Insights from in vitro exploration of factors influencing iron,zinc and provitamin A carotenoid bioaccessibility and intestinal absorption from cereals

Developments in genetics, agronomics and processing has positioned staple cereals as important sources of iron, zinc and provitamin A (pVA) carotenoids for nutritionally vulnerable populations. Significant effort has been placed on understanding the bioavailability of these micronutrients from cereal foods, including the exploration of underlying mechanisms by which their bioavailability can be modified. While micronutrient bioavailability is preferably assessed in clinical trials, relevant in vitro digestion and intestinal cell culture models have been applied to study effects of genetic, agronomic, post-harvest and food processing on micronutrient bioavailability. This review (1) critically assesses the application of in vitro models in the exploration of mechanisms associated with iron, zinc and provitamin A carotenoid bioaccessibility and intestinal absorption from cereal foods, and (2) identifies remaining gaps in order to frame future strategies to improve the nutritional impact of cereal foods.     

中国优控多环芳烃土壤污染特征及国内外生物可给性研究进展

多环芳烃(Polycyclic aromatic hydrocarbons,PAHs)是一类持久性有机污染物,容易在土壤中累积并且毒性显著,但PAHs在土壤固相上的吸附性较强,基于土壤污染总量暴露会导致高估人群健康风险。本研究基于2000-2020年间发表的123篇文献,总结了我国土壤中16种优先控制多环芳烃(∑16PAHs)的污染浓度分布和组成特征,介绍了11种常见的模拟PAHs生物可给性测试方法和主要影响因素,并总结了PAHs的生物可给性系数范围。研究结果表明土壤∑16PAHs最高与平均浓度分别为23 250 和1 314.7 μg.kg-1污染较为严重;近年来PAHs生物可给性测试方法主要基于生理原理提取法(PBET),在模拟消化过程和吸附剂等方面不断完善和改进,并且消化条件、土壤性质等因素对生物可给性结果影响较大。16种PAHs的生物可给性平均值范围为13.2%~72.4%,其中?和苯并[b]荧蒽的生物可给性较高,对∑16PAHs暴露产生贡献较高。本研究为开展土壤PAHs污染精细化风险评估研究提供重要理论参考依据。     

Large volume TENAX® extraction of the bioaccessible fraction of sediment-associated organic compounds for a subsequent effect-directed analysis

Background, Aims and Scope

Effect-directed analysis (EDA) is a powerful tool for the identification of key toxicants in complex environmental samples. In most cases, EDA is based on total extraction of organic contaminants, which may lead to an erroneous prioritisation with regard to hazard and risk. Bioaccessibility-directed extraction aims to discriminate between contaminants that take part in partitioning between sediment and biota in a relevant time frame and those that are enclosed in structures that do not allow rapid desorption. Standard protocols of targeted extraction of the rapidly desorbing, and thus bioaccessible, fraction using TENAX® are based only on small amounts of sediment. In order to obtain sufficient extract for subsequent biotesting, fractionation and structure elucidation, a large volume extraction technique needs to be developed applying one selected extraction time and excluding toxic procedural blanks.

Methods

Desorption behaviour of sediment contaminants was determined by combining consecutive extraction of sediment using TENAX® with a three-compartment desorption model. Time needed to remove the rapidly desorbing fraction, t_rap, was calculated to select a fixed extraction time for single extraction procedures. Up-scaling by about a factor of 125 provided a large volume extraction technique for EDA. Reproducibility and comparability to the small volume approach were analysed. TENAX® blanks and sediment extracts were tested for toxicity using Scenedesmus vacuolatus and Artemia salina as test organisms.

Results and Discussion

Desorption kinetics showed that 12 to 30% of sediment-associated pollutants were available for rapid desorption, while 70 to 90% of PAHs found in the sediment belong to the slowly and very slowly desorbing pool with very limited bioavailability. t_rap is compound dependent and covers a range of 2 to 18 h. A fixed extraction time of 24 h was selected as a time at which even the rapidly desorbing fraction of big hydrophobic compounds should be fully desorbed. High reproducibility of the large volume approach and good agreement with the small consecutive approach were found. Significant toxicity of procedural TENAX® blanks was found with Scenedesmus vacuolatus, which is in agreement with chemical analysis and could be reduced by pre-cleaning of TENAX® with Accelerated Solvent Extraction (ASE). Toxicity of blanks prior to ASE-clean up was about three orders of magnitude below the toxicity of sediment extracts.

