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Background: A highly soluble iron-casein complex has been developed for food fortification purposes with the aim to provide high iron bioavailability., Objective: We aimed to determine the iron bioavailability of the iron-casein complex relative to that of ferrous sulfate (control) when given with whole milk in healthy young women., Methods: A randomized comparator-controlled trial with a crossover design was conducted using the erythrocyte incorporation dual stable isotope (57Fe, 58Fe) technique. Iron absorption from the iron-casein complex was compared with that from ferrous sulfate in 21 healthy women aged 20-38 y with normal iron status., Results: Fractional iron absorption (geometric mean; -SD, +SD) from the iron-casein complex (3.4%; 1.4%, 5.4%) and from ferrous sulfate (3.9%; 1.7%, 6.1%) were not statistically different (P > 0.05). The relative bioavailability value of the iron-casein complex to ferrous sulfate was determined to be 0.87 (-1 SD, +1 SD: -0.90, +2.64)., Conclusions: The iron-casein complex has iron bioavailability comparable to that of ferrous sulfate in healthy young women. This trial was registered at www.anzctr.org.au as ACTRN12615000690550., (Copyright © American Society for Nutrition 2019.)

Background: Calcium inhibits and ascorbic acid (AA) enhances iron absorption from iron-fortified foods. Absorption efficiency depends on iron status, although the interaction is unclear., Objective: We investigated the ability of AA to overcome calcium-induced inhibition of iron absorption in children differing in iron status., Methods: The effect of calcium (0, 100, and 200 mg/test meal) on iron absorption in the absence and presence of AA (0, 42.5, and 85 mg/test meal) from a casein/whey-based drink fortified with ferrous sulfate was assessed in a series of randomized crossover studies both in iron-replete (IR) Indian schoolchildren and in children with iron deficiency anemia (IDA) (6-11 y; n = 14-16/group) by using stable isotopes., Results: In the absence of calcium and AA, iron absorption from the casein/whey-based drink was 20% lower in IR children than in children with IDA. The addition of calcium reduced mean iron absorption by 18-27%, with the effect being stronger for high added calcium (P < 0.01). AA at a 2:1 or 4:1 molar ratio enhanced iron absorption by a factor of 2-4 and greatly overcompensated for the inhibitory effect of calcium on iron absorption in a dose-dependent manner (P < 0.001). The dose-response effect tended to be stronger (P < 0.1) in the IDA group, and iron status was of far less influence on iron absorption than the enhancing effect of AA., Conclusion: When adding AA to iron-fortified milk products, care should be taken not to provide absorbable iron in excess of needs., (© 2014 American Society for Nutrition.)

The iron storage protein ferritin is a potential vehicle to enhance the iron content of biofortified crops. With the aim of evaluating the potential of ferritin iron in plant breeding, we used species-specific isotope dilution mass spectrometry to quantify ferritin iron in bean varieties with a wide range of total iron content. Zinc, phytic acid, and polyphenols were also measured. Total iron concentration in 21 bean varieties ranged from 32 to 115 ppm and was positively correlated with concentrations of zinc (P = 0.001) and nonferritin bound iron (P < 0.001). Ferritin iron ranged from 13% to 35% of total iron and increased only slightly in high iron beans (P = 0.007). Concentrations of nonferritin bound iron and phytic acid were correlated (P = 0.001), although phytic acid:iron molar ratio decreased with increasing iron concentration (P = 0.003). Most iron in high iron beans was present as nonferritin bound iron, which confirms our earlier finding showing that ferritin iron in beans was lower than previously published. As the range of ferritin iron content in beans is relatively narrow, there is less opportunity for breeders to breed for high ferritin. The relevance of these findings to the extent of iron absorption depends on resolving the question of whether ferritin iron is absorbed or not to a greater extent than nonferritin bound iron., (© 2014 Institute of Food Technologists®)

