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Use of amino acid profiles of preterm and term human milks in evaluating scoring patterns for routine protein quality assessment of infant formulas.

Reports on the amino acid composition of human milk vary considerably with respect to concentrations of sulfur amino acids. Often, analyses forego tryptophan determination. A complete analysis of protein and amino acid concentrations was performed on human milk samples (5-10 days postpartum) collected from mothers of preterm (gestations of 25-32 weeks) and term (gestations of > 36 weeks) infants. Careful attention was given to quantitate amino acids such as cysteine and tryptophan, which are vulnerable to acidic hydrolysis conditions. Differences in concentrations of total amino acids (expressed on protein basis) between preterm and term milks were small, despite the higher true protein content of preterm milk versus term milk (19.20 versus 12.60 g/L). The methionine + cyst(e)ine contents of term and preterm milks (3.72-3.84 g/100 g protein) were comparable with those reported in 1991 by the Food and Agricultural Organization/World Health Organization (FAO/WHO) for mature human milk (4.20 g/100 g protein) but higher than those reported in 1991 by the European Commission (2.9 g/100 g protein). The amino acid pattern of human milk obtained in this study confirms that the 1991 FAO/WHO amino acid scoring pattern for predicting protein quality of infant formulas is representative of the amino acid quality of both preterm and term human milks.

Amino Acids↗

Triglycerides, fatty acids, sterols, mono- and disaccharides and sugar alcohols in human milk and current types of infant formula milk.

OBJECTIVE: To investigate differences in the fatty acid composition, sterols, minor carbohydrates and sugar alcohols between human and formula milk. DESIGN: We analyzed the concentrations of triglycerides, sterols, di- and monosaccharides and sugar alcohols, as well as the fatty acid composition of 10 currently available types of formula milk for term babies. Results were compared with mature human milk from 99 exclusively breast-feeding Dutch women, who collected 24-hour samples in the second week (n = 99), sixth week (n = 99) and 3 months (n = 25) after delivery. Infant formula milk data were considered different if they fell outside the mean +/- 2s.d. range of corresponding human milk data. RESULTS: The triglyceride concentrations in human milk were lower than those of the formula milk, possibly due to an incomplete collection of fat-rich hindmilk. Formula milks tended towards a higher proportion of medium chain fatty acids and lower proportions of longer-chain polyunsaturated fatty acids. Formulas had cholesterol concentrations 3-35 times lower, and much higher phytosterol concentrations, compared with the human milk. In the formula milk types the glucose, sorbitol and myoinositol concentrations were generally lower, whereas the fucose and erythreitol concentrations were in the lower mean +/- 2s.d. human milk range. The galactose concentrations in the formulas were generally higher. CONCLUSIONS: Formula milk and human milk differ considerably in fatty acid composition and concentrations of cholesterol, phytosterols, monosaccharides and sugar alcohols. The biological consequences of these differences in composition are uncertain.

Carbohydrates↗

Animal models in food allergy: assessment of allergenicity and preventive activity of infant formulas.

Food allergies occur in about 5-10% of the overall infant and small-child population. Cow's milk protein allergy (CMPA) is the most common in young infants, with a 2-4% incidence. When breastfeeding is not possible, hypoallergenic (HA) cow's milk based formulas are usually given during the first months of life for prevention of CMPA. Depending on primary (sensitization) or secondary (triggering) prevention, the requested quality of HA formulas may be different. Besides in vitro methods, in vivo and ex vivo animal models are helpful in assessing residual allergenicity and the preventive effect of HA formulas. The sensitizing capacity of a formula can be examined by either the parenteral rat (IgE), the guinea pig (IgG1a mediated) or the oral mouse (IgE) models. The triggering IgE mediated allergenicity is tested by a parenteral rat model with oral gavage for intestinal mast cell protease (RMCPII) release. These animal models are also used for testing the oral tolerance inducing capacities of formulas. Together with cellular in vitro assays, animal models are very helpful in predicting allergenicity and the tolerogenic potential of HA infant formulas.

Animals↗

Determination of biotin by high-performance liquid chromatography in infant formula, medical nutritional products, and vitamin premixes.

A solid-phase extraction sample preparation procedure was developed for use with a high-performance liquid chromatography (HPLC) method for biotin analysis. The HPLC method used a reversed-phase C18 column; chromatography run time was 8.5 min. After eluting from the column, biotin went through postcolumn reaction to form a conjugate with streptavidin-fluorescein isothiocyanate, which was then detected by a fluorescence detector. This method was tested with infant formula, medical nutritional products, and vitamin premix samples.

Biotin↗

Assessment of the efficacious dose of arachidonic and docosahexaenoic acids in preterm infant formulas: fatty acid composition of erythrocyte membrane lipids.

