Nutrient Requirements and Interactions

Vitamin A and b-Carotene Can Improve Nonheme Iron Absorption from Rice, Wheat and Corn by Humans1,2
Mara Nieves Garca-Casal,* Miguel Layrisse,*3 Liseti Solano, Mara Adela Baron, Franklin Arguello, Daisy Llovera, Jose Ramrez,* Irene Leets* and Eleonora Tropper*
*Centro de Medicina Experimental, Laboratorio de Fisiopatologa, Instituto Venezolano de Investigaciones Cientcas (IVIC), Caracas 1020A, Venezuela; Unidad de Investigaciones en Nutricion, Universidad de Carabobo, Valencia, Venezuela; and Universidad Central de Venezuela, Caracas, Venezuela
ABSTRACT After the rapid decrease in the prevalence of iron deciency and iron-deciency anemia in the Venezuelan population when a national program for fortication of ours with iron and vitamins was instituted, we studied micronutrient interactions in Venezuelan diets. One hundred human adults were fed three cereal-based diets, labelled with either 59Fe or 55Fe in six studies. Each diet contained different concentrations of vitamin A (from 0.37 to 2.78 mmol/100 g cereal) or b-carotene (from 0.58 to 2.06 mmol/100 g cereal). The presence of vitamin A increased iron absorption up to twofold for rice, 0.8-fold for wheat and 1.4-fold for corn. b-carotene increased absorption more than threefold for rice and 1.8-fold for wheat and corn, suggesting that both compounds prevented the inhibitory effect of phytates on iron absorption. Increasing the doses of vitamin A or b-carotene did not further signicantly increase iron absorption. We measured the iron remaining in solution performing in vitro studies in which the pH of solutions was adjusted from 2 to 6 in the presence of vitamin A or b-carotene. All of the iron from ferrous fumarate was soluble after changing the pH of the solution containing 3.4 mmol of b-carotene to 6.0. Vitamin A was less effective. However, 78 { 18% of iron was soluble in the presence of 3.3 mmol of vitamin A, whereas with no vitamin addition, only 26 { 13% of iron was soluble (0.05). Vitamin A and b-carotene may form a complex with iron, keeping it soluble in the intestinal lumen and preventing the inhibitory effect of phytates and polyphenols on iron absorption. J. Nutr. 128: 646650, 1998. KEY WORDS: iron vitamin A b-carotene humans phytates

Vitamin A is a vital nutrient for cellular differentiation, vision, bone growth, reproduction and integrity of the immune system (Olson 1984). It is also essential for erythropoiesis. Deciency of this vitamin results in anemia in humans and animals that is reversed only by vitamin A supplementation (Bloem et al. 1989, Hodges et al. 1978, MejB a 1986, MejB a and Arroyave 1982, MejB a and Chew 1988, MejB a et al. 1979). b-carotene is the most abundant provitamin A in foods. Approximately 1050% of the total b-carotene consumed is absorbed in the gastrointestinal tract and within the intestinal wall is partially converted into vitamin A. The efciency of b-carotene absorption decreases as intake increases and conversion to vitamin A is regulated by the vitamin A status of the individual. b-carotene accumulation is not toxic, so it is considered a safe source of vitamin A (Roche Laboratories 1994, Wang 1994). After reporting the results on iron availability from diets consumed by different socioeconomic strata of the Venezuelan

population (Taylor et al. 1995), we continued studies of micronutrient interactions on iron ab sorption, especially since 1993, when the fortication of precooked corn and white wheat ours with iron and vitamins was started (Layrisse et al. 1996). The rst experiments already were presented in two workshops (GarcB a-Casal and Layrisse 1996, Layrisse and GarcB aCasal 1997); and published (Layrisse et al 1997, Layrisse and GarcB a 1997). The studies showed that addition of vitamin A (as retinol palmitate) and iron to corn or wheat ours used to prepare a typical Venezuelan breakfast (basal breakfast) containing corn or wheat bread, cheese, margarine and coffee or tea, diminished the inhibitory effect of phytates and polyphenols on iron absorption. We examined the effects of increasing doses of vitamin A and b-carotene on nonheme iron absorption. In addition, solubility changes of various iron compounds at pH 2 and 6 with and without vitamin A and b-carotene are presented. MATERIALS AND METHODS

Supported in part by CONICIT, Caracas, Venezuela. 2 The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 USC section 1734 solely to indicate this fact. 3 To whom correspondence should be addressed.