Ferritin is a hollow sphere protein composed of 24 subunits that can store up to 4500 iron atoms in its inner cavity. It is mainly found in the liver and spleen but also in serum at trace levels. Serum ferritin is considered as the best single indicator in assessing body iron stores except liver or bone marrow biopsy. However, it is confounded by other disease conditions. Ferritin bound iron (FBI) and ferritin saturation have been suggested as more robust biomarkers. The current techniques for FBI determination are limited by low antibody specificity, low instrument sensitivity and possible analyte losses during sample preparation. The need for a highly sensitive and reliable method is widely recognized. Here we describe a novel technique to detect serum FBI using species-specific isotope dilution mass spectrometry (SS-IDMS). [(57)Fe]-ferritin was produced by biosynthesis and in vitro labeling with the (57)Fe spike in the form of [(57)Fe]-citrate after cell lysis and heat treatment. [(57)Fe]-ferritin for sample spiking was further purified by fast liquid protein chromatography. Serum ferritin and added [(57)Fe]-ferritin were separated from other iron species by ultrafiltration followed by isotopic analysis of FBI using negative thermal ionization mass spectrometry. Repeatability of our assay is 8% with an absolute detection limit of 18 ng FBI in the sample. As compared to other speciation techniques, SS-IDMS offers maximum control over sample losses and species conversion during analysis. The described technique may therefore serve as a reference technique for clinical applications of FBI as a new biomarker for assessing body iron status.

Brain iron accumulation is supposed to play a central role in neurodegeneration by inducing oxidative stress. Currently it is unknown to which extent iron entering brain over lifetime exchanges with body iron or if uptake of iron is unidirectional without significant efflux from brain. To study brain iron dynamics in vivo, up to three stable isotope tracers were fed continuously with a standard rodent diet up to 5 months to healthy adult male Wistar rats (n = 8) in a staggered design. Brain iron uptake was found to be bi-directional but iron influx and efflux were unbalanced leading inevitably to brain iron accumulation over time. Brain iron turnover was found to be very low at a half-life of ca. 9 months for tracer iron entering brain. Observed tracer accumulation in brain iron can be extrapolated to an increase of brain iron by ca. 30% in the healthy rats from early adulthood to the end of their lives. In contrast to current beliefs that brain uptake of dietary iron is negligible during adulthood following short-term radiotracer studies, our long-term feeding experiments point to a possible role of the diet in brain iron accumulation and, subsequently, neurodegeneration.

We recently showed in an animal model that iron isotopic composition varies substantially between different organs. For instance, iron in ferritin-rich organs--such as the major storage tissues liver, spleen, and bone marrow--contain a larger fraction of the heavy iron isotopes compared with other tissues, including blood. As a consequence, partitioning of body iron into red blood cells and storage compartments should be reflected in the isotopic pattern of blood iron. To confirm this hypothesis, we monitored blood iron isotope patterns in iron-overloaded subjects undergoing phlebotomy treatment by multicollector inductively coupled plasma mass spectrometry. We found that bloodletting and consequential replacement of lost blood iron by storage iron led to a substantial increase of the heavy isotope fraction in the blood. The progress of iron depletion therapy and blood loss was quantitatively traceable by isotopic shifts of as much as +1‰ in δ((56)Fe). These results show that--together with iron absorption efficiency--partitioning of iron between blood and iron storage tissues is an important determinant of blood iron isotopic patterns, which could make blood iron isotopic composition the first composite measure of iron metabolism in humans.

Ferritin is nature's predominant iron storage protein. The molecule consists of a hollow protein shell composed of 24 subunits which is capable of storing up to 4500 iron atoms per molecule. Recently, this protein has been identified as a target molecule for increasing iron content in plant staple foods in order to combat dietary iron deficiency, a major public health problem in developing countries. Here, we present a novel technique for quantification of ferritin-bound iron in edible plant seeds using species-specific isotope dilution mass spectrometry (IDMS) by means of a biosynthetically produced (57)Fe-labeled ferritin spike and negative thermal ionization mass spectrometry (NTIMS). Native plant ferritin and added spike ferritin were extracted in 20 mM Tris buffer (pH 7.4) and separated by anion exchange chromatography (DEAE Sepharose), followed by isotopic analysis by thermal ionization mass spectrometry. The chosen IDMS approach was critically evaluated by assessing the (i) efficiency of analyte extraction, (ii) identical behavior of spike and analyte, and (iii) potential iron isotope exchange with natural iron. Repeatabilities that can be achieved are on the order of <5% RSD for quintuplicate analyses at an absolute detection limit of 60 ng of ferritin-bound iron for plant seeds. Studies in six different legumes revealed ferritin-iron contents ranging from 15% of total iron in red kidney beans up to 69% in lentils.

