Detection of methylphenidate in equine hair using liquid chromatography–high‐resolution mass spectrometry

Benjamin C. Moeller, Luis Flores, Amel Clifford, Gwendolyne Alarcio, Mary Mosburg, Rick M. Arthur

Research output: Contribution to journalArticlepeer-review


Methylphenidate is a powerful central nervous system stimulant with a high potential for abuse in horse racing. The detection of methylphenidate use is of interest to horse racing authorities for both prior to and during competition. The use of hair as an alternative sampling matrix for equine anti‐doping has increased as the number of detectable compounds has expanded. Our laboratory developed a liquid chromatography–high‐resolution mass spectrometry method to detect the presence of methylphenidate in submitted samples. Briefly, hair was decontaminated, cut, and pulverized prior to liquid–liquid extraction in basic conditions before introduction to the LC‐MS system. Instrumental analysis was conducted using a Thermo Q Exactive mass spectrometer using parallel reaction monitoring using a stepped collision energy to obtain sufficient product ions for qualitative identification. The method was validated and limits of quantitation, linearity, matrix effects, recovery, accuracy, and precision were determined. The method has been applied to confirm the presence of methylphenidate in official samples submitted by racing authorities.

Original languageEnglish (US)
Article number5798
Issue number19
StatePublished - Oct 1 2021


  • Anti‐doping
  • Hair
  • Horse
  • Liquid chromatography–mass spectrometry
  • Methylphenidate

ASJC Scopus subject areas

  • Analytical Chemistry
  • Chemistry (miscellaneous)
  • Molecular Medicine
  • Pharmaceutical Science
  • Drug Discovery
  • Physical and Theoretical Chemistry
  • Organic Chemistry


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