Effect of Zinc Sulfate Fortificant on Iron Absorption from Low Extraction Wheat Flour Co-Fortified with Ferrous Sulfate

The co-fortification of wheat flour with iron (Fe) and zinc (Zn) is a strategy used to prevent these deficiencies in the population. Given that Zn could interact negatively with Fe, the objective was to assess the effect of Zn on Fe absorption from bread prepared with wheat flour fortified with Fe and graded levels of Zn fortificant. Twelve women aged 30–43 years, with contraception and a negative pregnancy test, participated in the study. They received on four different days, after an overnight fast, 100 g of bread made with wheat flour (70 % extraction) fortified with 30 mg Fe/kg as ferrous sulfate (A) or prepared with the same Fe-fortified flour but with graded levels of Zn, as zinc sulfate: 30 mg/kg (B), 60 mg/kg (C), or 90 mg/kg (D). Fe radioisotopes (⁵⁹Fe and ⁵⁵Fe) of high specific activity were used as tracers and Fe absorption iron was measured by the incorporation of radioactive Fe into erythrocytes. Results: The geometric mean and range of ±1 SD of Fe absorption were: A?=?19.8 % (10.5–37.2 %), B?=?18.5 % (10.2–33.4 %), C?=?17.7 % (7.7–38.7 %), and D?=?11.2 % (6.2–20.3 %), respectively; ANOVA for repeated measures F?=?5.14, p?<?0.01 (Scheffè’s post hoc test: A vs D and B vs D, p?<?0.05). We can conclude that Fe is well absorbed from low extraction flour fortified with 30 mg/kg of Fe, as ferrous sulfate, and up to 60 mg/kg of Zn, as Zn sulfate. A statistically significant reduction of Fe absorption was observed at a Zn fortification level of 90 mg Zn/kg.     

A Combination of a Dairy Product Fermented by Lactobacilli and Galactooligosaccharides Shows Additive Effects on Mineral Balances in Growing Rats with Hypochlorhydria Induced by a Proton Pump Inhibitor

This study aimed to investigate the effects of a combination of a dairy product fermented by lactobacilli (DFL) and galactooligosaccharides (GOS) on mineral balances in growing rats with hypochlorhydria induced by a proton pump inhibitor (PPI). Three-week-old male rats were assigned to receive one of six diets: a control diet, control diets containing 1.6 or 5.0 % GOS, a DFL diet and DFL diets containing 1.6 or 5.0 % GOS for 9 days. From day 5 of the feeding period, half of the rats fed with control diets were subcutaneously administered with saline, whereas the remaining rats were administered with PPI for 5 days. Calcium (Ca), phosphorus (P), magnesium (Mg), iron (Fe) and zinc (Zn) balances were determined from days 6 to 9. PPI administration significantly decreased the apparent absorption of Ca and Fe and increased urinary P excretion, resulting in decreased Ca, Fe and P retention. GOS dose-dependently increased the apparent absorption of Ca, Mg and Fe and urinary Mg excretion and decreased urinary P excretion. DFL significantly increased the apparent absorption of Ca and Mg and urinary Mg excretion. The combination of DFL and GOS additively affected these parameters, resulting in increased Ca, P and Fe retention, and it further increased the apparent absorption and retention of Zn at 5.0 % GOS. In conclusion, the combination of DFL and GOS improves Ca, P and Fe retention in an additive manner and increases the Zn retention in growing rats with hypochlorhydria induced by PPI.     

