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This research aims at determining some free amino acids in amino acid-based infant formulas and amino acid-modified medical foods for inborn errors of metabolism to prove their quality. A method based on high-performance liquid chromatography and diode array detection was developed and validated. Then, overall uncertainty was estimated by the bottom-up approach. Applying the weighted least squares regression method suggested good linearity with coefficient of determinations ≥ 0.9960. The limits of detection were calculated between 0.01 and 0.28 μg/mL. The most repetitive recovery values were obtained in the range of 91-108%, with RSDs ≤ 15%. The expanded uncertainties were below 20% for most amino acids. The contributions of linear regression and repeatability are two main factors in estimating overall uncertainty. The results offer this method as a simple and easy procedure for analyzing free amino acids in seven powdered medical foods designed for phenylketonuria, maple syrup urine disease, methylmalonic, and propionic acidemia., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

2- and 3-monochloropropanediol esters (MCPDEs) are most commonly formed as process-induced contaminants during the refinement of vegetable oils used for food production. 'In vivo' hydrolysis of 3-MCPDEs releases the potential carcinogen 3-monochloropropanediol (3-MCPD). Levels of MCPDEs in infant formula are of particular concern, as refined oils are commonly used as main fat ingredients. For this study, infant formula samples (powders, liquid concentrates and ready-to-feed infant formula samples) from the Canadian market were purchased and analysed in 2015 (35 samples) and 2019 (33 samples). MCPDE concentrations (expressed as free MCPD equivalents) were examined through an indirect analytical approach, applying acid-catalysed ester cleavage and using cyclohexanone as derivatising agent. Labelled diesters were used as internal standards. 2015 Survey data were analysed by gas chromatography-mass spectrometry (GC-MS) in selected ion monitoring mode (SIM). 2019 Survey data were analysed with an updated method using GC-MS/MS in multiple reaction monitoring modes (MRM). In 2015, levels in reconstituted formula ranging from 3.7 ng/g to 111 ng/g for 3-MCPD and 2.2 ng/g to 56.2 ng/g for 2-MCPD were found. In 2019, levels ranging from 3.9 ng/g to 74.8 ng/g for 3-MCPD and 1.0 ng/g to 33.9 ng/g for 2-MCPD were found. A significantly reduced mean of combined MCPDEs was observed between 2015 and 2019 data (64.5 ng/g, standard deviation (SD) 8.6 ng/g in 2015 to 31.8 ng/g, SD 5.6 ng/g in 2019, p -value = 0.024). For the majority of manufacturers, the data comparison among brand products over time shows decreased levels of MCPDEs. Occurrence data of MCPDEs, including data from previously published surveys (2012/2013), were also compared and a temporal trend was established.

Background: While high protein intake during infancy may increase obesity risk, low qualities and quantities of protein contribute to undernutrition. This study aimed to investigate the impact of the amount and source of protein on infant growth during complementary feeding (CF) in a country where under- and overnutrition co-exist as the so-called the double burden of malnutrition., Methods: A multicenter, prospective cohort was conducted. Healthy term infants were enrolled with dietary and anthropometric assessments at 6, 9 and 12 months (M). Blood samples were collected at 12M for IGF-1, IGFBP-3 and insulin analyses., Results: A total of 145 infants were enrolled (49.7% female). Animal source foods (ASFs) were the main protein source and showed a positive, dose-response relationship with weight-for-age, weight-for-length and BMI z-scores after adjusting for potential confounders. However, dairy protein had a greater impact on those parameters than non-dairy ASFs, while plant-based protein had no effect. These findings were supported by higher levels of IGF-1, IGFBP-3 and insulin following a higher intake of dairy protein. None of the protein sources were associated with linear growth., Conclusions: This study showed the distinctive impact of different protein sources during CF on infant growth. A high intake of dairy protein, mainly from infant formula, had a greater impact on weight gain and growth-related hormones.

