NaFe3+EDTA as a food fortificant: influence on zinc, calcium and copper metabolism in the rat

The general acceptance of NaFe3+EDTA for food Fe fortification has been partly restricted by concern over the influence of EDTA on the metabolism of other nutritionally important trace elements and minerals. We have investigated the influence of NaFe3+EDTA, and of increasing dietary levels of Na2EDTA, on Zn, Cu and Ca metabolism in rats fed on Zn-sufficient and Zn-deficient soya-bean-isolate- based diets. With the Zn-deficient diets, changing the dietary Fe compound from FeSO4 to NaFe3+- EBTA significantly (P < 0·05) increased mean apparent Zn absorption from 50·2 to 67·4%. urinary Zn excretion from 2·0 to 4·0% of intake, and Zn retention from 48·2 to 63·4%. Increasing the dietary EDTA level to 1000 mg/kg further increased Zn absorption to 78·1%, urinary Zn excretion to 15·6% of intake and Zn retention to 62·5%. Increased Zn retention was accompanied by a significant increase in weight gain indicating that the extra Zn was available for normal metabolic processes. With rats fed on the Zn-sufficient diet, NaFe3+EDTA and Na2EDTA similarly increased the absorption, urinary excretion and retention of Zn but to a lesser extent. NaFe3+EDTA, however, had no influence on the absorption, urinary excretion and retention of Cu and Ca, and additional Na2EDTA caused only minor increases in Cu absorption and retention and in the urinary excretion of Ca. We conclude that using NaFe3+EDTA as a food fortificant would have no detrimental effect on the metabolism of Zn, Cu and Ca and, in some situations, could improve Zn absorption and retention from low-bioavailability diet

Fractional magnesium absorption is significantly lower in human subjects from a meal served with an oxalate-rich vegetable, spinach, as compared with a meal served with kale, a vegetable with a low oxalate content

The aim of the present study was to evaluate Mg absorption from a test meal served with an oxalate-rich vegetable, spinach, as compared with a test meal served with a vegetable with a low oxalate content, kale. Mg absorption was measured by a stable-isotope technique based on extrinsic labelling of the test meals and faecal monitoring of the excreted isotope labels. Nine healthy adults participated in the study. The test meals were based on 100g phytate-free white bread, served with 300g spinach (6·6mmol oxalate; 0·7mmol 25Mg label added, 5·0mmol total Mg) or 300g kale (0·1mmol oxalate; 1·2mmol 26Mg label added, 4·8mmol total Mg). The test meals were served on days 1 and 3, at breakfast and lunch, using a cross-over design. The results from the present study demonstrated that apparent Mg absorption was significantly lower from the meal served with spinach (26·7 (sd 10·4) %) than the meal served with kale (36·5 (sd 11·8) %) (P=0·01). However, the lower fractional apparent Mg absorption from the test meal served with spinach can be assumed to be, at least partly, counterbalanced by the higher native Mg content of spinach as compared with kale. Although based on indirect evidence, i.e. not based on an evaluation of added (or removed) oxalic acid, the difference in Mg absorption observed in the present study is attributed to the difference in oxalic acid content between the two vegetable

Iron absorption from experimental infant formulas based on pea (Pisum sativum)-protein isolate: the effect of phytic acid and ascorbic acid

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 (

Erythrocyte incorporation of iron by infants: iron bioavailability from a low-iron infant formula and an evaluation of the usefulness of correcting erythrocyte incorporation values, using a reference dose or plasma ferritin concentrations

Bioavailability of iron (Fe) from a low-Fe infant formula was determined by erythrocyte incorporation of 58Fe 14 d after administration in ten healthy, non-Fe-deficient infants. Two feeding protocols were compared, with each infant acting as his/her own control. At 140 and 154 d of age, infants were fed 1000 g of 58Fe-labelled formula (1·44 mg total Fe/1000 g) as six feeds over 24 h (Protocol A) or as two feeds/day on three consecutive days (Protocol B). A water solution with 57Fe and ascorbic acid was given separately as a reference dose in both study protocols. Erythrocyte incorporation of 58Fe and 57Fe was determined by thermal ionisation mass spectrometry. Geometric mean 58Fe incorporation was 7·6 % (range 3·3-13·5 %) with Protocol A as compared to 10·6 % (range 6·7-18·6 %) with Protocol B (P=0·05); paired t test. Inter-individual variability of 58Fe was not reduced by correcting for the incorporation of 57Fe from the reference dose, or by correcting for plasma ferritin concentration. Fractional erythrocyte incorporation of Fe from low-Fe infant formula was in the same range as our earlier published data on erythrocyte incorporation of Fe from human milk extrinsically labelled with 58Fe (). The methodological evaluations included in this study clearly indicate the importance of using standardised study protocols when evaluating Fe bioavailability in infants. Corrections of erythrocyte incorporation data based on plasma ferritin or erythrocyte incorporation of Fe from a reference dose were not found to be usefu