Endogenous iron excretion

The effects of supplemental oral (0, 40, and 400 ppm) and parenteral iron (0 and 2.72 mg Fe iv given initially as a single dose) on iron absorption, excretion, and retention were determined in 30 rats. Endogenous fecal iron excretion was determined by the radioisotope dilution technique after im injection of 80 kBq Fe-59, using blood and certain body tissues as reference sources for the estimation of the specific activity (Bq Fe-59/micrograms Fe) of endogenous iron. The basal diet contained 3.6 ppm Fe. Fe(III)-hydroxide-polymaltose was used as the sole iron source in oral, iv, and im iron treatments. Iron balance as determined from day 14 to 20 of the experiment was not significantly affected by iv iron administration. Nevertheless, a temporarily reduced retention should have occurred, since differences in final body iron contents were lower than 2.72 mg, as compared to the respective untreated groups. Apparent iron absorption and iron retention increased with surplus oral iron, and the efficiency rates were highest with adequate iron supply (40 ppm). True absorption rates of iron were similar without any, and with 40 ppm Fe amounting 40 to 50% of the intake. In the iron deficient rats, half of the actually absorbed iron (about 16 micrograms/d) was lost by endogenous fecal re-excretion, and another 3 micrograms/d by the urinary route. Endogenous loss with feces and with urine increased with further oral iron supply, but at a considerably lower rate as total fecal excretion. Parenterally administered iron did not affect endogenous loss at all. The results indicate that endogenous excretion cannot be regarded as a means to eliminate excessive iron, and might actually be an inevitable loss.     

Mineral supplementation of white wheat flour is necessary to maintain adequate mineral status and bone characteristics in rats

This experiment was designed to compare the effect of ingestion of a wheat flours on mineral status and bone characteristics in rats. White flour was tested either without further mineral supplementation or with Mg, Fe, Zn and Cu supplementation. The flour diets were compared to a control purified diet. Four groups of 10 male Wistar rats each were fed one of the experimental diets for 6 wk and mineral status and tissue retention as well as bone characteristics were determined. As expected, mineral intake, except for calcium, was significantly lesser in rats fed the white flour diet than in the other groups. The rats fed the white flour diet had the lowest food intake, weight gain, fecal excretion and intestinal fermentation. The most important result was that Mg and Fe status were drastically lower in rats fed the white flour diet than in those fed whole flour or control diets. The status of these both elements were significantly improved by the mineral supplementation of white flour. There were no major significant differences between mineral-supplemented white flour and whole flour groups in mineral status. Furthermore, bone mineral densities (total, metaphyseal and diphyseal) were significantly lower in rats fed white flour diet compared to the other diet groups, while no significant difference was observed between the mineral-supplemented white flour, whole flour or control diet groups. In conclusion, the present work shows clearly the importance of mineral-supplementation of white wheat flour to sustain an adequate intake of minerals. Our results indicate also that the whole wheat flour did not negatively alter mineral bioavailability, in comparison to mineral supplemented white flour. Clinical studies are still needed to confirm these rat results in human.     

An improved multi-element measurement of mineral absorption in the piglet utilizing the fecal monitoring technique

The fecal monitoring technique for measuring the absorption of Zn, Mn, Se, and Fe was studied in eight male piglets (mean±SEM birthweight (bw)=1695±50 g) using high resolution gamma spectrometry. Four d old piglets were fed a complete liquid milk diet for five d prior to the orogastric administration of an isotope dose (⁷⁵Se,⁵⁴Mn,⁵⁹Fe, and⁶⁵Zn) equilibrated with the liquid milk diet.⁵¹CrCl₃ was used as a fecal marker but was found to be partially absorbed. Stool samples were collected daily for 15 d, counted, and then the daily fecal excretion was calculated. Results indicate that endogenous excretion for each of the isotopes was not constant but decreased exponentially with time. The pattern of endogenous excretion varied between elements. An improved method for calculating the endogenous excretion was therefore developed. This method is based on the pattern of endogenous excretion in three-four d old male piglets (mean±SEM bw=2060±75 g) injected intravenously with the same isotopes and on the level of endogenous excretion in orally fed animals in the postabsorptive phase of excretion. These findings have important implications for the estimation of endogenous excretion in future fecal monitoring absorption studies in order to minimize underestimation of true absorption.     