Background: Since the publication of Standard Method Performance Requirements (SMPR®) for vitamin D in infant formula (SMPR 2011.004) by AOAC INTERNATIONAL, revised vitamin D limits have been recommended by the European Food Safety Authority (EFSA) for infant formula and adopted in Commission Delegated Regulation (EU) 2019/828. The vitamin D range introduced, 2-2.5 μg/100 kcal, is significantly narrower than previous limits specified by Codex Standard 72-1981 and requires lower method reproducibility metrics to adequately assess regulatory compliance. The narrower limits for vitamin D present a significant challenge for current-generation reference analytical methods that comply with SMPR 2011.004., Objective: We evaluate the impact of Delegated Regulation (EU) 2019/828 on the demonstrated performance of AOAC Method 2016.05/ISO 20636:2018 to assess the likelihood that vitamin D results produced by the method would be found outside the EU limits when testing infant formula that is compliant as manufactured., Methods: AOAC Method 2016.05/ISO 20636:2018, specifically data generated during multi-laboratory study, was used as a basis for statistical evaluation of the impact of the narrower EU vitamin D limits., Results: The review of AOAC Method 2016.05/ISO 20636:2018 method performance against the vitamin D regulatory range introduced in (EU) 2019/828 indicates methods capable of performing in alignment with SMPR 2011.004 are likely to produce results that fail to meet EU requirements., Conclusions: Our assessment illustrates the high probability that a well-manufactured product with vitamin D levels within the EU regulatory range would fail to meet the regulatory requirements due to analytical method variability when tested using fit-for-purpose methods. Further, required method performance cannot be expected with the future development of new methods. To avoid this, consideration should be given to aligning proposed regulatory limits with method performance metrics of current-generation compendial methods., Highlights: Current, state-of-the-art methods cannot consistently verify infant formula product compliance for vitamin D in accordance with (EU) 2019/828., (© The Author(s) 2022. Published by Oxford University Press on behalf of AOAC INTERNATIONAL. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Children are highly vulnerable to chemical exposure. Thus, metal and metalloid in infant formulas are a concern, although studies in this regard are still relatively scarce. Thus, the presence of aluminum, arsenic, cadmium, tin, mercury, lead, and uranium was investigated in infant formulas marketed in Brazil by inductively coupled plasma mass spectrometry, and the Target Hazard Quotients (THQ) and Target Cancer Risk (TCR) were calculated in to assess the potential risk of toxicity for children who consume these products continuously. Aluminum ranging from 0.432 ± 0.049 to 1.241 ± 0.113 mg·kg -1 , arsenic from 0.012 ± 0.009 to 0.034 ± 0.006 mg·kg -1 , and tin from 0.007 ± 0.003 to 0.095 ± 0.024 mg·kg -1 were the major elements, while cadmium and uranium were present at the lowest concentrations. According to the THQ, arsenic contents in infant formulas showed a THQ > 1, indicating potential health risk concerns for newborns or children. Minimal carcinogenic risks were observed for the elements considered carcinogenic. Metabolic and nutritional interactions are also discussed. This study indicates the need to improve infant formula surveillance concerning contamination by potentially toxic and carcinogenic elements.

In this study, solid-phase extraction (SPE) combined with the dispersive liquid-liquid microextraction based on novel hydrophobic deep eutectic solvent (DLLME - DES) has been developed as an ultra-pre-concentration technique for the extraction of aflatoxin M 1 (AFM1) in milk-based baby food (MBBF) and infant formula milk (IFM) samples followed by HPLC combined with fluorescence detection (HPLC - FL). In addition, carcinogenic and non-carcinogenic risk assessment was performed by health-related risk factors including liver cancer risk (LCR), margin of exposure (MOE) and target hazard quotient (THQ) were calculated using the mean of AFM1 in different infant food samples. The results of the study showed that the mean of AFM1 was statistically significant different between various brands and types of IFM and MBBF. The results of the study showed that the percentage of positive samples higher than the allowable limit of AFM1 in 36 samples of domestic infant formula milk (DIFM), 24 samples of imported infant formula milk (IIFM), 36 samples of domestic milk-based baby food (DMBBF) and 18 samples of imported milk-based baby food (IMBBF) were 41.6, 12.5, 66.7 and 33.3%, respectively. In addition, estimated values for health risk-related factors including LCR, MOE and THQ indicated that for most infants less than one-year-old were higher than the acceptable levels. Based on the results, it can be concluded that the quality of IFM and MBBF consumed in Iran in terms of AFM1 is poor. Therefore, it is necessary to take appropriate measures to reduce the amount of AFM1 in DIFM and DMBBF, and in addition, the IIFM and IMBBF should be controlled qualitatively before supplying the market.