A comparison of iron absorption in adults and infants consuming identical infant formulas

Fe absorption was estimated in adults and infants from the erythrocyte incorporation of Fe isotopes added to infant formula. Fe absorption was measured in adults using radioisotopes, and in infants with a stable-isotope technique. In adults, the geometric mean Fe absorption from a ready-to-feed soya formula with its native phytic acid content was 2·4%. This increased to 6·0% (P < 0·05) after almost complete dephytinization. In infants, mean Fe absorption values were 3·9 and 8·7% respectively from the same products (P <0·05). In adults, mean Fe absorption from a spray-dried soya formula containing 110mg ascorbic acid/I was 4·1%, increasing to 5·3% (P < 0·05) when ascorbic acid was doubled to 220 mg/l. In infants, mean Fe absorption values were 5·7 and 9·5% (P < 0·05) from the same products. Mean Fe absorption from a milk-based formula was 6·5% in adults compared with 6·7% in infants. All meals in the adult and infant studies were fed using an identical meal size of 217g. Increasing the meal size threefold in adults did not change fractional Fe absorption. Mean Fe absorption values for each meal were lower in adults than in infants, but the relative inhibitory effect of phytic acid and the enhancing effect of ascorbic acid were similar. We conclude that Fe absorption studies in adults can be used to assess the influence of enhancers and inhibitors of Fe absorption in infant formulas fed to infants. Further studies, however, are required to extend these findings to weaning foods and complete meal

Zinc and calcium apparent absorption from an infant cereal: a stable isotope study in healthy infants

Fractional apparent absorption of Zn and Ca from a wheat-milk-based infant cereal was studied in six healthy infants (18-30 weeks old). Mineral absorption was measured by a stable-isotope technique based on faecal excretion of the isotopes. Each test meal (40 g cereal) was extrinsically labelled with 70Zn and 42Ca before intake. All faecal material passed during the 21 d following intake of the labelled test meal was collected on trace-element-free nappies. Individual stool samples were analysed for their content of ‘OZn and 42Ca by thermal ionization mass spectrometry. Apparent absorption was calculated as intake minus total faecal excretion of the isotopes over 68-92 h after administration. The fractional apparent absorption values for Zn and Ca were 33.9 (SD 164) % (range 19.2-639 %) and 53.5 (SD 12.6) %) (range 36.7-71.7 %) respectively. Re-excretion of absorbed 70Zn (> 68-92 h to 21 d after intake of the labelled meal) was 044 (SD 038) %] of administered dose while only one infant re-excreted detectable amounts of 42Ca (1.74%) of administered dose). The analysis of individual stool samples confirmed that 72 h is a sufficient time period for complete collections of non-absorbed isotopes in faecal material from infants during the weaning period and that re-excretion of initially absorbed 70Zn and 42Ca (> 68-92 h to 21 d after intake of the labelled meal) is negligibl

A double stable isotope technique for measuring iron absorption in infants

A stable isotope technique has been developed which uses 57Fe and 58Fe as labels and which enables the simultaneous measurement of Fe absorption from two test meals in infants. The method was evaluated by measuring Fe absorption from a commercial whey-adjusted infant formula in nine healthy infants aged 13-25 weeks. Each infant was fed 210 ml formula, labelled with either 57Fe or 58Fe, on four consecutive mornings, in random order. The total Fe content in each feed was 2.5 mg Fe; either as 2.5 mg 57Fe, or 0 6 mg 58Fe plus 1.9 mg Fe with normal isotopic composition. Isotopic enrichment of Fe in erythrocytes was measured by thermal ionization mass spectrometry 14 d after the last administration, and Fe absorption was calculated based on isotope ratio shifts, total circulating Fe and intake of each isotope. Geometric mean absorption for the 57Fe and 58Fe labels was 6.72 and 658% respectively, and the absorption of the two isotopes was not significantly different (Student's paired t test). By this technique, paired comparisons of Fe absorption can be obtained and systematic studies of the influence of dietary factors on Fe absorption during infancy can he conducte

