2020
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Abstract: Stanozolol is still the most commonly used illicit anabolic‐androgenic steroid (AAS) in professional sports. Therefore, accurate and fast analysis and long detection windows are of great interest in the field of antidoping analysis. In this work, a very simple, fast, and highly sensitive online solid‐phase extraction method coupled with liquid chromatography–high‐resolution tandem mass spectrometry (HPLC‐HRMSMS) for the analysis of stanozolol‐N‐glucuronides was developed. This fully validated procedure is char… Show more

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“…Hence, a considerable number of studies conducted during the past 12 months focused on the identification of new diagnostic (long‐term) metabolites, synthesis of reference material, 56,57 and implementation into routine doping controls, with four recent projects investigating 17α‐methyl steroids. For instance, Göschl et al reported on the rapid and facile detection and confirmation of stanozolol‐ N ‐glucuronides in human urine by means of urine dilution followed by online‐SPE and LC‐HRMS/MS analysis 58 . As little as 25 μl of urine are injected onto a phenyl‐hexyl extraction column (10 × 3 mm, 2.6 μm particle size), and after loading, the retained analytes are flushed onto a C‐18 analytical column (100 × 2.1 mm, 2.6 μm particle size).…”
Section: Anabolic Agentsmentioning
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“…Hence, a considerable number of studies conducted during the past 12 months focused on the identification of new diagnostic (long‐term) metabolites, synthesis of reference material, 56,57 and implementation into routine doping controls, with four recent projects investigating 17α‐methyl steroids. For instance, Göschl et al reported on the rapid and facile detection and confirmation of stanozolol‐ N ‐glucuronides in human urine by means of urine dilution followed by online‐SPE and LC‐HRMS/MS analysis 58 . As little as 25 μl of urine are injected onto a phenyl‐hexyl extraction column (10 × 3 mm, 2.6 μm particle size), and after loading, the retained analytes are flushed onto a C‐18 analytical column (100 × 2.1 mm, 2.6 μm particle size).…”
Section: Anabolic Agentsmentioning
“…Very recently, an online turbulent flow sample preparation method coupled with UHPLC-high resolution mass spectrometry (HRMS) for the analysis of 10 mycotoxins in human urine after glucuronidase treatment was published [25]. Also, for the analysis of target compounds like metabolites of anabolic-androgenic steroids [26] or pesticide biomarkers [27], as well as for doping control analysis [28] in urine samples, online SPE approaches proved to be suitable.…”
Section: Introductionmentioning
“…4 In the following 35 years, many other techniques, primarily based on mass spectrometric techniques coupled to on either gas (GC-MS) or liquid chromatography (LC-MS), for analyzing a large number of different stanozolol metabolites, were published. [5][6][7][8][9][10][11][12][13][14][15][16][17][18][19][20] In general, the traditional approach for the simultaneous analysis of several different steroids is to perform enzymatic hydrolysis to cleave highly polar phase-II conjugates, like glucuronic acids and sulfates, followed by liquid-liquid extraction and the analysis of remaining phase-I metabolites and parent molecules with GC-or LC-MS. 21,22 For the measurement with GC-MS, the analytes are additionally derivatized with N-methyl-N-trimethylsilyltrifluoroacetamide (MSTFA) to reduce their polarity. This kind of approach is the gold standard nowadays and is commonly performed by anti-doping laboratories worldwide for the routine initial testing procedure (ITP), often including the detection of stanozolol parent or phase-I metabolites.…”
Section: Introductionmentioning
“…This approach was optimized by placing a fully automated online solid-phase extraction (SPE) procedure upstream of the analytical measurement with LC-HRMS. 18 Next to the aspect of saving time and resources by direct analysis of phase-II conjugates, no enzymatic hydrolysis step using, for example, β-glucuronidase from Escherichia coli is required. Consequently, issues like incomplete or inhibited hydrolysis to yield phase-I metabolites, as necessary for GC-MS methods, are no longer relevant.…”
Section: Introductionmentioning
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