High-performance liquid chromatography/tandem mass spectrometry: its use for the identification of stanozolol and its major metabolites in human and equine urine.
Abstract: A screening procedure for the anabolic steroid stanozolol in human and equine urine was developed based on enzymatic hydrolysis, liquid-liquid extraction and reversed-phase liquid chromatography combined on-line with tandem mass spectrometry. The column effluent was introduced into the atmospheric pressure ionization source of a triple-quadrupole mass spectrometer via a heated pneumatic nebulizer liquid chromatograph/mass spectrometer interface. Abundant protonated molecular ions were generated by corona discharge ionization. Confirmation of stanozolol and several of its hydroxylated and dihydroxylated metabolites isolated from both human and equine urine was accomplished by collision-induced dissociation of their parent ions. Interpretation of the daughter ion mass spectra gave valuable information for the structural elucidation of the detected metabolites. Using the selected reaction monitoring detection mode the presence of the urinary excretion products could be monitored in equine urine up to one day and in human urine for several days after oral administration of stanozolol. Microbore high-performance liquid chromatography/ion spray mass spectrometry of an ion-pair extract enabled the direct detection of intact sulfoconjugated hydroxy-metabolites in human urine.
Publication Date: 1990-01-01 PubMed ID: 2306548DOI: 10.1002/bms.1200190106Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This research article focuses on the development of a procedure that utilizes high-performance liquid chromatography and tandem mass spectrometry for identifying stanozolol, an anabolic steroid, and its primary metabolites in human and horse urine.
Introduction
- The study aimed to establish a screening method for the steroid stanozolol, widely used for performance enhancement, in both human and equine urine.
- It employed processes of enzymatic hydrolysis, liquid-liquid extraction, and reversed-phase liquid chromatography together with tandem mass spectrometry to achieve this goal.
Methodology
- The column effluent was inducted into the atmospheric pressure ionization source of a triple-quadrupole mass spectrometer via a heated pneumatic nebulizer liquid chromatograph and mass spectrometer interface.
- The use of corona discharge ionization produced plenty of protonated molecular ions.
Findings
- Confirmation of stanozolol and several of its hydroxylated and dihydroxylated metabolites isolated from both human and equine urine was attained by collision-induced dissociation of the parent ions.
- The interpretation of the daughter ion mass spectra delivered valuable data for the structural elucidation of the detected metabolites.
- With the help of the chosen reaction monitoring detection mode, the presence of the urinary excretion products could be monitored in equine urine up to one day and in human urine for several days after oral administration of stanozolol.
Additional Findings
- Microbore high-performance liquid chromatography and ion spray mass spectrometry of an ion-pair extract facilitated the direct detection of intact sulfoconjugated hydroxy-metabolites in human urine.
Conclusion
- The screening procedure outlined in the paper offers a reliable method for detecting the misuse of the steroid stanozolol in both humans and horses.
- It could help in the fair conduct of various sports and in human health monitoring where steroid misuse could be a contributing factor to health issues.
Cite This Article
APA
Mück WM, Henion JD.
(1990).
High-performance liquid chromatography/tandem mass spectrometry: its use for the identification of stanozolol and its major metabolites in human and equine urine.
Biomed Environ Mass Spectrom, 19(1), 37-51.
https://doi.org/10.1002/bms.1200190106 Publication
Researcher Affiliations
- Equine Drug Testing and Toxicology Program, New York State College of Veterinary Medicine, Cornell University, Ithaca 14850.
MeSH Terms
- Animals
- Chromatography, High Pressure Liquid
- Horses
- Humans
- Indicators and Reagents
- Male
- Mass Spectrometry
- Reference Standards
- Stanozolol / urine
Citations
This article has been cited 3 times.- Perkins JR, Parker CE, Tomer KB. Nanoscale separations combined with electrospray ionization mass spectrometry: Sulfonamide determination.. J Am Soc Mass Spectrom 1992 Feb;3(2):139-49.
- Schaefercor WH, Dixon F. Effect of high-performance liquid chromatography mobile phase components on sensitivity in negative atmospheric pressure chemical ionization liquid chromatography-mass spectrometry.. J Am Soc Mass Spectrom 1996 Oct;7(10):1059-69.
- Aebersold R, Bures EJ, Namchuk M, Goghari MH, Shushan B, Covey TC. Design, synthesis, and characterization of a protein sequencing reagent yielding amino acid derivatives with enhanced detectability by mass spectrometry.. Protein Sci 1992 Apr;1(4):494-503.
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