Douwina Bosscher

Availabilities of calcium, iron, and zinc from dairy infant formulas is affected by soluble dietary fibers and modified starch fractions.

OBJECTIVE Insoluble dietary fiber is a known inhibitor of mineral absorption, whereas the effects of soluble dietary fibers (including prebiotics) are less known. The aim was to study calcium, iron, and zinc availabilities from dairy infant formulas supplemented with soluble dietary fibers and modified starches in vitro. METHODS Dairy infant formulas were supplemented with soluble dietary fibers (3%, dry wt) and modified starches (16% pregelatinized rice starch and 1.9% maltodextrin, dry wt) and kept in a well-controlled and defined environment in vitro. Pooled mature human milk was used as the reference standard. RESULTS Calcium availability from standard formula was elevated by 30% after inulin supplementation (17.2%), whereas locust bean gum (11.9%) and high esterified pectin (11.7%) reduced availability by approximately 10%. Iron availability from standard formula was increased by pregelatinized rice starch (3.8%), whereas availability was reduced in the following order: high esterified pectin (2.3%), oligofructose (2.2%), and low esterified pectin (2.1%). Zinc availability was highest after the addition of pregelatinized rice starch (13.5%) but lowest with the addition of locust bean gum (6.8%) and maltodextrin (5.4%). CONCLUSIONS This study showed that addition of soluble dietary fiber affects calcium, iron, and zinc availabilities in positive (inulin) and negative ways, depending on the type of the dietary fiber used.

Effect of thickening agents, based on soluble dietary fiber, on the availability of calcium, iron, and zinc from infant formulas.

Although it is accepted that dietary fiber probably is not needed in the diets of infants younger than 1 y, babies suffering from regurgitation are often fed with infant formulas thickened with considerable amounts of fiber. The effect of increasing amounts of alginic acid, locust-bean gum, and guar gum was studied from casein and whey-based infant formulas. A dialysis in vitro method with preliminary intraluminal digestion, adapted to the conditions of infants younger than 6 mo, was used. Human milk was used as the reference standard. Elemental contents of samples and dialysates were determined by atomic absorption spectrometry. Soluble dietary fiber inhibited mineral availability more in casein than in whey-based formulas. Mineral availabilities from casein- and whey-based formulas supplemented with 0.42 g of locust-bean gum/100 mL were 9.4% (0.7) and 10.4% (0.6) for calcium (P < 0.05), 0.32% (0.08) and 1.45% (0.17) for iron (P < 0.05), and 3.2% (0.2) and 5.6% (0.5) for zinc (P < 0.05), respectively. Calcium availability from the whey formula decreased in the presence of each fiber source, especially guar gum and alginic acid. Supplementing 2 g of alginic acid-based agents per 100 mL depressed calcium availability from 13.3% (1.2) to 5.3% (0.3; P < 0.05). With respect to iron and zinc, availabilities increased from 1.28% (0.28) to 6.05% (0.96; P < 0.05) and from 6.7% (0.6) to 10.2% (1.0), respectively, with the addition of 2 g of alginic acid (P < 0.05). Both gums lowered iron and zinc availabilities, and guar gum affected iron availability more severely than locust-bean gum did. Iron availabilities were 1.45% (0.17) from formula thickened with locust-bean gum (0.42 g/100 mL) and 0.92% (0.15) from formula thickened with guar gum (P < 0.05). Adding thickening agents based on soluble dietary fiber to traditional infant formulas probably affects calcium, iron, and zinc availability in various ways.

Thickening infant formula with digestible and indigestible carbohydrate: availability of calcium, iron, and zinc in vitro.

