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Determination of pesticides in soy-based infant formula using liquid chromatography with electrospray ionization tandem mass spectrometry.

A sensitive method using liquid chromatography with electrospray ionization tandem mass spectrometry (LC/ESI-MS/MS) was developed and validated to quantify and confirm 13 pesticides, including aldicarb sulfoxide, aldicarb sulfone, oxamyl, methomyl, formetanate, 3-hydroxycarbofuran, carbendazim, thiabendazole, aldicarb, propoxur, carbofuran, carbaryl, and methiocarb, in soy-based infant formula. Data acquisition under MS/MS was achieved by applying multiple reaction monitoring of 2 fragment ion transitions to provide a high degree of sensitivity and selectivity for both quantitation and confirmation. Different approaches to constructing calibration curves were compared and discussed to address issues of the extraction efficiency or recovery, and matrix effects. Matrix-matched standard calibration curves with the use of isoprocarb as an internal standard were finally used to achieve the best accuracy of the method. Under most circumstances, recoveries of 13 pesticides, spiked at 5.0, 25.0, and 45.0 microg/kg, were close to 100%. The method detection limits (signal-to-noise ratio > or =3:1; microg/kg) of 13 pesticides were 0.2 for thiabendazole and methiocarb, 0.6 for aldicarb, and 0.1 for the others.

Aldicarb↗

Monoclonal and polyclonal antibodies against casein components of cow milk for evaluation of residual antigenic activity in 'hypoallergenic' infant formulas.

BACKGROUND: Hydrolysed casein and whey protein formulas have been developed with the aim of preventing sensitization in infants at high risk of cow milk allergy. Subsequently these products have also been used for treatment of children with cow milk allergy. However, severe reactions have occurred in some allergic infants fed with these formulas raising doubts about their absolute safety and suggest the need for developing in vitro techniques for detection of eventual residual allergenic activity in such preparations. OBJECTIVES: Our purpose was to evaluate the usefulness of monoclonal and polyclonal antibodies against casein components (alpha, beta and kappa casein) as reagents for the detection of the residual antigenic activity of casein components in several hydrolysed formulas. METHODS: The monoclonal and polyclonal antibodies were produced according to standard procedures by immunizing female Balb/c mice with casein fraction (a mixture of alpha, beta and kappa casein). ELISA assays were developed to test the specificity of the antibodies and to detect and evaluate the amount of residual antigenic activity of the casein components in hydrolysed formulas. RESULTS: Use of polyclonal antiserum specific for casein allowed detection of residual antigenic activity of casein components in all partial hydrolysates and in the two extensive whey protein hydrolysates in the amounts ranging from 0.05 to 0.67% of total protein. No such activity was detectable in either the two extensive casein hydrolysates tested or the aminoacid based formula. The polyclonal antiserum proved to be more suitable than monoclonals for detecting residual antigenic activity in the hydrolysates. The monoclonal antibodies were directed against epitopes expressed on different casein components. CONCLUSIONS: In this study the ELISA inhibition assay with polyclonal antibodies specific for casein components of cow milk proved to be a sensitive method for estimating residual antigenicity in the hydrolysed formulas commercially available for infants with cow milk allergy suggesting their potential application for the quality control of hypoallergenic infant formulas.

Animals↗

Fluoride content of infant formulas and market milk in Turkey.

The aim of the present study was to test the fluoride contents of infant formulas and market milk in Turkey. Fifteen formulas and nine market milks were prepared according to manufacturers' instructions with deionized water. Fluoride contents were analyzed with a spectrophotometer (CADAS 50S) and results were obtained as ppm (microgF/ml). Mean fluoride content of the formulas was 0.101 ppm F and of the milks was 0.08 ppm F. Formulas reconstituted with water containing < 0.3 ppm F do not provide a daily fluoride intake at any age. To decrease dental caries among children in Turkey, fluoride supplements could be prescribed at minimal dosages according to the ADA fluoride schedule.

Animals↗

Population-based study of the developmental outcome of children exposed to chloride-deficient infant formula.

