title: The application of reporter gene assays for the detection of endocrine disruptors in sport supplements.
abstract: The increasing availability and use of sports supplements is of concern as highlighted by a number of studies reporting endocrine disruptor contamination in such products. The health food supplement market, including sport supplements, is growing across the Developed World. Therefore, the need to ensure the quality and safety of sport supplements for the consumer is essential. The development and validation of two reporter gene assays coupled with solid phase sample preparation enabling the detection of estrogenic and androgenic constituents in sport supplements is reported. Both assays were shown to be of high sensitivity with the estrogen and androgen reporter gene assays having an EC(50) of 0.01 ng mL(-1) and 0.16 ng mL(-1) respectively. The developed assays were applied in a survey of 63 sport supplements samples obtained across the Island of Ireland with an additional seven reference samples previously investigated using LC-MS/MS. Androgen and estrogen bio-activity was found in 71% of the investigated samples. Bio-activity profiling was further broken down into agonists, partial agonists and antagonists. Supplements (13) with the strongest estrogenic bio-activity were chosen for further investigation. LC-MS/MS analysis of these samples determined the presence of phytoestrogens in seven of them. Supplements (38) with androgen bio-activity were also selected for further investigation. Androgen agonist bio-activity was detected in 12 supplements, antagonistic bio-activity was detected in 16 and partial antagonistic bio-activity was detected in 10. A further group of supplements (7) did not present androgenic bio-activity when tested alone but enhanced the androgenic agonist bio-activity of dihydrotestosterone when combined. The developed assays offer advantages in detection of known, unknown and low-level mixtures of endocrine disruptors over existing analytical screening techniques. For the detection and identification of constituent hormonally active compounds the combination of biological and physio-chemical techniques is optimal.
authors: Institute of Agri-Food and Land Use, School of Biological Sciences, Queen's University Belfast, Belfast, Northern Ireland, United Kingdom.
authors: Monika
authors: M
authors: Plotan
authors: Christopher T
authors: CT
authors: Elliott
authors: Marie Louise
authors: ML
authors: Scippo
authors: Marc
authors: M
authors: Muller
authors: Jean-Philippe
authors: JP
authors: Antignac
authors: Edward
authors: E
authors: Malone
authors: Toine F H
authors: TF
authors: Bovee
authors: Samuel
authors: S
authors: Mitchell
authors: Lisa
authors: L
authors: Connolly
mesh_terms: D000726:Androgen Antagonists / Q000032:analysis / Q000302:isolation & purification; D000728:Androgens / Q000819:agonists / Q000032:analysis / Q000302:isolation & purification; D001681:Biological Assay / Q000379:methods*; D002460:Cell Line; D019587:Dietary Supplements / Q000032:analysis*; D052244:Endocrine Disruptors / Q000032:analysis* / Q000302:isolation & purification; D004965:Estrogen Antagonists / Q000032:analysis / Q000302:isolation & purification; D020847:Estrogen Receptor Modulators / Q000032:analysis / Q000302:isolation & purification; D017930:Genes, Reporter; D006801:Humans; D011944:Receptors, Androgen / Q000235:genetics / Q000378:metabolism; D011960:Receptors, Estrogen / Q000235:genetics / Q000378:metabolism; D052616:Solid Phase Extraction / Q000379:methods
chemical_list: D000726:Androgen Antagonists; D000728:Androgens; D052244:Endocrine Disruptors; D004965:Estrogen Antagonists; D020847:Estrogen Receptor Modulators; D011944:Receptors, Androgen; D011960:Receptors, Estrogen