Identification of hormone esters in injection site in muscle tissues by LC/MS/MS

Abstract

The detection of hormone abuse for growth promotion in food animal production is a global concern. Initial testing for hormones in Canada was directed at the compounds approved for use in beef cattle, melengestrol acetate, trenbolone acetate and zeranol, and the banned compound diethylstilbestrol (DES). No hormonal growth promoters are approved for use in veal production in Canada. However, instances of use of trenbolone and clenbuterol were detected in Canada in the 1990s. During the development of a new analytical method for testosterone and progesterone, there were reports of suspicious injection sites being found in veal calves. Upon implementation of the method, analysis of investigative samples revealed significant residues of testosterone in some injection sites. To prove that the source of these residues was exogenous, a fully validated method for hormone esters was developed to confirm the presence of exogenous hormones in these injection sites. The QUECHERS model was employed in methods development and resulted in a simple, effective extraction technique that consisted of sample pre-homogenization, liquid/liquid partitioning, extract dilution, filtration and use of LC/MS/MS to provide detection selectivity. The result was an adaptable MS/MS confirmation technique that meets the needs of Canadian regulatory authorities to confirm the misuse of injectable testosterone, and potentially other hormones, in food animal production.

Quantification of steroids and endocrine disrupting chemicals in rat ovaries by LC-MS/MS for reproductive toxicology assessment

“…Therefore, it can now be successfully applied to animal tissues [19]. However, while the "QuEChERS" sample preparation has been extensively developed for chemical residue analysis, it was rarely used for the extraction of hormones [20].…”

Rapid confirmatory method for the determination of sixteen synthetic growth promoters and bisphenol A in bovine milk using dispersive solid-phase extraction and liquid chromatography–tandem mass spectrometry

“…For control at retail level, as well as for imported products however only edible matrices are available for testing such as muscle, fat and milk. There are quite a number of methods detailed in the literature for the analysis of these types of substances in muscle and fat [6][7][8][9][10][11][12][13][14][15][16][17][18][19]. However, as pointed out by Noppe et al in 2008, it was surprising that very few papers have been published within the framework of residue analysis that address the determination of steroid hormones in milk [20].…”

Confirmation and 3D profiling of anabolic steroid esters in injection sites using imaging desorption electrospray ionisation (DESI) mass spectrometry

“…Because of the very high concentrations of forbidden substances in injection sites and preparations seized at the farm, a simple extraction followed by proton NMR ( 1 H-NMR) analysis is feasible. Based on NMR or highresolution MS results, it can be decided which further sample preparation and LC-MS or GC-MS analysis steps are needed (Nielen et al 2001;Lommen et al 2002;De Wasch et al 2003;Costain et al 2008). NMR analysis is generally considered as expensive, and sample pre-treatment is needed.…”