In 1978 and 1979 a large number of US children were fed chloride-deficient soy-based infant formula. A representative sample of such children was identified in a southern county through a mailing to the homes of 3639 first- and second-grade children in the public schools. Of the 2329 (64%) who responded, 56 reported use of deficient formula and were invited to have developmental testing by one of four study psychologists at their school. Of the 310 users of other soy formulas, 112 were selected for testing as matched controls on the basis of their sex, feeding history, age, birth weight, and socioeconomic status (as indicated by school attended). After exclusions and refusals, 42 children who used deficient formula and 66 control children were tested using the McCarthy Scales of Children's Abilities. Examiners were unaware of the child's history of formula use. The mean General Cognitive Index was 102.8 in those using deficient formula and 105.4 in controls. After adjustment for demographic differences the children who used chloride-deficient formula were found to average 4.9 points less than the controls (P = .04, 1-tailed). The largest difference was in the Quantitative subscale (P = .005). These data show a statistically significant although small effect of chloride-deficient formula on the long-term developmental outcome of exposed children; however, further study of these results is needed for full confirmation.

Child↗

Effect of infant formuLas on blood and tissue cholesterol, bone calcium, and body composition in weanling pigs.

Weanling pigs were fed four commercial infant formulas to determine effects on blood and tissue cholesterol parameters and on body composition. Two milk protein (MP)-based formulas and two soy protein isolate (SPI)-based isocaloric formulas were fed in concentrated liquid form for 32 days. A commercial sow's milk replacer fed to a fifty group of pigs served as a control diet. Pigs fed SPI-based formula had significantly less cholesterol in the plasma than did pigs fed MP-based formula. Whole-body and adipose tissue cholesterol concentrations were greatest in pigs fed formulas containing a relatively high concentration of polyunsaturated fatty acids (PUFA). Liver cholesterol concentration was inversely related to plasma cholesterol concentrations. Bone calcium, measured as percentage of dry, fat-free bone (femur), and whole-carcass ash were significantly less in pigs fed SPI-based formula than in pigs fed MP-based formula. Similar growth and development were observed in pigs fed SPI-based or MP-based formulas; however, pigs fed SPI-based formulas had significantly less bone calcium.

Animals↗

The association of progesterone, infant formula use and pacifier use with the return of menstruation in breastfeeding women: a prospective cohort study.

OBJECTIVES: This study sought to explore hormonal and postnatal factors, which are associated with the return of menstruation in breastfeeding mothers. DESIGN: A prospective cohort study in Bristol, UK. Ninety-one breastfeeding mothers of mixed parity were recruited at community-based antenatal clinics. Eighty-five mothers completed all stages and informed us about the return of menstruation. Data were collected from interviews and questionnaires antenatally, immediately postnatally, and 1 week, 4 weeks and 6 months later. Blood samples were taken at all time points except at 6 months. Associations between hormone levels, postnatal factors and the return of menstruation were explored using univariate and multivariate statistical survival analysis. RESULTS: Sixty-five (76%) mothers were still breastfeeding at 6 months and 46 (54%) of these were fully breastfeeding (not giving formula milk); 21 (32%) mothers started menstruating in the first 6 months whilst they were still breastfeeding and 14 were fully breastfeeding at 6 months. Survival analysis modelling showed that formula use (RR=4.27; 95% CI=1.89, 9.67), pacifier (dummy) use (RR=3.26; 95% CI=1.35, 7.84) and lower postnatal progesterone levels (P=0.006) each made a significant contribution to the chance of earlier resumption of menstruation. Prolactin and oestradiol levels showed no associations with the return of menstruation. CONCLUSIONS: Higher postnatal progesterone levels are associated with delayed menstruation; while the use of pacifiers and infant formula milk are associated with an earlier return to menstruation.

Adult↗

Differences in protein digestibility and quality of liquid concentrate and powder forms of milk-based infant formulas fed to rats.

Protein and amino acid digestibility and protein quality of liquid concentrate and/or powder forms of infant formulas were studied by rat balance and growth methods. Casein plus methionine (control) and eight formulas were fed to weanling rats as the sole source of protein in diets containing 8% protein (nitrogen X 6.25). Values for true digestibility of protein, lysine, methionine, or cystine (85-92%) in liquid concentrates were up to 13% lower than those in powders. Similarly, the 2-wk relative protein-efficiency ratio values (64-85%) or the relative net protein ratio values (78-94%) of liquid concentrates were up to 25% lower than those for powders. Lower levels of bioavailable lysine and methionine plus cystine in liquid concentrates compared with powders (prepared by the same manufacturer) would suggest that inferior protein quality of liquid concentrates may be due to more heat treatment involved in their preparation.

Amino Acids, Essential↗

Fluorescence, browning index, and color in infant formulas during storage.

