Iron dissociates from the NaFeEDTA complex prior to or during intestinal absorption in rats

Sodium iron ethylenediaminetetraacetate (NaFeEDTA) has superior iron bioavailability especially in foods containing iron absorption inhibitors. However, mechanisms involved in the absorption and subsequent partitioning of iron complexed with EDTA are poorly understood. Our objectives were to compare retention and tissue distribution of iron administered to rats either as FeSO4 or NaFeEDTA, either orally (OR) or subcutaneously (SC). Weanling rats were fed semipurified diets supplemented with either FeSO4 or NaFeEDTA for 7 days. They were then given a meal containing 59Fe-labeled FeSO4 or NaFeEDTA, or they were injected SC with these two forms of radiolabeled Fe. 59Fe retention was measured by whole body counting. Urine was collected and counted at 24 h intervals throughout the counting period. Tissue samples were analyzed for nonheme iron and 59Fe activity. Absorption of iron from FeSO4 or NaFeEDTA was similar (57.7 and 53.4%, respectively). Seventy-seven percent of the injected Na59FeEDTA was excreted in the urine within 24 h, whereas only 0.5, 0.8, and 1.4% of the injected 59FeSO4, oral 59FeSO4, and oral Na59FeEDTA, respectively, was excreted in the urine. The nonheme iron content was lower in the liver and spleen, by 56.8 and 28.4%, respectively, among rats consuming the NaFeEDTA diet as compared to rats fed FeSO4. We conclude that iron is dissociated from EDTA prior to or during intestinal absorption and that some fraction of the dissociated EDTA is absorbed separately from the iron.     

Tissue iron distribution and urinary mineral excretion vary depending on the form of iron (FeSO4 or NaFeEDTA) and the route of administration (oral or subcutaneous) in rats given high doses of iron

Sodium iron ethylenediaminetetraacetate (NaFeEDTA) has considerable promise as an iron fortificant in food. However, effects of administering high levels of NaFeEDTA on tissue iron distribution and mineral excretion are not well understood. The objectives of this study were to assess nonheme iron distribution in the body and urinary excretion of Ca, Mg, Cu, Fe, and Zn after daily administration of high levels of iron to rats over 21 days. Iron was either given orally with food or injected subcutaneously, as either FeSO 4 or NaFeEDTA. Selected tissues were collected for nonheme iron analysis. Estimated total body nonheme iron levels were similar in rats fed NaFeEDTA or FeSO 4, but the tissue distribution was different: it was 53% lower in the liver and 86% higher in the kidneys among rats fed NaFeEDTA than among those fed FeSO 4. In contrast, body nonheme iron was 3.2-fold higher in rats injected with FeSO 4 than in rats injected with NaFeEDTA. Administering NaFeEDTA orally elevated urinary Cu, Fe, and Zn excretion compared with FeSO 4 (1.41-, 11.9-, and 13.9-fold higher, respectively). We conclude that iron is dissociated from the EDTA complex prior to or during intestinal absorption. A portion of intact FeEDTA may be absorbed via a paracellular route at high levels of intake but is mostly excreted in the urine. Metal-free EDTA may be absorbed and cause elevated urinary excretion of Fe, Cu, and Zn.     

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.     

Studies on water transport through the sweet cherry fruit surface. 7. Fe3+ and Al3+ reduce conductance for water uptake

