Chemical composition, antioxidant activity and bioaccessibility studies in phenolic extracts of two Hericium wild edible species

Mushrooms are rich sources of bioactive compounds such as phenolic acids. When ingested, these molecules have to be released from the matrix to be transformed/absorbed by the organism, so that they can exert their bioactivity. Several in vitro methodologies have been developed in order to evaluate the bioavailability of bioactive compounds. Herein, two Hericium species were analyzed for their chemical composition and antioxidant activity. Furthermore, an in vitro digestion of the mushrooms and mushroom phenolic extracts was performed, and the digested samples were also submitted to antioxidant activity evaluation in order to evaluate the bioaccessibility of the phenolic acids identified in the samples. Hericium species showed similar chemical profiles (except for tocopherols), varying only in the concentration of the compounds. The phenolic extracts revealed higher antioxidant activity than the in vitro digested samples, meaning that this process decrease the antioxidant properties of the extract/mushroom. Nevertheless, phenolic acids were found in the digested samples, meaning that those molecules are bioaccessible.

Effect of dietary fiber on the bioaccessibility of phenolic compounds of mango, papaya and pineapple fruits by an in vitro digestion model

The presence of dietary fiber (DF) in the food matrix of some tropical fruits plays an important role in the release and absorption of its bioactive compounds, such as phenolic compounds (PCs). The aim of this study was to evaluate the effect of the DF fractions in mango cv. ‘Ataulfo’, papaya cv. ‘Maradol’ and pineapple cv. ‘Esmeralda’, on the bioaccessibility of their PCs and antioxidant capacity (AOXC) under an in vitro digestion model. The highest PCs content and AOXC was found in mango (274.30 mg GAE/100 g FW), followed by papaya (212 mg GAE//100 g FW), and pineapple (107.63 mg GAE/100 g FW), respectively. About 50% of the total PCs in all fruits was released at gastric phase, increasing closer to 60% at intestinal phase in mango and pineapple. However, the highest content of PCs associated to DF was found in mango (2.48 mg GAE/100 g FW) compared with papaya DF fractions (0.96 GAE/100 g FW) and pineapple (0.52 GAE/100 g FW). The presence of DF in mango, papaya and pineapple did not represent a major limitation on the bioaccessibility of its PCs according to the in vitro digestion model used in this study.

In vitro bioaccessibility and gut biotransformation of polyphenols present in the water-insoluble cocoa fraction

Scope: Cocoa, especially the water-insoluble cocoa fraction (WICF), is a rich source of polyphenols. In this study, sequential in vitro digestion of the WICF with gastrointestinal enzymes as well as its bacterial fermentation in a human colonic model system were carried out to investigate bioaccessibility and biotransformation of WICF polyphenols, respectively. Methods and results: The yield of each enzymatic digestion step and the total antioxidant capacity (TAC) were measured and solubilized phenols were characterized by MS/MS. Fermentation of WICF and the effect on the gut microbiota, SCFA production and metabolism of polyphenols was analyzed. In vitro digestion solubilized 38.6% of WICF with pronase and Viscozyme L treatments releasing 51% of the total phenols from the insoluble material. This release of phenols does not determine a reduction in the total antioxidant capacity of the digestion-resistant material. In the colonic model WICF significantly increased of bifidobacteria and lactobacilli as well as butyrate production. Flavanols were converted into phenolic acids by the microbiota following a concentration gradient resulting in high concentrations of 3-hydroxyphenylpropionic acid (3-HPP) in the last gut compartment. Conclusion: Data showed that WICF may exert antioxidant action through the gastrointestinal tract despite its polyphenols being still bound to macromolecules and having prebiotic activity.

Colon extended physiologically based extraction test (CE-PBET) increases bioaccessibility of soil-bound PAH

Assessment of the risk to human health posed by contaminated land may be seriously overestimated if reliant on total pollutant concentration. In vitro extraction tests, such as the physiologically based extraction test (PBET), imitate the physicochemical conditions of the human gastro-intestinal tract and offer a more practicable alternative for routine testing purposes. However, even though passage through the colon accounts for approximately 80% of the transit time through the human digestive tract and the typical contents of the colon in vivo are a carbohydrate-rich aqueous medium with the potential to promote desorption of organic pollutants, PBET comprises stomach and small intestine compartments only. Through addition of an eight-hour colon compartment to PBET and use of a carbohydrate-rich fed-state medium we demonstrated that colon-extended PBET (CE-PBET) in- creased assessments of soil-bound PAH bioaccessibility by up to 50% in laboratory soils and a factor of 4 in field soils. We attribute this increased bioaccessibility to a combination of the additional extraction time and the presence of carbohydrates in the colon compartment, both of which favor PAH desorption from soil. We propose that future assessments of the bioaccessibility of organic pollutants in soils using physiologically based extraction tests should have a colon compartment as in CE-PBET.

