A quantitative method for the simultaneous detection of SR9009 and SR9011 in equine plasma using liquid chromatography-electrospray ionization-tandem mass spectrometry.
Abstract: SR9009 and SR9011 are metabolic modulators pharmacologically targeting REV-ERB receptors as synthetic agonists. A liquid chromatography-tandem mass spectrometry method for the detection of SR9009 and SR9011 in equine plasma was developed and validated. Plasma samples were pretreated by protein precipitation with methanol and were loaded onto an ACQUITY ultra performance liquid chromatography high-strength silica C18 column (2.1 × 150 mm, 1.8 μm) for chromatographic separation. The mobile phase consisted of 5-mM ammonium formate (pH 3.0) in distilled water and 0.1% formic acid in acetonitrile, and a gradient elution was used at a flow rate of 0.25 ml/min. For the mass spectrometry detection, the selected reaction monitoring mode was used with transitions of 438.2 → 124.9 for SR9009, 479.2 → 125.1 for SR9011, and 292.2 → 109.1 for the internal standard (testosterone-d3) in the positive ionization mode. The linearity, lower limit of quantification, intra- and inter-day precision, accuracy, matrix effect, recovery, and stability were evaluated. The method was found to be accurate and reproducible for the quantitation of SR9009 and SR9011. The developed method was successfully applied to plasma samples of thoroughbreds injected intramuscularly with SR9009.
© 2022 John Wiley & Sons Ltd.
Publication Date: 2022-04-19 PubMed ID: 35396832DOI: 10.1002/dta.3271Google Scholar: Lookup
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- Journal Article
- Analytical Methods
- Biochemistry
- Clinical Study
- Equine Health
- Equine Science
- High-performance Liquid Chromatography (HPLC)
- Horses
- Intramuscular Administration
- Laboratory Methods
- Metabolic Health
- Metabolism
- Pharmaceuticals
- Pharmacology
- Physiology
- Plasma
- Thoroughbreds
- Veterinary Care
- Veterinary Medicine
- Veterinary Research
Summary
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The study revolves around the development and validation of a method for detecting the presence of metabolic modulators SR9009 and SR9011 in horse plasma. The method employs liquid chromatography and mass spectrometry, demonstrating both accuracy and reproducibility in its applications.
Research background and objective
- SR9009 and SR9011 are metabolic modulators that function as synthetic agonists by pharmacologically targetting REV-ERB receptors.
- The objective of this study was to devise a method for detecting these modulators in equine plasma, importantly for thoroughbreds, known for racing and performance events.
Methodology
- The researchers used a method known as liquid chromatography-tandem mass spectrometry.
- Plasma samples were pretreated by protein precipitation with methanol. They were then loaded onto an ACQUITY ultra-performance liquid chromatography high-strength silica C18 column for chromatographic separation.
- The mobile phase consisted of 5-mM ammonium formate in distilled water and 0.1% formic acid in acetonitrile. A gradient elution was administered at a flow rate of 0.25 ml/min.
Mode of Detection
- Mass spectrometry detection was utilized in this study, specifically the selected reaction monitoring mode.
- The transitions were 438.2 → 124.9 for SR9009, 479.2 → 125.1 for SR9011, and 292.2 → 109.1 for the internal standard (testosterone-d3) during positive ionization mode.
Validation and Performance Evaluation
- The criteria used for performance evaluation included linearity, the lower limit of quantification, intra- and inter-day precision, accuracy, matrix effect, recovery, and stability.
- The method proved to be accurate and reproducible for the quantification of SR9009 and SR9011.
Practical Application
- The developed method was effectively applied to plasma samples of thoroughbred horses injected intramuscularly with SR9009.
- The ability to accurately detect the presence of SR9009 and SR9011 in equine plasma could potentially assist in ensuring fair play in equine sports by checking for unauthorised use of these performance modulators.
Cite This Article
APA
Kwak YB, Yu J, Yoo HH.
(2022).
A quantitative method for the simultaneous detection of SR9009 and SR9011 in equine plasma using liquid chromatography-electrospray ionization-tandem mass spectrometry.
Drug Test Anal, 14(8), 1532-1538.
https://doi.org/10.1002/dta.3271 Publication
Researcher Affiliations
- Korea Racing Authority, Gwachon, Republic of Korea.
- Institute of Pharmaceutical Science and Technology and College of Pharmacy, Hanyang University, Ansan, Republic of Korea.
- Korea Racing Authority, Gwachon, Republic of Korea.
- Institute of Pharmaceutical Science and Technology and College of Pharmacy, Hanyang University, Ansan, Republic of Korea.
MeSH Terms
- Animals
- Chromatography, High Pressure Liquid / methods
- Chromatography, Liquid
- Horses
- Pyrrolidines
- Reproducibility of Results
- Spectrometry, Mass, Electrospray Ionization / methods
- Tandem Mass Spectrometry / methods
- Thiophenes
References
This article includes 13 references
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Citations
This article has been cited 1 times.- Lu Z, Shen H, Li X, Li H, You W, Wang Z, Chen G. Circadian Rhythm Regulator REV-ERBα Attenuates Neuroapoptosis in Early Brain Injury After Experimental Subarachnoid Hemorrhage in Rats.. Neurochem Res 2023 Jun;48(6):1925-1944.
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