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Drug testing and analysis2020; 12(8); 1065-1077; doi: 10.1002/dta.2864

Quantification and validation of nine nonsteroidal anti-inflammatory drugs in equine urine using gas chromatography-mass spectrometry for doping control.

Abstract: Nonsteroidal anti-inflammatory drugs (NSAIDs) are commonly used in therapeutic doses in human and veterinary medicine for the treatment of inflammation, pain, and fever. A method for the simultaneous determination of nine NSAIDs, known as therapeutic prohibited substances, in equine urine was developed and fully validated according to the European Commission Decision 2002/657/EC and Association of Official Racing Chemists criteria. The validation was performed for naproxen, flunixin, ketoprofen, diclofenac, eltenac, meclofenamic acid, phenylbutazone, vedaprofen, and carprofen in equine urine in accordance with the International Screening Limits (ISL) regulated by International Federation of Horseracing Authorities. After basic hydrolysis, samples were extracted with a C18 cartridge using automated solid-phase extraction. Several derivatization reagents were investigated, and trimethylphenylammonium hydroxide/methanol (20/80, v/v) was selected. Analyses were carried out using gas chromatography-mass spectrometry with selected ion monitoring mode, but the method can be applied to a large number of analytes. The within-laboratory reproducibility was not more than 12.8% (≤15%), and mean relative recoveries ranged from 91.1% to 104.1% for inter-day and intra-day precision. The decision limits (CCα) and detection capabilities (CCβ) were evaluated at concentrations near the ISL for each therapeutic substance. The validation results demonstrated that the method is highly reproducible, easily applicable, and suitable for the analysis of some NSAIDs in equine urine that have not been previously published. Finally, the method was also applied to known positive samples.
© 2020 John Wiley & Sons, Ltd.
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Publication Date: 2020-06-28 PubMed ID: 32449838DOI: 10.1002/dta.2864Google 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.

This research article outlines a newly developed method for detecting nine nonsteroidal anti-inflammatory drugs (NSAIDs) in equine urine, aiming to enhance doping control in horse racing. The method was validated according to international regulations and shows high reliability and applicability.

Study Objective and Method Development

  • The research aimed to develop and validate a method for identifying nine NSAIDs in horses’ urine. These substances, used for pain and inflammation relief, are considered therapeutic prohibited substances in equine sports like horse racing.
  • The NSAIDs considered in the study are naproxen, flunixin, ketoprofen, diclofenac, eltenac, meclofenamic acid, phenylbutazone, vedaprofen, and carprofen.
  • Validation of the method followed the guidelines by the European Commission Decision 2002/657/EC and the Association of Official Racing Chemists, ensuring the study’s credibility and consistency with international standards.
  • The study employed a scientific process that involved basic hydrolysis and sample extraction using a C18 cartridge and automated solid-phase extraction.

Selection of Derivatization Reagents and Analysis

  • Different derivatization reagents were tested and the researchers selected a mixture of trimethylphenylammonium hydroxide and methanol for further analyses.
  • The detection of NSAIDs was done using gas chromatography-mass spectrometry with selected ion monitoring mode. This technique allows for a large number of analytes to be considered, expanding its applicability.
  • The study highlighted that the developed method could analyze some NSAIDs in equine urine that have not been previously published, marking a significant contribution to the field.

Reproducibility and Precision of the Method

  • The research presented a notable degree of reproducibility within the laboratory setting, with a maximum variation of 12.8% (a figure less than or equal to 15%, which is deemed acceptable).
  • Mean relative recoveries, which measure the method’s inter-day and intra-day precision, ranged between 91.1% and 104.1%, showing high accuracy.
  • The decision limits (CCα) and detection capabilities (CCβ) were evaluated near the International Screening Limits (ISL) for each therapeutic substance, providing a realistic assessment of their detection potential.

Practical Application of the Method

  • The developed method was tested on known positive samples, demonstrating its practical applicability.
  • The results of the study indicated that the method not only holds high reproducibility but can also easily be applied in different settings, making it a reliable tool for doping control in horse racing.

Cite This Article

APA
Göktaş EF, Kabil E, Arıöz F. (2020). Quantification and validation of nine nonsteroidal anti-inflammatory drugs in equine urine using gas chromatography-mass spectrometry for doping control. Drug Test Anal, 12(8), 1065-1077. https://doi.org/10.1002/dta.2864

Publication

Drug testing and analysis
ISSN: 1942-7611
NlmUniqueID: 101483449
Country: England
Language: English
Volume: 12
Issue: 8
Pages: 1065-1077

Researcher Affiliations

Göktaş, Eylem Funda
  • Doping Control Laboratory, Istanbul Pendik Veterinary Control Institute, Istanbul, Turkey.
Kabil, Erol
  • Doping Control Laboratory, Istanbul Pendik Veterinary Control Institute, Istanbul, Turkey.
Arıöz, Filiz
  • İstanbul, Turkey.

MeSH Terms

  • Animals
  • Anti-Inflammatory Agents, Non-Steroidal / urine
  • Doping in Sports / prevention & control
  • Gas Chromatography-Mass Spectrometry / methods
  • Gas Chromatography-Mass Spectrometry / veterinary
  • Horses
  • Reproducibility of Results
  • Solid Phase Extraction

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Citations

This article has been cited 1 times.
  1. Zhao J, Wang Y, Liu B. Doping Detection Based on the Nanoscale: Biosensing Mechanisms and Applications of Two-Dimensional Materials. Biosensors (Basel) 2025 Apr 3;15(4).
    doi: 10.3390/bios15040227pubmed: 40277541google scholar: lookup
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