One hundred and four adult subjects (27 men and 77 women from Valencia City, Venezuela) voluntarily participated in this study. The Committee for the Protection of Human Subjects of the Venezuelan Institute for Scientic Research approved the studies. The subjects

0022-3166/98 $3.00 1997 American Society for Nutritional Sciences. Manuscript received 4 June 1997. Initial revision completed 3 July 1997. Revision accepted 12 November 1997.

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TABLE 1
Composition of meals administered to human subjects participating in iron absorption studies
Meals2 Study Subjects n 1 2 3 4 5 6 17 17 20 11 20 19 Rice Corn Wheat Rice Corn Wheat (59Fe) (59Fe) (59Fe) (59Fe) (59Fe) (59Fe) 0.55 mmol (55Fe) 0.37 mmol (55Fe) 1.10 mmol (55Fe) 1.51 mmol (55Fe) 0.61 mmol (55Fe) 0.66 mmol (55Fe) Vitamin A Vitamin A Vitamin A Vitamin A Vitamin A Vitamin A 1.51 mmol Vitamin A (59Fe) 0.61 mmol Vitamin A (59Fe) 0.66 mmol Vitamin A (59Fe) 0.58 mmol b-carotene / coffee (59Fe) 0.67 mmol b-carotene (59Fe) 0.85 mmol b-carotene (59Fe) 2.78 mmol Vitamin A (55Fe) 0.73 mmol Vitamin A (55Fe) 1.51 mmol Vitamin A (55Fe) 0.95 mmol b-carotene (55Fe) 1.53 mmol b-carotene / coffee (55Fe) 2.06 mmol b-carotene / coffee (55Fe) Test Food1 1 2 3 4

1 Rice test: 100 g polished rice / 10 g margarine; corn test: 100 g precooked corn our / 50 g cheese / 10 g margarine; wheat test: 100 g white wheat our / 50 g cheese / 10 g margarine. 2 Parentheses enclose supplemented isotope.

were selected from a low socioeconomic stratum in a section of a Venezuelan city. They were in apparent good health, but some of the women had moderate iron-deciency anemia (hemoglobin 120 g/L). Blood (30 ml) was taken from each subject in a tube with ethylenediamine tetraacetic acid (EDTA) and kept refrigerated at 4 C until used to measure hemoglobin concentration (Crosby et al. 1954), serum iron (International Committee for Standardization in Hematology 1978), unsaturated iron bin ding capacity (International Committee for Standardization in Hematology 1978), serum ferritin concentration (Flowers et al. 1986) and radioactivity measurements (see next section). Three cereals were tested: corn, wheat and rice. The corn test contained bread from 100 g nonfortied precooked corn our and 5 mg iron as ferrous fumarate; the our was given with 10 g margarine and 50 g cheese. The wheat test contained a bread prepared from 100 g of commercial white wheat our enriched with 0.15 mg thiamine, 0.2 mg riboavin, 2 mg niacin and 2 mg iron as ferrous fumarate. It also contained 10 g margarine and 50 g cheese. The rice test contained 100 g polished rice and was administered with 10 g margarine. Corn, wheat and rice tests were given alone or with different amounts of vitamin A or b-carotene. The vitamin A used was watersoluble retinol palmitate, and b-carotene was a 10% water-soluble powder. Both products kindly were supplied by Roche Laboratories of Venezuela (Caracas). Absorption studies. Six studies were performed. Each included 20 randomly selected subjects. The studies were designed to determine iron absorption from each test given alone in meal 1, and with different amounts of vitamin A, b-carotene or coffee in meals 24 (Table 1). Each subject was included in only one study. Four meals were given to each individual. Radiolabeled iron was added to the water to prepare the cereal. Each meal contained either 59Fe or 55Fe (Table 1). The rst meal in each study was the corn, wheat or rice test given alone and administered after an overnight fast. The second meal was given in the afternoon of the same day. Blood was drawn 15 d later to determine the hematological prole of the subjects and measure the radioactivity of blood samples. The subjects were fed again in the morning and in the afternoon of d 15 with meals 3 and 4, and blood was drawn again on d 30 to measure the radioactivity in blood and serum ferritin. Duplicate 10 ml blood samples and triplicate samples of the radioactive foods were prepared for scintillation counting using the technique of Dern and Hart (1961a and 1961b). Iron absorption from food was calculated from the radioactivity in the subjects blood using

