title: Detection of glucocorticoid bioactivity in bovine urine samples using a reporter gene assay.
abstract: 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 low-level 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 EC(50) 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 11beta-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.
authors.affiliation: The Institute of Agri-Food and Land Use, School of Biological Sciences, Queens University Belfast, Belfast BT9 5AG, Northern Ireland, UK.
authors.forename: Lisa
authors.initials: L
authors.lastname: Connolly
authors.forename: Kai
authors.initials: K
authors.lastname: Cai
authors.forename: Edwige
authors.initials: E
authors.lastname: Van der Heiden
authors.forename: Marie-Louise
authors.initials: ML
authors.lastname: Scippo
authors.forename: Marc
authors.initials: M
authors.lastname: Muller
authors.forename: Jonathan
authors.initials: J
authors.lastname: Tarbin
authors.forename: Chris
authors.initials: C
authors.lastname: Elliott
mesh_terms: D045930:Anabolic Agents / Q000302:isolation & purification / Q000652:urine*; D000818:Animals; D001681:Biological Assay / Q000379:methods*; D002417:Cattle; D045744:Cell Line, Tumor; D002851:Chromatography, High Pressure Liquid; D017930:Genes, Reporter; D005938:Glucocorticoids / Q000302:isolation & purification / Q000652:urine*; D006801:Humans; D015813:Substance Abuse Detection / Q000379:methods*
chemical_list: D045930:Anabolic Agents; D005938:Glucocorticoids