Biofortification of staple foods with iron in the form of ferritin-iron is a promising approach to fighting iron-deficiency anemia in developing countries. However, contradictory results regarding iron bioavailability to humans from ferritin are not yet fully clarified. Furthermore, the question has been raised whether ferritin can potentially survive gastric passage intact and be absorbed via a ferritin-specific uptake mechanism. We studied changes of ferritin-iron and protein during cooking and in vitro gastric digestion. Water soluble, native ferritin-iron, measured in different legumes, represented 18% (soybeans) up to maximally 42% (peas) of total seed iron. Ferritin-iron was no longer detectable after boiling the legumes for 50 min in excess water. When the same cooking treatment was applied to recombinant bean ferritin propagated in Escherichia coli, some ferritin-iron remained measurable. During in vitro gastric digestion of recombinant bean ferritin and red kidney bean extract, ferritin-iron was fully released from the protein and dissolved at pH 2. Stability tests at varying pH at 37 degrees C showed that the release of ferritin-iron starts at pH 5 and is complete at pH 2. We concluded that ferritin-iron is efficiently released from the ferritin molecule during cooking and at gastric pH and that it should be absorbed as efficiently as all other nonheme iron in food.

Meat is a well-known enhancer of iron absorption, yet the molecular entity mediating the effect remains obscure. Recently published data indicate that highly acidic sulfated glycosaminoglycans (GAG) from fish and chicken muscle are effective stimulants of iron uptake in Caco-2 cells. Two fully randomized stable isotope studies with crossover design were performed in a group of 16 apparently healthy young women to assess the effect of purified sulfated and unsulfated GAG on human iron absorption. Iron absorption was measured on the basis of erythrocyte incorporation of (57)Fe or (58)Fe 14 d after the administration of labeled semisynthetic meals (SSM) based on egg albumin, corn oil, maltodextrin, and water. The meals were consumed with or without added sodium hyaluronate (NaH, 300 mg) or chondroitin sulfate (CS, 360 mg) as representative unsulfated and sulfated GAG, respectively. The level of GAG added was 3 times (NaH) to about 10 times (CS), the amount expected to be present in 150 g beef muscle. Geometric mean iron absorption from SSM containing NaH (21.2%) or CS (19.4%) did not differ from that of SSM without GAG (19.5 and 20.3%, respectively). NaH and CS at those levels do not affect human nonheme iron absorption.

Chili and turmeric are common spices in indigenous diets in tropical regions. Being rich in phenolic compounds, they would be expected to bind iron (Fe)(3) in the intestine and inhibit Fe absorption in humans. Three experiments were conducted in healthy young women (n = 10/study) to assess the effect of chili and turmeric on Fe absorption from a rice-based meal containing vegetables and iron fortified fish sauce in vivo. Iron absorption was determined by erythrocyte incorporation of stable isotope labels ((57)Fe/(58)Fe) using a randomized crossover design. Addition of freeze-dried chili (4.2 g dry powder, 25 mg polyphenols as gallic acid equivalents) reduced Fe absorption from the meal by 38% (6.0% with chili vs. 9.7% without chili, P = 0.0017). Turmeric (0.5 g dry powder, 50 mg polyphenols as gallic acid equivalents) did not inhibit iron absorption (P = 0.91). A possible effect of chili on gastric acid secretion was indirectly assessed by comparing Fe absorption from acid soluble [(57)Fe]-ferric pyrophosphate relative to water soluble [(58)Fe]-ferrous sulfate from the same meal in the presence and absence of chili. Chili did not enhance gastric acid secretion. Relative Fe bioavailability of ferric pyrophosphate was 5.4% in presence of chili and 6.4% in absence of chili (P = 0.47). Despite the much higher amount of phenolics in the turmeric meal, it did not affect iron absorption. We conclude that both phenol quality and quantity determine the inhibitory effect of phenolic compounds on iron absorption.