Free and total fluorescent compounds, browning index, and color formation were measured in milk-based powdered infant formulas (IF) during 2 years of storage at 20 and 37 degrees C. The excitation spectra from 415 nm emission show three peaks (ex lambda1 = 270 nm, lambda2 = 325/315 nm, lambda3 = 350 nm) and from 347 nm excitation two emission peaks (415 and 520 nm), and no wavelength shifts were observed. Temperature and time of storage exert in general no significant effect on the development of fluorescence emission intensity and browning index. However, an important increase in pentodilysine was recorded-probably because of the iron and ascorbic acid contents of the samples-as well as in browning index in adapted IF. In both IF a color increase (deltaE) throughout storage was observed, this increase being greater in samples stored at 37 degrees C than in those stored at 20 degrees C. The increase in color with time fitted a linear regression model. Color appeared to be an indicator of sufficient sensitivity to measure the effect of temperature or storage time.

Color↗

Trace element absorption in infants as a foundation to setting upper limits for trace elements in infant formulas.

The bioavailability of the trace elements iron, zinc, copper and manganese from human milk is high compared to cow's milk and infant formulas. This high bioavailability may be explained by the presence of lactoferrin in human milk, which may facilitate iron and manganese uptake via an intestinal receptor for this protein. High concentrations of ascorbate and citrate may also facilitate uptake of trace elements from human milk and milk formulas, while a high concentration of casein in cow's milk and cow's milk formulas may limit trace element absorption from these diets. Trace element absorption from soy formula is low, mostly due to the presence of phytate but possibly also due to some protein fraction. Trace elements sharing absorptive pathways compete for uptake, and imbalances in the ratios between trace elements (Fe/Zn, Zn/Cu, Fe/Mn) in formulas may impair trace element absorption. These factors need to be taken into consideration when setting upper limits for trace elements in formulas. With our present knowledge, an upper limit for iron of 14 mg/l, for zinc, 12 mg/l, copper, 1.2 mg/l, and manganese, 0.6 mg/l are suggested. The capacity of infants to homeostatically adapt to varying intakes of trace elements needs to be further evaluated.

Female↗

History and current status of infant formulas.

Formulas that approximate the composition of human milk are widely used as the sole source of nutrients for infants during the first months of life. Such formulas contain milk, soy, or meal proteins, or protein hydrolysates together with those forms of fat carbohydrate, vitamins, and minerals shown to be bioavailable to the infant. Legislation concerning nutrient composition and quality assurance of products labeled as infant formulas has been enacted by Congress. Governmental and industrial standards also help to assure microbiological safety and stability. Problems of nutrient composition and adverse reactions have occurred during the evolution and development of infant formulas. Problems concerning nutrient deficiency are now almost unknown, but adverse reactions of individuals to specific formula ingredients continue. Accurate information on composition and related nutritional properties of infant formulas is essential for their effective use and for the avoidance of undesirable complications.

Animals↗

Liquid chromatographic determination of vitamin K1 trans- and cis-isomers in infant formula.

The normal phase liquid chromatographic (LC) method for determination of trans- and cis-isomers of vitamin K1 (phylloquinone) in infant formula described here uses an Apex silica column, isocratic elution, and UV absorption detection at 254 nm. Vitamin K1 is extracted quantitatively from the product matrix by pretreating the as-fed liquid with concentrated ammonium hydroxide and methanol, and then extracting it with a 2:1 mixture of dichloromethane and isooctane. The extract is cleaned up by silica open-column chromatography and concentrated for LC analysis. For trans-vitamin K1, the method precision is less than or equal to 3.3% RSD (relative standard deviation), and the spike recovery is 98 +/- 4%. For cis-vitamin K1, the precision is less than or equal to 12% RSD, determined at levels near the detection limit, and the spike recovery is 95 +/- 9%. The detection limit is 0.3 ng for both isomers at signal/noise = 3.

Chromatography, Liquid↗

PCR and oligonucleotide array for detection of Enterobacter sakazakii in infant formula.

Enterobacter sakazakii has been implicated in a several form of neonatal meningitis with a high mortality rate. In the present study, the species-specific PCR and oligonucleotide array assays were developed to detect the 16S-23S rDNA internal transcribed spacer (ITS) of E. sakazakii. Two pairs of specific PCR primers and 10 oligonucleotide probes were designed by sequencing the ITS of six strains of E. sakazakii and BLAST of GenBank. The specificity and efficiency of the PCR and oligonucleotide array methods were tested against a panel of numerous strains from 88 different bacterial strains. All of the E. sakazakii strains generated positive signal, and no cross-reaction was observed with non-E. sakazakii strains in the PCR and oligonucleotide array detections based on ITS sequences. Sensitivity of the detections is 1.3 CFU/100 g infant formula with the selective enrichment. Both of the PCR and oligonucleotide array procedures take only 48 h including the enrichment culture, whereas the conventional methods required at least 5 days. This study demonstrated that both of the pathogenic detections are time-saved and reliable.

