Combining traditional dietary assessment methods with novel metabolomics techniques: present efforts by the Food Biomarker Alliance

FFQ, food diaries and 24 h recall methods represent the most commonly used dietary assessment tools in human studies on nutrition and health, but food intake biomarkers are assumed to provide a more objective reflection of intake. Unfortunately, very few of these biomarkers are sufficiently validated. This review provides an overview of food intake biomarker research and highlights present research efforts of the Joint Programming Initiative 'A Healthy Diet for a Healthy Life' (JPI-HDHL) Food Biomarkers Alliance (FoodBAll). In order to identify novel food intake biomarkers, the focus is on new food metabolomics techniques that allow the quantification of up to thousands of metabolites simultaneously, which may be applied in intervention and observational studies. As biomarkers are often influenced by various other factors than the food under investigation, FoodBAll developed a food intake biomarker quality and validity score aiming to assist the systematic evaluation of novel biomarkers. Moreover, to evaluate the applicability of nutritional biomarkers, studies are presently also focusing on associations between food intake biomarkers and diet-related disease risk. In order to be successful in these metabolomics studies, knowledge about available electronic metabolomics resources is necessary and further developments of these resources are essential. Ultimately, present efforts in this research area aim to advance quality control of traditional dietary assessment methods, advance compliance evaluation in nutritional intervention studies, and increase the significance of observational studies by investigating associations between nutrition and health. Keywords: Dietary assessment; Food intake biomarkers; Food metabolome; Metabolomics

Total antioxidant capacity as a tool to assess redox status: critical view and experimental data

The measure of antioxidant capacity (AC) considers the cumulative action of all the antioxidants present in plasma and body fluids, thus providing an integrated parameter rather than the simple sum of measurable antioxidants. The capacity of known and unknown antioxidants and their synergistic interaction is therefore assessed, thus giving an insight into the delicate balance in vivo between oxidants and antioxidants. Measuring plasma AC may help in the evaluation of physiological, environmental, and nutritional factors of the redox status in humans. Determining plasma AC may help to identify conditions affecting oxidative status in vivo (e.g., exposure to reactive oxygen species and antioxidant supplementation). Moreover, changes in the plasma AC after supplementation with galenic antioxidants or with antioxidant-rich foods may provide information on the absorption and bioavailability of nutritional compounds. Consequently, this review discusses the rationale, interpretation, confounding factors, measurement limits, and human applications of the measure of plasma AC

SCREENING GROWTH CONDITIONS TO INCREASE ANTHOCYANINS AND GLUCOSINOLATES CONTENT IN BROCCOLI SPROUTS

Broccoli (Brassicaceae) are an excellent dietary source of micronutrients (vitamins and minerals) and health-promoting phytochemicals (glucosinolates and phenolic compounds). Glucosinolates are a class of organic compounds derived from glucose and different aminoacids, and characterized by the high number of different side chains. Anthocyanins represent the largest group of natural, water-soluble plant pigments, and approximately 400 anthocyanins have been determined. Glucosinolates and anthocyanins protects plants against various biotic and abiotic stresses and possess antioxidant and antinflammatory activity in humans. In addition, glucosinolates have gained much attention due to the anticarcinogenic activity of isothiocyanates, their major hydrolysis products, molecules able to induce phase 2 detoxication enzymes and protect animals against chemically induced cancer (Zhang, 2012). Young broccoli plant are an especially good source of glucosinolates and anthocyanins and several studies report that growth and environmental conditions could affect the profiles of these secondary plant metabolites (Perez-Balibrea et al., 2008). We previously developed a rapid and sensitive LC-MS (MRM) method for direct and simultaneous quantification of 14 glucosinolates in broccoli sprouts. In this study, the effect of different abiotic stresses and hormones involved in plant defence responses of the accumulation of glucosinolates and anthocyanins in broccoli sprouts was investigated. Quantitative results show that all treatments modify glucosinolates and anthocyanins profile. In particular, light grown broccoli sprouts treated with sucrose show glucosinolates and anthocyanins levels double than control. This work was ,supported by Italian Ministry of Agriculture, Food & Forestry (MiPAAF) grant "NUTRIGEA" (DM 30281 23112/2009)

Untargeted Metabolomics Reveals Predominant Alterations in Lipid Metabolism Following Light Exposure in Broccoli Sprouts

The consumption of vegetables belonging to the family Brassicaceae (e.g., broccoli and cauliflower) is linked to a reduced incidence of cancer and cardiovascular diseases. The molecular composition of such plants is strongly affected by growing conditions. Here we developed an unbiased metabolomics approach to investigate the effect of light and dark exposure on the metabolome of broccoli sprouts and we applied such an approach to provide a bird’s-eye view of the overall metabolic response after light exposure. Broccoli seeds were germinated and grown hydroponically for five days in total darkness or with a light/dark photoperiod (16 h light/8 h dark cycle). We used an ultra-performance liquid-chromatography system coupled to an ion-mobility, time-of-flight mass spectrometer to profile the large array of metabolites present in the sprouts. Differences at the metabolite level between groups were analyzed using multivariate statistical analyses, including principal component analysis and correlation analysis. Altered metabolites were identified by searching publicly available and in-house databases. Metabolite pathway analyses were used to support the identification of subtle but significant changes among groups of related metabolites that may have gone unnoticed with conventional approaches. Besides the chlorophyll pathway, light exposure activated the biosynthesis and metabolism of sterol lipids, prenol lipids, and polyunsaturated lipids, which are essential for the photosynthetic machinery. Our results also revealed that light exposure increased the levels of polyketides, including flavonoids, and oxylipins, which play essential roles in the plant’s developmental processes and defense mechanism against herbivores. This study highlights the significant contribution of light exposure to the ultimate metabolic phenotype, which might affect the cellular physiology and nutritional value of broccoli sprouts. Furthermore, this study highlights the potential of an unbiased omics approach for the comprehensive study of the metabolism

Improvement of the nutraceutical quality of broccoli sprouts by elicitation

Epidemiological studies show an inverse association between Brassica consumption and chronic diseases. Phytochemicals are thought to be beneficial for human health and therefore responsible for this protective effect. Increasing their levels into Brassica food is considered an expedient nutritional strategy that can be achieved through the manipulation of growth conditions by elicitors. In this work we systematically evaluated the influence of treatment with different elicitors (sucrose, mannitol, NaCl, 1-aminocyclopropane-l-carboxylic acid, salicylic acid and methyl jasmonate) on the phytochemical composition of broccoli sprouts. The content of total and single glucosinolates, total phenolic compounds, total flavonoids, total anthocyanins, vitamin C and E and β-carotene was assessed. The exposure to different elicitors produced concentration- and elicitor-dependent specific changes in the content of all the phytochemicals considered. Sucrose, identified as the most effective elicitor by principal component analysis, induced a significant increase of total and specific glucosinolates, vitamin C, total anthocyanins and polyphenols. Sucrose is likely to represent an effective tool to increase the nutritional value of broccoli sprouts