The effects of the chloride salts LiCl, CaCl(2), MgCl(2), AlCl(3), EuCl(3), and FeCl(3) and the iron salts FeCl(2), FeCl(3), Fe(NO(3))(3), FeSO(4), and Fe(2)(SO(4))(3) on water conductance of exocarp segments (ES) and rates of water uptake into detached sweet cherry fruit (Prunus avium L. cv. Adriana, Early Rivers, Namare, Namosa, and Sam) were studied. ES were excised from the cheek of mature fruit and mounted in stainless steel diffusion cell; water penetration was monitored gravimetrically from donor solutions containing the above mineral salts into a PEG 6000 (osmolality = 1.14 osM, pH 4.8, 25 degrees C) receiver solution. Conductance of ES was calculated from the amount of water taken up per unit of surface area and time by dividing by the gradient in water activity across ES. LiCl, CaCl(2), MgCl(2), FeCl(2), and FeSO(4) had no significant effect on conductance, but AlCl(3), FeCl(3), Fe(NO(3))(3), and Fe(2)(SO(4))(3) significantly reduced conductance compared to water only as a donor. Also, EuCl(3) lowered conductance; however, this effect was not always significant. Effects of salts on water conductance of ES and rates of water uptake into detached fruit were closely related (R 2 = 0.97***). Upon application of an FeCl(3)-containing donor conductance decreased instantaneously. FeCl(3) concentrations of <6.6 x 10(-)(4) M had no effect on conductance, but concentrations at or above this threshold decreased conductance. FeCl(3) lowered water conductance at a receiver pH of 4.8, but not at pH < or =2.6. The effect of FeCl(3) on conductance was largest in cv. Namare and smallest in cv. Adriana. There was no significant effect of FeCl(3) on conductance for transpiration. Formation of aluminum and iron oxides and hydroxides in the exocarp as a result of a pH gradient between donor and receiver solution is discussed as the potential mechanism for Fe(3+) and Al(3+) reducing conductance for water uptake.     

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.     

Remobilization of iron from primary leaves of bean plants grown at various iron levels

In order to study the effect of different growth rates of the shoot apex, i.e. shoot demand, on the remobilization of iron (Fe) from mature (primary) leaves, bean (Phaseolus vulgaris L.) plants were precultured with 8x10^‐5 M FeEDTA for four days. Thereafter, plants were grown for another six days at various levels of Fe (0.0, 1.0, and 10.0μM FeEDTA), and simultaneously treated with or without shading of one primary leaf. Dry weight increment of the shoot apex decreased with decreased Fe in the nutrient solution. Shading of one primary leaf decreased total dry weight of plants irrespective of Fe supply, but increased the dry weight of the shoot apex of plants supplied without Fe or with only 1.0μM Fe. In these plants, the concentration of chlorophyll and Fe in the shoot apex corresponded with the treatment effects on dry weight of the shoot apex. Shading induced senescence of the shaded leaf, decreased the content of “active Fe”; (extractable in dilute acid), and also enhanced the remobilization of Fe and copper (Cu) from the shaded leaf. The remobilization of Fe from primary leaves was not related to the severity of chlorosis in the shoot apex (the Fe demand of sink tissue), indicating that only a certain fraction of the total Fe in mature leaves can be remobilized.     

Distribution pattern of root‐supplied 59iron in iron‐sufficient and iron‐deficient bean plants

In order to study the iron (Fe) distribution pattern in bean plants with different Fe nutritional status, french bean (Phaseolus vulgaris L.) seedlings were precultured in a complete nutrient solution with 8x10^‐5 M FeEDTA for five days. Thereafter, plants were further supplied with 8x10^‐5 M FeEDTA (Fe‐sufficient) or with only 2x10^‐6 M FeEDTA (Fe‐deficient) for another eight days. At this stage, the Fe‐deficient plants had much lower chlorophyll contents and lower dry weight of the leaves but higher reducing capacity of the roots compared with the Fe‐sufficient plants. For studies on short‐term distribution of Fe, the Fe‐sufficient plants were supplied 8x10^‐5 M ⁵⁹FeEDTA (specific activity 9.9 GBq/mol) and the Fe‐deficient plants 1x10⁶ M ⁵⁹FeEDTA (specific activity 98.8 GBq/mol). The plants were harvested after 4 and 24 hours. Despite a much lower supply of ⁵⁹FeEDTA/(factor 80), the Fe‐deficient plants took up significantly more ⁵⁹Fe but translocated less to the shoots (14.6% after 24 h) compared with the Fe‐sufficient plants (29.4% after 24 h). However, regardless of the Fe nutritional status of the plants, the majority of ⁵⁹Fe was translocated in the primary leaves. Our results demonstrate a similar distribution patterns of root‐derived ⁵⁹Fe in the shoots of Fe‐sufficient and Fe‐deficient plants, and thus, no preferential direct translocation of Fe to the shoot apex in the Fe‐deficient plants.     

