Application of stable carbon isotope analysis to the detection of 17β-estradiol administration to cattle

The use of anabolic agents in food producing animals is prohibited within the EU since 1988 (96/22/EC directive). The control of the illegal use of natural steroid hormones in cattle is still an exciting analytical challenge as far as no definitive method and non-ambiguous analytical criteria are available. The ability of gas chromatography/combustion/isotope ratio mass spectrometry (GC/C/IRMS) to demonstrate the administration of 17 beta-estradiol to bovine has been investigated in this paper. By comparison of C-13/C-12 isotopic ratio of main urinary estradiol metabolite, i.e. 17 alpha-estradiol, with two endogenous reference compounds (ERCs), i.e. dehydroepiandrosterone (DHEA) and 5-androstene-3 beta,17 alpha-diol, the differentiation of estradiol metabolite origin, either endogenous or exogenous, has been proved to be achievable. After treatment, the delta C-31(VPDB)-values of 17 alpha-estradiol reached -27 parts per thousand to -29 parts per thousand, whereas delta C-13(VPDB)-values of DHEA remained between -13 parts per thousand and -20 parts per thousand depending on the diet, maize and grass, respectively. A significant difference of delta C-13(VPDB) between ERCs and 17 alpha-estradiol was measurable over a period of 2 weeks after estradiol ester administration to the animal. (c) 2005 Elsevier B.V. All rights reserved.