estimated blood volume based on sex, weight and height (Nadler et al. 1962). The protocol for the administration of radioactive food in the morning after an overnight fast and the afternoon of the same day was based on experiments previously published (Taylor et al. 1995). Four-hour intervals between meals are sufcient for iron absorption studies. Chemical analysis. The total iron concentration of foods was determined by the digestion method (Bothwell et al. 1979), phytates by the method of Haug and Lantzsch (1983), tannate by the method of Price and Butler (1977), vitamin A by the method of Strohecker and Heming (1965) modied by Covenin (1989) and b-carotene by the method of Strohecker and Heming (1967). Phytate concentrations in precooked corn our, wheat our and rice were 257, 161 and 170 mg/100 g, respectively. The mean tannin concentration in coffee was 2500 mg/100 g. In studies in which coffee was included, each subject consumed coffee prepared by the infusion of 8 g of ground coffee beans. The mean iron concentration was 6 mg/100 g in the fortied precooked corn our, 3 mg/100 g in white wheat our, and 4 mg/100 g in boiled rice. The molar ratio of iron to phytate was 0.29 in precooked corn our, 0.23 in the wheat our and 0.30 in boiled rice.

TABLE 2
Changes in added Vitamin A and b-carotene centration during preparation and cooking of meals1
After cooking procedures Initially added Rice test Corn test Wheat test

mmol
Vitamin A 1.1 (1000) 2.2 (2000) 4.4 (4000) 1.7 (1000) 3.4 (2000)

mmol per 100 g cereal

0.55 (496) 1.51 (1376) 2.78 (2526) 0.58 (342) 0.95 (558) 0.37 (338) 0.61 (554) 0.73 (661) 0.67 (395) 1.53 (902) 1.1 (1000)2 0.66 (600) 1.51 (1372) 0.85 (500) 2.06 (1211)

b-carotene

1 Values are means, n 3; numbers in parentheses are in IU. 2 Vitamin A was not added to the dough. It was dissolved in water

and administered, freshly prepared, as a beverage.

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TABLE 3
Effect of increasing doses of vitamin A on iron absorption in humans from rice, corn and wheat tests1
Iron absorption Subjects and gender2 Hemoglobin g/L Rice, study 1 6 M, 11 F Corn, study 2 6 M, 11 F Wheat, study 3 3 M, 17 F 130 { 2 29 { 2 31 { 1 4.2a { 1.2 135 { 5 27 { 2 28 { 1 5.8a { 1.2 126 { 4 23 { 2 29 { 1 3.9a { 1.1 Serum transferrin saturation % Serum ferritin Meal 13 Meal 2 Meal 3 Meal 4

mg/L

Vitamin A administered, mmol 0.55 8.4b,c { 1.1 0.37 9.8b { 1.2 1.51 7.5b { 1.2 0.61 13.9c { 1.1 0.66 7.1b { 1.1 2.78 11.7c { 1.2

0.73 11.8b,c { 1.1 1.51 7.5b { 1.1

1.10 7.4b { 1.1

1 Values are means { SEM. Means in a row with dissimilar letters are signicantly different, P 0.05. Vitamin A was administered as a beverage while eating the bread in the wheat study, meal 2. 2 M, male; F, female. 3 No Vitamin A was added to any meal 1.

