Discrimination of different feed additives and poly-herbal formulations based on their untargeted phytochemical profiles

Introduction: Feed additives represents a valid tool in animal nutrition to improve animal performance and livestock productivity under a sustainable perspective; however, there is a paucity of information about their comprehensive metabolomic and bioactive profiles. Objective: In this study, we tested the ability of an untargeted metabolomics approach to discriminate nine commercial feed additives and unique blends of botanical extracts used in both ruminant and non-ruminant nutrition, according to their phytochemical profiles and different in vitro bioactive properties. Methods: An ultra-high-performance liquid chromatography coupled with Orbitrap mass spectrometry and multivariate statistics were combined to search for potential markers, in order to better discriminate the different commercial samples. Results: Several phytochemicals were identified, namely alkaloids, phenolics, organosulfurs, and terpenoids. The polyherbal formulation Zigbir was the best source of phytochemicals, accounting for a cumulative total content of phytochemicals equal to 3.03 mg Eq./g, being particularly abundant in terpenoids, stilbenes, phenolic acids, and small-molecular-weight phenolics. Multivariate statistics allowed to group the different products in 2 bioactive subclusters. The diterpenoid andrographolide recorded the highest abundance in Zigbir and Sangrovit. The most predictive biomarkers were: piperine, isoquercitrin, 6-methylthiohexyldesulfoglucosinolate, 6-methylumbelliferone, benzoic acid, (+)-(1R,2R)-1,2-diphenylethane-1,2-diol, and piperitenone. Flavonoids were highly correlated with both in vitro antioxidant and enzyme inhibition assays. Conclusions: Our findings provide new insights into the comprehensive phytochemical composition of commercial feed additives and blend of botanical extracts used for both ruminant and non-ruminant nutrition. A great importance of polyphenols in relation to the biological activities was detected.