Background: Although ferric pyrophosphate is a promising compound for iron fortification of foods, few data are available on the effect of food matrices, processing, and ascorbic acid on its bioavailability., Objective: We compared the relative bioavailability (RBV) of ferrous sulfate in an experimental form of micronized dispersible ferric pyrophosphate (MDFP) in a wheat-milk infant cereal given with and without ascorbic acid with the RBV of MDFP from a processed and unprocessed rice meal., Design: A crossover design was used to measure iron absorption in young women (n = 26) from test meals fortified with isotopically labeled [57Fe]-MDFP and [58Fe]-ferrous sulfate, based on erythrocyte incorporation of stable isotope labels 14 d later., Results: Geometric mean iron absorption from the wheat-based meal fortified with MDFP was 2.0% and that from the meal fortified with ferrous sulfate was 3.2% (RBV = 62). The addition of ascorbic acid at a molar ratio of 4:1 to iron increased iron absorption from MDFP to 5.8% and that from ferrous sulfate to 14.8% (RBV = 39). In the rice meals, mean iron absorption from MDFP added to the rice at the time of feeding was 1.7%, and that from ferrous sulfate was 11.6% (RBV = 15). The mean iron absorption from MDFP extruded into artificial rice grains was 3.0% and that from ferrous sulfate in unprocessed rice was 12.6% (RBV = 24). Sixteen of 26 subjects were iron deficient. Iron status was a highly significant predictor of the RBV of MDFP (P < 0.001)., Conclusion: RBV of the experimental MDFP varied markedly with food matrix and iron status. Assigning a single RBV value to poorly soluble compounds may be of limited value in evaluating their suitability for food fortification.

Background: Nonheme-iron absorption requires an acidic milieu. Reduced gastric acid output as a consequence of Helicobacter pylori infection could be an important limiting factor for iron absorption., Objective: We measured gastric acid output and iron absorption from a non-water-soluble iron compound (ferrous fumarate) and a water-soluble iron compound (ferrous sulfate) in children with and without H. pylori infection., Design: Gastric acid output was quantified before (basal acid output, or BAO) and after pentagastrin stimulation (stimulated acid output, or SAO) in 2-5-y-old children with iron deficiency anemia who were (n = 13) or were not (n = 12) infected with H. pylori. Iron absorption was measured by using a double-stable-isotope technique. H. pylori-infected children were studied before and after eradication therapy., Results: BAO and SAO were significantly lower in the H. pylori-infected children (0.2 +/- 0.2 and 1.6 +/- 0.9 mmol/h, respectively) than in the uninfected children (0.9 +/- 0.7 and 3.1 +/- 0.9 mmol/h, respectively; P = 0.01 and P < 0.005). BAO and SAO improved to 0.8 +/- 1.3 and 3.3 +/- 2.4 mmol/h, respectively, after therapy. Iron absorption from ferrous sulfate was significantly greater than that from ferrous fumarate both before (geometric : 19.7% compared with 5.3%; P < 0.0001) and after (22.5% compared with 6.4%; P < 0.0001) treatment in H. pylori-infected children. Corresponding values for uninfected children were 15.6% and 5.4%, respectively (P < 0.001; n = 12)., Conclusions: Iron absorption from ferrous fumarate was significantly lower than that from ferrous sulfate in both H. pylori-infected and uninfected Bangladeshi children. Treatment of H. pylori infection improved gastric acid output but did not significantly influence iron absorption. The efficacy of ferrous fumarate in iron fortification programs to prevent iron deficiency in young children should be evaluated.

The effects of added ascorbic acid and particle size on iron absorption from ferric pyrophosphate were evaluated in adult women (9-10 women/study) based on erythrocyte incorporation of iron stable isotopes (57Fe or 58Fe) 14 days after administration. Three separate studies were made with test meals of iron-fortified infant cereal (5 mg iron/meal) and the results are presented as geometric means and relative bioavailability values (RBV, FeSO4 = 100%). The results of study 1 showed that iron absorption was significantly lower from ferric pyrophosphate (mean particle size 8.5 microm) than from FeSO4 in meals without ascorbic acid (0.9 vs. 2.6%, p < 0.0001, RBV 36%) and in the same meals with ascorbic acid added at a 4:1 molar ratio relative to fortification iron (2.3 vs. 9.7%, p < 0.0001, RBV 23%). Ascorbic acid increased iron absorption from ferric pyrophosphate slightly less (2.6-fold) than from FeSO4 (3.7-fold) (p < 0.05). In studies 2 and 3, RBV of ferric pyrophosphate with an average particle size of 6.7 microm and 12.5 pm was not significantly different at 52 and 42% (p > 0.05), respectively. In conclusion, the addition of ascorbic acid increased fractional iron absorption from ferric pyrophosphate significantly, but to a lesser extent than from FeSO4. Decreasing the mean particle size to 6.7 microm did not significantly increase iron absorption from ferric pyrophosphate.