Effect of Increasing Levels of Zinc Fortificant on the Iron Absorption of Bread Co-Fortified with Iron and Zinc Consumed with a Black Tea

Iron (Fe) and zinc’s (Zn) interaction at the absorptive level can have an effect on the success of co-fortification of wheat flour with both minerals on iron deficiency prevention. The aim of the study was to determine the effect of increasing levels of zinc fortificant on the iron absorption of bread co-fortified with iron and zinc consumed with a black tea. Twelve women aged 33–42 years participated in the study. They received on four different days 200 mL of black tea and 100 g of bread made with wheat flour (70 % extraction) fortified with either 30 mg Fe/kg alone, as ferrous sulfate (A), or with the same Fe-fortified flour, but with graded levels of Zn, as zinc sulfate: 30 mg/kg (B), 60 mg/kg (C), or 90 mg/kg (D). Fe radioisotopes (⁵⁹Fe and ⁵⁵Fe) of high specific activity were used as tracers, and Fe absorption iron was measured by the incorporation of radioactive Fe into erythrocytes. The geometric mean and range of ±1 SD of Fe absorption were as follows: A?=?6.5 % (2.2–19.3 %), B?=?4.6 % (1.0–21.0 %), C?=?2.1 % (0.9–4.9 %), and D?=?2.2 % (0.7–6.6 %), respectively; ANOVA for repeated measures F?=?10.9, p?<?0.001 (Scheffè’s post hoc test: A vs. C, A vs. D, B vs. C, and B vs. D; p?<?0.05). We can conclude that Fe absorption of bread made from low-extraction flour fortified with 30 mg/kg of Fe, as ferrous sulfate, and co-fortified with zinc, as zinc sulfate consumed with black tea is significantly decreased at a zinc fortification level of ≥60 mg/kg flour.     

A histidine supplement and regulation of the zinc status in Swiss random mice.

The effects of histidine on the zinc status are controversial. In mice, we studied the effects of a moderate histidine supplement on the regulation of the zinc status using subcutaneously administered 65Zn. In animals fed a zinc-adequate diet, histidine supplement did not cause changes in the zinc status (zinc concentrations, 65Zn tissue distribution, and tissue specific activities). Neither effects on the regulation of the zinc status (65Zn retention, excretion and biological half-life) could be demonstrated. However, the combination of a low zinc diet and moderate histidine supplementation caused changes in the regulation of the zinc status (lower 65Zn retention, associated with increased fecal excretion and a shorter biological half-life), aggravating the dietary deficiency (lower bone zinc, a shift in the 65Zn tissue distribution). Reviewing the literature, it seems that only a molar histidine/zinc ration of 2,000 or higher will cause zinc deficiency.     

Change in apparent and true absorption and retention of dietary zinc with age in rats

Two groups of 16 rats each were fed the same diet with 12.9 ppm Zn. Nine days after each animal was injected with⁶⁵Zn for assessing fecal zinc of endogenous origin, zinc intake and excretion were determined for a six-day period at the age of about five (group I) and nine (II) weeks. At mean growth rates of 5.1 and 5.2 g/day, food consumption per gram of gain was 2.01 g in group I vs 2.86 g in II. Overall, zinc retention amounted to 21 vs 25 μg Zn/g of gain. Apparent absorption averaged 92 vs 74% of Zn intake (132 vs 189 μg/day), while true absorption averaged 98 vs 92%. It was concluded that endogenous fecal zinc excretion was limited to the indispensable loss (F _em) in group I (7 μg/day), while it exceeded this minimum loss in group II (33 μg/day). True retention, which reflected total zinc utilization (true absorption times metabolic efficiency), was derived from apparent absorption plusF _em (11 μg/day for group II according to the greater metabolic body size of the rats). It averaged 98% of Zn intake in group I vs 80% in group II. The mean metabolic efficiency was 100% vs 87%. The conclusion was that these marked differences between age groups in utilizing the dietary zinc reflected the efficient homeostatic adjustments in absorption and endogenous excretion of zinc to the respective zinc supply status.     