The mean tannin content in 8 g of coffee powder was 200 mg. The molar ratio of iron to tannin content in the corn test was 1.0, 0.50 in the wheat test, and 0.66 in the rice test. Because of the long distance between the laboratory and the place were the experiments were performed, the meals were administered to the subjects 1 d after the meals were prepared. Variations in added vitamin A and b-carotene content due to cooking procedures are shown in Table 2. In the case of the administration of 1.1 mmol (1000 IU) of vitamin A, the wheat bread was prepared separately, and this amount of vitamin A was dissolved in 20 ml water and drunk while eating the baked bread. In a previous experiment, we demonstrated that 80% of the vitamin A is lost during preparation of the dough and baking procedures due to the effect of yeast and the prolonged heat (Layrisse et al. 1997). Solubility of iron compounds at pH 2 and pH 6. The effect of vitamin A and b-carotene on iron solubility at pH 2 and 6 was measured. Four iron compounds were tested: ferrous sulfate, ferrous fumarate, an iron-amino acid chelate (Ferrochel) and Ferric-sodium EDTA. Ferrous sulfate was studied because it is a reference compound for iron studies. Fumarate is the salt used in Venezuela for enrichment of corn and wheat ours. Ferric-sodiumEDTA and Ferrochel (Al-

bion Laboratories, Cleareld, UT) were tested due to their stability at alkaline pH. Iron solutions from each of the compounds mentioned, containing 5 mg of iron, were prepared in 0.1 mol/L HCl containing 0, 0.55, 1.1, 1.65, 2.2 and 3.3 mmol (0, 500, 1000, 1500, 2000 and 3000 IU) of vitamin A or 0, 0,85, 1.70, 2.55, 3,40 and 5.10 mmol (0, 500, 1000, 1500, 2000 and 3000 IU) of b-carotene. A 2-ml aliquot was taken to measure soluble iron at pH 2, and to the remaining solution, the pH was adjusted to 6 with careful addition of NaOH. After standing 10 min at room temperature, duplicate 1-ml aliquots from the top of the solution were taken, and iron was measured for all iron compounds with different concentrations of vitamin A and b-carotene at pH 2 and 6 by the digestion method (Bothwell et al. 1979). Statistical analysis. Paired t test was performed for iron absorption studies comparing all meals within each study. It also was used to compare iron solubility at pH 2 and 6 in presence of vitamin A or b-carotene.

RESULTS Iron absorption studies. The effect of increasing doses of vitamin A on nonheme iron absorption from meals containing

TABLE 4
Iron absorption from meals containing rice, corn or wheat given alone or administered with Vitamin A or b-carotene1
Iron absorption Subjects and gender2 Hemoglobin g/L Serum transferrin saturation % Serum ferritin Meal 1 Meal 2 Meal 3 Meal 4

mg/L

Vitamin A

b-Carotene mmol

Rice, study 4 3 M, 8 F Corn, study 5 6 M, 14 F Wheat, study 6 3 M, 16 F 132 { 11 25 { 2 18 { 1 3.0a { 1.1 130 { 11 30 { 2 24 { 1 3.0a { 1.1 129 { 11 28 { 2 21 { 1 2.1a { 1.1

1.51 4.6b { 1.1 0.61 6.6b { 1.2 0.66 5.5b { 1.1 8.3c

0.58 6.4b,c { 1.1 0.67 8.5b { 1.1 8.8c

0.95 { 1.1 1.53 6.3b { 1.2 2.06 8.4c { 1.1

0.85 { 1.1

1 Values are means { SEM. Means in a row with dissimilar letters are signicantly different, P 0.05. Coffee infusion was administered with meal 3 for the rice meal and with meal 4 for the other two studies.

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TABLE 5
Effects of vitamin A content and pH on in vitro iron solubility
Ferrous fumarate pH Vitamin A2 Fe mg 3.87 0.99 1.32 1.75 1.90 2.01 3.0 { { { { { { { 0.1b 0.2a 0.3a 0.2a 0.2a 0.1a 0.5b 100 25.6 34.1 45.2 49.1 51.9 77.5 Soluble iron % { { { { { { { 2.6 5.2 7.8 5.2 5.2 2.6 12.9 6.18 2.24 3.33 3.85 3.88 4.18 5.47 Fe mg { { { { { { { 0.4b 0.2a 0.1a 0.4a 0.2a 0.1a 0.3b Ferrous sulfate Soluble iron % 100 36.2 53.9 62.3 62.8 67.6 88.5 { { { { { { { 6.5 3.2 1.6 6.5 3.2 1.6 4.9

mmol
2 6 6 6 6 6 6 0.55 (500) 1.10 (1000) 1.65 (1500) 2.20 (2000) 3.30 (3000)