Ferric pyrophosphate is a water-insoluble Fe compound used to fortify infant cereals and chocolate-drink powders as it causes no organoleptic changes to the food vehicle. However, it is only of low absorption in man. Recently, an innovative ferric pyrophosphate has been developed (Sunactive Fe trade mark ) based on small-particle-size ferric pyrophosphate (average size 0.3 microm) mixed with emulsifiers, so that it remains in suspension in liquid products. The aim of the present studies was to compare Fe absorption of micronised, dispersible ferric pyrophosphate (Sunactive Fe trade mark ) with that of ferrous sulfate in an infant cereal and a yoghurt drink. Two separate Fe absorption studies were made in adult women (ten women/study). Fe absorption was based on the erythrocyte incorporation of stable isotopes ((57)Fe and (58)Fe) 14 d after the intake of labelled test meals of infant cereal (study 1) or yoghurt drink (study 2). Each test meal was fortified with 5 mg Fe as ferrous sulfate or micronised, dispersible ferric pyrophosphate. Results are presented as geometric means. There was no statistically significant difference between Fe absorption from micronised, dispersible ferric pyrophosphate- and ferrous sulfate-fortified infant cereal (3.4 and 4.1 % respectively; P=0.24) and yoghurt drink (3.9 and 4.2 % respectively; P=0.72). The results of the present studies show that micronised, dispersible ferric pyrophosphate is as well absorbed as ferrous sulfate in adults. The high relative Fe bioavailability of micronised, dispersible ferric pyrophosphate indicates the potential usefulness of this compound for food fortification.

Retinyl palmitate added to Fe-fortified maize bread has been reported to enhance Fe absorption in adult Venezuelan subjects but not in Western Europeans. It is not known to what extent these results were influenced by differences in vitamin A status of the study subjects. The objective of the present study was to evaluate the influence of retinyl palmitate added to Fe-fortified maize porridge on erythrocyte incorporation of Fe in children with vitamin A deficiency, before and after vitamin A supplementation. Erythrocyte incorporation of Fe-stable isotopes was measured 14 d after intake of maize porridge (2.0 mg Fe added as ferrous sulfate) with and without added retinyl palmitate (3.5 micromol; 3300 IU). The study was repeated 3 weeks after vitamin A supplementation (intake of a single dose of 210 micromol retinyl palmitate; 'vitamin A capsule'). Vitamin A status was evaluated by the modified relative dose-response (MRDR) technique. Retinyl palmitate added to the test meal reduced the geometric mean erythrocyte incorporation of Fe at baseline from 4.0 to 2.6 % (P=0.008, n 13; paired t test). At 3 weeks after vitamin A supplementation, geometric mean erythrocyte incorporation was 1.9 and 2.3 % respectively from the test meal with and without added retinyl palmitate (P=0.283). Mean dehydroretinol:retinol molar ratios were 0.156 and 0.125 before and after intake of the single dose of 210 micromol retinyl palmitate; 'vitamin A capsule' (P=0.15). In conclusion, retinyl palmitate added to the labelled test meals significantly decreased erythrocyte incorporation of Fe in children with vitamin A deficiency at baseline but had no statistically significant effect 3 weeks after vitamin A supplementation. The difference in response to retinyl palmitate added to Fe-fortified maize porridge on erythrocyte incorporation of Fe before and after intake of the vitamin A capsule indicates, indirectly, changes in vitamin A status not measurable by the MRDR technique. The lack of conclusive data on the effect of retinyl palmitate on Fe absorption indicates the complexity of the interactions between vitamin A status, dietary vitamin A and Fe metabolism.

