Detection of testosterone propionate administration in horse hair samples.
Abstract: A sensitive and specific method has been developed to detect semi-quantitatively testosterone in horse hair samples. The method involved a washing step with sodium dodecylsulfate aqueous solution. The mane and tail hair samples (100mg) were dissolved in 1 mL of sodium hydroxide for 15 min at 95 degrees C in the presence of d3-boldenone used as internal standard. The next three steps involved diethyl ether extraction and a solid phase extraction on Isolute C18 (EC) cartridges eluted with methanol. The residue was derivatized by adding 100 microL of acetonitrile and 30 microL of PFPA then incubating for 15 min at 60 degrees C. After evaporation, 30 microL of hexane was added and 2.5 microL was injected into the column (a bonded phase fused silica capillary column DB5MS, 30 m x 0.25 mm i.d. x 0.25 microm film thickness) of a Trace GC chromatograph. In order to improve the sensitivity of the method, damping gas flow has been optimized. Testosterone was identified in MS(2) full scan mode on the Polaris Q instrument. The assay was capable of detecting less than 1 pg mg(-1). The recovery was close to 90%. The analysis of tail and mane samples collected from a gelding horse having received a single dose of testosterone propionate (1 mg kg(-1)) showed the presence of testosterone in the range of 1-6 pg mg(-1) in hair collected during 5 months after administration.
Publication Date: 2007-03-14 PubMed ID: 17383946DOI: 10.1016/j.jchromb.2007.02.046Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research presents a new method to semi-quantitatively detect testosterone presence in horse hair samples. The technique has high sensitivity and specificity, with an ability to detect less than 1 pg mg(-1) and a recovery rate near 90%.
Methodology
- The process begins with a washing step using a sodium dodecylsulfate aqueous solution. This helps in cleaning the hair samples.
- The mane and tail hair samples, weighing approximately 100mg each, are then dissolved in 1 mL of sodium hydroxide. The samples are treated for 15 minutes at a temperature of 95 degrees Celsius. This step uses an internal standard d3-boldenone.
- Following the dissolution, the sample undergoes diethyl ether extraction and a solid phase extraction on Isolute C18 (EC) cartridges, eluted with methanol. This step is aimed at the extraction of any remaining impurities from the sample.
- The residuary sample is then derivatized with the addition of 100 microL of acetonitrile and 30 microL of PFPA. This is incubated for 15 min at 60 degrees C. The primary goal here is to separate the ions of interest from the rest.
- After evaporation, 30 microL of hexane is added and a small volume (2.5 microL) is injected into the column of a Trace GC chromatograph. The column has specific dimensions and the chromatograph helps in the separation of components of the sample.
- The detection of testosterone takes place through MS(2) full scan mode on the Polaris Q instrument, improving the sensitivity of the method.
Results
- The methodology was found to be highly sensitive, capable of detecting testosterone levels below 1 pg mg(-1).
- It had a high recovery rate, close to 90%.
- The technique was tested on real-world samples – mane and tail hair from a gelding horse that had been given a single dose of testosterone propionate. The presence of testosterone was detected in the range of 1-6 pg mg(-1) in hair collected over the course of five months post-administration.
Conclusion
- This proposed method provides an efficient way to detect testosterone in horse hair, potentially serving as a detection tool for illicit use of performance-enhancing drugs in equine sports.
Cite This Article
APA
Boyer S, Garcia P, Popot MA, Steiner V, Lesieur M.
(2007).
Detection of testosterone propionate administration in horse hair samples.
J Chromatogr B Analyt Technol Biomed Life Sci, 852(1-2), 684-688.
https://doi.org/10.1016/j.jchromb.2007.02.046 Publication
Researcher Affiliations
- LCH 15 rue de Paradis, 91370 Verrières le Buisson, France.
MeSH Terms
- Animals
- Doping in Sports
- Gas Chromatography-Mass Spectrometry / methods
- Hair / chemistry
- Horses
- Substance Abuse Detection / methods
- Testosterone Propionate / administration & dosage
- Testosterone Propionate / analysis
Citations
This article has been cited 4 times.- Medill SA, Janz DM, McLoughlin PD. Hair Cortisol Concentrations in Feral Horses and the Influence of Physiological and Social Factors. Animals (Basel) 2023 Jun 27;13(13).
- Madry MM, Spycher BS, Kupper J, Fuerst A, Baumgartner MR, Kraemer T, Naegeli H. Long-term monitoring of opioid, sedative and anti-inflammatory drugs in horse hair using a selective and sensitive LC-MS/MS procedure. BMC Vet Res 2016 Jun 1;12:84.
- Keckeis K, Lepschy M, Schöpper H, Moser L, Troxler J, Palme R. Hair cortisol: a parameter of chronic stress? Insights from a radiometabolism study in guinea pigs. J Comp Physiol B 2012 Oct;182(7):985-96.
- Shah I, Hakeem MK, Alraeesi A, Barker J. Innovative Detection of Testosterone Esters in Camel Hair: Unravelling the Mysteries of Dromedary Endocrinology. Molecules 2023 Dec 22;29(1).
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