Detection of glucocorticoid bioactivity in bovine urine samples using a reporter gene assay

The illegal use of anabolic substances in the meat producing industry is an ongoing problem
due to the continual production of new synthetic compounds and/or the practice of lowlevel
cocktail administration to avoid detection by the surveillance schemes of EU member
states National Plan surveillance systems.
We present a highly sensitive reporter gene assay and sample extraction procedure based
on a two step solid phase extraction and high performance liquid chromatography, developed
for the detection of glucocorticoid abuse in bovine urine. The assay is capable of
detecting compounds with glucocorticoid bioactivity and is extremely sensitive with an
EC50 of 0.79 ngmL−1 for dexamethasone. New or unknown compounds with glucocorticoid
bioactivity and low-level cocktail mixtures are detectable by this assay.
Cross-reactivity data for a range of 11 -hydroxyglucocorticoids has been provided. This
assay shows low interference from the 11-keto prohormones and other steroidal hormones.
The assay may be suitable for application in other matrices such as hair. In conclusion this
screening assay offers advantages over existing analytical techniques.