Background: Vitamin A and beta-carotene were recently reported to enhance iron absorption by counteracting the inhibitory effect of phytic acid in cereal-based meals and of polyphenol-containing beverages on nonheme-iron absorption in humans., Objective: Our objective was to further evaluate the influence of vitamin A on iron absorption., Design: Iron absorption from corn bread with or without added vitamin A (retinyl palmitate) was determined in 5 studies in young adult human subjects by using either a stable-isotope method (2 studies) or a radioisotope technique (3 studies). Iron absorption was measured by erythrocyte incorporation of the isotopic labels and by whole-body retention of (59)Fe. Corn bread was served with water (studies 1 and 3) or coffee (studies 2, 4, and 5). The studies differed in the amounts and chemical forms of added tracer and fortification iron. The possibility of methodologic artifacts in earlier investigations was evaluated., Results: No effect of vitamin A on iron absorption from the test meals was identified in the individual studies by using paired Student's t test. A slightly negative effect of vitamin A on iron absorption was found with the use of analysis of variance., Conclusions: The previously reported findings of a positive effect of vitamin A on nonheme-iron absorption in humans was not confirmed. Incomplete isotopic equilibration of the tracer with native iron in the meal or with fortification iron cannot explain the previous findings. However, the present study does not exclude the possibility that suboptimal vitamin A status influences the effect of dietary vitamin A on iron absorption.

Isotopic analysis of human blood and liver and muscle tissue indicates that each individual bears a long-term iron (Fe) isotope signature in the blood. Blood and tissue differ slightly in isotopic composition and are depleted by up to 2.6 per mil in 56Fe relative to 54Fe when compared to dietary Fe. The 56Fe/54Fe isotope ratio in the blood of males is, on average, lower by 0.3 per mil than that of females. These results suggest that Fe isotope effects in the blood reflect differences in intestinal Fe absorption between individuals and genotypes.

Non-transferrin bound iron (NTBI) in serum can induce oxidative stress, even when present in minute amounts, due to its ability to catalyze the formation of reactive oxygen species (ROSs). Techniques to accurately quantify NTBI are needed in order to study its nature and possible health effects. To date, no gold standard method for serum NTBI quantification has been established. Existing methods yielded highly divergent results in previous interlaboratory comparisons when compared to each other. Here, we present the development and validation of a novel assay for serum NTBI quantification using isotope dilution mass spectrometry (IDMS) and negative thermal ionization mass spectrometry (NTI-MS) for isotopic analysis. NTBI is not a single species in serum but the fraction of iron not shielded by proteins from chemical reaction. In our assay, an isotopically labelled 57Fe(III) spike chelated by nitrilotriacetic acid (NTA·57Fespk) is mixed with blood during sampling to allow instant capture of NTBI by NTA while vacant binding sites of transferrin are being saturated with the 57Fe spike. NTBI is separated from the serum by ultrafiltration and quantified using an isotopically enriched 58Fe spike. Systematic evaluation of iron binding to NTA revealed potential artifacts in previous assays. Iron supplementation (65 mg elemental iron as ferrous fumarate) induced detectable concentrations of NTBI in serum in four out of six apparently healthy, adult males of Asian ethnicity (0.21 ± 0.14 μmol L−1; p < 0.05). [ABSTRACT FROM AUTHOR]

Purpose: The main purpose of this study was to establish bioavailability data in humans for the new (Fe) fortification compound ferrous ammonium phosphate (FAP), which was specially developed for fortification of difficult-to-fortify foods where soluble Fe compounds cannot be used due to their negative impact on product stability. Methods: A double-blind, randomized clinical trial with cross-over design was conducted to obtain bioavailability data for FAP in humans. In this trial, Fe absorption from FAP-fortified full-cream milk powder was compared to that from ferric pyrophosphate (FPP) and ferrous sulfate. Fe absorption was determined in 38 young women using the erythrocyte incorporation dual stable isotope technique (Fe, Fe). Results: Geometric mean Fe absorption from ferrous sulfate, FAP and FPP was 10.4, 7.4 and 3.3 %, respectively. Fe from FAP was significantly better absorbed from milk than Fe from FPP ( p < 0.0001). Fe absorption from FAP was significantly lower than Fe absorption from ferrous sulfate, which was used as water-soluble reference compound ( p = 0.0002). Absorption ratios of FAP and FPP relative to ferrous sulfate as a measure of relative bioavailability were 0.71 and 0.32, respectively. Conclusions: The results of the present studies show that replacing FPP with FAP in full-cream milk could significantly improve iron bioavailability. [ABSTRACT FROM AUTHOR]