1 Values are means { SEM, n 5. Means in a column with dissimilar letters are signicantly different, P 0.05. Initial iron content: 5 mg. 2 Values in parentheses are in IU.

rice, corn and wheat is shown in Table 3. Iron absorption from the meal given alone was signicantly lower than when vitamin A was included. There were no signicant differences in iron absorption from rice meals containing from 0.55 to 2.78 mmol (4962526 IU) vitamin A. The same patterns of iron absorption were observed from the tests containing pre cooked corn our and white wheat our. In study 4, iron absorption from the rice meal containing vitamin A (meal 2, Table 4), was more than double the absorption from the meal given alone. In the absorption test in which the meal contained 0.58 and 0.95 mmol (342 and 558 IU) bcarotene, iron absorption was more than two and three times greater than the meal given alone. The same pattern of iron absorption was observed for corn and wheat tests. The addition of 8 g of coffee powder as a beverage to meal 3 for rice test (0.58 mmol b-carotene) and meal 4 for corn and wheat tests (1.53 and 2.06 mmol b-carotene, respectively) (Table 4), produced no difference in iron absorption compared with meals with lower or higher content of b-carotene and without coffee, but the values were still signicantly higher than those obtained when the meal alone was consumed. Iron solubility with pH changes. Changes in iron solubility when pH is raised from 2 to 6 in presence of vitamin A are shown in Table 5. For ferrous fumarate, pH increase produced a 75% decrease in iron solubility. However, when vitamin A was added and the pH raised to 6, iron solubility in-

creased as a function of vitamin concentration. Iron solubility increased in 20% with 1.1 mmol (100 IU) of vitamin A, and when 3.3 mmol (3000 IU) was added, solubility was 80%. Ferrous sulfate showed a similar pattern. Iron solubility decreased in 64% when pH was raised to 6. When vitamin A was added iron solubility increased, and at 3.3 mmol (3000 IU), iron solubility was near 90%. When b-carotene was added to ferrous fumarate solutions at pH 2 and then raised to pH 6, virtually all of iron remained in solution when 3.4 mmol (2000 IU) or 5.1 mmol (3000 IU) of b-carotene was added (Table 6). For ferrous sulfate, 92% of iron remained in solution at pH 6 when 5.1 mmol (3000 IU) of b-carotene was added, whereas only 36% was soluble at pH 6 without any provitamin addition. Differences in percentage of soluble iron are not statistically signicant between ferrous fumarate and ferrous sulfate for the same content of vitamin A or b-carotene, except for 0.55 mmol vitamin A or 0.85 mmol b-carotene (P 0.05). It was not possible to observe an effect of vitamin A or bcarotene on iron solubility for EDTA and Ferrochel because iron is 100% soluble at pH 6 without any vitamin addition. DISCUSSION Vitamin A content is reduced to less than half during cooking procedures, but this amount of vitamin A signicantly

TABLE 6
Effects of b-carotene content and pH on in vitro iron solubility
Ferrous fumarate pH Ferrous sulfate Fe mg 2.6 2.6 5.3 2.6 2.6 2.6 2.6 6.25 2.24 2.57 3.17 3.57 4.96 5.74 { { { { { { { 0.1e 0.3a 0.1a 0.2b 0.2c 0.1d 0.1e Soluble iron % 100 35.8 41.1 50.7 57.1 79.4 91.8 { { { { { { { 1.6 4.8 1.6 3.2 3.2 1.6 1.6

b-Carotene2 mmol

Fe mg 3.78 0.99 1.00 1.96 3.18 3.54 3.81 { { { { { { { 0.1d 0.1a 0.2a 0.1b 0.1c 0.1d 0.1d

Soluble iron % 100 26.2 26.5 51.9 84.1 93.7 100 { { { { { { {

2 6 6 6 6 6 6

0.85 (500) 1.70 (1000) 2.55 (1500) 3.40 (2000) 5.10 (3000)

1 Values are means { SEM, n 5. Values in a column with dissimilar letters are signicantly different, P 0.05. Initial iron content: 5 mg. 2 Values in parentheses are in IU.

