Iron uptake by Caco-2 cells from NaFeEDTA and FeSO4: Effects of ascorbic acid, pH, and a Fe(II) chelating agent

Sodium iron(III) ethylenediaminetetraacetate (NaFeEDTA) has considerable promise as an iron fortificant because of its high bioavailability in foods containing iron absorption inhibitors. In this study, uptakes of iron from NaFeEDTA, FeSO4, and FeCl3 by Caco-2 cells were compared in the absence or presence of ascorbic acid (AA), an iron absorption enhancer; at selected pH levels; and in the absence or presence of an iron absorption inhibitor, bathophenanthroline disulfonic acid (BPDS). Ferritin formation in the cells was used as the indicator of iron uptake. Uptake from all three Fe sources was similar in the absence of AA. Adding AA at a 5:1 molar excess as compared to Fe increased uptake by 5.4-, 5.1-, and 2.8-fold for FeSO4, FeCl3, and NaFeEDTA, respectively. The smaller effect of AA on uptake from NaFeEDTA may be related to the higher solubility of NaFeEDTA and/or the strong binding affinity of EDTA for Fe3+, which may prevent AA and duodenal cytochrome b from effectively reducing EDTA-bound Fe. Uptake was inversely related to the pH of the media over a range of 5.8-7.2. Because uptake by DMT-1 is proton-coupled, the inverse relationship between pH and Fe uptake in all three iron sources suggests that they all follow the DMT-1 pathway into the cell. Adding BPDS to the media inhibited uptake from all three iron compounds equally. Because BPDS binds Fe2+ but not Fe3+ and because only Fe2+ is transported by DMT-1, the finding that BPDS inhibited uptake from NaFeEDTA suggests that at least some iron dissociates from EDTA and is reduced just as simple inorganic iron at the brush border membrane of the enterocyte. Taken together, these results suggest that uptake of iron from NaFeEDTA by intestinal enterocytes is regulated similarly to uptake from iron salts.     

Inhibition of iron uptake by phytic acid, tannic acid, and ZnCl2: studies using an in vitro digestion/Caco-2 cell model.

The objective of this study was to document the effects of phytic acid, tannic acid, and zinc on iron uptake in an in vitro digestion/Caco-2 cell culture model. The effects of phytic acid and tannic acid on iron uptake were measured at increasing molar ratios of FeCl3 to phytic acid or tannic acid. Maximal inhibition of iron uptake by phytic acid occurred at a 1:10 ratio of Fe to phytic acid. Dialyzable Fe decreased with a low Fe to phytic acid ratio but increased with Fe:phytic acid ratios greater than 1:3 indicating that more iron was soluble at higher phytic acid levels but less available. As in human studies, heme iron was less inhibited by phytic acid than nonheme iron. Tannic acid was a more potent inhibitor of nonheme iron uptake, as maximal inhibition (97.5%) of iron uptake occurred at a ratio of 1:1 or less. The addition of ZnCl2 to the digest at ratios of 1:0.5 and 1:1 decreased iron uptake by 57 and 80%, respectively. Overall, the results agree qualitatively with studies in humans and demonstrate the relative effects of these compounds on iron uptake in this model system. This study provides key information for determining iron availability under more complex meal conditions.     

Kinetics of Fe(III) reduction by ascorbic acid in aqueous solutions

The reaction of Fe(III) and ascorbic acid (AA) in food products and digestive tracts affects the efficiency and uptake of these two nutrients. We investigated the kinetics of Fe(III) reduction by AA at pH 5 and 6 in a model system at 25 degrees C. The results indicate that the reduction of Fe(III) by AA is of zero order with respect to AA. The reaction order with respect to Fe(III) cannot be represented by a simple kinetic model at pH 5 or 6. The major stage of the reduction (about 80%, stoichiometrically), however, could be represented by a general equation of -d[Fe(III)]/dt = k[Fe(III)],(1. 811) where k is a rate constant and [Fe(III)] is the total ferric concentration. The rate constant decreased 1 order of magnitude as pH increased from 5 to 6. Aging of Fe(III) solution slows its reduction rate at pH 6 but not at pH 5.     

