Analysis of β-agonists by HPLC/ESI-MS(n) in horse doping control.

This research article presents a sensitive method for detecting nine β(2) agonists (a type of medication used for respiratory disorders) in horse urine. The method combines solid-phase extraction and Liquid Chromatography/Electrospray Ionization/Multistage Mass Spectrometry (LC/ESI-MS(n)), capable of detecting these substances even at very low concentrations. It is intended for use both in initial screening and confirmatory analysis.

Introduction and Methodology

• The research centered around developing a new method to accurately detect nine specific β(2) agonists, which are typically used in the treatment of respiratory disorders, in horse urine. The agonists in question were cimaterol, clenbuterol, fenoterol, formoterol, mabuterol, terbutaline, ractopamine, salbutamol and salmeterol.
• To accomplish this, the research team used a combination of Solid-Phase Extraction (SPE) and Liquid Chromatography/Electrospray Ionization/Multistage Mass Spectrometry (LC/ESI-MS(n)). SPE was used to isolate the β(2) agonists from the horse urine, while LC/ESI-MS(n), performed on a quadrupole linear ion trap mass analyser, allowed for precise detection and analysis of the isolated compounds.

Results

• The combination of extraction efficiency of Solid-Phase Extraction and the sensitivity and selectivity of LC/ESI-MS(n) allowed for the detection of these β(2) agonists even at very low levels (picograms per milliliter).
• The research team conducted administration studies on two of the β(2) agonists — fenoterol and formoterol. The outcome demonstrated that their detection is possible even after these compounds have been inhaled, proving their method’s sensitivity.

Conclusion

• Due to the method’s sensitivity, it could be effectively used for both initial screening and confirmatory analysis in doping control in equestrian sport. Such an application could contribute significantly to the maintenance of fair competition standards and horse welfare.