Ferritin is the major iron storage protein in the biosphere. Iron stores of an organism are commonly assessed by measuring the concentration of the protein shell of the molecule in fluids and tissues. The amount of ferritin-bound iron, the more desirable information, still remains inaccessible owing to the lack of suitable techniques. Iron saturation of ferritin is highly variable, with a maximum capacity of 4,500 iron atoms per molecule. This study describes the direct isotopic labeling of a complex metalloprotein in vivo by biosynthesis, in order to measure ferritin-bound iron by isotope dilution mass spectrometry. [57Fe]ferritin was produced by cloning and overexpressing the Phaseolus vulgaris ferritin gene pfe in Escherichia coli in the presence of 57FeCl2. Recombinant ferritin was purified in a fully assembled form and contained approximately 1,000 iron atoms per molecule at an isotopic enrichment of more than 95% 57Fe. We did not find any evidence of species conversion of the isotopic label for at least 5 months of storage at −20 °C. Transfer efficiency of enriched iron into [57Fe]ferritin of 20% was sufficient to be economically feasible. Negligible amounts of non-ferritin-bound iron in the purified [57Fe]ferritin solution allows for use of this spike for quantification of ferritin-bound iron by isotope dilution mass spectrometry. [ABSTRACT FROM AUTHOR]

Meat is a well-known enhancer of iron absorption, yet the molecular entity mediating the effect remains obscure. Recently published data indicate that highly acidic sulfated glycosaminoglycans (GAG) from fish and chicken muscle are effective stimulants of iron uptake in Caco-2 cells. Two fully randomized stable isotope studies with crossover design were performed in a group of 16 apparently healthy young women to assess the effect of purified sulfated and unsulfated GAG on human iron absorption. Iron absorption was measured on the basis of erythrocyte incorporation of 57Fe or 58Fe 14 d after the administration of labeled semisynthetic meals (SSM( based on egg albumin, corn oil, maltodextrin, and water. The meals were consumed with or without added sodium hyaluronate (NaH, 300 mg) or chondroitin sulfate (CS, 360 mg) as representative unsulfated and sulfated GAG, respectively. The level of GAG added was 3 times (NaH) to about 10 times (CS), the amount expected to be present in 150 g beef muscle. Geometric mean iron absorption from SSM containing NaH (21.2%( or CS (19.4%) did not differ from that of SSM without GAG (19.5 and 20.3%, respectively). NaH and CS at those levels do not affect human nonheme iron absorption. [ABSTRACT FROM AUTHOR]

Abstract: Mass-dependent variations in isotopic composition are known since decades for the light elements such as hydrogen, carbon or oxygen. Multicollector-inductively coupled plasma mass spectrometry (MC-ICP-MS) and double-spike thermal ionization mass spectrometry (TIMS) permit us now to resolve small variations in isotopic composition even for the heavier elements such as iron. Recent studies on the iron isotopic composition of human blood and dietary iron sources have shown that lighter iron isotopes are enriched along the food chain and that each individual bears a certain iron isotopic signature in blood. To make use of this finding in biomedical research, underlying mechanisms of isotope fractionation by the human body need to be understood. In this paper available iron isotope data for biological samples are discussed within the context of isotope fractionation concepts and fundamental aspects of human iron metabolism. This includes evaluation of new data for body tissues which show that blood and muscle tissue have a similar iron isotopic composition while heavier iron isotopes are concentrated in the liver. This new observation is in agreement with our earlier hypothesis of a preferential absorption of lighter iron isotopes by the human body. Possible mechanisms for inducing an iron isotope effect at the cellular and molecular level during iron uptake are presented and the potential of iron isotope effects in human blood as a long-term measure of dietary iron absorption is discussed. [Copyright &y& Elsevier]

