Comprehensive screening of acidic and neutral drugs in equine plasma by liquid chromatography-tandem mass spectrometry.
Abstract: A multi-target high-throughput liquid chromatography-tandem mass spectrometry (LC-MS-MS) method for the detection of low ppt to low ppb levels of anabolic steroids, corticosteroids, anti-diabetics, and non-steroidal anti-inflammatory drugs (NSAIDs) in equine plasma was developed for the purpose of doping control. Plasma samples were first deproteinated by addition of trichloroacetic acid. Drugs were then extracted by solid-phase extraction (SPE) using Bond Elut Certify cartridges, and the extracts were analysed by a triple-quadrupole/linear ion trap LC-MS-MS instrument in positive electrospray ionization (+ESI) mode with selected reaction monitoring (SRM) scan function. Chromatographic separation of the targeted drugs was achieved using a reverse phase 3.3 cm L x 2.1 mm ID, 3 microm particle size LC column with gradient elution. Plasma samples fortified with 66 targeted drugs including betamethasone, boldione, capsaicin, flunisolide, gestrinone, gliclazide, 17alpha-hydroxyprogesterone hexanoate, isoflupredone and triamcinolone acetonide, etc. at low ppt to low ppb levels could be consistently detected. No significant matrix interference was observed at the retention time of the targeted ion transitions when blank plasma samples were analysed. The method has been validated for its extraction recoveries, precision and sensitivity, and is used regularly in the authors' laboratory to screen for the presence of these drugs in plasma samples from racehorses.
Publication Date: 2007-11-17 PubMed ID: 18054785DOI: 10.1016/j.chroma.2007.11.022Google Scholar: Lookup
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- Journal Article
Summary
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The research outlines the development of an efficient method for detecting a range of drugs, from anabolic steroids to NSAIDs, in equine plasma. This was done in an effort to improve anti-doping control in horse racing.
Introduction
- The researchers set out to devise an effective high-throughput method for drug detection in the plasma of racing horses. This was motivated by a requirement for strict doping control in equestrian sports.
- We are informed that a range of drugs were targeted in the study. These include anabolic steroids, corticosteroids, anti-diabetics, and NSAIDs.
Method
- The method the team developed involves some specific techniques in chromatography. Firstly, they removed proteins from plasma samples using trichloroacetic acid.
- They used solid-phase extraction (SPE) to extract drugs from these deproteinated samples, employing Bond Elut Certify cartridges for this task.
- These extracts were then examined using a triple-quadrupole/linear ion trap LC-MS-MS instrument. This analysis was carried out in positive electrospray ionization (+ESI) mode, with selected reaction monitoring (SRM) scan function.
- The procedure also involved chromatographic separation of the drugs being targeted. An LC column with gradient elution was used for this task. This made it possible to consistently detect varying concentrations of these drugs, from low ppt to low ppb levels.
Results
- The researchers observed no significant matrix interference at the retention time of the targeted ion transitions.
- Additionally, the method demonstrated the capacity to consistently detect plasma samples fortified with 66 different types of drugs within the low ppt to low ppb range. These include betamethasone, boldione, capsaicin, flunisolide, gestrinone, gliclazide, isoflupredone, and many others.
- All of this indicates that the newly developed method is reliable in detecting a wide range of drugs.
Conclusion
- The researchers performed validating tests on the proposed method. The results demonstrated that it had good extraction recoveries, precision, and sensitivity.
- The method has since been regularly used in the researchers’ lab for screening racehorse plasma samples for the presence of the said drugs.
Cite This Article
APA
Yu NH, Ho EN, Tang FP, Wan TS, Wong AS.
(2007).
Comprehensive screening of acidic and neutral drugs in equine plasma by liquid chromatography-tandem mass spectrometry.
J Chromatogr A, 1189(1-2), 426-434.
https://doi.org/10.1016/j.chroma.2007.11.022 Publication
Researcher Affiliations
- Racing Laboratory, The Hong Kong Jockey Club, Sha Tin Racecourse, Sha Tin, N.T., Hong Kong, China.
MeSH Terms
- Anabolic Agents / blood
- Animals
- Chromatography, Liquid / methods
- Horses
- Hypoglycemic Agents / blood
- Reproducibility of Results
- Substance Abuse Detection / methods
- Tandem Mass Spectrometry / methods
Citations
This article has been cited 2 times.- Aleman M, McCue PM, Chigerwe M, Madigan JE. Plasma concentrations of steroid precursors, steroids, neuroactive steroids, and neurosteroids in healthy neonatal foals from birth to 7 days of age. J Vet Intern Med 2019 Sep;33(5):2286-2293.
- Talari R, Varshosaz J, Mostafavi SA, Nokhodchi A. Gliclazide microcrystals prepared by two methods of in situ micronization: pharmacokinetic studies in diabetic and normal rats. AAPS PharmSciTech 2010 Jun;11(2):786-92.
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