Iron bioavailability of hemoglobin from soy root nodules using a Caco-2 cell culture model

Heme iron has been identified in many plant sources-most commonly in the root nodules of leguminous plants, such as soy. Our objective was to test the effectiveness of soy root nodule (SRN) and purified soy hemoglobin (LHb) in improving iron bioavailability using an in vitro Caco-2 cell model, with ferritin response as the bioavailability index. We assessed bioavailability of iron from LHb (either partially purified (LHbA) or purified (LHbD)) with and without food matrix and compared it with that from bovine hemoglobin (BHb), ferrous sulfate (FeSO4), or SRN. Bioavailability of each treatment was normalized to 100% of the FeSO4 treatment. When iron sources were tested alone (100 ug iron/mL), ferritin synthesis by LHbD and BHb were 19% (P > 0.05) and 113% (P < 0.001) higher than FeSO4, respectively. However, when iron sources were used for fortification of maize tortillas (50 ppm), LHbA and BHb showed similar bioavailability, being 27% (P < 0.05) and 33% (P < 0.05) higher than FeSO4. Heat treatment had no effect on heme iron but had a significant reduction on FeSO4 bioavailability. Adding heme (LHbA) iron with nonheme (FeSO4) had no enhancement on nonheme iron absorption. Our data suggest that heme iron from plant sources may be a novel value-added product that can provide highly bioavailable iron as a food fortificant.     

缺铁水稻根表铁膜对硒的转运和吸收的影响

Under anaerobic conditions, ferric hydroxide deposits on the surface of rice roots and affects uptake and translocation of certain nutrients. In the present study, rice plants were cultured in Fe-deficient or sufficient solutions and placed in a medium containing selenium （Se） for 2 h. Then, FeSO4 was added at the various concentrations of 0, 10, 40, or 70 mg L-1 to induce varying levels of iron plaque on the root surfaces and subsequent uptake of Se was monitored. The uptake of Se was inhibited by the iron plaque, with the effect proportional to the amount of plaque induced. The activity of cysteine synthase was decreased with increasing amounts of iron plaque on the roots. This may be the important reason for iron plaque inhibition of Se translocation. At each level of iron plaque, Fe-deficient rice had more Se than Fe-sufficient rice. Furthermore, with plaque induced by 20 mg Fe L-1, plants from Fe-deficient media accumulated more Se than those from Fe-sufficient media, as the Se concentration was increased from 10 to 30 or 50 mg L^-1. We found that phytosiderophores, highly effective iron chelating agents, could desorb selenite from ferrihydrite. Root exudates of the Fe-deficient rice, especially phytosiderophores in the exudates, could enhance Se uptake by rice plants with iron plaque.     

Iron Sources Effects on Growth,Physiological Parameters and Nutrition of Cacao

Productivity and sustainability of cacao (Theobroma cacao L.) in tropical soils are affected by levels of iron. Information is lacking on the cacao response to various sources of iron (Fe). A greenhouse experiment was conducted to evaluate the effects of five iron sources iron sulfate heptahydrate, ferric ethylenediamine-N,N’-bis(2-hydroxyphenylacetic acid), ferric diethylenetriaminepentaacetic acid, ferric ethylenediaminetetraacetic acid, fiesta herbicide (FeSO₄ · 7H₂O, FeEDDHA, FeDTPA, FeEDTA,) at 10 mg Fe kg^?1 soil on growth, photosynthesis, content of photosynthetic pigments and starch and macro- and micronutrient nutrition of cacao. The various iron sources had significant effects on shoot and root dry biomass accumulation, leaf chlorophyll a and b content, carotenoid levels, SPAD index and P_N. These parameters were significantly correlated with concentration, uptake, influx, and transport and use efficiency of Fe. In cacao net photosynthesis, stomatal conductance, internal carbon dioxide (CO₂)_, and transpiration in leaf level responded differently to the sources of Fe. Invariably, macro and micronutrient uptake, influx, transport, and use efficiency showed differential responses to sources of iron but significant effects were only observed for copper (Cu), Fe, manganese (Mn), and zinc (Zn). Overall, FeDTPA, FeEDTA and FeHEDTA could be the best sources of Fe in improving, growth, photosynthesis and macro and micro nutrition of cacao.     

