Quantification of glucuronidated and sulfated steroids in human urine by ultra-high pressure liquid chromatography quadrupole time-of-flight mass spectrometry

Badoud, Flavia; Grata, Elia; Boccard, Julien; Guillarme, Davy; Veuthey, Jean-Luc; Rudaz, Serge; Saugy, Martial

doi:10.1007/s00216-011-4779-8

Cited by 97Doping in SportsForensic Toxicology and Drug AnalysisMetabolomics and Mass Spectrometry Studies

Quantification of glucuronidated and sulfated steroids in human urine by ultra-high pressure liquid chromatography quadrupole time-of-flight mass spectrometry

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Badoud, Flavia, Grata, Elia, Boccard, Julien, Guillarme, Davy, Veuthey, Jean-Luc, Rudaz, Serge, Saugy, Martial·2011·Analytical and Bioanalytical Chemistry

DOI 10.1007/s00216-011-4779-8

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Abstract

The urinary steroid profile is constituted by anabolic androgenic steroids, including testosterone and its relatives, that are extensively metabolized into phase II sulfated or glucuronidated steroids. The use of liquid chromatography coupled to mass spectrometry (LC-MS) is an issue for the direct analysis of conjugated steroids, which can be used as urinary markers of exogenous steroid administration in doping analysis, without hydrolysis of the conjugated moiety. In this study, a sensitive and selective ultra high-pressure liquid chromatography coupled to quadrupole time-of-flight mass spectrometer (UHPLC-QTOF-MS) method was developed to quantify major urinary metabolites simultaneously after testosterone intake. The sample preparation of the urine (1 mL) was performed by solid-phase extraction on Oasis HLB sorbent using a 96-well plate format. The conjugated steroids were analyzed by UHPLC-QTOF-MS(E) with a single-gradient elution of 36 min (including re-equilibration time) in the negative electrospray ionization mode. MS(E) analysis involved parallel alternating acquisitions of both low- and high-collision energy functions. The method was validated and applied to samples collected from a clinical study performed with a group of healthy human volunteers who had taken testosterone, which were compared with samples from a placebo group. Quantitative results were also compared to GC-MS and LC-MS/MS measurements, and the correlations between data were found appropriate. The acquisition of full mass spectra over the entire mass range with QTOF mass analyzers gives promise of the opportunity to extend the steroid profile to a higher number of conjugated steroids.

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To date only a limited number of methods for the simultaneous measurement of free, sulfated and glucuronidated androgens have been established, although significant progress has been made.
“but to date only a limited number of methods for the simultaneous measurement of free, sulfated and glucuronidated androgens have been established, although significant progress has been made (Badoud et al., 2011, Doue et al., 2015, Galuska et al., 2013).”
From Intracrine androgen biosynthesis, metabolism and action revisited (2018) · cites this paper as Badoud et al., 2011

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Authors: Badoud, F., Grata, E., Boccard, J., Guillarme, D., Veuthey, J.-L., Rudaz, S., & Saugy, M.
Affiliations: University of Geneva (CH)
University Centre of Legal Medicine (CH)
Corresponding author: Saugy, Martial
ORCID: Badoud, Flavia
Boccard, Julien
Guillarme, Davy
Veuthey, Jean-Luc
Rudaz, Serge
Saugy, Martial
Venue: Analytical and Bioanalytical Chemistry
Publisher: Springer Science and Business Media LLC
Published: 2011-03-06
Volume: 400
Issue: 2
Pages: 503-516
ISSN: 1618-2642 (ISSN-L)
1618-2650
DOI: 10.1007/s00216-011-4779-8
OpenAlex: W2092125746
Cited by: 97
References: 38
Citations by year: 2026: 1 · 2025: 2 · 2024: 4 · 2023: 4 · 2022: 4 · 2021: 4 · 2020: 8 · 2019: 6 · 2018: 8 · 2017: 7 · 2016: 7 · 2015: 9 · 2014: 5 · 2013: 10 · 2012: 13
FWCI: 35.40
Type: journal article
Language: en
Open access: bronze
doi.org
Licence: www.springer.com/tdm (tdm)
Locations: Analytical and Bioanalytical Chemistry (publishedVersion)
PubMed (publishedVersion)
SERVAL (Université de Lausanne) (publishedVersion, other-oa)
Archive ouverte UNIGE (University of Geneva) (publishedVersion, other-oa)
reroDoc Digital Library (submittedVersion)
IRIS (submittedVersion)
Archive ouverte UNIGE (University of Geneva) (submittedVersion, cc-by)
Topics: Social Sciences › Sociology and Political Science › Doping in Sports
Life Sciences › Pharmacology, Toxicology and Pharmaceutics › Toxicology › Forensic Toxicology and Drug Analysis
Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Metabolomics and Mass Spectrometry Studies
Keywords: steroid sulfates, urinary steroid metabolites, testosterone, anabolic steroids, doping analysis, solid-phase extraction, UHPLC-Q-TOF-MS, method validation
MeSH terms: Chromatography, High Pressure Liquid, Doping in Sports, Humans, Mass Spectrometry, Steroids, Molecular Structure, Anabolic Agents

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Quantification of glucuronidated and sulfated steroids in human urine by ultra-high pressure liquid chromatography quadrupole time-of-flight mass spectrometry

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