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Covenin, N. V. de A. (1989) Determinacion de vitamina A. Caracas, Verezuela. Dern, J. R. & Hart, W. L. (1961a) Studies with doubly labeled iron. I. Simultaneous liquid scintillation counting isotopes of 55Fe and 59Fe as ferrous perchlorate. J. Lab. Clin. Med. 57: 322330. Dern, J. R. & Hart, W. L. (1961b) Studies with doubly labeled iron. II. Separation of iron from blood samples and preparation of ferrous perchlorate for liquid scintillation counting. J. Lab. Clin. Med. 57: 460467. Flowers, C. A., Kuizon, M., Beard, S. L., Skikne, B. S., Covell, A. & Cook, J. D. (1986) A serum ferritin assay for prevalence studies of iron deciency. Am. J. Hematol. 23: 141151. GarcB a-Casal, M. & Layrisse M. (1996) Forticacion del hierro de los alimentos y su impacto sobre la deciencia de hierro. In: Anemia por deciencia de hierro en la region Andina. Denicion y estrategias de intervencion (Berger, J., San Miguel, J., Arze, R., Fernandez, E., and Aguanyo, V., eds) pp 193 200. Orstom, La Paz, Bolivia. Haug, W. & Lantzsch, H. J. (1983) Sensitive method for the rapid deter mination of phytate in cereals and cereal products. J. Sci. Food. Agric. 34: 1423 1426. Hodges, R., Sauberlich, H., Ganham, J., Wallace, D., Rucker, R., MejB a, L. & Mohanram, M. (1978) Hematopoietic studies in vitamin A deciency. Am. J. Clin. Nutr. 31: 876885. International Committee for Standardization in Hematology (1978) Recommendation for measurement of serum iron in human blood. Br. J. Haematol. 38: 291294. International Committee for Standardization in Hematology (1978) The measurement of total and saturated iron-binding capacity in serum. Br. J. Haematol. 38: 281290. Layrisse, M., Chaves, J. F., Mendez-Castellano, H., Bosch, V., Tropper, E., Bas tardo, B. & Gonzalez E. (1996) Early response to the impact of iron fortication in the Venezuelan population. Am. J. Clin Nutr. 64: 903907. Layrisse, M. & GarcB a-Casal, M. (1997) Strategies for the prevention of iron deciency through foods in the household. Taller: desnutricion oculta en Lat inoamerica. Deciencia de Hierro. CESNI, Nutr. Rev. 55: 233239. Layrisse, M. & GarcB a, M. N. La respuesta a la forticacion con hierro en la pobla cion venezolana. Revista de Salud Publica de Mexico (in press). Layrisse, M., GarcB a-Casal. M., Solano, L., Baron, M., Arguello, F., Llovera, D., RamB rez, J., Leets, I. & Tropper, E. (1997) The role of vitamin A on the inhibitors of nonheme iron absorption: preliminary results, J. Nutr. Biochem. 8: 6167. MejB a, L. A. (1986) La deciencia de la vitamina A como factor d anemia nutrici onal. In: Vitaminas, agentes nutritivos y terapeuticos. (C. Rozo and M Mamome., eds.), pp. 6574. Ediciones Doyoma S.A. Barcelona, Spain. MejB a, L. and Arroyave, G. (1982) The effect of vitamin A fortication of sugar on iron metabolism in preschool children in Guatemala. Am. J. Clin. Nutr. 36: 8793. MejB a, L. A. & Chew, V. (1988) Hematological effect of supplementing iron with vitamin A alone and in combination with iron. Am. J. Clin. Nutr. 48: 595600. Mejia, L. A., Hodges, E. & Rucker R. B. (1979) Role of Vitamin A in the absorption, retention and distribution of iron in the rat. J. Nutr. 109: 129137. Nadler, S. B., Hidalgo, J. U. & Bloch, T. (1962) The Tulane table of blood volume in normal men. Surgery (St Louis) 51: 224232. Olson, J. (1984) Vitamin A. In: Handbook of Vitamins. Nutritional, Biochemical and Clinical Aspects (Machlin, L. J., ed.), vol. 28, pp. 145. Marcel Dekker, New York, NY. Price, M. L. & Butler, L. (1977) Rapid visual estimation and spectrophotometric determination of tannin content of sorghum grain. J. Agric. Food. Chem. 25: 12681273. Roche Laboratories (1994). b-Carotene. In: Vitamins. Basics. Basel, Switzerland. F. Hoffmann-La Roche, pp. 58. Strohecker, R. & Heming, H. (1965) Vitamins assay test method. Verlag Chem. CMBV, Weinheim. Strohecker, R. & Heming, H. (1967) Analisis de vitaminas (Paz, ed.). pp. 73 79. Montalvo, Madrid, Spain. Taylor, P., MartB nez-Torres, C., Mendez-Castellano, H., Jaffe, W., Lopez de Blanco, M., Landaeta Jimenez, M., Leets, I., Tropper, E., RamB rez, J., GarcB a-Casal, M. N. & Layrisse, M. (1995) Iron bioavailability from diets consumed by different socioeconomic strata of the venezuelan population. J. Nutr. 125: 18601868. Wang, X. D. (1994) Absorption and metabolism of beta-carotene. J. Am. Coll. Nutr. 13: 314325.

