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Área de trabajo: Control de Dopaje.

Título: Análisis sensitivo y robusto de esteroides anabólicos, ésteres de esteroides y otras sustancias prohibidas en plasma equino.

Título original: Sensitive and robust analysis of anabolic steroids, steroid esters and other banned substances in equine plasma.

Autor: Bob Gray, Eleanor Menzies,James Scarth, Sport and Specialized Analytical Services, LGC Ltd, Fordham, UK

 

Introduction

Anabolic steroids and related substances, such as anabolic steroid esters, are important compounds for equine doping control laboratories. Traditionally, the detection of these compounds has been undertaken by the analysis of urine samples using gas chromatography  of anabolic steroids, steroid esters and other banned substances in equine plasma . Recently there has been a move towards alternative sample matrices, such as blood and hair, and this has coincided with a shift towards the increased use of liquid chromatography mass spectrometry (LC-MS) as an analytical technique. LC-MS offers significant advantages in sensitivity and throughput. Methods for the detection of these banned substances necessitate excellent sensitivity to detect the very low concentrations of drug present and robustness to ensure the successful analysis of a high number of samples in short timeframes.

 

Experimental

Sample Preparation

Equine plasma samples were augmented with an internal standard mix (testosterone-D3, testosterone propionate-D3, testosterone phenylpropionate-D3, and testosterone decanoate-D3), pH adjusted with 0.1 M NaOH and extracted with 4 mL of methyl tert-butyl ether/ ethyl acetate (1:1). The organic layer was evaporated, and the residue was reconstituted in a 100 mM solution of methoxyamine hydrochloride in methanol/water (80:20). The sample was heated to 100 °C for 60 minutes to form methyloxime derivatives. A 10 µL aliquot of derivatized sample was analyzed by LC-MS.

 

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