Title
Detection of hormonal anabolic compounds in calf urine and unverified growth-promoting preparations: Application of the AR-LUX bioassay for screening and determination of androgenic activity
Author
Blankvoort, B.M.G.
Aarts, J.M.M.J.G.
Schilt, R.
Geerdink, P.
Spenkelink, B.
Rodenburg, R.J.T.
Centraal Instituut voor Voedingsonderzoek TNO
Publication year
2003
Abstract
Despite a ban by the European Union, the use of anabolic steroids and repartitioning agents in cattle is still occasionally observed. Due to continuing improvements in analytical techniques, very low detection limits for individual compounds have been achieved. In response to these developments, cocktails composed of several steroids have been applied, thus hampering detection due to lower levels of the individual compounds. Bioassays capable of measuring the integrated effect of cocktails might therefore provide valuable additional tools in controlling the use of illegal anabolics. We investigated the feasibility of using the AR-LUX assay to detect the presence in cattle urine of growth promoters that exert their effects via androgen response elements (AREs). The AR-LUX assay is based on a human cell line featuring a luciferase reporter gene under transcriptional control of an authenticated ARE. Several column purification and liquid/liquid extraction methods were investigated to optimize the efficiency of anabolic compounds extraction and minimize cytotoxic effects of the urine matrix. The AR-LUX assay was found to be applicable to the detection of anabolic steroids excreted in urine samples with a discriminatory power similar to that of GC-MS analysis. Finally, some liquid products probably destined for growth-promoting purposes confiscated outside the Netherlands were analyzed. Although common chemical-analytical methods did not detect any anabolic steroids in these samples, the presence of compounds activating ARE-mediated gene expression was clearly established.
Subject
Biology
Analytical research
Anabolic agent
Androgen
Boldenone
Drug residue
Luciferase
Androgen receptor mediated luciferase expression system
Animal
Animal experiment
Bioassay
Body growth
Cattle
Cell line
Conference paper
Controlled study
Cytotoxicity
Drug determination
Enzyme assay
European Union
Female
Food contamination
Gene expression
Human
Human cell
Liquid liquid extraction
Male
Methodology
Nonhuman
Reporter gene
Tumor cell line
Urine
Urine level
Anabolic Agents
Animals
Biological Assay
Cattle
Cell Line, Tumor
Drug Residues
Female
Food Contamination
Humans
Luciferases
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http://resolver.tudelft.nl/uuid:64aee72f-7c30-47f0-ac7f-6d62a7b16f37
DOI
https://doi.org/10.1039/b309642d
TNO identifier
42740
ISSN
0003-2654
Source
Analyst, 128, 1373-1381
Bibliographical note
Correspondence Address: Aarts, J.M.M.J.G.; Dept. Agrotechnology and Food Sci., Sub-department of Toxicology, Wageningen University, Wageningen, Netherlands Chemicals/CAS: boldenone, 846-48-0; Anabolic Agents; Luciferases, EC 1.13.12.-
Document type
article