In vitro assessment of the bioaccessibility of brominated flame retardants in indoor dust using a colon extended model of the human gastrointestinal tract

An in vitro colon extended physiologically based extraction test (CEPBET) which incorporates human gastrointestinal tract (GIT) parameters (including pH and chemistry, solid-to-fluid ratio, mixing and emptying rates) was applied for the first time to study the bioaccessibility of brominated flame retardants (BFRs) from the 3 main GIT compartments (stomach, small intestine and colon) following ingestion of indoor dust. Results revealed the bioaccessibility of γ-HBCD (72%) was less than that for α- and β-isomers (92% and 80% respectively) which may be attributed to the lower aqueous solubility of the γ-isomer (2 μg L−1) compared to the α- and β-isomers (45 and 15 μg L−1 respectively). No significant change in the enantiomeric fractions of HBCDs was observed in any of the studied samples. However, this does not completely exclude the possibility of in vivo enantioselective absorption of HBCDs, as the GIT cell lining and bacterial flora – which may act enantioselectively – are not included in the current CE-PBET model. While TBBP-A was almost completely (94%) bioaccessible, BDE-209 was the least (14%) bioaccessible of the studied BFRs. Bioaccessibility of tri-hepta BDEs ranged from 32–58%. No decrease in the bioaccessibility with increasing level of bromination was observed in the studied PBDEs.

Is there sufficient 'sink' in current bioaccessibility determinations of organic pollutants in soils?

Bioaccessibility tests can be used to improve contaminated land risk assessments. For organic pollutants a ‘sink’ is required within these tests to better mimic their desorption under the physiological conditions prevailing in the intestinal tract, where a steep diffusion gradient for the removal of organic pollutants from the soil matrix would exist. This is currently ignored in most PBET systems. By combining the CEPBET bioaccessibility test with an infinite sink, the removal of PAH from spiked solutions was monitored. Less than 10% of spiked PAH remained in the stomach media after 1 h, 10% by 4 h in the small intestine compartment and c.15% after 16 h in the colon. The addition of the infinite sink increased bioaccessibility estimates for field soils by a factor of 1.2–2.8, confirming its importance for robust PBET tests. TOC or BC were not the only factors controlling desorption of the PAH from the soils.

Towards a unified approach for the determination of the bioaccessibility of organic pollutants

Bioaccessibility studies have been widely used as a research tool to determine the potential human exposure to ingested contaminants. More recently they have been practically applied for soil borne toxic elements. This paper reviews the application of bioaccessibility tests across a range of organic pollutants and contaminated matrices. Important factors are reported to be: the physiological relevance of the test, the components in the gut media, the size fraction chosen for the test and whether it contains a sorptive sink. The bioaccessibility is also a function of the composition of the matrix (e.g. organic carbon content of soils) and the physico-chemical characteristics of the pollutant under test. Despite the widespread use of these tests, there are a large number of formats used and very few validation studies with animal models. We propose a unified format for a bioaccessibility test for organic pollutants. The robustness of this test should first be confirmed through inter laboratory comparison, then tested in-vivo.

Polymeric Nanoparticles as Oral Delivery Systems for Encapsulation and Release of Polyphenolic Compounds: Impact on Quercetin Antioxidant Activity & Bioaccessibility

A delivery system containing polymeric (Eudragit) nanoparticles has been developed for encapsulation and controlled release of bioactive flavonoids (quercetin). Nanoparticles were fabricated using a solvent displacement method. Particle size, morphology, and charge were measured by light scattering, electron microscopy and zeta-potential. Encapsulation efficiency (EE) and release profiles were determined using electrochemical methods. Molecular interactions within the particle matrix were characterized by X-ray diffraction, differential scanning calorimetry, and infrared spectroscopy. Antioxidant properties of free and encapsulated quercetin were analyzed by TBARS and fluorescence spectroscopy. Bioaccessibility of quercetin was evaluated using an in vitro digestion model. Relatively small (d a parts per thousand aEuro parts per thousand 370 nm) anionic polymeric nanoparticles were formed containing quercetin in a non-crystalline form (EE a parts per thousand aEuro parts per thousand 67 %). The main interaction between quercetin and Eudragit was hydrogen bonding. Encapsulated quercetin remained stable during 6 months storage and maintained its antioxidant activity. Quercetin bioaccessibility within simulated small intestinal conditions was improved by encapsulation. The knowledge obtained from this study will facilitate the rational design and fabrication of polymeric nanoparticles as oral delivery systems for encapsulation, protection, and release of bioactive compounds.

