Detection of gonadotropin-releasing hormone and its analogues in equine and canine urine by high-resolution data-independent acquisition.
Abstract: Gonadotropin-releasing hormone (GnRH) and its synthetic analogues are considered banned substances by the racing industry. GnRH is used as a pharmaceutical to regulate the female oestrous cycle, but the hormone is also thought to increase the production of testosterone in male animals. Using liquid chromatography in conjunction with high-resolution mass spectrometry (LC-HRMS) and data-independent acquisition (DIA), a method is presented for the detection of intact and truncated peptides of GnRH and its analogues down to the low picogram level in equine urine. The study of the catabolism of GnRH and analogues in plasma, combined with the analysis of urine from administration studies, reveals a common pattern of peptide catabolites that can be used to guide the design of MS-based screens for this class of drugs. This culminated in the successful detection of the peptide in two out-of-competition canine urine samples.
© 2023 John Wiley & Sons Ltd.
Publication Date: 2023-05-03 PubMed ID: 37085977DOI: 10.1002/dta.3486Google Scholar: Lookup
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- Journal Article
Summary
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The study introduces a new method to detect gonadotropin-releasing hormone (GnRH) and its synthetic analogues in horse and dog urine, which are believed to be misused to enhance performance in racing animals.
Objective of the Study
- The main purpose of the study was to develop and demonstrate a new analytical procedure that can accurately detect the traces of GnRH and its synthetic analogues in the urine of horses and dogs. GnRH is often used to manipulate the oestrous cycle in females, and is believed to increase testosterone production in males, and hence, is considered a banned substance in animal racing.
Methodology
- The researchers used liquid chromatography in combination with high-resolution mass spectrometry (LC-HRMS) and data-independent acquisition (DIA) to establish this method.
- They also studied the catabolism of GnRH in plasma and combined it with the analysis of urine from relevant administration studies. This enabled them to observe a common pattern of peptide catabolites.
- The mentioned pattern can guide in the design of mass spectrometry (MS)-based screening processes for detecting this class of drugs.
Findings
- The researchers successfully detected these substances at very low concentrations, on a picogram level, in equine urine.
- Furthermore, they were able to identify these peptides in two out-of-competition canine urine samples, further demonstrating the effectiveness of the developed detection method.
Significance
- The findings of this study offer a promising new technique to detect the misuse of GnRH and its analogues in racing animals, giving authorities a more precise and reliable tool in ensuring fair competition and the well-being of racing animals.
Cite This Article
APA
Timms M, Botteon A, Manos C, Griffin J, Levina V, Steel R.
(2023).
Detection of gonadotropin-releasing hormone and its analogues in equine and canine urine by high-resolution data-independent acquisition.
Drug Test Anal, 15(8), 896-908.
https://doi.org/10.1002/dta.3486 Publication
Researcher Affiliations
- Racing Analytical Services Ltd, Flemington, Victoria, Australia.
- Biological Research Unit, Racing Analytical Services Limited, Flemington, Victoria, Australia.
- Racing Analytical Services Ltd, Flemington, Victoria, Australia.
- Biological Research Unit, Racing Analytical Services Limited, Flemington, Victoria, Australia.
- Racing Analytical Services Ltd, Flemington, Victoria, Australia.
- Biological Research Unit, Racing Analytical Services Limited, Flemington, Victoria, Australia.
- Racing Analytical Services Ltd, Flemington, Victoria, Australia.
- Biological Research Unit, Racing Analytical Services Limited, Flemington, Victoria, Australia.
- Racing Analytical Services Ltd, Flemington, Victoria, Australia.
- Biological Research Unit, Racing Analytical Services Limited, Flemington, Victoria, Australia.
- Department of Biochemistry and Molecular Biology, Biomedicine Discovery Institute, Faculty of Medicine, Nursing and Health Sciences, Monash University, Clayton, Victoria, Australia.
- Racing Analytical Services Ltd, Flemington, Victoria, Australia.
- Biological Research Unit, Racing Analytical Services Limited, Flemington, Victoria, Australia.
MeSH Terms
- Animals
- Male
- Female
- Horses
- Dogs
- Gonadotropin-Releasing Hormone
- Mass Spectrometry
- Chromatography, Liquid / methods
- Testosterone
- Substance Abuse Detection / veterinary
- Substance Abuse Detection / methods
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