Generation and processing of urinary and plasmatic metabolomic fingerprints to reveal an illegal administration of recombinant equine growth hormone from LC-HRMS measurements

Abstract

Growth hormones are proteins produced by the anterior pituitary gland responsible for bone and tissue growth through their effects on carbohydrates, lipids and proteins metabolisms. Despite strict regulations banning the use of recombinant equine growth hormone, this substance is suspected to be misused to improve the horse physical performances. In order to check whether the regulation is fulfilled or not, controls are organized and a new analytical screening tool potentially able to detect such abuse was investigated in this paper. An untargeted metabolomics approach, based on liquid chromatography coupled to high resolution mass spectrometry, was developed and applied to characterize and compare horse urinary and plasmatic metabolic profiles upon reGH administrations. After minimal sample preparation, biological fluids were analyzed by LC-ESI(±)-Q-TOF. Data processing was performed by XCMS software and multivariate data analysis applied to the generated data set allowed building OPLS models to discriminate control versus treated populations. Results demonstrated significant metabolic modifications consecutively to the reGH treatment. A comparative study between urinary and plasmatic signatures was performed to evaluate the resulting metabolomic models and to asses their respective interests in the scope of real application for screening reGH administration.

Challenges in detecting substances for equine anti‐doping

“…[131] Using chemometric models, modifications of horse metabolism upon reGH administration were highlighted thanks to fingerprinting analyses acquired in both ionization modes. Plasma proved to be interesting for the prediction of unknown sample and urine more promising for long term detection.…”

Natural Hormones in Food-Producing Animals:Legal Measurements and Analytical Implications

“…Metabolomics is based on detecting small molecules and excluding big biopolymers such as proteins, generating this way a large set of descriptors characteristic of the biological matrix under investigation in different experimental groups. Metabolomic approaches have already shown to be affective in relation to different biochemical processes, such as drug toxicity and diseases (Brindle et al, 2002;Coen et al, 2003;Lindon et al, 2004;Vallejo et al, 2009), prediction of gender (Lutz et al, 2006), nutritional effects studies (Wang et al, 2005) and even doping control in horses (Kieken et al, 2011) or the clenbuterol use in cattle (Courant et al, 2009). Basically, the general principle of metabolomic studies is the characterization of the biological system or sample in question through the generation of metabolomic fingerprints.…”

Structure elucidation and HPLC-MS/MS determination of a potential biomarker for estradiol administration in cattle

“…Metabolomics is based on detecting small molecules and excluding big biopolymers such as proteins, generating this way a large set of descriptors characteristic of the biological matrix under investigation in different experimental groups [20]. Last generation of highresolution devices are being used progressively in this field because of their high performance in terms of sensitivity and specificity [21][22][23][24][25][26][27]. However, given the large amount of data generated during metabolomics acquisitions, specifically dedicated bioinformatics tools such as XCMS software are required for processing and analyzing this huge volume of complex data.…”