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Untargeted Metabolomics approaches for Identifying Common Urinary Metabolites After Consumption of Olea europaea and Lippia citriodora Leaf Extracts
Antonio Segura-Carretero
University of Granada
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Create a topicOlea europaea (OE) and Lippia citriodora (LC) are two botanical matrices well-known for their similar bioactive properties. Although they both share common families of phenolic compounds, their specific chemical compositions differ significantly. As the compounds from these plants undergo metabolic transformation in the body, particularly through phase I and phase II pathways, this study hypothesizes that common metabolite derivatives may arise, potentially contributing to their shared bioactive effects. Therefore, this study aims to identify common urinary metabolites derived from the intake of OE and LC leaf extracts. To achieve this, an acute double-blind dietary intervention was conducted with twenty-five volunteers, divided into three groups: OE (n=8), LC (n=8), and placebo (n=9). Urine samples were collected, before supplementation and afterwards at various intervals, and were analyzed using an untargeted metabolomic approach based on HPLC-ESI-QTOF-MS. Following data processing with MZmine and R packages, 18,823 molecular features were detected. Filtering steps and statistical analysis allowed the identification of exogenous metabolites derived from the extracts, revealing 580 and 424 significant features associated with the intake of OE and LC extracts, respectively. Notably, 129 molecular features were common to both matrices. Identification of significant molecular features was performed by comparing MS and MS/MS data with open-access metabolomics databases and existing literature. A total of 56 common features could be annotated, with phenolic acids derivatives (e.g., diferulic acid, homovanillic alcohol glucuronide etc.) as one of the most prominent families of bioavailable metabolites. In general, phase II metabolites, particularly sulfates and glucuronides, were the most predominant type of annotated metabolites. These findings underscore the potential of untargeted metabolomics in detecting bioavailable metabolites in urine after supplementation of bioactive extracts, enhancing the understanding of the bioactive mechanisms that could be useful for developing applications based on them, such as functional foods, nutraceuticals, or cosmeceuticals.
Emilio Alvarez Parrilla
First of all, congratulations for your work. I was wondering if you could tell me the amount of extract (and the equivalents of phenolic compounds) given to each participant
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Patricia Fernández-Moreno
Thank you very much for your comment and question, Emilio. It is a very good point. The extracts were encapsulated at a ratio of 500 mg of extract per capsule. The Total Phenolic Content (TPC) was measured by the Folin–Ciocalteu method using gallic acid as a reference compound for the standard curve. After performing the measurements in triplicate, the extract of Olea europaea leaves showed a TPC of 216 ± 28 mg Gallic Acid Equivalent/g Dry Extract, while the extract of Lippia citriodora showed a value of 344 ± 15 mg Gallic Acid Equivalent/g Dry Extract. If you have any further questions, please do not hesitate to reach out.
Emilio Alvarez Parrilla
Thanks for you responce I will be waiting for the published paper. Best regards