Infants who accustomed to consume human milk can hardly adapt to the odor of infant formula in a short time and prefer the odor of human milk. In this study, the sensory-directed analysis was used to investigate the odor differences between human milk and infant formula. Aroma extraction dilution analysis (AEDA) results showed that carbonyl compounds and alcohols were the most important components with the higher dilution factors (FD) in human milk and infant formula. There were 14 key aroma active compounds (OAV ≥ 1) in human milk, like octanal, linalool, benzaldehyde, and furfural, while 11 in infant formula, like hexanal, 1-octen-3-one, (E)-2-octenal, and octanal. The aroma recombination and omission experiment further revealed that compounds such as (E)-2-decenal, linalool, 2-furanmethanol, 2-pentylfuran, (E,E)-2,4-heptadienal, nonanal, (E)-2-nonenal, and 1-octen-3-one were the major reason for the odor difference between human milk and infant formula., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Objective: To explore the contamination characteristics of chloropropanol esters and glycidyl esters in infant formulas sold in Beijing in 2021, and to evaluate the exposure risk of chloropropanol esters and glycidyl esters for infants and toldders aged 0-36 months old., Methods: The contents of chloropropanol esters and glycidyl esters in infant formula samples were determined using gas chromatography-mass spectrometry with deuterated internal standards. Combined with the recommended consumption of infant formulas, the exposure level of chloropropanol esters and glycidyl esters in infants and toddlers aged 0-36 months was calculated., Results: The detection rate of 3-chloropropane-1, 2-diol esters(3-MCPDE), 2-chloropropane-1, 3-diol esters(2-MCPDE) and glycidyl esters in infant formulas were 98.6%, 97.1% and 95.7%, respectively. The average contents of 3-MCPDE, 2-MCPDE and glycidyl esters were 44.54, 15.65 and 12.65 μg/kg. For infant of each age groups, the daily intakes of 3-MCPDE via infant formulas by each infant groups were 0.28-0.90 μg/(kg BW), which were all lower than the tolerable daily intake(TDI, 2 μg/(kg BW));the daily intakes of 2-MCPDE via infant formulas by each infant groups were 0.10-0.29 μg/(kg BW);the exposure levels of glycidyl were 0.08-0.22 μg/(kg BW), and the margin of exposure(MOE) values were all higher than 25 000., Conclusion: Chloropropanol esters and glycidyl esters in infant formulas sold in Beijing from 2021 were less polluted and their intake was within the safe range.

