Comprehensive LC-HRMS metabolomics analyses for the estimation of environmental inputs of altrenogest in pig breeding.

Altrenogest (ALT), a synthetic progestogen, is used in pig farming for estrus synchronization in gilts. Residues of ALT and its metabolites may reach the aquatic environment via the spread of liquid manure and may present a risk for fish and other higher aquatic organisms due to its endocrine disrupting potential. A pilot study was conducted in which spot urine samples from ALT-treated and non-medicated gilts were collected. We applied LC-HRMS analysis to perform targeted analysis of ALT and known metabolites as well as non-targeted metabolomics analyses to find previously unknown metabolites. The targeted investigation showed that glucuronide conjugates of ALT and its photo-isomerization product are main urinary metabolites of ALT in gilts. Furthermore, an unknown isomerization product of ALT was observed at trace level, whereas ALT and ALT sulfate were not found. The chemometric analysis of non-targeted data revealed a clear difference between ALT-treated gilts and control animals. Furthermore, a hydroxylated ALT glucuronide was identified as highly significant in the ALT-treated group. Additional biomarker annotation and pathway mapping revealed changes in the metabolism of ALT-treated animals which can be explained by ALT's hormonal action. This study illustrates the exceptional potential of LC-HRMS and metabolomics for the detection of potentially new environmental contaminants with high biological activity. Further advantages of the method described are the sampling during routine breeding conditions, a relatively small number of animals required and no particular stress for the animals.

Comprehensive Metabolomics Analysis of Nontargeted LC-HRMS Data Provides Valuable Insights Regarding the Origin of Veterinary Drug Residues.

Liquid chromatography coupled with high resolution mass spectrometry (LC-HRMS) complements standard triple-quadrupole mass spectrometry in veterinary drug residue control. LC-HRMS offers the opportunity for nontargeted screening for metabolites and biomarkers representing metabolic changes. In this work, the feasibility of a nontargeted metabolomics approach based on LC-HRMS data (LC-Q-Orbitrap and LC-Q-TOF) to distinguish between porcine muscle tissue from infected animals and from healthy animals is demonstrated. The differences arise from various compounds associated with metabolic changes in infected animals. Two new biomarker candidates have been identified: tripeptide prolyphenylalanylglycine and a lysophosphatidylcholine derivative. For the first time, a bivariate data analysis procedure is described that may be used to evaluate whether the presence of antibiotic residues points to a therapeutic application or may be the result of a contamination during sampling and/or analysis.