Absorption,endogenous excretion,and balance of zinc in growing rats on diets with various sugars replacing starch

The effect of partially replacing starch for various sugars on the apparent and true absorption, endogenous excretion, and balance of zinc was investigated in a study with growing rats. Six groups of five or six animals with an initial live weight of 39.4 +/- 2.7 g were fed diets that had the same Zn content (22 mg/kg), but differed in the sugar content: 1. Starch only (56%); 2. Glucose (15%); 3. Fructose (15%); 4. Sucrose (30%); 5. Galactose (15%); and 6. Lactose (30%). At the start of a 15-d fecal and urinary collection period, each animal was given an intramuscular injection of 380 kBq 65Zn for estimating endogenous Zn excretion by isotope dilution. The ratio of the specific activity of fecal Zn (after 12 d) to that of urinary Zn (after 9 d) was applied to reflect the ratio of endogenous to total fecal Zn collected from day 10 to 15. This ratio averaged 0.59, without significant differences among treatments. For this period, apparent and true absorption averaged 87.1 and 94.7% of Zn intake, respectively, and did not significantly differ among diets. Urinary excretion of 65Zn and of stable zinc by the galactose-fed rats was markedly higher than that by the other animals. Their Zn balance was, per unit weight gain, comparable with that of the other groups (30.7 vs 28.2 to 30.2 micrograms/g).     

Long-term effects of iron: Zinc interactions on growth in rats

The influence of iron (Fe) on the bioavailability and functional status of zinc (Zn) was studied in young rats using metabolic balances and tissue dosages, which were compared to growth. Diets supplied adequate intakes of Fe (45 and 300 mg/kg diet) and Zn (14 and 45 mg/kg) for 2 mo. Two metabolic balance determinations were performed that were correlated for Zn and Fe during the first and the last weeks of the study. A significant effect of Fe supply, but not of Zn was displayed on Fe absorption; both Fe and Zn diet concentrations had a significant influence on Zn absorption. Fe and Zn organ contents were significantly correlated with the amount absorbed during the two metabolic balances. There was a positive correlation between liver and muscle Fe and Fe absorption, and Fe absorption and muscle Zn, as well as a negative one with liver Zn; a positive correlation was displayed between Zn absorption and Zn organ content. No correlation was found between Zn absorption and Fe tissue content. Growth was correlated with Zn, but not with Fe absorption during both balances. A positive correlation was displayed between growth and Zn liver content, and a negative one with Fe liver content. Care must be taken to give growing subjects balanced diets or supplementation, since the negative interactions between these trace elements are likely to persist as long as the diet is given.     

Guar crispbread in the diabetic diet.

Nine diabetic patients who were receiving various treatments supplemented their normal home diets (two patients) or metabolic ward diets (seven patients) with guar crispbread for five days. Their mean urinary glucose excretion fell significantly by 38% during the last two days. A significant fall in fasting blood glucose concentration of 1.1 +/- 0.4 mmol/1 (19.8 +/- 7.2 mg/100 ml) was seen only in those who took guar after the control period. Over eight weeks'' treatment insulin dosage was reduced by 21% in five patients, and home testing showed that glycosuria was reduced by 68% in six patients. Guar crispbread is likely to be a useful adjunct to diabetic treatment irrespective of the type of treatment or insulin dosage used.     