不同供磷水平对饭豆体内铁有效性的影响

采用溶液培养试验研究了低铁条件下（1 μmol/L FeEDTA）不同供磷水平P 3、30和300 μmol/L对饭豆叶绿素含量、生物量、铁含量以及质外体铁的影响。结果表明,饭豆叶片叶绿素含量及根系干重均随磷处理浓度的增加而显著降低; 低磷处理的植株地上部的铁含量明显高于中磷和高磷处理。随着供磷水平的增加,地上部和根系总铁量的比值呈降低趋势,说明铁由根系向地上部的转运显著减少,从而加剧了植株缺铁症状。进一步分析发现,低磷处理的根系质外体铁含量显著低于中磷和高磷处理。说明在铁吸收过程中,供磷水平增加促使铁在根系质外体空间中的固定,不利于根系中的铁转运至地上部,这可能是磷是对铁产生拮抗作用造成植物铁营养不利的原因之一。     

Enrichment of higher molecular weight fractions in inulin

Inulin (general formulas GFn and Fm, with G = anhydroglucose and F = anhydrofructose) naturally occurs as a homologous series of oligo- and polysaccharides with different chain lengths. For reasons of growing interest in the food and pet food industries, the short chain inulins have to be separated from their long chain analogues because their properties (digestibility, prebiotic activity and health promoting potential, caloric value, sweetening power, water binding capacity, etc.) differ substantially. To study these properties in relation to the number average degree of polymerization (DPn), ultrafiltration, specific crystallization from aqueous solution, and precipitation from solvent/water mixtures were used to enrich native chicory and dahlia inulin in the higher molecular weight fractions. Depending on the membrane module used, the DPn of chicory inulin (DPn = 8.1) and dahlia inulin (DPn = 29) could be increased by ultrafiltration to a maximum value of, respectively, 22 and 43. With crystallization from aqueous solutions (25 degrees C), similar results were obtained but at a much higher yield. Finally, long chain inulin could be precipitated from aqueous solutions in the presence of high concentrations of methanol, ethanol, and acetone. Acetone demonstrated to be the best solvent system to increase the DPn, followed by ethanol and methanol. However, for safety reasons and food purposes, ethanol was evaluated to be the best choice. With ethanol, the DPn could be raised to 25 for chicory inulin and up to 40 for dahlia inulin.     

Iron tissue storage and hemoglobin levels of deficient rats repleted with iron bound to the caseinophosphopeptide 1-25 of beta-casein.

Caseinophosphopeptides (CPP) issued from enzyme digestion of caseins bind cations and keep them soluble in the digestive tract. They could be used as ligands to improve iron (Fe) bioavailability. Fe-deficient young rats were repleted with Fe (40 or 200 mg/kg of diet) bound either to the beta-CN (1-25) of beta-casein or to whole beta-casein or as FeSO(4). A control pair-fed group was given 200 mg of Fe (FeSO(4))/kg of diet for 6 weeks. After repletion, hemoglobin concentration of the control group was reached only by the ) animals fed 200 mg of Fe/kg; beta-CN (1-25) bound Fe (40 and 200 mg) produced higher Fe liver and spleen stores than FeSO(4). Binding Fe to the whole, nonhydrolyzed beta-casein gave results intermediate between the other experimental groups. Binding Fe to phosphoserine residues of low molecular weight CPP improved its ability to cure anemia and to restore iron tissue stores, as compared to Fe bound to the whole casein and to inorganic salts.     