BACKGROUND Regurgitation is common in infants and is usually due to gastroesophageal reflux. Often parental reassurance and dietary management by feeding thickened formulas are the only therapeutic steps necessary. Adding fibers may interfere with the absorption of micronutrients. METHODS A continuous-flow dialysis in vitro method with a preliminary intraluminal digestive phase, modified to simulate the conditions of infants less than 6 months of age and children from 6 months of age on, was used to study the availability of calcium, iron, and zinc from thickened and nonthickened first-and second-age infant formulas. Pooled mature human milk was used as the reference standard. The elemental content of the samples and dialysate fractions of calcium, iron, and zinc after digestion was determined by atomic absorption spectrometry. RESULTS In human milk, calcium, iron, and zinc were highly available for absorptive processes. Availability of calcium, iron, and zinc from nonthickened first- and second-age infant formulas tends to be significantly better than in the corresponding products thickened with locust bean gum. Thickening infant formulas with pregelatinized rice starch, however, does not affect the availability of calcium, iron, and zinc. CONCLUSIONS It appears that human milk provides optimal conditions for the availability of calcium, iron, and zinc. Availability of calcium, iron, and zinc seems to lower when infant formulas are thickened with indigestible carbohydrates, whereas it does not by adding digestible carbohydrates.

In vitro availability of calcium, iron, and zinc from first-age infant formulae and human milk.

Background Variation in the bioavailability of calcium (Ca), iron (Fe), and zinc (Zn) occurs because of interactions of food components in the gastrointestinal microenvironment. Bioavailability is preferably determined by in vivo tests, but these are expensive, labor-intensive, time consuming, and often unethical. As an alternative, in vitro methods can be used to predict bioavailability of nutrients from foodstuffs. Methods A continuous-flow dialysis model with preliminary intraluminal digestive phase, adapted to the gastrointestinal conditions of infants younger than 6 months, was used. Human milk was the reference standard. Ca, Fe, and Zn content of samples and dialysates after digestion were analyzed by atomic absorption spectrometry. Results Ca availability is similar in human milk (13.1% ± 0.8%), whey (13.3% ± 1.2%), and soy-based formulae (13.0% ± 1.2%;P > 0.05), and higher in casein-predominant formula (21.2% ± 0.6%;P < 0.05). Availability of Fe is highest in human milk (8.12% ± 0.27%;P < 0.05). Fe availability in whey (1.28% ± 0.28%) and soy formulae (1.48% ± 0.28%) is similar (P > 0.05), but availability is lower in casein-predominant formula (0.48% ± 0.22%;P < 0.05). Zn availability is also highest in human milk (13.1% ± 0.7%;P < 0.05). However, Zn availability is similar in whey (6.7% ± 0.6%) and casein formulae (8.5% ± 1.6%;P > 0.05), but lower in soy formula (2.3% ± 0.4%;P < 0.05). Conclusions Our observations are in agreement with previous data from in vivo studies in term infants. This in vitro procedure is an inexpensive, simple, rapid, and reliable method that predicts the bioavailability of Ca, Fe, and Zn in foods.

A method for **in vitro** determination of calcium, iron and zinc availability from first-age infant formula and human milk

A method for in vitro determination of available calcium, iron and zinc content from infant food after digestion was evaluated. This method introduced an intraluminal digestive phase, adapted to the gastrointestinal conditions of infants younger than 6 months of age, prior to continuous flow dialysis of the resultant gastric digest. Precautions handling the method were discussed and enzymatic parameters were defined. Ruggedness of the method was determined from the availability of calcium, iron and zinc at different gastrointestinal conditions. Availability of all three elements was higher at gastric pH of 2 (20.0 +/- 1.1% for calcium, 4.06 +/- 0.66% for iron and 17.5 +/- 1.3% for zinc), than from the normal procedure (pH 4) (15.6 +/- 1.2% for calcium, 1.18 +/- 0.26% for iron and 8.2 +/- 0.9% for zinc). At pH 5, however, calcium availability appeared to be lower (11.7 +/- 1.0%) (P < 0.05). The intestinal pH also had a major influence on the availability. At low intestinal pH (5.5), availability was 40.5 +/- 2.3% for calcium, 3.01 +/- 0.58% for iron and 26.8 +/- 1.8% for zinc, which was higher compared with the normal procedure (P < 0.05). Moreover, other factors, such as digestion time, mixing and filtration pressure, also affected the availability. Recovery tests yielded mean values of 94 +/- 3% for calcium, 109 +/- 9% for iron and 106 +/- 4% for zinc. Mean intra- and inter-batch precision of the availability procedure was 4.1 CV% and 6.6 CV% for calcium, 14.5 CV% and 19.2 CV% for iron, and 4.0 CV% and 13.6 CV% for zinc. The method provides adequate accuracy, acceptable precision and good recovery. It offers the advantage of being simple, rapid and inexpensive, since it takes only 1 day to run the whole availability procedure (including four replicates per sample), and the low costs of the dialysis equipment. It can therefore be considered as suitable for predicting the availability of essential elements from foods used during the first months of infancy.