increased iron absorption from rice, corn and wheat meals compared with these meals given alone. It also shows that the increase in doses from 0.37 to 2.78 mmol (338 to 2526 IU) did not produce any further signicant increase. b-carotene has the same effect on iron absorption than vitamin A. In the case of rice meal, it was shown that 0.95 mmol (558 IU) of b-carotene increases iron absorption more than threefold compared with the meal given alone. Absorption from the other two cereals showed similar behavior. It seems that b-carotene also prevents the inhibitory effect of polyphenols on iron absorption. When coffee was administered with the test meals containing b-carotene, iron absorption did not show a signicant decrease as expected. On the contrary, it increased from one- to twofold compared with the meal without b-carotene or coffee. Solubility tests demonstrated that vitamin A and b-carotene are capable of solubilizing iron at pH 6. It seems that bcarotene is more efcient that vitamin A at least for ferrous fumarate. This is may be due to a greater stability of b-carotene to storage, handling and pH changes. Results from solubility experiments as well as human absorption studies show an important role for vitamin A and bcarotene in improving iron absorption especially from foods with a high content of inhibitors, which are the staple foods of many countries worldwide. The unexpected behavior of vitamin A and b-carotene of improving iron absorption requires further studies to discover the mechanism for this peculiar reaction. A previous publication (Layrisse et al. 1997) showed that spectrophotometric results, elution patterns from high-performance liquid chromatography (HPLC) and solubility of iron with vitamin A at pH 6 suggest that vitamin A binds iron liberated during the digestive process and forms a complex that acts as chelating agent preventing the inhibitory effect of phytates and polyphenols on nonheme iron absorption. This hypothesis agrees with the results of Mejia (1986), who demonstrated interactions between vitamin A and iron metabolisms. It also supported the dramatic reduction of the prevalence of iron deciency after 1 y of an iron-fortication program in Venezuelan population, in which one food vehicle, precooked corn our, was fortied with vitamin A (Layrisse et al. 1996). Because iron and vitamin A deciencies are the two main deciencies in many developing populations (Bloem et al. 1989, MejB a and Arroyave 1982), it is recommended that both micronutrient should be used in fortication programs. LITERATURE CITED
Bloem, M. W., Wedel M., Egger, R., Speek A. J. & Shrisjver, J. (1989) Iron metabolism and vitamin A deciency in children in the Northwest Thailand. Am. J. Clin. Nutr. 50: 332338. Bothwell, T. H., Charlton, R. W., Cook, J. D. & Finch, C. A. (1979) Iron metabolism in man. Blackwell Scientic Publications, Oxford. Crosby, W. H., Munn, J. L. & Furth, F. W. (1954) Standardizing a method for clinical hemoglobinometry. U S Armed Forces Med. J. 5: 693703.

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