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Validated UHPLC-MS-MS method for rapid analysis of capsaicin and dihydrocapsaicin in equine plasma for doping control.
Abstract: A method involving ultra high-performance liquid chromatography-tandem mass spectrometry was developed and validated for the analysis of capsaicin and dihydrocapsaicin in equine plasma. The analytes were recovered from plasma by liquid-liquid extraction using methyl tert-butyl ether and separated on a sub-2 micron column. The mobile phase was composed of 2 mM ammonium formate and methanol. A triple quadrupole mass spectrometer was used to detect the analytes in positive electrospray ionization mode with selected reaction monitoring. The limits of detection, quantification and confirmation for both analytes were 0.5, 1.0 and 2.5 pg/mL, respectively. The linear dynamic range of quantification was 1.0-1,000 pg/mL. During storage, both analytes in equine plasma were unstable at room temperature but stable at -20 and -70°C. The retention time and product ion ratios were employed as the criteria for confirmation of the presence of the analytes in plasma. The total analysis time was 2 min. The method is fast, selectively sensitive, reproducible, reliable and fully validated.
Publication Date: 2013-01-11 PubMed ID: 23316028DOI: 10.1093/jat/bks098Google Scholar: Lookup The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
- Validation Study
- Analytical Methods
- Diagnosis
- Diagnostic Technique
- Disease control
- Disease Diagnosis
- Disease Management
- Disease Treatment
- Doping
- Drug
- Equine Diseases
- Equine Health
- Equine Science
- High-performance Liquid Chromatography (HPLC)
- Horses
- Laboratory Methods
- Pharmaceuticals
- Plasma
- Veterinary Medicine
- Veterinary Research
- Veterinary Science
Summary
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The research article is about a method that has been developed and validated for detecting capsaicin and dihydrocapsaicin in horse’s blood quickly and effectively. This method uses ultra high-performance liquid chromatography and mass spectrometry, and is designed for use in doping control.
Research Overview
- This study was aimed at developing and validating a method to swiftly and accurately analyze capsaicin and dihydrocapsaicin in equine plasma using ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS-MS). These substances are derived from chili peppers and can be abused in horse racing to mask pain and enhance performance, a process known as “doping”.
- The process involves recovering the substance from the blood using a process known as liquid-liquid extraction. After extraction, they were then separated on a specially designed column.
Methodology
- The column used was a sub-2 micron column and the substances were separated using a mobile phase composed of 2 mM ammonium formate and methanol.
- A triple quadrupole mass spectrometer was used to detect the substances using positive electrospray ionization mode with selected reaction monitoring.
- The limits of detection, quantification, and confirmation for both substances were set at 0.5, 1.0, and 2.5 pg/mL, respectively.
- The range within which the substance levels can be quantified was between 1.0-1,000 pg/mL.
Stability and Confirmation of Analytes
- The study found that capsaicin and dihydrocapsaicin were unstable at room temperature during storage. However, they remained stable at temperatures of -20 and -70°C.
- The presence of the substances in the plasma was confirmed using the retention time and product ion ratios.
Conclusion
- The duration of the total analysis was only 2 minutes, making it a time-efficient method.
- The researchers concluded that the method was fast, selectively sensitive, reproducible, reliable, and fully validated for the intended purpose.
Cite This Article
APA
You Y, Uboh CE, Soma LR, Guan F, Taylor D, Li X, Liu Y, Chen J.
(2013).
Validated UHPLC-MS-MS method for rapid analysis of capsaicin and dihydrocapsaicin in equine plasma for doping control.
J Anal Toxicol, 37(2), 122-132.
https://doi.org/10.1093/jat/bks098 Publication
Researcher Affiliations
- Department of Clinical Studies, School of Veterinary Medicine, University of Pennsylvania, New Bolton Center Campus, Kennett Square, PA 19348, USA.
MeSH Terms
- Animals
- Capsaicin / analogs & derivatives
- Capsaicin / blood
- Chromatography, High Pressure Liquid
- Doping in Sports
- Female
- Horses
- Limit of Detection
- Male
- Performance-Enhancing Substances / analysis
- Substance Abuse Detection / methods
- Tandem Mass Spectrometry / methods
Citations
This article has been cited 5 times.- Rodríguez-Palazón MC, Arroyo-Manzanares N, Viñas P, López-García I, Hernández-Córdoba M, Campillo N. Dispersive magnetic solid-phase extraction for capsaicinoid compounds in human serum using LC-HRMS: targeted and non-targeted approaches. Anal Bioanal Chem 2023 May;415(11):2133-2145.
- Zamljen T, Medic A, Hudina M, Veberic R, Slatnar A. Biostimulatory Effects of Amino Acids on Phenylalanine Ammonia Lyase, Capsaicin Synthase, and Peroxidase Activities in Capsicum baccatum L. Biology (Basel) 2022 Apr 27;11(5).
- Liu Z, Yu S, Xu S, Zhao B, Xu W. Ultrasensitive Detection of Capsaicin in Oil for Fast Identification of Illegal Cooking Oil by SERRS. ACS Omega 2017 Nov 30;2(11):8401-8406.
- Zak A, Siwinska N, Slowikowska M, Borowicz H, Szpot P, Zawadzki M, Niedzwiedz A. The detection of capsaicin and dihydrocapsaicin in horse serum following long-term local administration. BMC Vet Res 2018 Jun 19;14(1):193.
- Harlan R, Zhang H. Targeted proteomics: a bridge between discovery and validation. Expert Rev Proteomics 2014 Dec;11(6):657-61.
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