Iron absorption from ferrous fumarate in adult women is influenced by ascorbic acid but not by Na2EDTA

Ascorbic acid and Na2EDTA enhance Fe absorption from the water-soluble Fe compound FeSO4 but their effect on poorly water-soluble Fe compounds such as ferrous fumarate is less well established. In the present study, the effects of ascorbic acid and Na2EDTA on Fe absorption from ferrous fumarate were evaluated in adult women (ten women/study) from the erythrocyte incorporation of Fe stable isotopes (57Fe or 58Fe) 14 d after administration. Two separate studies were made with test meals of Fe-fortified infant cereal (5 mg Fe/meal). Data were evaluated by paired t tests and the results are presented as geometric means. In study 1a, the comparison between Fe absorption from ferrous fumarate- and FeSO4-fortified cereal showed that adult women absorb Fe as well from ferrous fumarate as from FeSO4 (3·0 and 3·1 % respectively, P=0·85). After addition of Na2EDTA (Na2EDTA:fortification Fe molar ratio of 1:1), Fe absorption from FeSO4 was significantly higher than from ferrous fumarate (5·3 v. 3·3 % respectively, P<0·01; study 1b). In study 2, Fe absorption was compared from ferrous fumarate-fortified meals with and without ascorbic acid added at a 4:1 molar ratio (relative to fortification Fe) and the results showed that ascorbic acid increased Fe absorption from ferrous fumarate significantly (6·3 v. 10·4 %, P=0·02). The results of the present studies show that Fe absorption from ferrous fumarate is enhanced by ascorbic acid but not by Na2EDTA, thus emphasising that not all findings from Fe absorption studies made with FeSO4 can be extrapolated to Fe compounds with different solubility propertie

Zinc absorption in adult humans: the effect of protein sources added to liquid test meals

The influence of different protein sources on Zn absorption was evaluated in healthy adults by radioisotopic labelling of single meals, followed by whole-body retention measurements 14 d after intake. Semi-synthetic liquid diets were used for the evaluation of different animal-protein sources and dephytinized soyabean-protein isolate ( ··01 g phytic acid/kg). Zn absorption was measured in the same subjects from identical test meals containing no added protein. No statistically significant differences were found in the Zn absorption from test meals containing bovine whey, casein or egg albumen when compared with test meals without added protein. Bovine serum albumin (BSA) and soyabean-protein isolate (< ··01 g phytic acid/kg) significantly reduced the mean absorption of Zn from 45-49% (no added protein) to 38·0 (SD 10·9) (BSA, P < ··05) and 33·9 (SD 12·6)% (soyabean-protein isolate < ··01 g phytic acid/kg, P < ··01). These results demonstrate that Zn absorption is inhibited by certain protein sources, such as BSA and dephytinized soyabean-protein isolate, while other proteins have little or no effec

Comparison of urinary monitoring, faecal monitoring and erythrocyte analysis of stable isotope labels to determine magnesium absorption in human subjects

We have evaluated urinary monitoring and erythrocyte analysis to determine Mg absorption in human subjects as alternatives to the conventional technique of faecal monitoring by stable-isotope techniques. Ten healthy adults received 2·2 mmol 25Mg in water, together with wheat bread, followed 15 min later by intravenous injection of 0·6 mmol 26Mg (day 1). Brilliant blue and Yb (given on day 0 and day 1 respectively) served as qualitative and quantitative faecal markers. Urine was collected for 6 d after test meal intake. Complete collections of faeces were made until excretion of the second brilliant blue marker (given on day 7). Mg isotope ratios were determined by thermal ionisation-MS in urine and faeces and by inductively coupled plasma-MS in erythrocytes. Absorption was determined based on: (1) 6 d urine pools; (2) 24 h urine pools (collected 22-46 h after test meal intake); (3) erythrocytes from a blood sample drawn on day 14; (4) complete 6 d faecal pools; (5) faecal pools based on the first three consecutive stools after excretion of the first brilliant blue marker. Differences in mean Mg absorption (42 44 %) were statistically insignificant between techniques, except when based on 6 d urine pools for which the value was significantly lower (33 (sd 7) %, P=0·0003, ANOVA). The results indicate that Mg absorption can be determined from 24 h urine pools or erythrocytes obtained 14 d after test meal intake, an alternative method to the more time-consuming and labour-intense faecal monitoring. The choice of technique depends on practical and financial consideration