title: In vitro steroid profiling system for the evaluation of endocrine disruptors.
abstract: Endocrine disruptors (ED) are chemicals that affect various aspects of the endocrine system, often leading to the inhibition of steroidogenesis. Current chemical safety policies that restrict human exposure to such chemicals describe often time-consuming and costly methods for the evaluation of ED effects. We aimed to develop an effective tool for accurate phenotypic chemical toxicology studies. We developed an in vitro ED evaluation system using gas chromatography/mass spectrometry (GC/MS/MS) methods for metabolomic analysis of multi-marker profiles. Accounting for sample preparation and GC/MS/MS conditions, we established a screening method that allowed the simultaneous analysis of 17 steroids with good reproducibility and a linear calibration curve. Moreover, we applied the developed system to H295R human adrenocortical cells exposed to forskolin and prochloraz in accordance with the Organization for Economic Cooperation and Development (OECD) guidelines and observed dose-dependent variations in steroid profiles. While the OECD guidelines include only testosterone and 17β-estradiol, our system enabled a comprehensive and highly sensitive analysis of steroid profile alteration due to ED exposure. The application of our ED evaluation screen could be economical and provide novel insights into the hazards of ED exposure to the endocrine system.
authors.affiliation: Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka 565-0871, Japan.
authors.forename: Yosuke
authors.initials: Y
authors.lastname: Nakano
authors.affiliation: Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka 565-0871, Japan.
authors.forename: Toshiyuki
authors.initials: T
authors.lastname: Yamashita
authors.affiliation: Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka 565-0871, Japan.
authors.forename: Masashi
authors.initials: M
authors.lastname: Okuno
authors.affiliation: Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka 565-0871, Japan.
authors.forename: Eiichiro
authors.initials: E
authors.lastname: Fukusaki
authors.affiliation: Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka 565-0871, Japan; Division of Metabolomics, Research Center for Transomics Medicine, Medical Institute of Bioregulation, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan. Electronic address: bamba@bioreg.kyushu-u.ac.jp.
authors.forename: Takeshi
authors.initials: T
authors.lastname: Bamba
mesh_terms: D000302:Adrenal Cortex / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D002138:Calibration; D002460:Cell Line; D005576:Colforsin / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D052244:Endocrine Disruptors / Q000008:administration & dosage / Q000633:toxicity*; D004958:Estradiol / Q000032:analysis / Q000378:metabolism; D008401:Gas Chromatography-Mass Spectrometry / Q000191:economics / Q000379:methods; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology; D066298:In Vitro Techniques; D055432:Metabolomics / Q000191:economics / Q000379:methods*; D015203:Reproducibility of Results; D013256:Steroids / Q000032:analysis*; D053719:Tandem Mass Spectrometry / Q000191:economics / Q000379:methods; D013739:Testosterone / Q000032:analysis / Q000378:metabolism
chemical_list: D052244:Endocrine Disruptors; D007093:Imidazoles; D013256:Steroids; D005576:Colforsin; D013739:Testosterone; D004958:Estradiol; C045362:prochloraz
keywords: Endocrine disruptors; Gas chromatography/mass spectrometry; H295R cells; Metabolic profiling; Steroid