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Home / Equine Research Bank / Anal Bioanal Chem / A sensitive HPLC-MS/MS method for the detection, resolution ...
Analytical and bioanalytical chemistry2021; 413(8); 2147-2161; doi: 10.1007/s00216-021-03182-1

A sensitive HPLC-MS/MS method for the detection, resolution and quantitation of cathinone enantiomers in horse blood plasma and urine.

Abstract: Resolution of cathinone enantiomers in equine anti-doping analysis is becoming more important to distinguish the inadvertent ingestion of plant-based products from those of deliberate administration of designer synthetic analogs. With this in mind, a rapid and sensitive method was developed and validated for the detection, resolution and quantitative determination of cathinone enantiomers in horse blood plasma and urine. The analytes were recovered from the blood plasma and urine matrices by using a liquid-liquid extraction after adjusting the pH to 9. The recovered analytes were derivatized with N-(2,4-dinitro-5-fluorophenyl)-L-valinamide, a chiral derivatizing agent analogous to Marfey's reagent. The resulting diastereoisomers were baseline resolved under a reversed-phase liquid chromatographic condition. Derivatization of the analytes not only allowed the separation of the enantiomers using cost-effective traditional liquid chromatography conditions and reversed-phase columns but also increased the sensitivity, at least to an order of magnitude, when tandem mass spectrometry is used for the detection. A limit of detection of 0.05 ng/mL was achieved for cathinone enantiomers for both matrices. Acceptable intraday and interday precision and accuracy along with satisfactory dilution accuracy and precision were observed during the method validation. The method suitability was tested using the post administration urine samples collected after single doses of cathinone and ephedrine as single-enantiomeric form and methcathinone as racemic form. Finally, a proof of concept of the isomeric ratio in urine samples to distinguish the presence of cathinone as a result of accidental ingestion of plant-based product from that of an illicit use of a designer product is demonstrated. To the best of our knowledge, this is the first such work where cathinone enantiomers were resolved and quantified in horse blood plasma and urine at sub nanogram levels.
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Publication Date: 2021-01-31 PubMed ID: 33517480DOI: 10.1007/s00216-021-03182-1Google Scholar: Lookup
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  • Journal Article
  • Validation Study

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research developed and validated a rapid, highly sensitive method for detecting and quantifying specific substances (cathinone enantiomers) in horse blood and urine. This can help distinguish between accidental ingestion of certain plants and deliberate use of synthetic drugs.

Objective of the Research

  • The team aimed to devise a method for identifying and quantifying cathinone enantiomers in horse blood and urine.
  • They sought to differentiate between the unintentional consumption of cathinone-containing plants and the intentional administration of synthetic designer drugs in horses.

Methodology

  • The researchers recovered the analytes from the horse’s blood plasma and urine by carrying out a liquid-liquid extraction, after adjusting the pH to 9.
  • The recovered analytes were then chemically modified with N-(2,4-dinitro-5-fluorophenyl)-L-valinamide, a chiral derivatizing agent analogous to Marfey’s reagent.
  • Resulting diastereoisomers were then differentiated under specific liquid chromatographic conditions.
  • Through this method, the team was able to identify enantiomers, not just by using cost-effective traditional liquid chromatography methods and reversed-phase columns, but also by enhancing the sensitivity using tandem mass spectrometry for detection.

Results

  • The developed method succeeded in achieving limit of detection (LOD) of 0.05 ng/mL for cathinone enantiomers in horse blood and urine.
  • The process displayed acceptable intra-day and inter-day precision and accuracy, along with satisfactory dilution accuracy and precision.
  • The method’s appropriateness was tested using urine samples obtained post administration of cathinone and ephedrine (single-enantiomeric form) and methcathinone (racemic form).
  • The research also demonstrated a proof of concept of the isomeric ratio in urine samples, which can be used to distinguish between accidental ingestion of a plant-based product and illegal utilization of a designer product.

Significance and Novelty

  • This study is reported to be the first to involve resolution and quantification of cathinone enantiomers in horse blood plasma and urine at such minute levels.
  • The method developed can potentially be used as an effective and highly sensitive tool in equine anti-doping analysis.

Cite This Article

APA
Loganathan D, Yi R, Patel B, Zhang J, Kong N. (2021). A sensitive HPLC-MS/MS method for the detection, resolution and quantitation of cathinone enantiomers in horse blood plasma and urine. Anal Bioanal Chem, 413(8), 2147-2161. https://doi.org/10.1007/s00216-021-03182-1

Publication

Analytical and bioanalytical chemistry
ISSN: 1618-2650
NlmUniqueID: 101134327
Country: Germany
Language: English
Volume: 413
Issue: 8
Pages: 2147-2161

Researcher Affiliations

Loganathan, Devan
  • Forensic Equine Drug Testing, Bureau Veritas Laboratories, 8577 Commerce Court, Burnaby, BC, V5A 4N5, Canada. devanesan.loganathan@bureauveritas.com.
Yi, Rong
  • Forensic Equine Drug Testing, Bureau Veritas Laboratories, 8577 Commerce Court, Burnaby, BC, V5A 4N5, Canada.
Patel, Bhavesh
  • Forensic Equine Drug Testing, Bureau Veritas Laboratories, 8577 Commerce Court, Burnaby, BC, V5A 4N5, Canada.
Zhang, Julia
  • Forensic Equine Drug Testing, Bureau Veritas Laboratories, 8577 Commerce Court, Burnaby, BC, V5A 4N5, Canada.
Kong, Noel
  • Forensic Equine Drug Testing, Bureau Veritas Laboratories, 8577 Commerce Court, Burnaby, BC, V5A 4N5, Canada.

MeSH Terms

  • Alkaloids / analysis
  • Alkaloids / blood
  • Alkaloids / urine
  • Animals
  • Central Nervous System Stimulants / analysis
  • Central Nervous System Stimulants / blood
  • Central Nervous System Stimulants / urine
  • Chromatography, High Pressure Liquid / methods
  • Doping in Sports
  • Horses / blood
  • Horses / urine
  • Limit of Detection
  • Stereoisomerism
  • Substance Abuse Detection / methods
  • Tandem Mass Spectrometry / methods

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Citations

This article has been cited 3 times.
  1. Boroujerdi R, Paul R. Graphene-Based Electrochemical Sensors for Psychoactive Drugs.. Nanomaterials (Basel) 2022 Jun 30;12(13).
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