Determination of anabolic steroids in dried blood using microsampling and gas chromatography-tandem mass spectrometry: Application to a testosterone gel administration study

Determination of anabolic steroids in dried blood using microsampling and gas chromatography-tandem mass spectrometry: Application to a testosterone gel administration study / William Chih-Wei Chang, David A. Cowan, Christopher J. Walker, Nick Wojek, Alan D. Brailsford. - (Journal of Chromatography A 1628 (2020) 461445 (27 September)

  • PMID: 32822984
  • DOI: 10.1016/j.chroma.2020.461445

Abstract

Anabolic androgenic steroids (AAS) have been the most commonly abused substances taken by not only professional sportsmen but also recreational bodybuilders. The detection of micro-dose testosterone (T) misuse is particularly challenging as it possesses pseudo-endogenous origin and is sometimes impossible to be identified in urine samples. Dried blood (DB) obtained by finger pricking has been proven to be an alternative matrix for better correlating to physiological responses. Moreover, the introduction of the volumetric absorptive microsampling (VAMS) technology allows overcoming some major limitations of spotting blood onto a filter paper card. In this work, a fast and sensitive GC-MS/MS method was developed and validated for the quantification of AAS in DB collected by means of VAMS. T and the eight top abused synthetic AAS, namely nandrolone, boldenone, mesterolone, drostanolone, metenolone, metandienone, oxandrolone, and dehydrochloromethyl T were selected as the target analytes. The method based on VAMS exhibited good precision, accuracy as well as stability, and superior extraction recoveries over the punched DB spots reported in the literature. The chromatographic separation was achieved within 6.4 min and the detection limit is as little as 50 fg (i.e. able to detect 0.10 ng mL-1 in 20 μL of DB). Confirmed by forty real blood samples, the Deming regression and Bland-Altman analysis revealed that the VAMS DB could be employed for quantifying blood T level in agreement with using the serum specimen. The feasibility of the method was then successfully proven by the analysis of samples collected from a three-arm T administration trial. Our results highlighted that DB total T was a sensitive indicator for identifying transdermal micro-dosing of T. In the groups of receiving T gel administration, T concentrations could rise up to ten times higher than the baseline at 9 h after the application. As a future step, this approach is being expanded to a large cohort screening of bodybuilders at gym and ultimately may allow universal applications on monitoring sports drug misuse.

Parameters

Science
Study
Date
1 August 2020
People
Brailsford, Alan D.
Chang, William Chih-Wei
Cowan, David A.
Walker, Christopher J.
Wojek, Nick
Country
United Kingdom
Language
English
Other organisations
UK Anti-Doping (UKAD)
Laboratories
London, United Kingdom: Drug Control Centre
Analytical aspects
Blood testing method
Mass spectrometry analysis
Testing method development
Doping classes
S1. Anabolic Agents
Substances
Testosterone
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Abstract
Date generated
2 September 2020
Date of last modification
16 September 2020
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  • Legal Source
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  • Statistics
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  • ADRV
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  • Sport/IFs
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  • Laboratories
  • Analytical aspects
  • Doping classes
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  • Various
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