Gastrointestinal modelling and optimization of protein and phosphorus bioaccessibility by phytase in fish diets

Programa de doctorado: Acuicultura

Assessment of oral bioaccessibility of arsenic in playground soil in Madrid (Spain): A three-method comparison and implications for risk assessment

Three methodologies to assess As bioaccessibility were evaluated using playgroundsoil collected from 16 playgrounds in Madrid, Spain: two (Simplified Bioaccessibility Extraction Test: SBET, and hydrochloric acid-extraction: HCl) assess gastric-only bioaccessibility and the third (Physiologically Based Extraction Test: PBET) evaluates mouth–gastric–intestinal bioaccessibility. Aqua regia-extractable (pseudo total) As contents, which are routinely employed in riskassessments, were used as the reference to establish the following percentages of bioaccessibility: SBET – 63.1; HCl – 51.8; PBET – 41.6, the highest values associated with the gastric-only extractions. For Madridplaygroundsoils – characterised by a very uniform, weakly alkaline pH, and low Fe oxide and organic matter contents – the statistical analysis of the results indicates that, in contrast with other studies, the highest percentage of As in the samples was bound to carbonates and/or present as calcium arsenate. As opposed to the As bound to Fe oxides, this As is readily released in the gastric environment as the carbonate matrix is decomposed and calcium arsenate is dissolved, but some of it is subsequently sequestered in unavailable forms as the pH is raised to 5.5 to mimic intestinal conditions. The HCl extraction can be used as a simple and reliable (i.e. low residual standard error) proxy for the more expensive, time consuming, and error-prone PBET methodology. The HCl method would essentially halve the estimate of carcinogenic risk for children playing in Madridplaygroundsoils, providing a more representative value of associated risk than the pseudo-total concentrations used at present

The relationship between soil geochemistry and the bioaccessibility of trace elements in playground soil

A total of 32 samples of surficial soil were collected from 16 playground areas in Madrid (Spain), in order to investigate the importance of the geochemistry of the soil on subsequent bioaccessibility of trace elements. The in vitro bioaccessibility of As, Co, Cr, Cu, Ni, Pb and Zn was evaluated by means of two extraction processes that simulate the gastric environment and one that reproduces a gastric + intestinal digestion sequence. The results of the in vitro bioaccessibility were compared against aqua regia extractions (“total” concentration), and it was found that total concentrations of As, Cu, Pb and Zn were double those of bioaccessible values, whilst that of Cr was ten times higher. Whereas the results of the gastric + intestinal extraction were affected by a high uncertainty, both gastric methods offered very similar and consistent results, with bioaccessibilities following the order: As = Cu = Pb = Zn > Co > Ni > Cr, and ranging from 63 to 7 %. Selected soil properties including pH, organic matter, Fe and CaCO3 content were determined to assess their influence on trace element bioaccessibility, and it was found that Cu, Pb and Zn were predominantly bound to organic matter and, to a lesser extent, Fe oxides. The former fraction was readily accessible in the gastric solution, whereas Fe oxides seemed to recapture negatively charged chloride complexes of these elements in the gastric solution, lowering their bioaccessibility. The homogeneous pH of the playground soils included in the study does not influence trace element bioaccessibility to any significant extent except for Cr, where the very low gastric accessibility seems to be related to the strongly pH-dependent formation of complexes with organic matter. The results for As, which have been previously described and discussed in detail in Mingot et al. (Chemosphere 84: 1386–1391, 2011), indicate a high gastric bioaccessibility for this element as a consequence of its strong association with calcium carbonate and the ease with which these bonds are broken in the gastric solution. The calculation of risk assessments are therefore dependant on the methodology used and the specific environment they address. This has impacts on management strategies formulated to ensure that the most vulnerable of society, children, can live and play without adverse consequences to their health.

Bioaccessibility of heavy metals and risk assessment in urban gardens of Madrid

Human health problems and solutions. Urban gardening has spread worldwide in recent years as it enhances food security and selfsupply and promotes community integration. However urban soils are significantly enriched in trace elements relative to background levels. Exposure to the soil in urban gardens may therefore result in adverse health effects depending on the degree of contact during gardening, infant recreational activities and ingestion of vegetables grown in them. In order to evaluate this potential risk, 36 composite samples were collected from the top 20 cm of the soil of 6 urban gardens in Madrid. The aqua regia (pseudototal) and glycine-extractable (bioaccessible) concentrations of Co, Cr, Cu, Ni, Pb and Zn were determined by atomic absorption spectrophotometry. Additionally, pH, texture, Fe, Ca, and Mn concentrations, and organic matter and calcium carbonate contents were determined in all urban gardens and their influence on trace element bioaccessibility was analyzed.

Bioaccessibility of metals and human health risk assessment in community urban gardens

Pseudo-total (i.e. aqua regia extractable) and gastric-bioaccessible (i.e. glycine + HCl extractable) concentrations of Ca, Co, Cr, Cu, Fe, Mn, Ni, Pb and Zn were determined in a total of 48 samples collected from six community urban gardens of different characteristics in the city of Madrid (Spain). Calcium carbonate appears to be the soil property that determines the bioaccessibility of a majority of those elements, and the lack of influence of organic matter, pH and texture can be explained by their low levels in the samples (organic matter) or their narrow range of variation (pH and texture). A conservative risk assessment with bioaccessible concentrations in two scenarios, i.e. adult urban farmers and children playing in urban gardens, revealed acceptable levels of risk, but with large differences between urban gardens depending on their history of land use and their proximity to busy areas in the city center. Only in a worst-case scenario in which children who use urban gardens as recreational areas also eat the produce grown in them would the risk exceed the limits of acceptability