Detection of inhaled salbutamol in equine urine by ELISA and GC/MS2.
Abstract: Salbutamol is a beta-adrenergic agonist that is used in the treatment of asthma in humans and chronic obstructive pulmonary disease in horses. Because of its stimulating and growth promoting properties, it is prohibited by horse racing authorities. Recently a number of adapters (eg Equinehaler) have been designed, allowing the use of metered dose inhalers (MDI) approved for human use. However, information on detection times of salbutamol after administration of salbutamol in therapeutic doses by inhalation is lacking. In this study, 2 mg salbutamol (Ventolin) was administered to four standardbred mares via an MDI with an Equinehaler and urine was collected during 48 h. Quantification of salbutamol in horse urine was done via an overnight beta-agonist ELISA kit. Salbutamol was detected between 1 and 48 h post-administration. Relatively large interindividual variations in the total amount excreted during the first 12 h were noticed. The maximum urinary concentrations varied between 4.6 and 8.1 ng/mL. The total amount excreted within the first 12 h varied between 0.2 and 0.7% of the administered dose. For confirmatory analysis in doping control, a GC/MS(2) method was developed and validated. Analysis was performed on an ion trap instrument after solid phase extraction. The limit of detection was 0.25 ng/mL and was lower than in previously reported methods in human urine.
Copyright 2002 John Wiley & Sons, Ltd.
Publication Date: 2002-12-11 PubMed ID: 12474214DOI: 10.1002/bmc.194Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research article explores the detection of salbutamol, a drug used for asthma treatment in humans and lung disease treatment in horses, in horse urine post-administration through inhalation with the help of an Equinehaler and a human-appropriate metered dose inhaler. The amount of salbutamol excretion varies per horse and can be detected post-administration from 1 to 48 hours.
Research Objectives and Methods
- The research was primarily focused on detecting the presence and measuring the quantity of the drug salbutamol in horse urine. This is crucial, especially for horse racing events, as salbutamol can potentially enhance performance thus it is banned.
- The study involved administering 2 mg salbutamol (under the brand name Ventolin) to four standardbred mares through a metered dose inhaler (MDI) using an Equinehaler. These mares’ urine was then collected over a period of 48 hours.
- The researchers applied an overnight beta-agonist ELISA (Enzyme-Linked Immunosorbent Assay) kit for quantification of the salbutamol present in the equine urine.
Observations and Results
- Salbutamol was detected in the urine samples between 1 and 48 hours after it was administered to the mares.
- Significant interindividual variations were observed in the total amount of salbutamol excreted during the first 12 hours. The highest concentrations of salbutamol in the urine varied between 4.6 and 8.1 ng/mL.
- The total amount of salbutamol excreted within the first 12 hours ranged from 0.2 to 0.7% of the given dose.
Confirmatory Analysis
- A gas chromatography/mass spectrometry (GC/MS²) method was developed and validated for confirmatory analysis, which was crucial for accurate doping control. This analysis was conducted on an ion trap instrument post solid phase extraction.
- The limit of detection with this method was 0.25 ng/mL, a sensitivity lower than other previously reported methods used for detection in human urine, indicating this method’s superior sensitivity for detection in equine urine.
Cite This Article
APA
Eenoo PV, Delbeke FT.
(2002).
Detection of inhaled salbutamol in equine urine by ELISA and GC/MS2.
Biomed Chromatogr, 16(8), 513-516.
https://doi.org/10.1002/bmc.194 Publication
Researcher Affiliations
- Doping Control Unit, Department of Pharmacology, Pharmacy and Toxicology, University Ghent, Salisburylaan 133, B-9820 Merelbeke, Belgium.
MeSH Terms
- Adrenergic beta-Agonists / urine
- Albuterol / urine
- Animals
- Calibration
- Enzyme-Linked Immunosorbent Assay / methods
- Female
- Gas Chromatography-Mass Spectrometry / methods
- Horses / urine
- Reproducibility of Results
Citations
This article has been cited 2 times.- Tozaki T, Ohnuma A, Kikuchi M, Ishige T, Kakoi H, Hirota KI, Kusano K, Nagata SI. Microfluidic Quantitative PCR Detection of 12 Transgenes from Horse Plasma for Gene Doping Control. Genes (Basel) 2020 Apr 23;11(4).
- Tozaki T, Ohnuma A, Takasu M, Kikuchi M, Kakoi H, Hirota KI, Kusano K, Nagata SI. Droplet Digital PCR Detection of the Erythropoietin Transgene from Horse Plasma and Urine for Gene-Doping Control. Genes (Basel) 2019 Mar 21;10(3).
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