Stéphanie Prévost

Determination of a Large Set of β-Adrenergic Agonists in Animal Matrices Based on Ion Mobility and Mass Separations

While the coupling of traveling wave ion mobility spectrometry (TWIMS) and mass spectrometry is mainly reported for structural purposes, we studied its potential in enhancing compounds analysis such as growth promoters used in livestock animals at trace concentrations. β-Adrenergic agonists have been selected as model compounds since they exhibit a range of close physicochemical properties leading to analytical issues using classical approaches. In this paper, the potential of Synapt G2-S (Q-TWIM-TOF MS) has been investigated for sensitive and specific detection of a range of these synthetic phenethanolamines in various complex biological matrices (retina, meat, and urine) from bovine considered as relevant in the context of detecting β-adrenergic agonists use in animals. In particular, the specificity of the additional information provided by the TWIMS (i.e., collision cross section) together with the interest of the extra dimension of separation is discussed.

Use of isotope ratio mass spectrometry to differentiate between endogenous steroids and synthetic homologues in cattle: a review.

Although substantial technical advances have been achieved during the past decades to extend and facilitate the analysis of growth promoters in cattle, the detection of abuse of synthetic analogs of naturally occurring hormones has remained a challenging issue. When it became clear that the exogenous origin of steroid hormones could be traced based on the (13)C/(12)C isotope ratio of the substances, GC/C/IRMS has been successfully implemented to this aim since the end of the past century. However, due to the costly character of the instrumental setup, the susceptibility of the equipment to errors and the complex and time consuming sample preparation, this method is up until now only applied by a limited number of laboratories. In this review, the general principles as well as the practical application of GC/C/IRMS to differentiate between endogenous steroids and exogenously synthesized homologous compounds in cattle will be discussed in detail, and will be placed next to other existing and to be developed methods based on isotope ratio mass spectrometry. Finally, the link will be made with the field of sports doping, where GC/C/IRMS has been established within the World Anti-Doping Agency (WADA) approved methods as the official technique to differentiate between exogenous and endogenous steroids over the past few years.

Toward a criterion for suspect thiouracil administration in animal husbandry

Thyreostats are growth-promoters banned in Europe since 1981. The identification of thiouracil (TU) in animal biological matrices can, however, no longer be systematically interpreted as a consequence of illegal administration. Indeed, some experimental results have indicated a causal link between cruciferous-based diet and the presence of TU in urine of bovines. The present study aims at investigating, on a large scale (n > 1300), the natural occurrence of thiouracil in urine samples collected from different animal species. TU was identified in main breeding animal species: bovine, porcine and ovine. The natural distribution of TU allowed proposing threshold values to differentiate compliant from suspect urine samples. Suggested values are 5.7 and 9.1 µg l−1 in male adult bovines (6–24 months), 3.1 and 8.1 µg l−1 in female adult bovines (6–24 months), 7.3 and 17.7 µg l−1 in calves (<6 months), 3.9 and 8.8 µg l−1 in female bovines (>24 months), and 2.9 and 4.1 µg l−1 in porcines at a 95 and 99% confidence level, respectively.

Selective androgen receptor modulators: comparative excretion study of bicalutamide in bovine urine and faeces: Bicalutamide excretion in bovine urine and feces

Besides their development for therapeutic purposes, non-steroidal selective androgen receptor modulators (non-steroidal SARMs) are also known to impact growth-associated pathways as ligands of androgenic receptors (AR). They present a potential for abuse in sports and food-producing animals as an interesting alternative to anabolic androgenic steroids (AAS). These compounds are easily available and could therefore be (mis)used in livestock production as growth promoters. To prevent such practices, dedicated analytical strategies should be developed for specific and sensitive detection of these compounds in biological matrices. The present study focused on Bicalutamide, a non-steroidal SARM used in human treatment of non-metastatic prostate cancer because of its anti-androgenic activity exhibiting no anti-anabolic effects. To select the most appropriate matrix to be used for control purposes, different animal matrices (urine and faeces) have been investigated and SARM metabolism studied to highlight relevant metabolites of such treatments and establish associated detection time windows. The aim of this work was thus to compare the urinary and faecal eliminations of bicalutamide in a calf, and investigate phase I and II metabolites. The results in both matrices showed that bicalutamide was very rapidly and mainly excreted under its free form. The concentration levels were observed as higher in faeces (ppm) than urine (ppb); although both matrices were assessed as suitable for residue control. The metabolites found were consistent with hydroxylation (phase I reaction) combined or not with glucuronidation and sulfation (phase II reactions). Copyright

Analytical strategies to detect enobosarm administration in bovines

ABSTRACT Selective androgen receptor modulators (SARMs) are a novel class of androgen receptor ligands. They are intended to exhibit the same kind of effects as androgenic drugs, like anabolic steroids, but be much more selective in their action, targeting particular tissues without any undesirable effects on others. While the main applications of these synthetic substances are for therapeutic purposes, they also have a high potential for misuse in veterinary practice and the sporting world. In order to guarantee for consumers with food from animal origin that it is free from any residues of such compounds, analytical strategies are required to ensure safe food and also to enable fair trade between producers. In this context an animal experiment involving bovines administered with enobosarm was conducted to provide the study with biological matrices. Different animal matrices (urine and faeces) were investigated to select the most appropriate matrix for use for control purposes, in terms of metabolite relevance and detection time window. Based on ultra-high-pressure liquid chromatography (UHPLC) with tandem mass spectrometry (LC-MS/MS) this work highlighted the presence of sulfonated and glucuronated-conjugated forms of the molecule in the urine of treated animals. Enobosarm could be detected in urine up to 9 days after the administration when samples underwent phase II hydrolysis. Faeces was demonstrated to be the main matrix of excretion of enobosarm since values up to 500 times higher compared with urine could be detected for 21 days. There was no difference between the kinetic profiles when a deconjugation step was or was not was applied. GRAPHICAL ABSTRACT

Residues of medroxyprogesterone acetate detected in sows at a slaughterhouse, Madagascar

In Madagascar, little information about drug residues in animal products is available. However, recently, official veterinary services were informed about the misuse of human injectable contraceptives in pig farms as an alternative for chirurgical castration of adult sows before culling. We investigated pigs (n = 80) slaughtered in 7 Malagasy abattoirs and raised in 8 of the 22 Malagasy regions (1) to confirm the contamination of carcasses by anabolic hormones by using LC-MS/MS, (2) to identify the substances of concern and (3) to explore the consumers’ exposure to hormone residues. Medroxyprogesterone acetate was the only synthetic hormone detected in kidney fat. Samples positive with medroxyprogesterone acetate were observed in 66.7% of the districts investigated and in 87.5% of the surveyed regions, confirming its large misuse in livestock. Public awareness campaigns and control improvement among the animal production sector and among the Malagasy public health sector are therefore urgent.