Fenton's oxidation of food processing wastewater components. Kinetic modeling of protocatechuic acid degradation

The oxidation of protocatechuic acid (PA), a typical phenol-type compound present in food processing wastewater, has been carried out by means of Fenton's reagent. Both the H2O2 and Fe(II) initial concentrations increase the PA degradation rate. Temperature also enhances the PA conversion when raised from 283 to 313 K, a further increase to 323 K results in a lower PA removal. Increasing the PA initial concentration leads to a decrease of conversion values but an opposite effect in terms of removal rate. pH values in the range 3-4 resulted in the total inhibition of the oxidation process. Similar PA depletion rates were experienced regardless of the oxidation state of the catalyst (ferrous or ferric iron). Additionally, an attempt based on the classic Fenton's chemistry plus some other stages accounting for the Fe(II) regeneration from Fe(III) and the inefficient H2O2 decomposition was conducted to model the process.     

基于近红外光谱的沼液挥发性脂肪酸含量快速检测

挥发性脂肪酸（Volatile Fatty Acids,VFA）作为厌氧发酵过程的重要中间产物,其在厌氧反应器中的累积能够反映出产甲烷菌的不活跃状态或厌氧发酵条件的恶化。为了实现对农牧废弃物厌氧发酵进行过程分析和状态监控,将近红外光谱（Near Infrared Spectroscopy,NIRS）与偏最小二乘（Partial Least Squares,PLS）相结合构建玉米秸秆和畜禽粪便厌氧发酵液乙酸、丙酸和总酸含量快速检测模型。将竞争自适应重加权采样法（Competitive Adaptive Reweighted Sampling,CARS）与遗传模拟退火算法（Genetic Simulated Annealing algorithm,GSA）相结合构建CARS-GSA算法对沼液中的乙酸、丙酸和总酸进行特征波长优选,原始光谱数据1 557个波长点经预处理和波长优选后,得到乙酸、丙酸和总酸特征波长变量分别为135、101和245个,建立的回归模型验证决定系数分别为0.988、0.923和0.886,预测均方根误差（Root Mean Squared Error of Prediction,RMSEP）分别为0.111、0.120和0.727,相对分析误差分别为9.685、3.685和3.484,与全谱建模相比RMSEP分别减少了17.78%、15.49%和1.22%,能够满足农牧废弃物厌氧发酵过程发酵液中乙酸和丙酸含量的快速检测需求,基本满足总酸的检测需求。结果表明,通过构建CARS-GSA算法优选乙酸、丙酸和总酸的敏感波长变量,参与建模的波长点数量显著减少,有效降低了变量维度和模型复杂度,提升了回归模型检测精度和预测能力,为快速准确检测沼液VFA提供了新途径。     

Iron derivatives from casein hydrolysates as a potential source in the treatment of iron deficiency

The properties of an Fe(3+)-peptide complex containing 5.6% Fe, obtained by the reaction of ferric chloride with an enzymatic hydrolysate of casein, are described. The major site of iron binding corresponds primarily to the carboxylate groups and to a lesser extent to the peptide bonds. The Fe(3+)-peptide complex is insoluble at acid pH and completely soluble at neutral to alkaline pH. When soluble, the Fe(3+) is tightly bound to the complex peptide mixture but can be displaced and complexed by a low molecular weight ligand such as cysteine. Its efficacy in relation to iron sulfate was compared in rats. Both iron sources were administrated in Milli-Q water by gastric gavage to male Wistar rats (180-200 g) after an 18 h fast with water ad libitum. Fe(3+) from the Fe(3+)-peptide complex was transferred to the blood in a dose-dependent manner (1-8 mg of Fe/kg), and the serum iron levels were significantly higher (p < 0.001) than in a similar group of rats treated with iron sulfate. In the comparative kinetics experiments, the rats received 4 mg of Fe/kg. Both iron sources presented maximum absorption, as indicated by the elevation of serum iron levels, 30 min after administration, and the AUC(0)(-->2h) of the Fe(3+)-peptide complex was significantly higher (p < 0.05) than that observed with iron sulfate. The simultaneous administration of free peptides (0-192 mg) with the Fe(3+)-peptide complex or iron sulfate did not modify the extent of absorption of iron from both sources, suggesting that the absorption is due to the complex formed and probably not to exchange reactions in the gastrointestinal tract. In the hemoglobin repletion experiments carried out on newly weaned rats with anemia induced by a low-iron diet, supplementation of the diet with the the Fe(3+)-peptide complex was as efficient as supplementation with iron sulfate in the conversion from diet to hemoglobin iron. These results, taken together, suggest that the Fe(3+)-peptide complex is a potential compound for use as an iron source in biological situations.     