Metabolic engineering of bread wheat improves grain iron concentration and bioavailability

Bread wheat (Triticum aestivum L.) is cultivated on more land than any other crop and produces a fifth of the calories consumed by humans. Wheat endosperm is rich in starch yet contains low concentrations of dietary iron (Fe) and zinc (Zn). Biofortification is a micronutrient intervention aimed at increasing the density and bioavailability of essential vitamins and minerals in staple crops; Fe biofortification of wheat has proved challenging. In this study we employed constitutive expression (CE) of the rice (Oryza sativa L.) nicotianamine synthase 2 (OsNAS2) gene in bread wheat to up‐regulate biosynthesis of two low molecular weight metal chelators – nicotianamine (NA) and 2′‐deoxymugineic acid (DMA) – that play key roles in metal transport and nutrition. The CE‐OsNAS2 plants accumulated higher concentrations of grain Fe, Zn, NA and DMA and synchrotron X‐ray fluorescence microscopy (XFM) revealed enhanced localization of Fe and Zn in endosperm and crease tissues, respectively. Iron bioavailability was increased in white flour milled from field‐grown CE‐OsNAS2 grain and positively correlated with NA and DMA concentrations.     

Effect of boron as an antidote on dry matter intake, nutrient utilization and fluorine balance in buffalo (Bubalus bubalis) exposed to high fluoride ration

It is well known that excessive accumulation of fluorides can exert toxic effects on various tissues and organs so as to severely damage the health and production of animals. The aim of this study was to determine beneficial effect of boron on nutrient utilization in buffalo calves exposed to high fluoride (F) ration. For this purpose, we used three groups of four male Murrah buffalo calves (body weight 98-100 kg, aged 6-8 month) each. Control animal was given only basal diet and concentrate mixture. However, treatment I animals were fed basal diet, concentrate mixture, and F [as NaF, 60 ppm of dry matter (DM)]. The treatment II animals were fed basal diet, concentrate mixture, F (as NaF, 60 ppm of DM), and B (as sodium tetraborate, 140 ppm of DM). After 90 days of experimental feeding, a metabolism trial of 7 days duration was conducted to study the treatment effect on nutrient utilization of proximate nutrients, absorption, excretion, and retention of N, Ca, P, Fe, Zn, Cu, and F. Dietary F significantly (p < 0.05) depressed the dry matter intake and increased the apparent digestibility, absorption, and retention of F. However, boron supplementation significantly (p < 0.05) decreased the apparent digestibility, absorption, and retention of F and improved the dry matter intake, fecal excretion, and percent of absorbed F excreted via urine. Apparent digestibility of proximate nutrients (viz. DM, crude protein, crude fiber, ether extract, and nitrogen free extract) was unaffected on either F or F+B treatment. However, absorption and excretion of N, Ca, P, Fe, Zn, and Cu were affected significantly (p < 0.05) on F or F+B treatment. These findings suggest that fluoride-containing diet for short duration has effect on nutrient utilization, and boron at 140-ppm dose level, in general, antagonized the absorption and retention of F and also improved the feed intake in buffalo calves.     

Ingestion of difructose anhydride III enhances absorption and retention of calcium in healthy men

We examined the effects of a nondigestible disaccharide difructose anhydride III (DFAIII) on calcium absorption and retention by means of a human balance study of single-blind crossover design. Twelve healthy male subjects ingested 250 mg of shell powder as calcium carbonate (corresponding to 100 mg of calcium) with or without 1.0 g DFAIII three times a day for 13 d. In the last 4 d as a balance period, all urine and feces were collected and evaluated for calcium excretion. The apparent calcium absorption (mg/d) and rate of absorption (%) were higher, and those of retention were much higher, in the DFAIII group than in the control group. Furthermore, serum osteocalcin increased after the experimental period in the DFAIII group but not in the control group. These results indicate that DFAIII ingestion enhances intestinal calcium absorption, which might be beneficial for bone metabolism.     

Effect of exchangeable sodium percentage and presubmergence on yield and nutrition of rice under field conditions

Summary The effects of exchangeable sodium percentage (ESP) levels of 82, 72, 65 and 35 and 0, 15 and 30 days of presubmergence (submergence prior to the transplanting of rice) on yield and chemical composition of rice and availability of Fe, Mn, Zn and P in soil were studied factorially in a field experiment. Presubmergence increased rice yields at all ESP levels, the effect being more pronounced at high ESP's. Increasing ESP decreased yields and the contents of Ca, Mg, K, Fe, Mn, Zn and Cu but increased that of P and Na in the crop. Presubmergence enhanced absorption of all the above elements by the crop except P, K, Mg, Zn and Cu in the grain and decreased Na in grain and straw. Growing of rice under submerged conditions also facilitated the improvement of these soils. Effects of submergence and ESP on the availability of Fe, Mn, Zn and P in soil and their role in the nutrition of rice are discussed. The results suggest that 15 to 30 days presubmergence improved rice yields on a calcareous sodic soil of the Indo-Gangetic alluvial plain.     