Conclusions

For consideration of bioaccessibility in EDA, a large volume TENAX® extraction method was presented. Although several other solid phases can be used to extract the bioaccessible fraction, TENAX® has unique properties for depletive extraction of the rapidly desorbing fraction from large amounts of sediment. Toxicity and chemical blanks due to production residues are shortcomings of the method that can be overcome by accurate pre-cleaning, e.g. with ASE.

Recommendations and Perspectives

Higher purity of TENAX® guaranteed by the manufacturers would significantly enhance the applicability of the method. Using TENAX® instead of total extraction may improve key toxicant prioritisation by considering exposure and effect rather than effect only.

     

Influence of baking and in vitro digestion on steryl ferulates from wheat

So far, data on absorption and metabolism of steryl ferulates from edible sources is scarce. Therefore, the impact of enzyme-aided baking and in vitro digestion was examined in this study. Wheat flours and wheat breads were subjected to a static in vitro digestion model and changes in the contents of steryl ferulates and free sterols (possible hydrolysis products of steryl ferulates) were monitored. Baking degraded steryl ferulates at a similar rate in all types of breads (43–47%) compared to the corresponding flours, while baking induced changes in free sterols showed no clear pattern. In vitro digestion provoked five folds lower content of steryl ferulates in flours than in breads and it also resulted in significant free sterol accumulation. Interestingly, bioaccessibility (0.01–0.25%) was not influenced by the cereal matrix. The four steryl ferulate species, which were detected in wheat, showed similar hydrolysis rates during digestion. As baking had a significant impact on the steryl ferulate content of wheat, we suggest that both raw and processed sources should be considered further in vitro, animal or human trials, when studying the metabolic fate of steryl ferulates.     

Quinoa flavonoids and their bioaccessibility during in vitro gastrointestinal digestion

This research evaluated the effect of in vitro gastrointestinal digestion on the bioaccessibility of flavonoid compounds present in quinoa products (seeds, sprouts, and flakes) and changes in their antioxidant activity. Flavonoids that were present after treatment with gastric and intestinal enzymes were characterized using HPLC-ESI-MS³, and the antioxidant activity was measured using two different antioxidant assays. Seven out of eleven flavonoid compounds were found intact after in vitro digestion, and their concentration were significantly increased after in vitro gastrointestinal digestion. The increase in flavonoids led to an approximately 2-fold increase in the antioxidant activity. The main class of flavonoid compounds identified in quinoa were flavonols, mainly quercetin and kaempferol glycosides. The results suggest that flavonoid compounds from quinoa products, which are associated with health benefits, are bioaccessible after efficient extraction by the digestive enzymes, and are available for further absorption in the intestine.     

Analysis of bioaccessibility of campesterol,stigmasterol, and β-sitosterol in maize by in vitro digestion method

The content of campesterol, stigmasterol, and β-sitosterol in raw and cooked Big Flint Maize (BFM), Pop Corn Maize (PCM) and Red Maize (RM) were analyzed by using a chemical method and using in vitro digestion of cooked samples. RM contained 15.14, 8.36, and 72.19 mg/100 g campesterol, stigmasterol, and β-sitosterol, respectively, which were higher compared to in BFM and PCM. Samples cooking resulted in 6–48% increase in phytosterol content, while cooked RM had the highest phytosterol content. Using the in vitro digestion method, 44–61% reduction in phytosterol content was observed in contrast to the chemical method. The difference bioaccessible campesterol and β-sitosterol contents among varieties was insignificant, except for stigmasterol in RM. The bioaccessibility of campesterol was 44–56% higher than those of stigmasterol and β-sitosterol, and it was higher in BFM and PCM. In conclusion, phytosterol contents estimated in different maize samples using in vitro digestion can be another determinative for the ultimate content and reveals the necessity to consider with chemical method of determination.     

In vitro digestion kinetics and bioaccessibility of starch in cereal food products

The study of starch digestion in cereal-based products is essential since the extent and rate of hydrolysis affects the glycemic index associated with several food-related diseases. Besides, a unique matrix is usually studied and many variations of in vitro techniques (e.g. enzymes, pH, food:enzymes ratio) are selected, then comparison of results became difficult. The recently published INFOGEST in vitro static method with international consensus (Minekus et al., 2014) was applied to several cereal-based products to address whether it is suitable for analysis of starch hydrolysis kinetics. Bread, pasta and cookies were selected taking into account the analysis of different cereal matrixes, including gluten-free products. White bread presented the highest in vitro starch hydrolysis (87%) showing significant differences compared to gluten-free bread (76.5%). Refined flour sheeted pasta (72.6%), whole-wheat extruded pasta (92.0%) and gluten-free pasta (54.3%) showed differences in the extent and rate of hydrolysis. Cookie samples presented the lowest starch hydrolysis (∼45%). The starch availability was estimated by the dializability method, which measures the maltose equivalents dialyzed after simulating digestion. Starch dializability was 35%, 25% and 15% on average for bread, pasta and cookie samples respectively, with positive correlation with rapidly digested starch. The tested in vitro method allowed discriminating the effect of different processing techniques, product types and formulation of the three most common cereal-food matrixes in starch digestion.     