Beneficial effects of humic acid on micronutrient availability to wheat

Humic acid (HA) is a relatively stable product of organic matter decomposition and thus accumulates in environmental systems. Humic acid might benefit plant growth by chelating unavailable nutrients and buffering pH. We examined the effect of HA on growth and micronutrient uptake in wheat (Triticum aestivum L.) grown hydroponically. Four root-zone treatments were compared: (i) 25 micromoles synthetic chelate N-(4-hydroxyethyl)ethylenediaminetriacetic acid (C10H18N2O7) (HEDTA at 0.25 mM C); (ii) 25 micromoles synthetic chelate with 4-morpholineethanesulfonic acid (C6H13N4S) (MES at 5 mM C) pH buffer; (iii) HA at 1 mM C without synthetic chelate or buffer; and (iv) no synthetic chelate or buffer. Ample inorganic Fe (35 micromoles Fe3+) was supplied in all treatments. There was no statistically significant difference in total biomass or seed yield among treatments, but HA was effective at ameliorating the leaf interveinal chlorosis that occurred during early growth of the nonchelated treatment. Leaf-tissue Cu and Zn concentrations were lower in the HEDTA treatment relative to no chelate (NC), indicating HEDTA strongly complexed these nutrients, thus reducing their free ion activities and hence, bioavailability. Humic acid did not complex Zn as strongly and chemical equilibrium modeling supported these results. Titration tests indicated that HA was not an effective pH buffer at 1 mM C, and higher levels resulted in HA-Ca and HA-Mg flocculation in the nutrient solution.     

Organic acids influence iron uptake in the human epithelial cell line Caco-2

It has previously been suggested that organic acids enhance iron absorption. We have studied the effect of nine organic acids on the absorption of Fe(II) and Fe(III) in the human epithelial cell line Caco-2. The effect obtained was dose-dependent, and the greatest increase (43-fold) was observed for tartaric acid (4 mmol/L) on Fe(III) (10 micromol/L). Tartaric, malic, succinic, and fumaric acids enhanced Fe(II) and Fe(III) uptake. Citric and oxalic acid, on the other hand, inhibited Fe(II) uptake but enhanced Fe(III) uptake. Propionic and acetic acid increased the Fe(II) uptake, but had no effect on Fe(III) uptake. Our results show a correlation between absorption pattern and chemical structure; e.g. hydroxyl groups, in addition to carboxyls, were connected with a positive influence. The results may be important for elucidating factors affecting iron bioavailability in the small intestine and for the development of foods with improved iron bioavailability.     

Lactic acid decreases Fe(II) and Fe(III) retention but increases Fe(III) transepithelial transfer by Caco-2 cells

Lactic acid (LA) has been proposed to be an enhancer for dietary iron absorption, but contradictory results have also been reported. In the present study, fully differentiated Caco-2 cell monolayers were used to evaluate the effects of LA (1-50 mmol/L) on the cellular retention and transepithelial transport of soluble non-heme iron (as ferric nitrilotriacetate). Our data revealed a linear decline in Fe(III) retention with respect to the concentration of LA added. In the presence of 50 mmol/L LA, retention of Fe(III) and Fe(II) decreased 57% and 58%, respectively. In contrast, transfer of Fe(III) across the cell monolayer was doubled, while Fe(II) transfer across the cell monolayer decreased 35%. We conclude that LA reduces cellular retention and transepithelial transport of Fe(II) by Caco-2 cells in a dose-dependent manner. However, while LA also reduces retention of Fe(III) by Caco-2 cells, the transfer of Fe(III) across cell monolayers is enhanced, possibly due to effects on paracellular transport.     

