An innovative derivatization-free IC-MS/MS method for the detection of bisphosphonates in horse plasma.
Abstract: Bisphosphonates are prohibited drugs according to Article 6 of the International Agreement on Breeding, Racing and Wagering of the International Federation of Horseracing Authorities (IFHA) and the International Equestrian Federation (FEI). These compounds are used for the treatment of lameness, navicular and bone diseases in horses and are divided into two groups: non-nitrogen-containing bisphosphonate drugs (e.g. clodronic acid) and nitrogen-containing bisphosphonate drugs (e.g. zoledronic acid). Their hydrophilic properties and the high affinity for the bone matrix make the control of their use quite difficult. Current analytical strategies to detect such compounds often rely on a solid phase extraction (SPE) followed by detection by means of UHPLC-MS/MS after methylation with chemical reagents. To improve the analysis throughput and to eliminate the need for chemical derivatization, an innovative 96-well SPE followed by ion chromatography-mass spectrometry was developed. Analyses are conducted on an ICS-6000 HPIC system coupled to a TSQ Altis™ (Thermo Scientific™). The use of a 96-well SPE allowed 5-fold sample increase and a 6-fold throughput improvement. While preliminary results conducted on horse plasma exhibited similar performances to the method for the detection of non-nitrogen-containing bisphosphonates, the analytical performances of nitrogen-containing bisphosphonates were greatly improved.
© 2020 John Wiley & Sons, Ltd.
Publication Date: 2020-07-14 PubMed ID: 32615643DOI: 10.1002/dta.2892Google Scholar: Lookup
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- Journal Article
Summary
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The research introduces a new method for detecting prohibited bisphosphonate drugs in horse plasma, using an ion chromatography-mass spectrometry technique that eliminates the need for chemical derivatization, therefore increasing the analysis throughput five-fold while also, improving detection of nitrogen-containing bisphosphonates specifically.
Background
- Bisphosphonates are drugs typically used for the treatment of various bone diseases in horses. They are however, prohibited for use in horse racing and equestrian events, due to their potential performance-enhancing properties.
- Detection and control of bisphosphonate use in horses is challenging due to their hydrophilic properties and high affinity for bone matrix.
- Current detection methods often involve solid phase extraction (SPE) and detection through UHPLC-MS/MS after chemically inducing methylation.
Methodology
- The researchers proposed a novel approach aiming to improve the throughput of the detection process, by eliminating the need for chemical derivatization.
- A 96-well SPE technique was used followed by an ion chromatography-mass spectrometry process on an ICS-6000 HPIC system coupled with a TSQ Altis™.
Results and Conclusion
- The new method allowed for a 5-fold increase in sample size, along with a 6-fold improvement in throughput, hence, the effectiveness and speed of the test were significantly improved.
- While the new method showed similar results to the existing procedures in the detection of non-nitrogen-containing bisphosphonates, it showed greater effectiveness in detecting nitrogen-containing bisphosphonates.
Implications
- This research presents a faster and potentially more reliable method for controlling the use of bisphosphonate drugs in horses, specifically in the sporting and racing industries.
- The researchers’ methodology, due to its higher throughput, could provide a more efficient testing approach in drug control procedures, ensuring fair play in equestrian sports.
- Further studies could expand on this research by continuing to refine and develop this method, making it even more reliable and efficient.
Cite This Article
APA
Garcia P, Pinètre J, Morel S, Jaubert M, Deruy X, Perot I, Delcourt V, Loup B, Popot MA, Bailly-Chouriberry L.
(2020).
An innovative derivatization-free IC-MS/MS method for the detection of bisphosphonates in horse plasma.
Drug Test Anal, 12(10), 1452-1461.
https://doi.org/10.1002/dta.2892 Publication
Researcher Affiliations
- GIE LCH, Laboratoire des Courses Hippiques, Verrières le Buisson, France.
- GIE LCH, Laboratoire des Courses Hippiques, Verrières le Buisson, France.
- Thermo Fisher Scientific, Villebon sur Yvette, France.
- GIE LCH, Laboratoire des Courses Hippiques, Verrières le Buisson, France.
- Thermo Fisher Scientific, Villebon sur Yvette, France.
- GIE LCH, Laboratoire des Courses Hippiques, Verrières le Buisson, France.
- GIE LCH, Laboratoire des Courses Hippiques, Verrières le Buisson, France.
- GIE LCH, Laboratoire des Courses Hippiques, Verrières le Buisson, France.
- GIE LCH, Laboratoire des Courses Hippiques, Verrières le Buisson, France.
- GIE LCH, Laboratoire des Courses Hippiques, Verrières le Buisson, France.
MeSH Terms
- Animals
- Bone Density Conservation Agents / blood
- Chromatography, High Pressure Liquid
- Diphosphonates / blood
- Doping in Sports
- Horses / blood
- Solid Phase Extraction
- Tandem Mass Spectrometry
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Citations
This article has been cited 2 times.- Yi YX, Jin MC. [Advances in the application of ion chromatography-mass spectrometry in the fields of life and health]. Se Pu 2024 Oct;42(10):923-934.
- Ngere JB, Ebrahimi KH, Williams R, Pires E, Walsby-Tickle J, McCullagh JSO. Ion-Exchange Chromatography Coupled to Mass Spectrometry in Life Science, Environmental, and Medical Research. Anal Chem 2023 Jan 10;95(1):152-166.
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