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Home / Equine Research Bank / Drug Test Anal / Detection and confirmation of zilpaterol in equine hair usin...
Drug testing and analysis2021; 14(1); 31-38; doi: 10.1002/dta.3138

Detection and confirmation of zilpaterol in equine hair using liquid chromatography-mass spectrometry.

Abstract: Zilpaterol is a β -adrenergic agonist and a repartitioning agent that has a high potential for abuse in equine performance athletes. Analysis of zilpaterol in hair is an alternative sampling matrix that extends detection time periods beyond those found in urine or blood samples. Our laboratory has been screening for zilpaterol in hair for many years and recently detected and confirmed its presence in official samples. Accordingly, a liquid chromatography-mass spectrometry method was developed and validated to detect and confirm zilpaterol in equine hair. Briefly, equine hair was decontaminated, cut, and pulverized prior to disruption and liquid-liquid extraction in basic conditions. Following extraction, the sample was introduced to an Agilent 1260 HPLC and zilpaterol was separated using a reverse phase gradient with a total run time of 12.5 min. Following chromatographic separation, zilpaterol and its corresponding stable isotope labeled internal standard were introduced via positive mode electrospray ionization to a Thermo Q-Exactive Plus mass spectrometer and spectra collected using parallel reaction monitoring. The methodology was validated using in-house criteria including characterization of accuracy, precision, recovery, linear range, matrix effects, limit of detection, and limit of quantitation, and the method was found to be fit-for-purpose to confirm the presence of zilpaterol in equine hair. This methodology has been used to detect and confirm the presence of zilpaterol from out-of-competition hair samples submitted by regional racing authorities.
© 2021 John Wiley & Sons, Ltd.
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Publication Date: 2021-08-21 PubMed ID: 34355536DOI: 10.1002/dta.3138Google Scholar: Lookup
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  • Journal Article
  • Validation Study

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research paper provides a method for detecting zilpaterol, a substance potentially abused in equine performance athletes, in horse hair using liquid chromatography-mass spectrometry. The method is validated and proven effective for confirming the presence of zilpaterol in equine hair samples.

Objective of the Research

  • Despite multiple screening methods for zilpaterol in different matrices, the authors believed that analyzing it in hair could extend detection time periods beyond those offered by urine or blood samples.
  • The study was designed to develop and validate a liquid chromatography-mass spectrometry method for detecting and confirming the presence of zilpaterol in equine hair.

Methodology

  • Equine hair was first decontaminated, cut, and pulverized. Post this, it was disrupted and subjected to a liquid-liquid extraction in basic conditions.
  • The resultant extract was introduced to an Agilent 1260 HPLC and zilpaterol was separated using a reverse phase gradient. The total run time for this step was 12.5 minutes.
  • After chromatographic separation, zilpaterol and its stable isotope labeled internal standard were introduced to a Thermo Q-Exactive Plus mass spectrometer. Spectra were then collected using parallel reaction monitoring.

Validation of Methodology

  • The methodology was validated based on several in-house criteria: accuracy, precision, recovery, linear range, matrix effects, limit of detection, and limit of quantitation.
  • The method was found to be accurate, precise, and capable of recovering zilpaterol effectively. It detected and quantified zilpaterol across a linear range and showed minimal matrix effects.

Confirmation and Further Applications

  • The method was confirmed to be fit-for-purpose and could be used to confirm the presence of zilpaterol in equine hair.
  • It has been successfully used to detect and confirm the presence of zilpaterol from out-of-competition hair samples submitted by regional racing authorities.

Cite This Article

APA
Moeller BC, Clifford A, Emery RT, Alarcio G, Favro G, Arthur RM. (2021). Detection and confirmation of zilpaterol in equine hair using liquid chromatography-mass spectrometry. Drug Test Anal, 14(1), 31-38. https://doi.org/10.1002/dta.3138

Publication

Drug testing and analysis
ISSN: 1942-7611
NlmUniqueID: 101483449
Country: England
Language: English
Volume: 14
Issue: 1
Pages: 31-38

Researcher Affiliations

Moeller, Benjamin C
  • KL Maddy Equine Analytical Chemistry Laboratory, School of Veterinary Medicine, University of California, Davis, Davis, California, USA.
  • Department of Molecular Biosciences, School of Veterinary Medicine, University of California, Davis, Davis, California, USA.
Clifford, Amel
  • KL Maddy Equine Analytical Chemistry Laboratory, School of Veterinary Medicine, University of California, Davis, Davis, California, USA.
Emery, Richard T
  • KL Maddy Equine Analytical Chemistry Laboratory, School of Veterinary Medicine, University of California, Davis, Davis, California, USA.
Alarcio, Gwen
  • KL Maddy Equine Analytical Chemistry Laboratory, School of Veterinary Medicine, University of California, Davis, Davis, California, USA.
Favro, Gianna
  • KL Maddy Equine Analytical Chemistry Laboratory, School of Veterinary Medicine, University of California, Davis, Davis, California, USA.
Arthur, Rick M
  • School of Veterinary Medicine, University of California, Davis, Davis, California, USA.

MeSH Terms

  • Adrenergic beta-2 Receptor Agonists / analysis
  • Animals
  • Chromatography, High Pressure Liquid / methods
  • Chromatography, High Pressure Liquid / veterinary
  • Doping in Sports / prevention & control
  • Hair / chemistry
  • Horses
  • Limit of Detection
  • Liquid-Liquid Extraction / methods
  • Substance Abuse Detection / methods
  • Substance Abuse Detection / veterinary
  • Tandem Mass Spectrometry / methods
  • Tandem Mass Spectrometry / veterinary
  • Trimethylsilyl Compounds / analysis

Grant Funding

  • California Horse Racing Board

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