquid Chromatography Coupled with Qualitative and Quantitative High-Resolution Mass Spectrometry for the Evaluation of Endocrine Disrupting Chemicals in H295R Cells.
abstract: The screening of endocrine disrupting chemicals (EDCs) that may alter steroidogenesis represents a highly important field mainly due to the numerous pathologies, such as cancer, diabetes, obesity, osteoporosis, and infertility that have been related to impaired steroid-mediated regulation. The adrenal H295R cell model has been validated to study steroidogenesis by the Organization for Economic Co-operation and Development (OECD) guideline. However, this guideline focuses solely on testosterone and estradiol monitoring, hormones not typically produced by the adrenals, hence limiting possible in-depth mechanistic investigations. The present work proposes an untargeted steroidomic footprinting workflow based on ultra-high pressure liquid chromatography (UHPLC) coupled to high-resolution MS for the screening and mechanistic investigations of EDCs in H295R cell supernatants. A suspected EDC, triclocarban (TCC), used in detergents, cosmetics, and personal care products, was selected to demonstrate the efficiency of the reported methodology, allowing the simultaneous assessment of a steroidomic footprint and quantification of a selected subset of steroids in a single analysis. The effects of exposure to increasing TCC concentrations were assessed, and the selection of features with database matching followed by multivariate analysis has led to the selection of the most salient affected steroids. Using correlation analysis, 11 steroids were associated with a high, 18 with a medium, and 8 with a relatively low sensitivity behavior to TCC. Among the candidates, 13 identified steroids were simultaneously quantified, leading to the evaluation and localization of the disruption of steroidogenesis caused by TCC upstream of the formation of pregnenolone. The remaining candidates could be associated with a specific steroid class (progestogens and corticosteroids, or androgens) and represent a specific footprint of steroidogenesis disruption by TCC. This strategy was devised to be compatible with medium/high-throughput screening and could be useful for the mechanistic elucidation of EDCs.
authors.affiliation: †School of Pharmaceutical Sciences, University of Geneva and University of Lausanne, Geneva, Switzerland.|‡Department of Human Protein Sciences, Faculty of Medicine, University of Geneva, Geneva, Switzerland.|§Swiss Centre for Applied Human Toxicology (SCAHT), Universities of Basel and Geneva, Basel, Switzerland.
authors.forename: David
authors.initials: D
authors.lastname: Tonoli
authors.affiliation: §Swiss Centre for Applied Human Toxicology (SCAHT), Universities of Basel and Geneva, Basel, Switzerland.|∥Division of Molecular and Systems Toxicology, Department of Pharmaceutical Sciences, University of Basel, Basel, Switzerland.
authors.forename: Cornelia
authors.initials: C
authors.lastname: Fürstenberger
authors.affiliation: †School of Pharmaceutical Sciences, University of Geneva and University of Lausanne, Geneva, Switzerland.
authors.forename: Julien
authors.initials: J
authors.lastname: Boccard
authors.affiliation: ⊥Department of Genetic and Laboratory Medicine, Geneva University Hospitals, Geneva, Switzerland.
authors.forename: Denis
authors.initials: D
authors.lastname: Hochstrasser
authors.affiliation: †School of Pharmaceutical Sciences, University of Geneva and University of Lausanne, Geneva, Switzerland.|‡Department of Human Protein Sciences, Faculty of Medicine, University of Geneva, Geneva, Switzerland.|§Swiss Centre for Applied Human Toxicology (SCAHT), Universities of Basel and Geneva, Basel, Switzerland.
authors.forename: Fabienne
authors.initials: F
authors.lastname: Jeanneret
authors.affiliation: §Swiss Centre for Applied Human Toxicology (SCAHT), Universities of Basel and Geneva, Basel, Switzerland.|∥Division of Molecular and Systems Toxicology, Department of Pharmaceutical Sciences, University of Basel, Basel, Switzerland.
authors.forename: Alex
authors.initials: A
authors.lastname: Odermatt
authors.affiliation: †School of Pharmaceutical Sciences, University of Geneva and University of Lausanne, Geneva, Switzerland.|§Swiss Centre for Applied Human Toxicology (SCAHT), Universities of Basel and Geneva, Basel, Switzerland.
authors.forename: Serge
authors.initials: S
authors.lastname: Rudaz
mesh_terms: D002224:Carbanilides / Q000009:adverse effects*; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003358:Cosmetics / Q000009:adverse effects; D052244:Endocrine Disruptors / Q000009:adverse effects*; D006801:Humans; D013256:Steroids / Q000032:analysis / Q000378:metabolism*; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: D002224:Carbanilides; D003358:Cosmetics; D052244:Endocrine Disruptors; D013256:Steroids; C009540:triclocarban