Effect of bread baking on the bioavailability of hydrogen-reduced iron powder added to unenriched refined wheat flour

Elemental iron powders are widely used to fortify flour and other cereal products. Our objective was to test the hypothesis that baking enhances the bioavailability of elemental iron powders by oxidizing Fe(0) to Fe(2+) or Fe(3+). An in vitro digestion/Caco-2 cell culture model and a piglet model were used to measure bioavailability. Bread flour, either unfortified or fortified with hydrogen-reduced (HR) iron powder or FeSO(4) (300 mg Fe/kg flour), was baked into bread. For the in vitro studies, bread samples were treated with pepsin at pH 2, 3, 4, 5, 6, or 7 and subsequently incubated with pancreatic enzymes at pH 7 in a chamber positioned above monolayers of cultured Caco-2 cells. Ferritin formation in the cells was used as an index of iron bioavailability. Ferritin formation in cells fed HR Fe bread was similar to cells fed FeSO(4) bread when the peptic digestion was conducted at a pH 2 but lower when the peptic phase was conducted at pH 3, 4, 5, 6, or 7 (P < 0.05). Pig diets containing 35% dried bread were prepared and fed to cross-bred (Hampshire x Landrace x Yorkshire) anemic pigs in two studies. The rate of increase in hemoglobin Fe over the feeding period was used to calculate relative biological value (RBV), an index of iron bioavailability. In the first pig study, RBV of HR Fe added to flour prior to baking was 47.9% when compared to FeSO(4) fortified flour (P < 0.05). In the second pig study, a third treatment consisting of unfortified bread with HR iron added during diet mixing (after bread baking) was included. RBVs of the HR Fe diet (Fe added after baking) and HR Fe diet (Fe added before baking) were 40.1% and 53.5%, respectively, compared to the FeSO(4) diet. Differences in RBV between the HR Fe (before and after baking) and FeSO(4) (before baking) treatment groups were significant, but the difference between the before and after HR treatment groups was not significant. We conclude that bread baking does not enhance the bioavailability of elemental iron powders.     

Effect of different nitrogen‐forms and iron‐chelates on the development of stinging nettle

The development of stinging nettle (Urtica dioica L.) grown on culture solution containing with either ammonium or nitrate ions, or urea, was investigated under iron deficiency conditions, and with added FeEDTA or FeCto. Both seed‐cultured and vegetatively‐cultured stinging nettle plants produced normally developed green shoots when nitrate and 4 μM FeEDTA or FeCto were supplied. Stinging nettle plants were able to utilize Fe‐citrate, Fe‐ascorbate, and Fe‐malate effectively at the same concentration as well. When K3Fe(CN)6 was supplied, which is impermeable to the plasmalemma, and therefore is used to measure the reductive capacity of the roots, stinging nettle plants became chlorotic because the complex was stable at the pH of the culture solution. Urea did not induce chlorosis but inhibited growth. The plants died when ammonium was supplied as a sole N source. Applying bicarbonate and ammonium together prevented the plants from dying, but the plants became chlorotic. Total exclusion of iron from the culture solution resulted in iron‐deficiency stress reactions as has been described for other dicotyledonous plants (Strategy II).     

Analysis of Some Divalent Metal Contents in Tobacco Expressing the Exogenous Soybean Ferritin Gene

