In vivo enrichment of hepatic microsomes with isotopic iron.
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The absorption and endogenous excretion of iron in man was studied by monitoring the fecal excretion of a stable iron isotope (58Fe). The study was carried out for 12 healthy volunteers who were divided into two groups. Group I received 58Fe-labeled ferric ammonium citrate (III) (58FeAC) equivalent to 6 mg of iron as a control, and group II received a combination of 500 mg of vitamin C and 58FeAC. A new formula was used to calculate the 58Fe absorption ratio reflecting the pool of iron in the intestinal cells, and the ratio was compared with that obtained from Janghorbani's formula, which has been used as one of the common methods. As a result, the 58Fe absorption ratio in group II was statistically significantly higher than that of group I (34.4 +/- 6.1% vs. 15.0 +/- 5.5%, M +/- SD) using Janghorbani's formula. The similar absorption ratio (34.1 +/- 6.0% vs. 14.8 +/- 5.5%) was also obtained by our new formula. Our results confirmed the previous findings that the availability of iron is stimulated by the supplementation of vitamin C. Both formulae agreed in the absorption of iron, indicating that the endogenous excretion of iron (caused by the desquamated cells) in the intestine does not disguise the iron absorption.
Fecal excretion of a stable iron isotope (58Fe) in five young adult men consuming a diet enriched with the isotope added as an extrinsic tag was investigated as a possible tool for the study of iron absorption. Data are presented to illustrate rapidity and completeness of unabsorbed 58Fe excretion. Procedures are outlined to investigate the extent of 58Fe re-entry into the gastrointestinal tract of the absorbed 58Fe from the label with the cumulative fecal 58Fe method and its limitations are discussed. Two methods of estimating 58Fe absorption are discussed and compared.
The methodology of precise isotope abundance determinations of erythrocyte iron by fast atom bombardment mass spectrometry and signal averaging is established. For the determination of the 54Fe/56Fe ratio a relative precision of 0.5% and an absolute precision of 0.03% is achieved. After oral loading with 54Fe-enriched samples in the range between 5 and 25 mg per subject, the 14-day erythrocyte incorporation of 54Fe has been determined in five individuals, namely, two adults, two children, and one infant. In the two adults, the oral dose of 54Fe was simultaneously labeled with a trace amount of carrier-free 59Fe. In these double-isotope loading tests, a good agreement was observed between the absorption data determined on the basis of whole body retention of 59Fe and on the basis of the 54Fe erythrocyte incorporation. The stable isotope methodology applied allows measurement of the iron absorption using highly enriched 54Fe at a dose of 25 mg for an adult or at a dose of 5 mg for infants of about 1 year of age.
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Methodology for use of stable isotopes of iron (54Fe, 57Fe, and 58Fe) as biological tracers was developed. Tracers were quantitated by measurement of ion abundances with a quadrupole mass spectrometer. The volatility of the iron was enhanced by chelation with 2,4-pentanedione prior to analysis. Samples were introduced into the mass spectrometer via a direct inlet probe. Ion abundance ratios were calculated from integrated ion current measurements obtained for selected ions in the two-ligand fragment of the chelate. Stable isotope tracer concentrations were calculated from these ratios. A prodecure was developed for the formation and purification of serum iron chelates. The method was used to analyze iron standards and blood serum samples containing known amounts of added 58Fe. The disappearance from pony serum of injected 58Fe was used as an in vivo test of the method. It was estimated that a minimum of 1.5 mg of either 54Fe, 57Fe, or 58Fe would be required to label 1 g of natural iron at detectable levels. The methods has promise as an alternative to radioisotope tracer techniques for some applications involving human subjects.
Because of a possible hazard from the use of radioisotopes to determine iron absorption by infants, the use of stable isotopes for this purpose has much appeal. We have applied the method of inductively coupled plasma mass spectrometry (ICP/MS) to determine the mass ratio, 58Fe/57Fe, in blood before and after oral administration of 58Fe. From the increase in erythrocyte enrichment with 58Fe, we have calculated percentage absorption of iron. We have shown that the coefficient of variation of measured mass isotope ratio is 0.1-1.0%, depending on the conditions of the measurement. The method has been applied to a feasibility study involving four infants. Each infant was given 58Fe either as a single dose or as one dose on each of two consecutive days. Each dose provided 1.945 mg iron and 1.440 mg 58Fe. Samples of blood were obtained before isotope administration and at 14, 42, and 60 days thereafter. Isotopic analysis of the samples demonstrates that this approach results in a sufficiently large isotope enrichment to permit satisfactory measurement of iron availability. It is concluded that this new method is highly promising for studies of iron availability in infants and children.
The absorption of iron, copper, and zinc was determined in 22 women 19 to 25 years of age from the difference between intake and fecal output of the stable isotopes 58Fe, 65Cu, and 70Zn, as measured by neutron activation analysis. Of the 22 women, 14 were using oral contraceptive agents, and the other eight were not. Absorption in the group using oral contraceptive agents did not differ significantly from the group not using oral contraceptive agents. The overall iron absorption averaged 14%, copper 57%, and zinc 38%.
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An investigation of iron metabolism in a female patient volunteer by administration of stable iron isotopes as tracers was performed. The applied methodology had already been tested in rabbits in comparison with radioactive tracer technique. The subject under study was given 58Fe solution intravenously and about 45 min later 57Fe solution orally. Ten blood samples were drawn at different times within 522 min from injection. Single iron isotopes content in plasma samples was determined by proton nuclear activation. A Compton suppressor system was utilized to improve the detector limits. The characteristic parameters of iron plasma clearance and of iron intestinal absorption were determined.
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A study was made on the effect of various drinks on the absorption on non-heme iron. The drinks were taken with standard meals composed of a hamburger, string beans and mashed potatoes. In each series identical meals were served to the same subject either with water or with the drink under study, labelling the meals with two different radio-iron isotopes. A reduction in iron absorption was seen when serving tea (62 per cent) or coffee (35 per cent) with the meals. Orange juice increased the iron absorption (85 per cent). Pure alcohol and wine increased only slightly the percentage absorbed. Wine often has a high iron content, which increased significantly the amount of iron absorbed (three times). Milk and beer have no significant effect. Coca-Cola increased only slightly the absorption. The present studies clearly shows that the choice of drink drunk with a meal can markedly affect the absorption of non-heme iron.