A histidine supplement and regulation of the zinc status in Swiss random mice

The effects of histidine on the zinc status are controversial. In mice, we studied the effects of a moderate histidine supplement on the regulation of the zinc status using subcutaneously administered⁶⁵Zn. In animals fed a zinc-adequate diet, histidine supplement did not cause changes in the zinc status (zinc concentrations,⁶⁵Zn tissue distribution, and tissue specific activities). Neither effects on the regulation of the zinc status (⁶⁵Zn retention, excretion and biological half-life) could be demonstrated. However, the combination of a low zinc diet and moderate histidine supplementation caused changes in the regulation of the zinc status (lower⁶⁵Zn retention, associated with increased fecal excretion and a shorter biological half-life), aggravating the dietary deficiency (lower bone zinc, a shift in the⁶⁵Zn tissue distribution). Reviewing the literature, it seems that only a molar histidine/zinc ration of 2,000 or higher will cause zinc deficiency.     

Acute inhibition of iron bioavailability by zinc: studies in humans

Iron (Fe) and zinc (Zn) deficiencies constitute two of the most important nutritional and public health problems affecting developing countries. Combined supplementation or fortification with Zn and Fe are strategies that can be used to improve the Zn and Fe status of a population. However, there is concern about potential negative interactions between these two micronutrients due to a competitive binding to DMT1 and Zip14 transporter. Studies performed in humans have shown an inhibitory effect of Zn on Fe absorption when both minerals are given together as a solution in fasting conditions. We found that at low doses of iron (0.5 mg) the threshold for the inhibition of iron bioavailability was at a Zn:Fe wt/wt ratio ≥5.9:1, whereas at higher doses of Fe (10 mg) this inhibition occurred at 1:1 Zn:Fe wt/wt ratio. This differential response could be explained by the variation in the abundance of both cations as they compete for a limited number of shared transporters at the enterocyte. Conflicting results have been obtained when this interaction was studied in different food matrices. A negative interaction was not observed when Fe and Zn were provided in a composite hamburger meal, premature formula, human milk, or cow milk. A decrease on Fe absorption was observed in only 1 of 3 studies when Fe and Zn were supplied in wheat flour. The possibility of a negative interaction should be considered for supplementation or fortification programs with both microminerals.     

The effects of copper, manganese and zinc on plant growth and elemental accumulation in the manganese-hyperaccumulator Phytolacca americana

Synchrotron radiation X-ray fluorescence (SRXRF) and inductively coupled plasma mass spectrometry were used to estimate major, minor and trace elements in Cu-, Zn- and Mn-treated Phytolacca americana. The effects of the addition of Cu, Zn and Mn on morphological parameters, such as root length, shoot height, and fresh and dry weights of shoots and roots, were also examined. In addition, the activities of superoxide dismutase (SOD), ascorbate peroxidase (APX), guaiacol peroxidases (GPX) and catalase (CAT) and the expression of Fe-SOD, Cu/Zn-SOD, metallothionein-2 and glutathione S-transferase (GST) exposed to the highest amounts of Cu, Zn or Mn were detected. Our results confirmed the following: (1) Zn supplementation leads to chlorosis, disturbed elemental homeostasis and decreased concentrations of micro- and macroelements such as Fe, Mg, Mn, Ca and K. Cu competed with Fe, Mn and Zn uptake in plants supplemented with 25μM Cu. However, no antagonistic interactions took place between Cu, Zn, Mn and Fe uptake in plants supplemented with 100μM Cu. Mn supplementation at various concentrations had no negative effects on elemental deficits. Mn was co-located with high concentrations of Fe and Zn in mature leaves and the concentrations of macro elements were unchanged. (2) P. americana supplemented with increased concentrations of Zn and Cu exhibited lower biomass production and reduced plant growth. (3) When plants were supplemented with the highest Zn and Cu concentrations, symptoms of toxicity corresponded to decreased SOD or CAT activities and increased APX and GPX activities. However, Mn tolerance corresponded to increased SOD and CAT activities and decreased POD and APX activities. Our study revealed that heavy metals partially exert toxicity by disturbing the nutrient balance and modifying enzyme activities that induce damage in plants. However, P. americana has evolved hyper accumulating mechanisms to maintain elemental balance and redox homeostasis under excess Mn.     

