The extraction and untargeted UHPLC-HRMS analysis of endogenous steroids in serum samples is described in this protocol. The employed full-scan acquisition mode provides the adequate sensitivity to highlight the main endogenous steroids present in blood, including mineralocorticoids, progestogens, and androgens. Technical aspects for both chromatography and mass spectrometry are discussed in detail, together with a proposition of setup for sample sequence and data analysis. Furthermore, general comments are given to help the assessment of data quality and system performance.
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Sanderson JT (2006) The steroid hormone biosynthesis pathway as a target for endocrine-disrupting chemicals. Toxicol Sci 94:3–21CrossRefGoogle Scholar
Arukwe A (2008) Steroidogenic acute regulatory (StAR) protein and cholesterol side-chain cleavage (P450scc)-regulated steroidogenesis as an organ-specific molecular and cellular target for endocrine disrupting chemicals in fish. Cell Biol Toxicol 24:527–540CrossRefGoogle Scholar
Shackleton CHL (2012) Role of a disordered steroid metabolome in the elucidation of sterol and steroid biosynthesis. Lipids 47:1–12CrossRefGoogle Scholar
World Anti-Doping Agency (WADA), Montreal (2015) Anti-doping testing figures report. http://www.wada-ama.org. Accessed May 2017
Basaria S (2010) Androgen abuse in athletes: detection and consequences. J Clin Endocrinol Metab 95:1533–1543CrossRefGoogle Scholar
Büttner A, Thieme D (2010) Side effects of anabolic androgenic steroids: pathological findings and structure-activity relationships. Handb Exp Pharmacol 195:459–484CrossRefGoogle Scholar
Fanelli F, Belluomo I, Di Lallo VD et al (2011) Serum steroid profiling by isotopic dilution–liquid chromatography–mass spectrometry: comparison with current immunoassays and reference intervals in healthy adults. Steroids 76:244–253CrossRefGoogle Scholar
Rosner W, Auchus RJ, Azziz R et al (2007) Position statement: utility, limitations, and pitfalls in measuring testosterone: an endocrine society position statement. J Clin Endocrinol Metab 92:405–413CrossRefGoogle Scholar
Handelsman DJ, Wartofsky L (2013) Requirement for mass spectrometry sex steroid assays in the journal of clinical endocrinology and metabolism. J Clin Endocrinol Metab 98:3971–3973CrossRefGoogle Scholar
Wartofsky L, Handelsman DJ (2010) Standardization of hormonal assays for the 21st century. J Clin Endocrinol Metab 95:5141–5143CrossRefGoogle Scholar
World Anti-Doping Agency (WADA), Montreal (2016) endogenous anabolic androgenic steroids, measurement and reporting, technical document TD2016EAAS. http://www.wada-ama.org. Accessed May 2017
Sjovall J (2004) Fifty years with bile acids and steroids in health and disease. Lipids 39:703–722CrossRefGoogle Scholar
Randazzo GM, Tonoli D, Strajhar P et al (2017) Enhanced metabolite annotation via dynamic retention time prediction: steroidogenesis alterations as a case study. J Chromatogr B Analyt Technol Biomed Life Sci. 1071:11–18CrossRefGoogle Scholar