The nutritional requirements of preterm infants for the long chain polyunsaturated essential fatty acids, arachidonic acid (AA) and docosahexaenoic acid (DHA), have not been clearly defined. The present study evaluated preterm infants of less than 2.3 kg birth weight fed a commercial formula (Preemie SMA) devoid of AA and DHA and compared this control group with similar infant groups fed one of three formulas containing a range of 0.32 to 1.1% AA and 0.24 to 0.76% DHA. An analogous group of infants fed their mothers' breast milk and a breast milk fortifier (when indicated) was also studied. Erythrocyte membrane phospholipids were isolated from blood samples collected at 12 d of age and after a further 4 wk of feeding. Infants fed the formula without AA and DHA showed a reduction in AA level in erythrocyte phosphatidylcholine, and a reduced level of DHA in phosphatidylethanolamine in comparison with infants fed breast milk or infant formula containing AA and DHA. Supplementing infant formula with increasing levels of AA and DHA produced a clear dose response in the levels of AA and DHA found in erythrocyte membrane phospholipids. From comparison of membrane phospholipid fatty acid composition it appears that a formula level of 0.32-1.1% AA and 0.24-0.76% DHA provides sufficient levels of these fatty acids to achieve a similar fatty acid composition to that of infants fed human milk for most of the lipid fractions examined.

Analysis of Variance↗

Antigenicity of infant formulas: role of immature intestine on protein permeability.

Preliminary studies of 25 infants fed casein hydrolysate, soy-based or milk-based formulas were undertaken to determine the importance of age with respect to the relative antigenicity of formula proteins. Infants fed casein hydrolysate for the first three months had lower antibody titers (hemagglutinins) to subsequent milk-based or soy-based proteins than those given these formulas since birth. The clinical implications with regard to atopy and gastrointestinal protein intolerance are discussed. Soy protein is at least as antigenic as milk protein and should be used with caution in prophylaxis against possible dietary antigen/antibody-related disease.

Animals↗

Innovation in infant formula development: a reassessment of ribonucleotides in 2002.

Nucleotides play a significant role in many physiologic functions, ranging from the encoding of genetic information to signal transduction. Accumulating evidence from recent animal and clinical studies supports the original notion that nucleotides are semi-essential dietary nutrients. The development of a new technique, total potentially available nucleosides, to accurately quantify the nucleotide content in various biological fluids has allowed investigators to properly assess the importance of nucleotide function and availability in various conditions. Data from animal studies indicate that exogenous nucleotides produce beneficial gastrointestinal and immunologic effects, especially during times of rapid growth, when nucleotide availability may be low. Infant studies confirm these findings and are presented in this review. Regulatory agencies are currently using this information collectively to support the rationale for the supplementation of infant formulas with higher amounts of nucleotides.

Animals↗

High performance liquid chromatographic determination of vitamin D in fortified milks, margarine, and infant formulas.

Fortified milks were saponified overnight at room temperature with 1% ethanolic pyrogallol and KOH. The digest was extracted with hexane after adding water and ethanol, and the extract was washed consecutively with 5% KOH, water, and 55% aqueous ethanol to remove polar lipids. After evaporation, the residue was first chromatographed on a column of 5 micrometer silica. A fraction containing vitamin D was collected, evaporated, and rechromatographed on a reverse phase column for the separation and quantitation of vitamins D2 and D3. Recovery was 96-99% and the coefficient of variation was 3% (8 replicates). Infant formula was diluted and then saponified and extracted as in the analysis of milk. Margarine was saponified by shaking overnight with 1% ethanolic pyrogallol and 80% KOH. Water and ethanol were added to the digest before extraction. Extracts from formula and margarine were chromatographed as milk except, before HPLC, the extract was dissolved in isopropanol-hexane (1 + 99) and passed through 5 cm alumina in a Pasteur pipet, and the concentration of isopropanol in the first high performance liquid chromatographic (HPLC) solvent system was halved to improve the separation of vitamin D from other absorbing lipids. Usually several peaks were obtained during the final HPLC analysis, and the identification of vitamins D2 and D3 was less certain than in the analysis of milk. The coefficients of variation for formula and margarine were 6% (5 replicates) and 9% (6 replicates), respectively.

Animals↗

Polyamines in human and rat milk and in infant formulas.

Polyamines are ubiquitous compounds known to be involved in cell proliferation and differentiation in many tissues. Enteral administration of these compounds has been shown to produce effects in suckling and adult animals. Using HPLC techniques, we verified the presence of putrescine, spermidine, and spermine in human milk and quantitated their concentration in samples collected from the first week up to 4 mo of lactation. Mean values of these compounds ranged (per liter) from 0 to 615 nmol putrescine, from 73 to 3512 nmol spermidine, and from 722 to 4458 nmol spermine. Polyamine concentrations in infant formulas were dependent on the protein source, the particular polyamine, and the protein concentration of the formula. Concentrations of these three compounds in rat milk over the first 3 wk of lactation were higher than in human milk, with spermidine being the polyamine most elevated compared with human milk (almost 20-fold higher). An artificial formula used for the rearing of suckling rats contained trace to immeasurable amounts of polyamines. Our study identifies milk as one vehicle for polyamine delivery to the intestinal mucosa of suckling animals.