Osteopontin (OPN) is a multifunctional whey protein with numerous health benefits and is recommended to supplement Infant Milk Formula (IMF) with it. For the first time, the denaturation kinetics and storage stability of bovine OPN were studied in reconstituted IMF. The effect of thermal treatments (63-90 °C) and High Pressure Processing (HPP) were investigated. OPN showed higher retention (around 90%) after both LTLT (Low Temperature Long Time) and HTST (High Temperature Short Time) treatments, however higher temperature and longer treatment induced substantial denaturation indicating UHT may not suit OPN-added items. Significant denaturation occurred at higher pressure (>400 MPa), and more pronounced OPN denaturation was observed after HPP at elevated temperature. OPN remained stable during4 °Cstorage for 14 days. This investigation deduced thermal stability of OPN as α-Lac > OPN > LF, whereas the pressure resistance observed as α-Lac > LF > OPN > β-Lg., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Background: In 2018, the AOAC Stakeholder Panel on Strategic Food Analytical Methods approved Standard Method Performance Requirement (SMPR®) 2018.009, for lactose in low-lactose or lactose-free milk, milk products, and products containing dairy ingredients, establishing the minimum recommended performance characteristics to be addressed during the evaluation of methods. Subsequently, AOAC INTERNATIONAL opened a call for methods under the Official Method of AnalysisSM program with the aim of finding a candidate method for confirming compliance with regulatory standards and dispute resolution., Objective: A biosensor-based analytical method, BIOMILK 300 LAC, was developed by BIOLAN Microbiosensores S.L. (www.biolanmb.com) to rapidly, easily, and accurately quantify lactose in free or low-lactose dairy products. In response to the AOAC call for methods, BIOLAN performed a single laboratory validation of this method against SMPR 2018.009. Several different matrixes were tested, including: milk, sugary plain yogurt, fruit plain yogurt, flavored liquid yogurt, Greek yogurt, cream, soft cheese, infant formula, café latte, chocolate milk, and high-protein milk shake. Evaluated method parameters included: linearity, selectivity, matrix effect, recovery, accuracy, repeatability, intermediate reproducibility, robustness, reagent lot-to-lot consistency, and stability., Methods: The method is based on the use of the Biotest gold electrode together with the BIOMILK 300 biosensor reader, for the quantification of residual levels of lactose in dairy samples via an enzymatic recognition/electrochemical transduction system., Results: Assay linearity, applicability to different matrixes, recovery, and precision demonstrated that the method is fit for purpose. The method proved to be robust, consistent, and stable, under conditions detailed in the Instructions For Use guide., Conclusion: The overall results were within requirements stated by SMPR 2018.009 for low-lactose and lactose-free milk, milk products and products containing dairy ingredients., Highlights: The BIOMILK 300 LAC method enables the quantification of reduced levels of lactose in less than 5 min, without the requirement for expert technicians, toxic solvents or intricate procedures, maintaining a high degree of precision and accuracy in the results. BIOMILK 300 LAC was adopted as a First Action Official MethodSM by the Expert Review Panel of Low-lactose Methods in April 2020 after rigorous review., (© The Author(s) 2021. Published by Oxford University Press on behalf of AOAC INTERNATIONAL. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Infant milk formulas are designed to substitute human milk when breastfeeding is unavailable. In addition to human milk and milk-derived products, these formulas can be a vehicle of contaminants. In this work, a multiclass method based on the QuEChERS (quick, easy, cheap, effective, rugged, and safe) approach was developed for the simultaneous determination of contaminants (n = 45), including mycotoxins and veterinary drug residues, occurring in infant milk formulas. By using an ultra-high-performance liquid chromatography quadrupole-Orbitrap coupled with high-resolution mass spectrometry analysis (UHPLC-Q-Orbitrap HRMS; Thermo Fisher Scientific), further retrospective analysis of 337 contaminants, including pesticides, was achieved. The method was validated in accordance with European regulations and applied for the analysis of 54 infant milk samples. Risk assessment was also performed. Dexamethasone was detected in 16.6% of samples (range: 0.905-1.131 ng/mL), and procaine benzyl penicillin in 1 sample at a concentration of 0.295 ng/mL. Zearalenone was found in 55.5% of samples (range: 0.133-0.638 ng/mL) and α-zearalenol in 16.6% of samples (range: 1.534-10.408 ng/mL). Up to 49 pesticides, 11 veterinary drug residues, and 5 mycotoxins were tentatively identified via retrospective analysis based on the mass spectral library. These findings highlight the necessity of careful evaluation of contaminants in infant formulas, considering that they are intended for a vulnerable part of the population., (The Authors. Published by Elsevier Inc. and Fass Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Upon the European commission's request in 2013, Denmark initiated a survey of the levels of chlorate and perchlorate in agriculture products from the Danish market over a period of 4 years from 2014 to 2017. The results of the survey were used to guide decision making with a view to setting maximum levels