Confirmatory analysis of steroids in muscle using liquid chromatography–tandem mass spectrometry

Abstract

A method is described for screening and confirmation of synthetic and endogenous steroids in muscle tissue. The method is sensitive, selective, and rapid and the consumption of organic solvents is low, compared to previously published methods. The procedure involves hydrolysis, defattening with heptane and final clean up with SPE using C₁₈ cartridge. After filtration, the analytes are analysed by LC/MS/MS and quantification is performed using deuterated internal standards. Decision limits (CCα) varied from 0.02 to 0.33 µg kg⁻¹ and the detection capabilities (CCβ) were <0.50 µg kg⁻¹. The mean within-laboratory reproducibility ranged 5–22% (%RSD_IR). Endogenous steroids (e.g. testosterone, epitestosterone and androstenedione) have been included in the method, to provide an insight into their levels, as the presence of these steroids was detected several times during analysis of imported meat.

Keywords:

• chromatography, liquid chromatography/mass spectrometry
• method validation
• veterinary drug residues, anabolic steroids
• animal products, meat

Acknowledgements

We thank Lis Abildgaard Andersen, Maud Bering Andersen and Lene Gram Hansen for their skilful technical assistance with chemical analyses.