Cronobacter sakazakii↗

Effect of linoleic acid-rich infant formula feeding on brain synaptosomal lipid accretion and enzyme thermotropic behavior in the piglet.

The effects of a vegetable oil-based infant formula, virtually devoid of n-6 and n-3 long chain polyenoid fatty acids (LCP) and high in 18:2(n-6) and 18:2(n-6)/18:3(n-3) ratio, on brain synaptosome lipid composition and enzyme thermotropic behavior were studied in neonatal piglets. Term gestation piglets were fed either sow milk (SMF) or formula (FF) from birth for 5, 10, 15, or 25 days. Synaptosomal cholesterol, total lipid phosphorus, and phospholipid class composition did not differ between SMF and FF piglets. Synaptosomal fatty acid composition, however, was influenced by diet. The proportion of n-3 LCP, especially 22:6(n-3), was decreased, while the n-6 LCP, especially 22:4(n-6) and 22:5(n-6), were increased in FF compared to SMF piglets. These diet-related changes were most pronounced in the ethanolamine glycerophospholipid fraction and increased with the duration of feeding. FF thus reversed an apparent developmental increase in the synaptosomal n-3/n-6 LCP ratio. The monoene content, especially 18:1, was also reduced in the synaptosomes of FF compared to SMF pigs. FF had no effect on the activity of synaptosomal acetylcholinesterase. However, higher transition temperatures for this enzyme, indicating decreased membrane fluidity, were found in the FF compared to SMF piglets. The data suggest that exclusive feeding of proprietary formulae, devoid of LCP and high in 18:2(n-6) and/or the 18:2 (n-6)/18:3(n-3) ratio, may compromise normal fatty acid accretion and physical properties of brain synaptosomal membranes.

Acetylcholinesterase↗

Influence of a mineral water on the rheological characteristics of reconstituted infant formulas and diluted cows' milk.

A bottled spring water with a low mineral content was compared with tap water in the reconstitution and/or dilution of five different infant formulas and cows' milk. The osmolality, buffering power and renal solute load potential of the formulas reconstituted with the bottled water were all significantly lower than when tap water was used (P < 0.01). When the bottled water was used to dilute cows' milk, the morphology of milk casein precipitates (after addition of rennet) was finer and more dispersed than when tap water was used. For formula reconstitution and milk dilution, a benefit, in terms of solute/electrolyte balance, appears to be conferred on infants by the improved rheological characteristics of modified milks reconstituted or diluted with this bottled mineral water.

Analysis of Variance↗

Taurine analysis in milk and infant formulae by liquid chromatography: collaborative study.

A collaborative study was conducted on a liquid chromatographic (LC) method for determination of taurine in infant formula and milk powders. Twenty laboratories participated in the analysis of 8 blind duplicates over the range of approximately 3-60 mg/100 g sample. The method involved protein removal, conversion to the dansyl-derivative, and isocratic LC separation with UV and/or fluorescence detection. Following outlier treatment, overall mean RSDR has been estimated at 7.00% for supplemented products with a HORRAT value of 1.1. The poorer precision at endogenous levels establishes a lower limit of determination of about 5 mg/100 g. An overall mean RSDr:RSDR value of 0.7 for all products demonstrated acceptable performance.

Acetonitriles↗

Determination of vitamin K in milk and infant formulas by liquid chromatography: collaborative study.

A simple procedure for determination of vitamin K1 was developed for routine compliance monitoring of supplemented infant formula and measurement of endogenous levels in milk and milk powders. Samples are digested with lipase and extracted into hexane; and aliquot is evaporated, reconstituted in methanol, and analyzed by reversed-phase LC. Post-column zinc reduction of phylloquinone facilitates detection by fluorescence. The procedure was subjected to an AOAC collaborative study involving 8 materials, each in blind duplicate, across the range of 5-120 micrograms/100 g solids and including NIST 1846 reference material. Thirty-three laboratories returned valid data which were then statistically analyzed for outliers and precision parameters. Mean RSDR (%) was 6.53 (4.33-10.94), with a mean HORRAT value of 0.33 (0.23-0.43) and RSDr:RSDR ratio of 0.74. K1 isomers (cis and trans) were aggregated with conventional C18 columns, but may be selectively estimated with use of the C30 column.

Animals↗