Isolated glycosaminoglycans from cooked haddock enhance nonheme iron uptake by Caco-2 cells

This study continues previous research to confirm that glycosaminoglycans (GAGs) exert a positive effect on promoting iron uptake by Caco-2 cells. Cooked haddock was digested with papain, and GAGs were further purified on the basis of their sulfur content. Reverse phase chromatography (RP-HPLC) and digestion with chondroitinase ABC (Chase) (50 mU/mg) were used to approach the identification of the GAGs. FeCl 3 was mixed with the purified GAGs, and Fe uptake was measured by ferritin formation using an in vitro digestion/Caco-2 cell model. The identificative analyses suggest that chondroitin/dermatan sulfate-related structures promote Fe uptake by Caco-2 cells; however, this effect was lower (40%) than that observed with whole fish muscle. Chase eliminated the positive effect on Fe uptake. These results indicate that specific GAGs may contribute to the enhancing effect of meat on Fe absorption. Further in vivo studies addressing these aspects of the meat factor are needed.     

Comparison of iron uptake from reduced iron powder and FeSO4 using the Caco-2 cell model: effects of ascorbic acid, phytic acid, and pH

The reduced iron powder has considerable potential for use as an iron fortificant because it does not change organoleptically during storage or food preparation for cereal flour, and its bioavailability is scarcely influenced by iron absorption inhibitors in foods. The objective of this article is to study the effects of ascorbic acid, phytic acid, and pH on iron uptake from reduced iron powder (43 microm) and FeSO 4, and to compare iron bioavailability of reduced iron powders among four selected granularity levels. The cell ferritin formation is used as a marker of iron uptake. Obviously, iron uptake of reduced iron powder is increased with decreasing of powder granularity and is much lower than FeSO 4 when the size is above 43 microm, but significantly higher at 40-60 nm. In the presence of ascorbic acid or phytic acid, Caco-2 cell iron absorption from reduced iron powder (43 microm) is significantly higher than that from FeSO 4. And iron uptake of Caco-2 cells is decreased with increasing of pH from 5.5 to 7.5. Moreover, the decrease trend is more obvious for reduced iron powder than for FeSO 4. Our results indicated that iron bioavailability of reduced iron powder by intestinal enterocytes is similar to that of iron salts, and reduced iron powder is more excellent than FeSO 4 as food fortificant, especially at ultramicroscopic granularity.     

Short‐term effects of pH and aluminium on mineral nutrition in maize varieties differing in proton and aluminium tolerance

In short‐term (24 h) nutrient solution experiments, the influence of different proton (pH 6.0 and pH 4.3) and aluminium (Al) (0, 20, and 50 μM) concentrations on root and coleoptile elongation, dry weight, and the uptake of selected mineral nutrients was studied in maize (Zea mays L.) varieties that differ in acid soil tolerance under field conditions. The acid‐soil‐tolerant maize varieties, Adour 250 and C525M, proved to be hydrogen (H⁺) ion sensitive, but Al tolerant, while the acid soil tolerant variety BR201F was H⁺ tolerant but Al sensitive. The acid soil sensitive variety HS 7777 was affected by both H⁺ and Al toxicity. The proton‐induced inhibition of root elongation was closely related to the proton‐induced decrease of the specific absorption rates (SAR) of boron (B), iron (Fe), magnesium (Mg), calcium (Ca), and phosphorus (P). In contrast, only the specific absorption rate of B (SARB) was significantly correlated to the Al‐induced inhibition of root elongation. It is concluded, that alterations of nutrient uptake may play an important role in H⁺ toxicity, while at least after short‐term exposure to Al, alterations of Ca, Fe, Mg, or P uptake do not seem to be responsible for Al‐induced inhibition of root elongation. Further attention deserves the Al‐B interaction, moreover taking into account that a highly significant correlation between Al‐induced increase of callose concentration in root tips and Al‐induced decrease of SAR_B could be established.     