Animals↗

Exposure of infants to phyto-oestrogens from soy-based infant formula.

BACKGROUND: The isoflavones genistein, daidzein, and their glycosides, found in high concentrations in soybeans and soy-protein foods, may have beneficial effects in the prevention or treatment of many hormone-dependent diseases. Because these bioactive phyto-oestrogens possess a wide range of hormonal and non-hormonal activities, it has been suggested that adverse effects may occur in infants fed soy-based formulas. METHODS: To evaluate the extent of infant exposure to phyto-oestrogens from soy formula, the isoflavone composition of 25 randomly selected samples from five major brands of commercially available soy-based infant formulas were analysed, and the plasma concentrations of genistein and daidzein, and the intestinally derived metabolite, equol, were compared in 4-month-old infants fed exclusively soy-based infant formula (n = 7), cow-milk formula (n = 7), or human breast-milk (n = 7). FINDINGS: All of the soy formulas contained mainly glycosides of genistein and daidzein, and the total isoflavone content was similar among the five formulas analysed and was related to the proportion of soy isolate used in their manufacture. From the concentrations of isoflavones in these formulas (means 32-47 micrograms/mL), the typical daily volume of milk consumed, and average bodyweight, a 4-month-old infant fed soy formula would be exposed to 28-47 per day, or about 4.5-8.0 mg/kg bodyweight per day, of total isoflavones. Mean (SD) plasma concentrations of genistein and daidzein in the seven infants fed soy-based formulas were 684 (443) ng/mL and 295 (60) ng/mL, respectively, which was significantly greater (p < 0.05) than in the infants fed either cow-milk formulas (3.2 [0.7] and 2.1 [0.3] ng/mL), or human breast-milk (2.8 [0.7] and 1.4 [0.1] ng/mL), and an order of magnitude higher per bodyweight than typical plasma concentrations of adults consuming soy foods. INTERPRETATION: The daily exposure of infants to isoflavones in soy infant-formulas is 6-11 fold higher on a bodyweight basis than the dose that has hormonal effects in adults consuming soy foods. Circulating concentrations of isoflavones in the seven infants fed soy-based formula were 13000-22000 times higher than plasma oestradiol concentrations in early life, and may be sufficient to exert biological effects, whereas the contribution of isoflavones from breast-milk and cow-milk is negligible.

Animals↗

Predigestion of soybean proteins with immobilized trypsin for infant formula.

Soybean protein isolates of low soybean trypsin inhibitor (STI) residue were prepared by acidic precipitation of soybean flour water extracts (0.8-1.2%) at pH 5.0, followed by acidic washing at this pH and affinity adsorption of residual STI with immobilized trypsin on polystyrene anion-exchange resin GM 201. After heat treatment, soybean protein isolates were subjected to controlled hydrolysis with the immobilized trypsin. Then, the predigested soybean protein was prepared. The predigested soybean protein was free of STI activity, and its solubility at acidic pH range was greatly increased. Sedimentation test showed that it formed a much finer clot at pH 4.5 than that of untreated soybean protein. The pepsin digestibility index at pH 4.0 and chymotrypsin digestibility index at pH 8.0 were obviously improved. These results suggested that the predigested soybean protein prepared by this method may be used in infant formulas.

Chemical Precipitation↗

A perfusion reversed-phase chromatographic method for ultrarapid determination of soybean proteins in soybean infant formulas and soybean milks: method development and validation.

Perfusion reversed-phase high-performance liquid chromatography (perfusion RP-HPLC) is applied for the first time to the determination of soybean proteins in commercial samples prepared from soybean protein isolate. A linear binary gradient of acetonitrile-water-0.1% trifluoroacetic acid at a flow rate of 3 mL/min and a temperature of 60 degrees C is optimized to analyze soybean proteins in approximately 3 min. Direct injection of samples into the chromatographic system is possible by dissolving them in water. Quantitation of soybean proteins is achieved by the proposed method using a soybean protein isolate as an external standard. The method is validated and applied to the quantitation of soybean proteins in infant formulas and powdered and liquid soybean milks.

Calibration↗

Protein in infant formulas. Future aspects of development.