Bioavailability of calcium and zinc from cow's milk-based versus soya-based infant food

A modified continuous flow dialysis in vitro method was used to study the availability of calcium and zinc from cows milk-based versus soya-based infant food. First and second age infant formulae, milk for pre-schoolers and cows milk were studied. The elemental content of the samples and dialysate fractions was determined by flame atomic absorption spectrometry. The relative availability of calcium and zinc from the soya-based first age infant formula (calcium: 31.6 ± 0.5%, zinc: 17.8 ± 1.8%), from the soya-based second age infant formula (calcium: 46.1 ± 2.1%, zinc: 20.9 ± 1.4%) and from the soya milk for pre-schoolers (calcium: 32.6 ± 0.7%, zinc: 39.7 ± 2.1%) proved to be as good as or even better than from the first age infant formula (calcium: 21.9 ± 1.4%, zinc: 23.3 ± 4.1%), from the second age infant formula (calcium: 31.0 ± 2.5%, zinc: 14.6 ± 0.7%) and from the milk for pre-schoolers (calcium: 27.7 ± 0.6%, zinc: 23.7 ± 2.8%) based on cows milk. A similar relative availability from semi-skimmed c...

Do thickening properties of locust bean gum affect the amount of calcium, iron and zinc available for absorption from infant formula? In vitro studies.

Locust bean gum acts as a milk thickener in infant formula because of its high apparent viscosity. The effects of such thickening agents on metabolic and physiologic responses during infancy have not been clarified sufficiently. Due to the increased volume of the digest and the bulking and trapping effects, digestion and absorption of nutrients may be influenced in presence of locust bean gum. The central question addressed in this paper is whether the thickening properties of locust bean gum affect the availability of calcium, iron, and zinc. Increasing amounts of powdered locust bean gum were homogenised with infant formula and samples were diluted to 0.14, 0.27, 0.42 and 0.71 g/100 ml. Viscosity of the samples was measured by a Carrie-Med CSL100 rheometer. Available amounts of calcium, iron, and zinc were evaluated using a continuous-flow dialysis model with preliminary digestion. Elemental contents of samples and dialysates were analysed with atomic absorption spectrometry. The first set of experiments showed that addition of locust bean gum to infant formulas increased the viscosity of the luminal contents. Correlations between the locust bean gum concentration and the viscosity of the samples before and after gastric or intestinal digestion were highly significant (0.97). In the second set of experiments, the correlations between the locust bean gum concentration and the amounts of calcium trapped by the locust bean gum fraction also showed high significance (0.93). In the third experimental design, again strong correlations were found between the viscosity of the intestinal digest and the amounts of calcium trapped by the fibre fraction (0.90). For iron and zinc, no such relationships were found. From this experimental set-up it appears that locust bean gum influences calcium availability in infant formulas by means of its physical properties to act as thickening agent, rather than its chemical ability to form complexes as demonstrated earlier with respect to iron and zinc.

