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Discovering potential urinary biomarkers of tomato consumption using untargeted metabolomics

ABSTRACT: Tomatoes are one of the most consumed vegetables in the United States, making them an economically and nutritionally important crop. Previous epidemiological and intervention studies have suggested positive health outcomes associated with tomato consumption. To more conclusively understand the relationship between tomato intake and health benefits, an accurate measurement of tomato consumption is required. Current methods rely on self-report dietary assessments which suffer from recall bias or, plasma lycopene analysis which lacks reliability. Urine sampling allows an objective, non- invasive assessment of dietary intake, resulting in a more robust determination of consumption. Untargeted metabolomics allows for detection of differences in small molecules between groups without preconceived bias, allowing for several classes of compounds to be evaluated as potential biomarkers. The objective of this study was to discover putative urinary biomarkers of tomato consumption using untargeted metabolomics. Healthy subjects (n=11) were evaluated on two separate days: on each day they consumed a controlled diet with either red or tangerine tomato juice with breakfast. Two varieties of tomato juices were included in order to understand how variation in tomato source contributes to intake biomarkers observed. Urine was collected at baseline, 3, 6, 9, 12, and 24 hours. UHPLC-QTOF-MS was used to compare osmolality-normalized urine samples at each time point. Data was collected in both positive and negative modes for comprehensive coverage of metabolites. Raw data was processed using MZmine2. Univariate and multivariate statistical approaches were applied to understand which metabolites were significantly different between urine pre- and-post-tomato consumption. Feature lists were reduced to those that met all three of the following criteria: the compound was significantly different (p<0.05) at every timepoint compared to baseline; the compound had a fold change greater than 2 at every timepoint compared to baseline; the compound had a VIP score greater than 1 for the Partial Least Squares Regression model. We found 99 metabolites in ESI (+) and 46 metabolites in ESI (-) that met these criteria. Online databases were consulted as a first attempt at identifying unknown compounds. Data-independent MS/MS analysis was conducted to generate fragmentation patterns to support feature identification. Authentic standards for naringenin-4-O-glucuronide and naringenin-7-O-glucuronide confirmed their identities in both modes compared to pooled 6 hour urine. Previously reported compounds in urine after tomato consumption, N-caprylhistidinol and N-caprylhistidinol glucuronide were also identified in positive mode with a level 2 confidence, giving more support for future investigation into these compounds as biomarkers. Several compounds from the features of interest reduced lists could be tentatively identified as metabolites that had undergone phase II reactions prior to urinary excretion. Additional studies will need to be done to test the robustness, analytical performance, stability, and reliability of these compounds.

INSTRUMENT(S): Liquid Chromatography MS - negative - reverse phase, Liquid Chromatography MS - positive - reverse phase

PROVIDER: MTBLS2328 | MetaboLights | 2024-11-04

REPOSITORIES: MetaboLights

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Project description:PURPOSE: Validated biomarkers of food intake (BFIs) have recently been suggested as a useful tool to assess adherence to dietary guidelines or compliance in human dietary interventions. Although many new candidate biomarkers have emerged in the last decades for different foods from metabolic profiling studies, the number of comprehensively validated biomarkers of food intake is limited. Apples are among the most frequently consumed fruits and a rich source of polyphenols and fibers, an important mediator for their health-protective properties.METHODS: Using an untargeted metabolomics approach, we aimed to identify biomarkers of long-term apple intake and explore how apples impact on the human plasma and urine metabolite profiles. Forty mildly hypercholesterolemic volunteers consumed two whole apples or a sugar and energy-matched control beverage, daily for 8 weeks in a randomized, controlled, crossover intervention study. The metabolome in plasma and urine samples was analyzed via untargeted metabolomics.RESULTS: We found 61 urine and 9 plasma metabolites being statistically significant after the whole apple intake compared to the control beverage, including several polyphenol metabolites that could be used as BFIs. Furthermore, we identified several endogenous indole and phenylacetyl-glutamine microbial metabolites significantly increasing in urine after apple consumption. The multiomic dataset allowed exploration of the correlations between metabolites modulated significantly by the dietary intervention and fecal microbiota species at genus level, showing interesting interactions between Granulicatella genus and phenyl-acetic acid metabolites. Phloretin glucuronide and phloretin glucuronide sulfate appeared promising biomarkers of apple intake; however, robustness, reliability and stability data are needed for full BFI validation.CONCLUSION: The identified apple BFIs can be used in future studies to assess compliance and to explore their health effects after apple intake. Moreover, the identification of polyphenol microbial metabolites suggests that apple consumption mediates significant gut microbial metabolic activity which should be further explored.

Project description:It has been proposed that endogenously formed N-nitroso compounds (NOCs) are partly responsible for the link between red meat consumption and colorectal cancer (CRC) risk. As nitrite has been indicated as one of the critical factors in the formation of endogenous NOCs, it is of high importance to replace or reduce the nitrite levels in meat. Therefore, the PHYTOME project was initiated (Phytochemicals to reduce nitrite in meat products; www.phytome.eu), an EU funded research project aiming to develop innovative meat products in which the food additive sodium nitrite (E251) has been replaced by natural compounds originating from fruits and vegetables. A human dietary intervention study was conducted in which healthy subjects consumed 300 grams of meat for two weeks, in subsequent order: normal processed red meat, white meat, and red processed meat with standard or reduced levels of nitrite and added phytochemicals. Consumption of standard-nitrite PHYTOME meat products leads to a significant reduction in Apparent Total N-nitroso Compounds (ATNC) levels in faecal water, a surrogate marker of endogenously formed NOCs, as compared to the consumption of conventional processed red meat products. A reduction of nitrite in the PHYTOME meat lowered these levels even further. In addition, DNA strand breaks induced in ex-vivo faecal water exposed Caco-2 cells and O6-methyl-guanine adducts levels in colonic DNA were significantly higher after consumption of normal processed red meat as compared to white meat intake. PHYTOME meat intake resulted in reduced levels of these genotoxic markers, however, these were not statistically significant. Whole genome gene expression analyses in colonic tissue identified differentially expressed genes and genes associated with ATNC, which are related to molecular pathways which may explain cancer risk initiation after intake of processed red meat and cancer risk prevention after intake of the PHYTOME meat. Together these results indicate that addition of natural extracts to conventional processed red meat products results in reduced endogenous formation of NOCs, and may therefore contribute to a reduced risk of CRC, which is mechanistically supported by gene expression analyses.

			Action	DRS
	113_Hr0_RED_20.mzML	Mzml
	113_Hr0_TANG_63.mzML	Mzml
	113_Hr12_RED_53.mzML	Mzml
	113_Hr12_TANG_79.mzML	Mzml
	113_Hr24_RED_101.mzML	Mzml

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Discovering potential urinary biomarkers of tomato consumption using untargeted metabolomics

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