Abstract

T2 tobacco lines overexpressing soybean ferritin in the plastids (+TPs) or apoplasm (AFs) under the regulation of CaMV 35S promoter were grown on MS nutrient solution. After 1 month growth, statuses of six major divalent‐metals (Ca, Cu, Fe, Mg, Mn, and Zn) were measured in leaves and roots. Both +TPs and AFs showed enhanced growth (max. 1.7×) in leaves than the control line. The Fe contents in leaves of +TPs and AFs were significantly larger (1.9–2.8×) than that of the control line. The other metal contents in leaves of +TPs and AFs were almost the same as or less than those of the control line. In contrast to the result of leaves, the growth enhancement in roots was not clear in +TPs, but in AFs. Also, some of the non‐ferrous metal contents in roots of +TPs and AFs were dramatically increased compared with those of the control line (Mn, 1.9–10.4×; Zn, 1.6–2.3×), whereas the differences in content of Fe, Cu, Ca, and Mg were insignificant. These results demonstrated that the ferritin overexpression in apoplasm was as effective for inducing Fe accumulation as that in plastid. Under the normal metal‐balanced condition, even if the activation of Fe uptake related enzymes leads to the accumulation of non‐specific accumulation of divalent metal ions in roots, an Fe loading/unloading system and/or an internal translocator in xylem and phloem might specifically deliver Fe to the upper part of plants.     

Iron transport in plants: Future research in view of a plant nutritionist and a molecular biologist

Iron is attractive to plant physiologists since J. Sachs has proven in 1868 the essentiality and the possible leaf uptake of Fe. It lasted about 100 years before the principal processes for Fe mobilization in the rhizosphere were discovered and classified as two distinct strategies for Fe acquisition. During the 80's and 90's of the last century the uptake of Fe²⁺ and Fe^III-phytosiderophores by specific transporters in strategy I- and strategy II-plants, respectively, were postulated without any application of the new approaching molecular techniques. In the following decade, the various transporters for Fe uptake by roots, such as AtIRT1 in Arabidopsis or ZmYS1 in maize and their possible regulation were characterized. In the following years with fast developing molecular approaches further Fe trans ortsrs were genetically described with often only vague physiological functions. In view of a plant nutritionist, besides uptake processes by roots, the following transport processes within the respective target tissue have to be considered by molecular biologists in more detail: 1) radial transfer of Fe from the root cortex through the endodermis, 2) xylem loading in roots, 3) transfer of Fe from xylem to phloem via transfer cells, 4) phloem loading with Fe in source leaves and retranslocation to sink organs, and 5) remobilization and retranslocation via the phloem during senescence of perennial plants. The importance of these various specific transport processes for a well-regulated Fe homeostasis in plants and new strategies to identify and characterize proteins involved in Fe transport and homeostasis will be discussed.     

Haematological response to haem iron or ferrous sulphate mixed with refried black beans in moderately anaemic Guatemalan pre-school children

OBJECTIVE: Combating iron deficiency in toddlers with iron-fortified food has proved difficult in countries with phytate-rich diets. For this purpose, a new haem iron preparation was developed. The study compared changes in iron status after administration of refried beans with beans fortified with a haem iron preparation or ferrous sulphate (FeSO4). DESIGN: In a masked, stratified-randomised intervention trial, children received five 156-g cans of refried black beans per week for 10 consecutive weeks. The beans-only (control), FeSO4 and haem iron groups were offered a cumulative dose of 155 mg, 1625 mg and 1700 mg of iron from the bean intervention, respectively. Haemoglobin (Hb) and ferritin concentrations were determined at baseline and after 5 and 10 weeks. Compliance was examined weekly. SETTING: A low-income community in Guatemala City. SUBJECTS: One hundred and ten children aged 12-36 months with initial Hb values between 100 and 115 g l(-1). RESULTS: The cumulative intake of beans was approximately 80% of that offered, signifying an additional approximately 1300 mg of either haem or inorganic iron in the corresponding treatment groups over 10 weeks. Hb concentrations increased by the order of 7.3-11.4 g l(-1) during the intervention, but without significant differences across treatments. Average ferritin concentrations were unaffected by treatment assignment. However, post hoc analysis by subgroups of initial high ferritin and initial low ferritin found the Hb increments after 10 weeks in the haem iron group (13.1+/-7.7 g l(-1)) to be significantly greater than the respective increases (6.8+/-11.2 and 6.4+/-8.5 g l(-1)) in the inorganic iron and beans-only groups. CONCLUSIONS: Canned refried beans are a candidate vehicle for fortificant iron. Given the improved colour and organoleptic properties imparted by haem iron added to refried beans, its additional potential for benefiting the iron status of consumers with iron deficiency may recommend it over FeSO4.     