Background: Fish sauce and soy sauce have been suggested as food vehicles for iron fortification in Asia. NaFeEDTA is a potentially useful fortificant because it can be added to these condiments without causing precipitation during storage. Objectives: The objectives were to evaluate iron absorption from NaFeEDTA-fortified fish sauce and soy sauce against a reference fortificant (FeSO4), to compare iron absorption from NaFeEDTA-fortified fish sauce and soy sauce, and to evaluate the influence of fish sauce and soy sauce per se on iron absorption. Design: Five separate iron-absorption studies were made in adult women (10 women per study). Iron absorption was measured on the basis of erythrocyte incorporation of 57Fe or 58Fe 14 d after the intake of labeled meals of rice or rice and vegetables. Fish sauce or soy sauce (10 g) fortified with 5 mg Fe as NaFeEDTA or FeSO4 was fed with selected meals. The results are presented as geometric means. Results: Iron absorption from NaFeEDTA- and FeSO4-fortified fish sauce (3.3% and 3.1%, respectively) and soy sauce (6.1% and 5.6%, respectively) was not significantly different. No significant difference was observed when NaFeEDTA-fortified fish sauce and soy sauce were compared directly (6.7% and 7.9%, respectively). Soy sauce inhibited iron absorption from rice-based meals (8.5% without and 6.0% with soy sauce; P < 0.02), whereas fish sauce did not affect iron absorption significantly. Conclusion: The relatively high iron absorption from NaFeEDTA-fortified fish sauce and soy sauce and the acceptable organoleptic properties of NaFeEDTA indicate the potential usefulness of this iron fortificant in fish sauce and soy sauce fortification programs. [ABSTRACT FROM AUTHOR]

Background: Iron-fortified school breakfasts have been introduced in Peru to combat childhood iron deficiency. Objective: We evaluated whether iron absorption from a school breakfast meal was improved by increasing the ascorbic acid content or by adding an alternative enhancer of iron absorption, Na2EDTA. Design: In a crossover design, iron absorption from test meals was evaluated by erythrocyte incorporation of 58Fe and 57Fe. The test meals (wheat bread and a drink containing cereal, milk, and soy) contained 14 mg added Fe (as ferrous sulfate) including 2.0-2.6 mg 58Fe or 4.0-7.0 mg 57Fe. Results: Geometric mean iron absorption increased significantly from 5.1% to 8.2% after the molar ratio of ascorbic acid to fortification iron was increased from 0.6:1 to 1.6:1 (P < 0.01; n = 9). Geometric mean iron absorption increased significantly from 2.9% to 3.8%, from 2.2% to 3.5%, and from 2.4% to 3.7% after addition of Na2EDTA at molar ratios relative to fortification iron of 0.3:1, 0.7:1, and 1:1, respectively, compared with test meals containing no added enhancers (P < 0.01; n = 10 for all). Iron absorption after addition of ascorbic acid (molar ratio 0.6:1) was not significantly different from that after addition of Na2EDTA (molar ratio 0.7:1). Conclusions: Ascorbic acid and Na2EDTA did not differ significantly in their enhancing effects on iron absorption at molar ratios of 0.6:1 to 0.7:1 relative to fortification iron. Additional ascorbic acid (molar ratio 1.6:1) increased iron absorption significantly. Increasing the molar ratio of Na2EDTA to fortification iron from 2 0.3:1 to 1:1 had no effect on iron absorption. [ABSTRACT FROM AUTHOR]

Infant formula based on pea (Pisum sativum)-protein isolate has been suggested as an alternative to soyabean formula in countries where soyabean is not a native crop, or when soyabean protein cannot be used due to allergic reactions or intolerances. In the present study, Fe absorption from experimental infant formulas based on pea-protein isolate was measured in healthy non-anaemic young women. The influence of phytic acid and ascorbic acid on Fe absorption was evaluated, using a stable-isotope technique based on incorporation of Fe stable-isotope labels into erythrocytes 14 d after administration. Geometric mean Fe absorption increased from 20·7 (+1SD 41·6, -1sd 10·3) % to 33·1 (+1sd 58·6, -1sd 18·7) %; (P<0·0001; n 10) after enzymic degradation of virtually all phytic acid. Doubling the molar ratio Fe : ascorbic acid from 1 : 2·1 to 1 : 4·2 in the infant formula with native phytic acid content also increased Fe absorption significantly (P<0·0001; n 10); geometric mean Fe absorption increased from 14·8 (+1sd 32·1, -1sd 6·8) % to 22·1 (+1sd 47·2, -1sd 10·4) %. These results confirm the inhibitory and enhancing effects of phytic acid and ascorbic acid respectively on Fe absorption, but also indicate relatively high fractional Fe absorption from the pea-protein-based formulas. After adjusting for differences in Fe status, our data indicate that Fe absorption from dephytinised pea protein might be less inhibitory than dephytinised soyabean protein as measured in a previous study (). [ABSTRACT FROM PUBLISHER]