The Bioavailability of Copper and Mercury to the Common Nettle (Urtica Dioica) and the Earthworm Eisenia Fetida from Contaminated Dredge Spoil

The contaminants Hg and Cu, as well as Fe, Mn and K were sequentially extracted from upland disposed dredge spoil using DTPA and 10% nitric acid. Concentrations of these metals in aerial plant tissue and roots of Urtica dioica growing on the dredge spoil were also determined and used to correlate the biological absorption coefficients (BACs) and mobile element absorption coefficients (MACs) with soil extractable metals. DTPA extractions were most suitable for prediction of aerial plant tissue uptake of Cu, Mn and K whilst total Hg and Fe soil concentrations were correlated with plant root BACs and MACs. A laboratory bioassay using Eisenia fetida was also used to assess the potential biological uptake of the contaminants. Both Hg and Cu were accumulated by the worms, but interpretation of the results was hampered by the inherent difficulties of such active biomonitoring.     

Effects of phosphate carriers,iron, and indoleacetic acid on iron nutrition and productivity of peanut on a calcareous soil

Results of a field experiment designed to assess the effects of phosphate carriers, iron (Fe), and indoleacetic acid (IAA) on the Fe nutrition of peanut grown on a calcareous soil showed that single superphosphate (SSP) was more effective than diammonium phosphate (DAP) in improving Fe nutrition and chlorophyll synthesis. Increased phosphorus (P) and Fe contents of chlorotic leaves showing symptoms of Fe deficiency suggested that Fe, despite absorption and uptake, was subjected to inactivation, and that the Fe content per se was not the cause of the observed chlorosis. Better amelioration of chlorosis with the SSP treatment as compared with DAP indicated a role of sulphur (S) in preventing inactivation of Fe, possibly caused by excessive P accumulation. A foliar spray of Fe‐EDDHA corrected the chlorosis, but a ferric citrate foliar treatment did not. This further suggested that the mobility of Fe was impaired in chlorotic plants. An IAA foliar spray only also tended to improve Fe nutrition. Significant increase in peanut productivity was observed following improvement in Fe nutrition both with soil and foliar treatments.     

Nitrogen assimilation studies using 15N in soybean plants treated with imazethapyr, an inhibitor of branched-chain amino acid biosynthesis

The pattern of nitrogen assimilation in soybean plants treated with a herbicide that inhibits branched-chain amino acid biosynthesis was evaluated by (15)N isotopic analysis. The herbicide imazethapyr caused a strong decrease in nitrate uptake by roots, partly due to a reduced stomatal conductance. The inhibition of (15)N uptake was accompanied by a decrease in the (15)N content in the plant and, concomitantly, an inhibition of translocation to the shoot. Imazethapyr inhibited nitrate reductase activity in leaves and roots. Among all parameters studied, "de novo" synthesis of proteins was the first parameter of the N assimilation metabolism affected by the herbicide. These results show that this class of herbicides totally damages N metabolism and indicates a regulatory effect on N uptake and translocation that would be mediated by the increase in free amino acid pool provoked by the inhibition of branched-chain amino acid biosynthesis.     

Phenolic acids are absorbed from the rat stomach with different absorption rates

The intestinal absorption characteristics of phenolic acids (PAs) have been elucidated in terms of their affinity for the monocarboxylic acid transporter (MCT). Recently, the involvement of the stomach has been implicated in the absorption of polyphenols. The present work demonstrates that the gastric absorption efficiency of each PA is apparently different between various PAs. Various PAs with different affinities for MCT were administered (2.25 mumol) to rat stomach, and then the plasma concentration of the PA was measured. The plasma concentration of ferulic acid (FA) peaked 5 min after administration in the stomach. At 5 min after administration, the plasma concentration of each PA increased in the order: gallic acid = chlorogenic acid < caffeic acid < p-coumaric acid = FA. This order matches their respective affinity for MCT in Caco-2 cells, which we have demonstrated in previous studies. These results indicated that MCT might be involved in the gastric absorption of PAs, similar to the intestinal absorption.     

