Additional studies on nicotine exposure in horses: Accurate quantification and elimination profiles of potential biomarkers in plasma and urine.
Abstract: For the purpose of doping control, this is the first report of accurate quantification of four critical structural isomers of nicotine metabolites (trans-3'-hydroxycotinine, cis-3'-hydroxycotinine, 5'-hydroxycotinine, and N'-hydroxymethylnorcotinine) in equine plasma and urine for the establishment of their elimination profiles. Besides, the pharmacokinetic studies of trans-3'-hydroxycotinine and N'-hydroxymethylnorcotinine in equine plasma and urine are also presented for the first time. Methods: The accurate quantification methods of the aforementioned four structural isomers in horse plasma and urine were successfully developed and validated using the solid-phase extractions followed by liquid chromatography/tandem mass spectrometry analysis. Baseline chromatographic separation was achieved to completely differentiate these isomers, which shared the same selected reaction monitoring transition. Such methods were applied to post-administration samples obtained from the nicotine and tobacco leaf administration studies for the establishment of pharmacokinetic profiles. Results: N'-Hydroxymethylnorcotinine could be quantified for the longest period, ranging from 48 to 72 h in plasma and 96 h in urine after a single administration of 250 mg of nicotine and an equivalent amount of nicotine in tobacco leaves. In terms of detection, both N'-hydroxymethylnorcotinine and trans-3'-hydroxycotinine could be detected up to the last sample collection time point (96 h), indicating that they are the most appropriate biomarkers for nicotine exposure. Conclusions: N'-Hydroxymethylnorcotinine and trans-3'-hydroxycotinine were detected longest in plasma and urine samples after both nicotine and tobacco leaf administrations, and N'-hydroxymethylnorcotinine was deemed most appropriate as a monitoring target due to its relatively higher abundance and slower elimination rate. These two biomarkers could also be used to differentiate sample contamination by tobacco products and genuine nicotine exposure to horse regardless of intentionality.
© 2022 John Wiley & Sons Ltd.
Publication Date: 2022-09-14 PubMed ID: 36098053DOI: 10.1002/rcm.9396Google Scholar: Lookup
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- Journal Article
Summary
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This study reveals the first report on the accurate measurement of four specific metabolites of nicotine (trans-3′-hydroxycotinine, cis-3′-hydroxycotinine, 5′-hydroxycotinine, and N’-hydroxymethylnorcotinine) in horse plasma and urine for the intention of establishing their elimination profiles. The results will be used in doping control for horses.
Materials and Methods
- A solid-phase extraction follow-up by liquid chromatography/tandem mass spectrometric analysis was employed to conduct an accurate quantification of the four isomers of nicotine’s metabolites in horse plasma and urine.
- To differentiate completely these isomers, which share the same selected reaction monitoring transition, a baseline chromatographic separation was achieved.
- The methods developed in this study were applied to samples taken after nicotine or tobacco leaf administration to set up pharmacokinetic profiles.
Results and Findings
- The metabolite, N’-Hydroxymethylnorcotinine, could be quantified for the most extended period of 48 to 72 hours in plasma and 96 hours in urine after a 250mg administration of nicotine or an equivalent amount from tobacco leaves.
- Both N’-hydroxymethylnorcotinine and trans-3′-hydroxycotinine could be detected up to the final sample collection time point which was 96 hours post-administration, indicating that they are most suitable biomarkers for nicotine exposure.
Conclusion and Implications
- N’-Hydroxymethylnorcotinine and trans-3′-hydroxycotinine were detected longest in the plasma and urine samples after both nicotine and tobacco leaf administrations.
- With a relatively higher abundance and slower elimination rate, N’-hydroxymethylnorcotinine was considered the most suitable biomarker for monitoring.
- These two biomarkers can be applied in distinguishing sample contamination by tobacco products and genuine nicotine exposure to horses, regardless of intentionality.
- The study provides vital insights for doping controls in horse racing and similar sports events, as these biomarkers can help determine whether horses have been exposed to nicotine.
Cite This Article
APA
Ishii H, Shibuya M, Leung GN, Yamashita S, Nagata SI, Kushiro A, Sakai S, Toju K, Okada J, Kawasaki K, Kusano K, Kijima-Suda I.
(2022).
Additional studies on nicotine exposure in horses: Accurate quantification and elimination profiles of potential biomarkers in plasma and urine.
Rapid Commun Mass Spectrom, 36(23), e9396.
https://doi.org/10.1002/rcm.9396 Publication
Researcher Affiliations
- Drug Analysis Department, Laboratory of Racing Chemistry, Utsunomiya, Tochigi, Japan.
- Department of Pharmaceutical Sciences, Tohoku University Hospital, Sendai, Miyagi, Japan.
- Drug Analysis Department, Laboratory of Racing Chemistry, Utsunomiya, Tochigi, Japan.
- Drug Analysis Department, Laboratory of Racing Chemistry, Utsunomiya, Tochigi, Japan.
- Drug Analysis Department, Laboratory of Racing Chemistry, Utsunomiya, Tochigi, Japan.
- Genetic Analysis Department, Laboratory of Racing Chemistry, Utsunomiya, Tochigi, Japan.
- Equine Research Institute, Research Planning & Coordination Division, Japan Racing Association, Shimotsuke, Tochigi, Japan.
- Race Horse Hospital, Miho Training Center, Japan Racing Association, Inashiki-gun, Ibaraki, Japan.
- Race Horse Hospital, Miho Training Center, Japan Racing Association, Inashiki-gun, Ibaraki, Japan.
- Veterinarian Section, Equine Department, Japan Racing Association, Minato-ku, Tokyo, Japan.
- Veterinarian Section, Equine Department, Japan Racing Association, Minato-ku, Tokyo, Japan.
- Race Horse Hospital, Ritto Training Center, Japan Racing Association, Ritto, Shiga, Japan.
- Drug Analysis Department, Laboratory of Racing Chemistry, Utsunomiya, Tochigi, Japan.
MeSH Terms
- Horses
- Animals
- Nicotine / metabolism
- Chromatography, Liquid / methods
- Mass Spectrometry
- Solid Phase Extraction
- Biomarkers
Grant Funding
- 2018 / Japan Racing Association
References
This article includes 18 references
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