Carotenoid bioaccessibility from whole grain and decorticated yellow endosperm sorghum porridge

Sorghum is a staple crop and a potential dietary source of carotenoids in semi-arid regions of Africa, but information on the bioavailability of these pigments is limited. This study aimed at exploring the effects of agronomic manipulation on sorghum carotenoid contents at selected stages of kernel development and maturation and assessing carotenoid bioaccessibility from matured yellow-endosperm sorghum varieties (P88 and P1222), by comparing porridge made from sorghum whole and decorticated milled grains. Carotenoid content of sorghum milled fractions ranged from 2.90 to 7.22 mg/kg in P88 unbagged decorticated flour, at 50 and 30 days after half bloom (DAHB) respectively, to 9.87-13.69 mg/kg in bagged decorticated bran fractions in P88, at 50 and 30 DAHB respectively. Maize milled fractions were significantly (P < 0.05) higher in carotenoid content than all sorghum products. Bagging increased sorghum carotenoid content by 8-184% vs. unbagged panicles. Carotenoid bioaccessibility was generally higher from sorghum (63-81%) compared to maize (45-47%). Micellarization of xanthophylls (75%) was more efficient than carotenes (52%) in sorghum, while they were similar in maize (40-49%). These results suggest that the higher bioaccessibility of sorghum carotenoids combined with efforts to enhance sorghum carotenoid content may allow for sorghum to provide similar levels of bioaccessible carotenoid pigments as common yellow maize.     

Effect of different soaking conditions on inhibitory factors and bioaccessibility of iron and zinc in pearl millet

Pearl millet was decorticated to obtain a bran rich and endosperm rich fraction. The two fractions were soaked in solutions with varying pH. Pearl millet grains were germinated and steamed followed by decortication to obtain two fractions. It was observed that bran rich fractions contained high concentrations of iron, zinc, polyphenols, phytic acid, fibre and flavonoids. Soaking for short duration of 3 h did not result in major mineral losses but decreased the inhibitory factors which depended on the pH. Alkaline soaking decreased flavonoid content by 62.7% in the endosperm rich fraction, while acidic soaking decreased phytic acid content to the maximum in the bran rich fraction. Combination of treatments like germination and heat decreased the phytate content to the maximum in the endosperm rich fraction. Acidic conditions improved zinc bioaccessibility in the bran rich fraction (35%) and iron bioaccessibility (2.5%) in the endosperm rich fraction. Bran rich fraction from germinated grain also had enhanced bioaccessibility of both the minerals but comparatively lesser when compared to soaking under acidic conditions. Soaking the grain components under slightly less than neutral conditions also decreased some of the inhibitory factors and improved the zinc bioaccessibility to some extent in the bran rich fraction.     

Micellarisation of Carotenoids from Raw and Cooked Vegetables

The efficiency of carotenoid micellarisation from plant foods can be used as an effective tool for the initial screening of carotenoid bioavailability. Therefore, the objectives of the present study were to assess the effects of cooking on the micellarisation of beta-carotene, lycopene, beta-cryptoxanthin and lutein from courgette (zucchini), red pepper and tomato; and, to a minor extent, investigate uptake of lutein by Caco-2 cells from micellar fractions obtained from raw and cooked courgettes. Both raw and cooked vegetables were subjected to an in vitro digestion procedure. beta-Carotene levels were significantly decreased in the digesta from each vegetable after boiling, grilling, microwave-cooking, or steaming, however all of the cooking methods enhanced beta-carotene transfer to micelles. Carotenoid micellarisation ranged from 1.7% to 100% depending on the food, carotenoid, and the cooking method tested. Grilling and microwave-cooking were generally the most detrimental on the transfer of xanthophyll carotenoids, namely beta-cryptoxanthin, to the micelles. Caco-2 cells absorbed greater amounts of lutein from the micelles of microwave-cooked courgettes than those that were raw, boiled, grilled, or steamed. Depending on the cooking methods used, carotenoid retention as well as micellarisation varied for each carotenoid among the different vegetables and different carotenoids present in each particular food.