for intra-Union trade in 2013 and reviewing and updating the levels from 2020. The results of the Danish survey and the employed analytical method are documented in this paper. In addition to the 89 domestic samples, 30 samples of foreign origin were analysed for chlorate and perchlorate, together with ten samples of baby food and seven samples of infant formula collected from the Danish market. Samples were extracted using the Quick Polar Pesticides extraction method (QuPPe) and analysed on a triple quadrupole LC-MS/MS system. The levels of both chlorate and perchlorate detected in samples originating from Denmark were lower than the current MRLs except for a single sample of lettuce. Moreover, the percentage of foreign samples that were positive for chlorate and perchlorate residues was higher than the percentage of domestic samples. The reason for the generally low levels and low frequency of chlorate and perchlorate in Danish products is most likely linked to the fact that the water supply in Denmark is 100% unchlorinated ground water. The present study shows that residues of perchlorate and chlorate are found in Danish food products at a relatively low frequency and that the levels found are below the EU's MRLs implemented in 2020.

Bioactive ingredients for infant formula have been sought to reduce disparities in health outcomes between breastfed and formula-fed infants. Traditional food safety methodologies have limited ability to assess some bioactive ingredients. It is difficult to assess the effects of nutrition on the infant immune system because of coincident developmental adaptations to birth, establishment of the microbiome and introduction to solid foods, and perinatal environmental factors. An expert panel was convened to review information on immune system development published since the 2004 Institute of Medicine report on evaluating the safety of new infant formula ingredients and to recommend measurements that demonstrate the safety of bioactive ingredients intended for that use. Panel members participated in a 2-d virtual symposium in November 2020 and in follow-up discussions throughout early 2021. Key topics included identification of immune system endpoints from nutritional intervention studies, effects of human milk feeding and human milk substances on infant health outcomes, ontologic development of the infant immune system, and microbial influences on tolerance. The panel explored how "nonnormal" conditions such as preterm birth, allergy, and genetic disorders could help define developmental immune markers for healthy term infants. With consideration of breastfed infants as a reference, ensuring proper control groups, and attention to numerous potential confounders, the panel recommended a set of standard clinical endpoints including growth, response to vaccination, infection and other adverse effects related to inflammation, and allergy and atopic diseases. It compiled a set of candidate markers to characterize stereotypical patterns of immune system development during infancy, but absence of reference ranges, variability in methods and populations, and unreliability of individual markers to predict disease prevented the panel from including many markers as safety endpoints. The panel's findings and recommendations are applicable for industry, regulatory, and academic settings, and will inform safety assessments for immunomodulatory ingredients in foods besides infant formula., (© The Author(s) 2021. Published by Oxford University Press on behalf of the American Society for Nutrition.)

Background: β-Galactooligosaccharides (GOS) are typically used in infant formula and adult nutritionals as a source of nondigestible oligosaccharides, which may bring beneficial effects through modulation of the gut microbiota. However, suitable methods for the determination of GOS in products with a high background of lactose do not exist., Objective: The aim of this work was to develop a method suitable for the determination of GOS in infant formula and adult nutritionals and demonstrate suitability through single laboratory validation., Methods: Reducing oligosaccharides are labeled with 2-aminobenzamide (2AB), separated by hydrophilic interaction LC, and determined assuming that all oligosaccharides give an equimolar response in the detector. The same sample is analyzed a second time after treatment with β-galactosidase to remove GOS. The difference in the determined oligosaccharides between the two measurements will be the GOS content of the sample. The method was validated in a single laboratory on infant formula and adult nutritionals., Results: Recoveries were in the range 91.5-102%, relative standards of deviation (RSDr) were in the range 0.7-5.99%, and one sample had an RSDr of 8.30%. Except for the one sample with an RSDr of 8.30%, the performance is within the requirements outlined in the Standard Method Performance Requirements, which specifies recoveries in the range 90-110% and RSDr of below 6%., Conclusions: The method is suitable for the determination of GOS in infant formula and adult nutritionals., Highlights: A method has been developed which is suitable for the determination of GOS in products with a high background concentration of lactose (infant fromula and adult nutritionals). The method does not require access to the GOS ingredient used for the production of the finished product. It is also possible to separately quantify the amount of GOS containing three or more monomeric units in order to support dietary fibre analysis., (© AOAC INTERNATIONAL 2021.)

