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 17β-estradiol to bovine has been investigated in this paper. By comparison of 13C/12C isotopic ratio of main urinary estradiol metabolite, i.e. 17α-estradiol, with two endogenous reference compounds (ERCs), i.e. dehydroepiandrosterone (DHEA) and 5-androstene-3β,17α-diol, the differentiation of estradiol metabolite origin, either endogenous or exogenous, has been proved to be achievable. After treatment, the δ13CVPDB-values of 17α-estradiol reached −27‰ to −29‰, whereas δ13CVPDB-values of DHEA remained between −13‰ and −20‰ depending on the diet, maize and grass, respectively. A significant difference of δ13CVPDB between ERCs and 17α-estradiol was measurable over a period of 2 weeks after estradiol ester administration to the animal.