Selenium distribution in wheat grain, and the effect of postharvest processing on wheat selenium content

Selenium (Se) is an essential micronutrient for animals and humans, and wheat is a major dietary source of this element. It is improtant that postharvest processing losses of grain Se are minimized. This study, using grain dissection, milling with a Quadrumat mill, and baking and toasting studies, investigated the distribution of Se and other mineral nutrients in wheat grain and the effect of postharvest processing on their retention. The dissection study, although showing Se concentration to be highest in the embryo, confirmed (along with the milling study) previous findings that Se (which occurs mostly as selenomethionine in wheat grain) and S are more evenly distributed throughout the grain when compared to other mineral nutrients, and hence, lower proportions are removed in the milling residue. Postmilling processing did not affect Se concentration or content of wheat products in this study. No genotypic variability was observed for grain distribution of Se in the dissection and milling studies, in contrast to Cu, Fe., Mn, and Zn. This variability could be exploited in breeding for higher proportions of these nutrients in the endosperm to make white flour more nutritious. Further research could include grain dissection and milling studies using larger numbers of cultivars that have been grown together and a flour, extraction rate of around 70%     

Effect of Increasing Concentrations of Zinc on the Absorption of Iron from Iron-Fortified Milk

The cofortification of milk with iron (Fe) and zinc (Zn) is a strategy used to prevent these deficiencies during childhood. Given that Zn can negatively interact with iron in aqueous solutions, the objective of the present study was to determine the effect of Zn on Fe absorption of milk fortified with Fe and Zn. Twenty-eight women between 33 and 47?years of age, with contraception and a negative pregnancy test, participated in one of two absorption studies. They received on four different days, after an overnight fast, 200?mL of milk (26?% fat) fortified with 10?mg Fe/L, as (A) ferrous sulfate, or the same milk but with graded doses of added Zn, as Zn sulfate of (B) 5, (C) 10, and (D) 20?mg/L (study 1, n?=?15). In study 2 (n?=?13), subjects received the same milk formulations, but these were also fortified with ascorbic acid (70?mg/L). Milk was labeled with radioisotopes ⁵⁹Fe or ⁵⁵Fe, and the absorption of iron was measured by erythrocyte incorporation of radioactive Fe. The geometric mean and range of ±1 SD of Fe absorption in study 1 were as follows: formula A?=?6.0?% (2.8?C13.0?%); B?=?6.7?% (3.3?C13.6?%); C?=?5.4?% (2.2?C13.2?%); and D?=?5.2?% (2.8?C10.0?%) (ANOVA for repeated measures, not significant). For study 2, data are as follows: 8.2?% (3.6?C18.7?%); B?=?6.4?% (2.5?C16.4?%); C?=?7.7?% (3.2?C18.9?%); and D?=?5.2 (1.8?C14.8?%) (ANOVA for repeated measures, not significant). In conclusion, according to the results from this study, it appears that the addition of zinc up to 20?mg/L does not significantly inhibit iron absorption from milk fortified with 10?mg/L of iron.