Detection of Methylphenidate in Equine Hair Using Liquid Chromatography-High-Resolution Mass Spectrometry.
Abstract: 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.
Publication Date: 2021-09-24 PubMed ID: 34641342PubMed Central: PMC8510229DOI: 10.3390/molecules26195798Google Scholar: Lookup
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Summary
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The research paper presents a method developed for detecting a potent stimulant, methylphenidate, in horse hair samples through liquid chromatography interconnected with high-resolution mass spectrometry. This method is founded on meticulous preparation of the hair sample followed by sophisticated examination using a Thermo Q Exactive mass spectrometer.
Why Detect Methylphenidate?
- Methylphenidate is a potent stimulant that affects the central nervous system and is known for its high potential for misuse, particularly in competitive horse racing.
- Monitoring the presence of this drug both prior to and during competition is of major concern for horse racing authorities, in a bid to ensure lawful and ethical practice in the sport.
Advantages of Hair as a Sampling Medium
- Using hair for anti-doping testing in horses is becoming more popular amongst testers because of the growing range of compounds that can be spotted using this sampling matrix.
- Hair testing provides a wider window of detection compared to traditional blood or urine analysis.
Liquid Chromatography-High-Resolution Mass Spectrometry (LC-HRMS)
- Researchers developed a method involving liquid chromatography and high-resolution mass spectrometry (LC-HRMS) to trace any methylphenidate in hair samples submitted for testing.
- This process involves several steps, starting with the decontamination, chopping, and pulverizing of the hair sample. This is followed by liquid-liquid extraction under basic conditions before the resultant is subjected to the LC-MS system.
Validation of the Method
- The detection method was validated and several factors were determined, such as the limits of quantitation, linearity, matrix effects, recovery, accuracy, and precision.
- The validated method has been successfully put to use to confirm the presence of methylphenidate in official samples provided by racing authorities.
Cite This Article
APA
Moeller BC, Flores L, Clifford A, Alarcio G, Mosburg M, Arthur RM.
(2021).
Detection of Methylphenidate in Equine Hair Using Liquid Chromatography-High-Resolution Mass Spectrometry.
Molecules, 26(19), 5798.
https://doi.org/10.3390/molecules26195798 Publication
Researcher Affiliations
- KL Maddy Equine Analytical Chemistry Laboratory, School of Veterinary Medicine, University of California, Davis, CA 95616, USA.
- Department of Molecular Biosciences, School of Veterinary Medicine, University of California, Davis, CA 95616, USA.
- KL Maddy Equine Analytical Chemistry Laboratory, School of Veterinary Medicine, University of California, Davis, CA 95616, USA.
- KL Maddy Equine Analytical Chemistry Laboratory, School of Veterinary Medicine, University of California, Davis, CA 95616, USA.
- KL Maddy Equine Analytical Chemistry Laboratory, School of Veterinary Medicine, University of California, Davis, CA 95616, USA.
- KL Maddy Equine Analytical Chemistry Laboratory, School of Veterinary Medicine, University of California, Davis, CA 95616, USA.
- School of Veterinary Medicine, University of California, Davis, CA 95616, USA.
MeSH Terms
- Animals
- Chromatography, Liquid
- Hair / chemistry
- Horses
- Liquid-Liquid Extraction / methods
- Mass Spectrometry
- Methylphenidate / analysis
- Substance Abuse Detection / veterinary
Conflict of Interest Statement
The authors declare no conflict of interest.
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This article includes 21 references
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Citations
This article has been cited 1 times.- Jîtcă G, Mehelean IE, Maier AN, Jîtcă CM. Methylphenidate and Its Impact on Redox Balance and Behavior. J Xenobiot 2025 Sep 30;15(5).
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