Red grape juice inhibits iron availability: application of an in vitro digestion/caco-2 cell model

Adequate bioavailable Fe intake is essential for optimal growth and intellectual development of infants and children. Fruit juices are nutritious and popular drinks for infants and children and are known to contain Fe uptake inhibitors (e.g., polyphenolic compounds) and a dominant promoter, ascorbic acid. Ascorbic acid is naturally present in fruit juices and is added during processing to almost all juices found in supermarkets. With these facts taken into account, an in vitro digestion/Caco-2 cell culture model was developed to compare the effects of apple, pear, white grape, red grape, prune, grapefruir, and orange juices on iron bioavailability. In two series of experiments, juices from a local supermarket were combined with FeCl(3) or commercial infant cereal fortified with elemental iron and subject to simulated gastric and intestinal digestion. Caco-2 cell ferritin formation in response to exposure to the digests served as the measure of Fe uptake. The pear, apple, grapefruit, orange, and white grape juice significantly increased Fe bioavailability from FeCl(3). For the infant cereal studies, the apple, orange, pear, and white grape juices increased the Fe bioavailability of the infant cereal. In contrast, the red grape juice and prune juice had profound inhibitory effects on iron bioavailability. These inhibitory effects were likely due to high levels of polyphenolic compounds that bind and thereby prevent absorption of soluble Fe. These inhibitory compounds appeared to counteract the promotional effects of ascorbic acid as they were in considerable molar excess relative to ascorbic acid and Fe in the digest. From a nutritional standpoint, the results suggest that individuals in need of optimal Fe absorption should avoid red grape and prune juice or at least vary the types of juices consumed. Alternatively, individuals seeking to limit Fe uptake (e.g., hemochromatitics and astronauts) may be able to utilize red grape or prune juice as effective inhibitors of Fe uptake. Consumers should be aware that the compounds that inhibit Fe availability are also linked to anticancer benefits; thus, a dietary balance of the above juices may be optimal.     

Interactions between grape anthocyanins and pyruvic acid, with effect of pH and acid concentration on anthocyanin composition and color in model solutions.

The formation of vitisin A, an anthocyanin formed naturally in small quantities in maturing port wines, was studied in model wine solutions at a range of pH values (2.0-4.5) and pyruvate concentrations [molar ratios of pyruvic acid to total anthocyanins (PA/TA) ranging from 12.20 to 172.40]. Additionally, the effect of vitisin A formation on the color changes of these model wines was evaluated. Vitisin A was formed through the interaction between malvidin 3-glucoside and pyruvic acid, and vitisin A in acylated forms, having the 6-position of the sugar acylated with acetic acid (3-acetylvitisin A) and p-coumaric acid (3-p-coumarylvitisin A), formed through the interaction between pyruvic acid and malvidin 3-acetylglucoside and malvidin 3-p-coumarylglucoside, respectively; their identities were confirmed by spectral analysis and FABMS. The maximum formation of these new anthocyanin derivatives was at pH 2. 7-3.0, at the higher pyruvic acid concentration (PA/TA of 172.40 units). The vitisins A caused changes in the color of the solution and expressed about 11 times (pH 3) to 14 times (pH 2) more color than the normal anthocyanins. On aging, the model solutions changed from a bluish red, attributable to the main anthocyanins present, to a slightly more orange red, attributable to the vitisin compounds. The aged models containing vitisins A were all much redder than the more red-brown color of the models aged without pyruvic acid.     

Nutritional Composition of Iron,Zinc, Calcium,and Phosphorus in Wheat Grain Milling Fractions as Affected by Fertilizer Nitrogen Supply

Fe and Zn deficiencies are global nutritional problems. N supply could increase Fe and Zn concentrations in wheat grain. This study was conducted to determine the impacts of different N rates (0, 122, 174, and 300 kg/ha) on the distribution and speciation of Fe and Zn in wheat grain milling fractions under field conditions. Zn and protein concentrations were increased, whereas Fe was less affected in the flour fractions with increasing N rates. Further analysis with size‐exclusion chromatography coupled with inductively coupled plasma mass spectrometry revealed that Fe and Zn bound to low‐molecular‐weight (LMW) compounds in the flour fractions (probably Fe‐nicotianamine [NA], Fe‐deoxymugineic acid, or Zn‐NA) were less affected by increasing N supply, representing 3.5–10.9% of total Fe and 2.5–56.6% of total Zn. In the shorts fraction, LMW‐Fe was absent, and LMW‐Zn with higher N supply was over twice as high as that in control and 3–27 times as high as that in the other milling fractions. In the flour fractions, the molar ratios of phytic acid (PA)/Fe and PA/Zn (both less than 30.5) decreased, whereas soluble LMW‐Fe/Zn was not affected with increasing N rates.