Sensitive and robust method for anabolic agents in human urine by gas chromatography–triple quadrupole mass spectrometry

Delgadillo, Miguel A.; Garrostas, Lorena; Pozo, Óscar J.; Ventura, Rosa; Velasco, Benjamín; Segura, Jordi; Marcos, Josep

doi:10.1016/j.jchromb.2012.03.037

Cited by 26 publications

(28 citation statements)

References 19 publications

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“…Furthermore, detection of unaltered glucuronide metabolites METm1-G and METm2-3-G was up to 1 and 4 days, respectively. Using GC-MS/MS analysis after hydrolysis and derivatization, detection times of 4 and 6 days were described for METm1-G and METm2-G, respectively [6,36]. The developed methodology could distinguish the two different METm2-G isomers (METm2-3-G and METm2-17-G) which are observed in only one peak (corresponding to the phase I metabolite) using the GC-MS/MS procedure.…”

Section: Excretion Studies Of Methyltestosterone and Stanozololmentioning

confidence: 99%

Screening for anabolic steroids in sports: Analytical strategy based on the detection of phase I and phase II intact urinary metabolites by liquid chromatography tandem mass spectrometry

Balcells

Pozo

Esquivel

et al. 2015

Journal of Chromatography A

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In order to improve the detection capabilities of anabolic androgenic steroids (AAS) in sports, a liquid chromatography-tandem mass spectrometry (LC-MS/MS) screening method for the simultaneous detection of AAS phase I and phase II intact urinary metabolites (glucuronides and sulfates) was developed. A total of 36 metabolites (7 unconjugated; 19 glucuronides and 10 sulfates) corresponding to 15 of the most reported AAS were included. Analytes were extracted from urine using C18 cartridges. LC and MS conditions were studied in-depth to determine the most sensitive and selective conditions for each analyte. A selected reaction monitoring method was set up. The optimization of the experimental parameters for 13 metabolites not available as standards was performed using excretion study urines. Extraction recoveries were above 77% for all 23 validated analytes. Intra-day precision was lower than 21%, and LODs were in the range 0.25-4ng/mL for 18 of the 23 analytes. Matrix effect was evaluated using post column infusion and ranged from 92 to 147%. The method was successfully applied to excretion study urines of different exogenous AAS. The suitability of the strategy was demonstrated with methyltestosterone and stanozolol excretion study urines by achieving detection times of 22 and 21 days, respectively. The method is compliant with the World Antidoping Agency requirements for most of the studied compounds. It represents a cost-effective approach that improves the detection capabilities of AAS by increasing the sensitivity for some metabolites and by including recently described phase II long-term metabolites not detectable using the current screening strategy.

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Section: Excretion Studies Of Methyltestosterone and Stanozololmentioning

confidence: 99%

Screening for anabolic steroids in sports: Analytical strategy based on the detection of phase I and phase II intact urinary metabolites by liquid chromatography tandem mass spectrometry

Balcells

Pozo

Esquivel

et al. 2015

Journal of Chromatography A

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“…Thus, most analytes included in doping control analysis for the detection of T misuse are phase II metabolites. The detection of these metabolites in routine methods is performed by the release of phase I metabolites by enzymatic hydrolysis with β‐glucuronidase and the detection of the combined free and glucuronide conjugated steroidal fraction by gas chromatography (tandem) mass spectrometry (GC‐MS(/MS)) . Although liquid chromatography‐tandem mass spectrometry (LC‐MS/MS) methods allow for the direct detection of glucuronides, enzymatic hydrolysis and LC‐MS/MS detection of phase I metabolites is also a common approach for the detection of these metabolites .…”

Section: Introductionmentioning

confidence: 99%

Synthesis and characterization of 6β‐hydroxyandrosterone and 6β‐hydroxyetiocholanolone conjugated with glucuronic acid

Kotronoulas

Fabregat

Alfonso

et al. 2014

Drug Testing and Analysis

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The detection of testosterone (T) misuse by doping control laboratories is mainly based on monitoring urinary T phase I metabolites released after enzymatic hydrolysis of the corresponding phase II glucuronide metabolites by gas chromatography (tandem) mass spectrometry (GC-MS(/MS)) methods. However, this strategy fails to properly determine two recently reported phase II metabolites of T conjugated with glucuronic acid that remained mostly conjugated after the hydrolysis step. These metabolites were identified as glucuronides of 6β-hydroxyandrosterone (6β-OH-And) and 6β-hydroxyetiocholanolone (6β-OH-Etio) but their exact conjugation site remained undetermined. In this study, the four possible glucuronides of 6β-OH-And and 6β-OH-Etio were synthesized and characterized by nuclear magnetic resonance (NMR) spectroscopy. Moreover, their chromatographic properties and MS spectra were compared to those obtained for the urine samples collected after administration of T. Results confirmed that the recently reported metabolites were the 3α-glucuronides of 6β-OH-And and 6β-OH-Etio. The synthesis and the elucidation of the exact structure of the metabolites presented in this study are crucial steps for the development of analytical methods in order to explore their role in T metabolism and their potential usefulness as biomarkers of T misuse.

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“…Gas chromatography/mass spectrometry (GC/MS) has been extensively used to determine anabolic compounds in urine but a major drawback is the need of derivatization [8][9][10][11]. This task can be quite demanding for trenbolone since several derivatives are formed using its most usual derivatives agents due to epimerization of its keto group in position 3 unless quite specific conditions are employed [12,13].…”

Section: Introductionmentioning

confidence: 99%

Simultaneous determination of resorcylic acid lactones, β and α trenbolone and stilbenes in bovine urine by UHPLC/MS/MS

Fernández-Arauzo

Pimentel-Trapero

Hernández-Carrasquilla

2014

Journal of Chromatography B

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Sensitive and robust method for anabolic agents in human urine by gas chromatography–triple quadrupole mass spectrometry

Cited by 26 publications

References 19 publications

Screening for anabolic steroids in sports: Analytical strategy based on the detection of phase I and phase II intact urinary metabolites by liquid chromatography tandem mass spectrometry

Screening for anabolic steroids in sports: Analytical strategy based on the detection of phase I and phase II intact urinary metabolites by liquid chromatography tandem mass spectrometry

Synthesis and characterization of 6β‐hydroxyandrosterone and 6β‐hydroxyetiocholanolone conjugated with glucuronic acid

Simultaneous determination of resorcylic acid lactones, β and α trenbolone and stilbenes in bovine urine by UHPLC/MS/MS

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