Quantifying the human diet in the crosstalk between nutrition and health by multi-targeted metabolomics of food and microbiota-derived metabolites

Abstract

Background Metabolomics is a powerful tool for investigating the association between nutrition and health status. Although urine is commonly employed for studying the metabolism and transformation of food components, the use of blood samples could be preferable to gain new insights into the bioavailability of diet-derived compounds and their involvement in health.However, the chemical complexity of blood samples hinders the analysis of this biological fluid considerably, which makes the development of novel and comprehensive analytical methods mandatory.Methods In this work, we optimized a multi-targeted metabolomics platform for the quantitative and simultaneous analysis of 450 food-derived metabolites by ultra-high performance liquid chromatography coupled to tandem mass spectrometry. To handle the chemical complexity of blood samples, three complementary extraction methods were assayed and compared in terms of recovery, sensitivity, precision and matrix effects with the aim of maximizing metabolomics coverage: protein precipitation, reversed solid-phase extraction, and hybrid protein precipitation with solid-phase extraction-mediated phospholipid removal.Results After careful optimization of the extraction conditions, protein precipitation enabled the most efficient and highthroughput extraction of the food metabolome in plasma, although solid-phase extraction-based protocols provided complementary performance for the analysis of specific polyphenol classes. The developed method yielded accurate recovery rates with negligible matrix effects, and good linearity, as well as high sensitivity and precision for most of the analyzed metabolites.Conclusions The multi-targeted metabolomics platform optimized in this work enables the simultaneous detection and quantitation of 450 dietary metabolites in short-run times using small volumes of biological sample, which facilitates its application to epidemiological studies.

Crosstalk among intestinal barrier, gut microbiota and serum metabolome after a polyphenol-rich diet in older subjects with “leaky gut”: The MaPLE trial

“…After protein precipitation, 500 mL of supernatant were dried on vacuum and the residue was recovered with 100 mL of an 80/20 v/ v mixture of water/ACN, containing 0.5% v/v formic acid, 10 mM of ammonium formiate and 100 ppb of a mixture of 13 internal standards (glutamic acid 15 13 C, ferulic acid, 13 C 3 , epicatechin 13 C 3 , and taxifolin, SigmaeAldrich, Steinheim, Germany). Finally, after centrifugation at 10,000 rpm and 4 C for 5 min, samples were transferred to a 96-well plate and analysed using the targeted UHPLC-MS/MS method described in [18]. Briefly, an Agilent 1290 Infinity UHPLC system coupled to a Sciex QTRAP 6500 mass spectrometer was used for the analysis.…”

Mechanistic Insights into Alzheimer’s Disease Unveiled through the Investigation of Disturbances in Central Metabolites and Metabolic Pathways

“…However, it is recognized that the low sensitivity and low spectral resolution of NMR considerably reduce its coverage, thus impeding comprehensive metabolomics analysis [ 6 ]. On the other hand, although great efforts have been made to develop large-scale RPLC-MS metabolomics approaches [ 7 ], this analytical platform is usually limited to the determination of low and medium polarity metabolites, such as lipids (i.e., lipidomics) [ 8 ], aromatic amino acids and their microbiota derivatives [ 9 ], some nutrients and food-related metabolites (i.e., nutrimetabolomics) [ 10 , 11 ], and a few other metabolite classes. Accordingly, the efficient profiling of hydrophilic metabolites is still a methodological challenge.…”

Wine’s Phenolic Compounds and Health: A Pythagorean View

“…It is worth underscoring that we are making progress in the identification of metabolites, but—until recently—we mainly focused on the liver-derived ones [ 61 ]. The relatively recent discovery of microbiota-synthesized metabolites amplifies the list of potential biologically-active molecules produced by the body after the ingestion of (poly)phenol-rich foods [ 58 , 62 , 63 ].…”