Moving toward a more physiological model: application of mucin to refine the in vitro digestion/Caco-2 cell culture system

The objective of this study was to determine if a combination of commercially available mucin and an 8 microm microporous membrane insert can be used to replace the 15 kDa molecular weight cutoff (MWCO) dialysis membrane used in an established in vitro digestion/Caco-2 cell culture system. Although the current model with the 15 kDa membrane correlates well with human studies, use of mucin may improve the system as the mucus layer is suspected to play a physiological role in Fe absorption. Use of mucin may also enable more complete assessment of iron bioavailability from large molecular weight forms of Fe such as heme and ferritin Fe. A range of foods or Fe (i.e., FeCl(3) +/- ascorbic acid, cooked beef, red bean, white bean, soybean, horse spleen ferritin and plant-type ferritin) were subjected to in vitro digestion. In the presence of mucin, significantly more Fe was taken up from the heme Fe (86%) and ferritin (91%) samples and significantly less Fe was taken up from the white bean samples ( approximately 70%) relative to the 15 kDa membrane. The results indicated that the forms of iron interact with mucin. The mucus layer has a significant effect on Fe uptake. Further refinement and characterization of the mucin method is needed before it can be deemed to be a suitable replacement for the dialysis membrane.     

Rates of cuticular penetration of chelated Fe(III): role of humidity, concentration, adjuvants, temperature, and type of chelate

Time courses of cuticular penetration of FeCl3 and Fe(III) complexes of citric acid, EDTA, EDDHA (Sequestrene 138Fe), imidodisuccinic acid (IDHA), and ligninsulfonic acid (Natrel) were studied using astomatous cuticular membranes (CMs) isolated from Populus x canescens leaves. At 100% relative humidity, the Fe(III) chelates disappeared exponentially with time from the surface of the CMs; that is, penetration was a first-order process that can be described using rate constants or half-times of penetration (t(1/2)). Half-times ranged from 20 to 30 h. At 90% humidity, penetration rates were insignificant with the exception of Natrel, for which t(1/2) amounted to 58 h. Rate constants were independent of temperature (15, 25, and 35 degrees C). Permeability decreased with increasing Fe chelate concentration (IDHA and EDTA). At 100% humidity, half-times measured with FeIDHA were 11 h (2 mmol L(-1)), 17 h (10 mmol L(-1)) and 36 h (20 mmol L(-1)), respectively. In the presence of FeEDTA, penetration of CaCl2 was slowed greatly. Half-times for penetration of CaCl2, which were 1.9 h in the absence of FeEDTA, rose to 3.12 h in the presence of an equimolar concentration of EDTA and 13.3 h when the FeEDTA concentration was doubled. Hence, Fe chelates reduced permeability of CMs to CaCl2 and to the Fe chelates themselves. It is suggested that Fe chelates reduced the size of aqueous pores. This view is supported by the fact that rate constants for calcium salts were about 5 times higher than for Fe chelates with the same molecular weights. Adding Tween 20 (5 g L(-1)) as a humectant did not increase permeability to FeIDHA at 90% humidity and below, while addition of glycine betaine did. Penetration of FeCl3 applied at 5 g L(-1) (pH 1.5) was not a first order process as rate constants decreased rapidly with time. Only 2% of the dose penetrated during the first 2 h and less than that in the subsequent 8 h. Recovery was only 70%. This was attributed to the formation of insoluble Fe hydroxide precipitates on CMs. These results explain why in the past foliar application of Fe compounds had limited success. Inorganic Fe salts are instable and phytotoxic because of low pH, while Fe chelates penetrate slowly and 100% humidity is required for significant penetration rates. Concentrations as low as reasonably possible should be used. These physical facts are expected to apply to stomatous leaf surfaces as well, but absolute rates probably depend on leaf age and plant species. High humidity in stagnant air layers may favor penetration rates across stomatous leaf surfaces when humidity in bulk air is below 100%.