Silicon in Foods: Content and Bioavailability

The silicon content of various foodstuffs marketed in Belgium was measured by a validated graphite furnace absorption spectrometric method. Dietary intake has been identified as the major source of silicon. However, data on its bioavailability remain scarce and insufficient. In vitro methods can provide an indication of bioavailability in case of lacking in vivo data. Bioavailability of silicon from different foodstuffs was estimated using an in vitro continuous flow gastroduodenal simulation method. The major food sources of silicon were unrefined grains, cereal products and root vegetables. The availabilities of silicon from, meat, milk and beers were high, whereas low availability was observed for seafood and cereal products. Plotting the availability data versus the total elemental silicon content of the foods revealed an exponential inverse relationship. The inverse relationship between silicon content and silicon availability was found in all foods, with the exception of various silicon containing drinks. Nevertheless, food categories classified as major silicon sources in the diet still appear to provide the highest absolute amounts of available silicon per 100 g of food including breakfast cereals, bread and baking products, and beers.

Daily dietary fibre intake of children, 2 to 3 years of age, living in Antwerp, Belgium

Daily dietary fibre intake of toddlers living in Antwerp (Belgium) have been calculated by weighted food records with subsequent conversion using computerised food composition tables. Dietary fibre intakes were also measured by the duplicate portion technique to evaluate the intakes of total, soluble and insoluble dietary fibre. Calculated dietary fibre intake was 10 ± 3 g/d. Measured intakes of total, soluble and insoluble dietary fibre were 14.5 ± 2.8 g/d, 6.0 ± 1.9 g/d, and 8.6 ± 1.6 g/d. Values about the fibre intakes between both methods differed for 31.7%. Bread contributed for more than one third of the total dietary fibre intake. The contributions of the other food groups were 15.1% for fruits, 14.5% for starchy foods and 13.9% for vegetables. The intakes of dietary fibre from bread by the toddlers in this study could be considered as high, whilst intakes from fruits and vegetables might be increased.

Development and Validation of an in vitro Experimental GastroIntestinal Dialysis Model with Colon Phase to Study the Availability and Colonic Metabolisation of Polyphenolic Compounds

The biological effects of polyphenols depend on their mechanism of action in the body. This is affected by bioconversion by colon microbiota and absorption of colonic metabolites. We developed and validated an in vitro continuous flow dialysis model with colon phase (GastroIntestinal dialysis model with colon phase) to study the gastrointestinal metabolism and absorption of phenolic food constituents. Chlorogenic acid was used as model compound. The physiological conditions during gastrointestinal digestion were mimicked. A continuous flow dialysis system simulated the one-way absorption by passive diffusion from lumen to mucosa. The colon phase was developed using pooled faecal suspensions. Several methodological aspects including implementation of an anaerobic environment, adapted Wilkins Chalgren broth medium, 1.10(8) CFU/mL bacteria suspension as inoculum, pH adaptation to 5.8 and implementation of the dialysis system were conducted. Validation of the GastroIntestinal dialysis model with colon phase system showed a good recovery and precision (CV < 16 %). Availability of chlorogenic acid in the small intestinal phase (37 ± 3 %) of the GastroIntestinal dialysis model with colon phase is comparable with in vivo studies on ileostomy patients. In the colon phase, the human faecal microbiota deconjugated chlorogenic acid to caffeic acid, 3,4-dihydroxyphenyl propionic acid, 4-hydroxybenzoic acid, 3- or 4-hydroxyphenyl acetic acid, 2-methoxy-4-methylphenol and 3-phenylpropionic acid. The GastroIntestinal dialysis model with colon phase is a new, reliable gastrointestinal simulation system. It permits a fast and easy way to predict the availability of complex secondary metabolites, and to detect metabolites in an early stage after digestion. Isolation and identification of these metabolites may be used as references for in vivo bioavailability experiments and for investigating their bioactivity in in vitro experiments.