Cow's milk protein intolerant infants (CMPI), shifted to a cow's milk protein hydrolysate based formula, often experience an improvement in signs of malabsorption; on the other hand, they also present a pattern of watery stools. In an attempt to find high levels of some gastrointestinal hormones that might induce hypermotility, motilin and neurotensin levels were studied in infants on starting formula and in cow's milk protein intolerant infants, on a cow's milk protein hydrolysate based formula. In 12 infants on unmodified cow's milk based formula (starting formula), motilin levels were: mean 71.66 pmol/L (s: 17); neurotensin 26.53 (s: 10.9). In 19 cow's milk protein intolerant infants on a cow's milk protein hydrolysate based formula, motilin levels were: mean 163.65 pmol/L (s: 70.06) (p<0.05); neurotensin: 31.76 pmol/L (s: 15.03) (p>0.05). Motilin (but not neurotensin) is higher in cow's milk protein intolerant infants on a cow's milk protein hydrolysate based formula. We conclude that high motilin levels can induce a different pattern of motility, and can be a pathogenetic factor in the persistence of loose and watery stools in infants with CMPI on a protein hydrolysate based formula.

Body Height↗

Dietary intakes of selected elements from longitudinal 6-day duplicate diets for pregnant and nonpregnant subjects and elemental concentrations of breast milk and infant formula.

As part of a longitudinal investigation into mobilization of lead from the maternal skeleton during pregnancy and lactation, we have determined the daily intake of selected elements (hereafter called micronutrients) for various subjects and compared these intakes with recommended and/or published intakes, especially those of the United States, through the U.S. National Health and Nutrition Examination Survey (NHANES). We also sought to ascertain whether there was any seasonal effect in the diets. Six-day duplicate diets were collected from 15 pregnant and 16 nonpregnant migrants to Australia, 6 pregnant Australian control subjects, and 8 children of nonpregnant migrants (6 to 11 years). Samples of breast milk and infant formula were also analyzed. Blended samples were analyzed by inductively coupled plasma mass spectrometry for the elements Ca, Cu, Fe, Mg, P, K, Na, Zn, Ba, Sr, and Pb. Daily intakes of micronutrients were only about half of the daily intake estimated for non-Hispanic white females and infants in the U.S. NHANES III. Estimates of daily intakes from breast milk were also considerably lower for the migrant and Australian infants compared with the values extracted from tables of food composition and dietary recall for non-Hispanic white infants in the U.S. NHANES III. For example, Ca was a factor of approximately 3 times lower, Fe approximately 50, and Zn approximately 4. We consider our estimates a reliable indication of the daily intakes for several reasons, including the collection of up to nine quarterly collections of 6-day duplicate diets and retention of subjects in a longitudinal prospective study. The low intakes of the essential elements such as Ca, Fe, and Zn in all these population groups are of potential concern from a public health viewpoint.

Adult↗

Copper, iron, zinc, and manganese in dietary supplements, infant formulas, and ready-to-eat breakfast cereals.

High intakes of iron, zinc, or manganese can interfere with copper absorption. Thus, the purpose of this study was to determine whether the amounts and chemical forms of iron, zinc, manganese, and copper added to food products and nutritional supplements might pose a threat to copper status. More than 25% of the vitamin and mineral supplements examined contained no copper, 40% contained the poorly absorbed cupric oxide, and < 30% contained a highly bioavailable form of copper such as cupric sulfate or cupric chloride. Nearly 40% of the prenatal supplements examined contained both iron and zinc without a nutritionally significant amount of copper. More than 80% of the infant formulas examined had ratios of iron to copper exceeding 20:1, which is higher than the recommended ratios of 10-17:1. None of the 40 ready-to-eat breakfast cereals examined were fortified with copper or manganese although 50% of these cereals contained > or = 25% of the reference daily intake for both iron and zinc. Copper availability could be improved by reformulation of several food products and supplements.

Biological Availability↗

Betaine and homocysteine concentrations in infant formulae and breast milk.

BACKGROUND: Early diagnosis and treatment of homocystinuria with methionine-free or low methionine formulae significantly improve prognosis in children found by newborn screening and treated soon after birth. Betaine (Bet) supplementation is also an effective strategy for dietary treatment of homocystinuria. However, previous reports on diet therapy have only examined methionine and cysteine concentrations but not those of Bet and homocysteine (Hcy) in infant diets. METHODS: We measured Bet and Hcy concentrations in three methionine-free formulae, five infant starter formulae, and 33 samples of human breast milk from 10 mothers. RESULTS: In methionine-free formulae, Hcy but not Bet was isolated. However, in human breast milk and infant starter formulae, both Bet and Hcy were detected. However, the Bet concentration was not sufficient for the treatment. CONCLUSIONS: Our study suggests that methionine-free formulae should have Hcy eliminated and be fortified with Bet to the concentration of 150 mg/dL for the treatment of homocystinuria.

Animals↗