Detection of inhaled clenbuterol in horse urine by GC/MS2.
Abstract: Clenbuterol, a beta-adrenergic agonist, is used in the treatment of recurrent airway obstruction in horses. It is prohibited by horse racing authorities, because of its stimulating and growth-promoting properties. However, information on detection times of clenbuterol after administration by nebulization is lacking. In this study, a fast, sensitive quantitative GC-MS(2) method for the detection of clenbuterol in urine was developed. Alkaline liquid-liquid extraction was followed by derivatization to a cyclic methyl boronate derivative and analysis on a Finnigan MAT GCQ instrument. Method validation showed good linearity in the range 0.1-2.0 ng/mL, excellent repeatability and specificity. The limit of quantitative detection of the method was 0.1 ng/ml. Different instrumental parameters of the ion trap mass spectrometer were changed to increase the number of diagnostic ions for the cyclic methyl boronate derivative of clenbuterol. The influence of these changes and their applicability within the requirements and the criteria for mass spectrometry set by the responsible regulatory bodies are discussed. Clenbuterol was administered via nebulization to five standardbred mares (0.4 micro g/kg body weight). Analysis of the urine samples resulted in the detection of clenbuterol, as early as 2 h post administration and for up to 36 h post treatment. Generally, maximum urinary concentrations of 1.2 ng/mL were reached after -6-9 h.
Copyright 2002 John Wiley & Sons, Ltd.
Publication Date: 2002-10-16 PubMed ID: 12378561DOI: 10.1002/bmc.188Google Scholar: Lookup
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- Journal Article
Summary
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This research paper discusses the development of a sensitive and rapid testing method to detect the presence of the drug clenbuterol in horse urine, which could be relevant to enforcing regulations in horse racing where the drug is prohibited.
Background
- Clenbuterol is a beta-adrenergic agonist, typically used to treat recurring airway blockages in horses.
- Despite its medical uses, there’s been an effort to curtail its misuse in horse racing, as it can also stimulate and promote growth in horses.
- In this context, the researchers developed a tool to detect when horses have inhaled dose of the drug via nebulization, where previously there was no information on how long the drug could be detected that way.
The Method
- The researchers developed a fast and sensitive testing process that could detect the presence of clenbuterol in horse urine.
- This involved first extracting the drug from the urine, then using a chemical process to create a derivative of the drug that the researchers could reliably detect when using a mass spectrometer.
- The spot detection of the drug was capably determined, with the limit set to 0.1 ng/mL.
- The researchers also tinkered with different parameters on the mass spectrometer to further improve the sensitivity of the detection.
The Experiment
- After having developed the method, they then performed an experiment where five standardbred mares were administered a clenbuterol dose via nebulization.
- Urine samples were then taken at different intervals and then examined for the presence of the drug.
- Results shown clenbuterol was detectable as early as 2 hours after administration and remained detectable for up to 36 hours after the treatment.
- Fark test resulted that its maximum concentration in the urine usually hit around 6 to 9 hours after dosage.
Conclusion
- This study offers a method for detecting clenbuterol in horse urine post-inhalation, potentially providing improvements to testing procedures and fairness in horse racing regulations enforcement.
- The developed method was found to be fast, reliable, and capable of detecting small quantities of the drug.
Cite This Article
APA
Van Eenoo P, Delbeke FT, Deprez P.
(2002).
Detection of inhaled clenbuterol in horse urine by GC/MS2.
Biomed Chromatogr, 16(7), 475-481.
https://doi.org/10.1002/bmc.188 Publication
Researcher Affiliations
- Doping Control Unit, Department of Pharmacology, Pharmacy and Toxicology, Ghent University, Salisburylaan 133, B-9820 Merelbeke, Belgium.
MeSH Terms
- Administration, Inhalation
- Adrenergic beta-Agonists / administration & dosage
- Adrenergic beta-Agonists / urine
- Animals
- Clenbuterol / administration & dosage
- Clenbuterol / urine
- Female
- Gas Chromatography-Mass Spectrometry / methods
- Horses
- Reproducibility of Results
- Sensitivity and Specificity
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