Background-Aim: Aflatoxin M 1 (AFM 1 ) contaminates milk and dairy products that are widely consumed foods. Therefore, there is increased concern for human exposure to the toxic action and the negative health effects that AFM 1 presents. The aim of this study was to assess the contamination levels with AFM 1 , of dairy products available in the Greek market (Thessaly), and also to estimate the relevant nutritional exposure., Methods: A total of 52 infant milk, 25 Feta cheese and 32 pasteurized milk samples were randomly selected from the market. The determination of AFM 1 was based on indirect immunoenzymatic ELISA method with two analytical packages; namely Tecna (Italy) and Prognosis Biotech (Greece), for comparison purposes, strictly following the manufacturer's instructions. The estimated daily intake (EDI) of AFM 1 through dairy consumption and the relevant hazard index (HI) were calculated., Results: The levels of AFM 1 , in all samples tested, were below the tolerable maximum level of 50 ng/L for milk and 25 ng/L for infant formulas (EC Regulation 1881/2006), while concentrations were very low (0.23-9.38 ng/L for infant formula, 0.54-4.09 ng/L for cheese and 0.20-17.84 ng/L for pasteurized milk). The exposure assessment revealed that infants, toddlers and children were the most exposed groups in AFM 1 via milk and formula consumption (0.65, 0.35 and 0.35 respectively). The HI was also noticed higher in those ages, with the maximum value to reach 1.90. Though, for feta cheese the exposure was calculated as very low (EDI: 0.01 and HI: 0.05). Comparing the two analytical packages used, no statistically significant difference was found (p-value >0.05)., Conclusions: Milk and dairy products tested were found safe for consumption, regarding the AFM 1 levels. From a public health perspective, the results are satisfactory, given that the population exposure to AFM 1 , based on the nutrient profile, was rather limited., (Copyright © 2021 European Society for Clinical Nutrition and Metabolism. Published by Elsevier Ltd. All rights reserved.)

Ready-to-feed liquid infant formulas (IF) were subjected to direct (D) or indirect (ID) ultra-high-temperature (UHT) treatment and then stored at 40 °C under aseptic conditions for 60-120 days simulating global transportation which accelerates the Maillard reaction. Low pasteurized and unstored IF (LP) was included as a control for the UHT treatments. Simulated infant in vitro digestion was conducted. SDS-PAGE indicated that protein aggregate formation correlated with thermal treatment, being greatest after 60 days of storage. Limited protein digestion was observed after pepsin treatment for 2 h. Beta-lactoglobulin (β-Lg), alpha-lactalbumin (α-La) and protein aggregates remained undigested after 2 h of pepsin digestion in LP and D, but less β-Lg and α-La remained in ID. The digestion of β-Lg and α-La was enhanced in D and ID stored for 60 days, but aggregates remained undigested. After pepsin and pancreatin digestion, large amounts of β-Lg remained undigested in the LP, but digestion increased after UHT treatment (ID > D) and increased further after storage for 60 and 120 days, indicating that heat treatment and storage facilitate the digestion of unaggregated proteins. No aggregates remained after pancreatin digestion of LP, D, ID and D stored for 60 days, but were present in ID stored for 60 days. Aggregates were mainly disulphide-linked, but dityrosine linkages were detected in D and ID stored for 120 days. LC-MS/MS indicated limited proteolysis arising from endogenous milk proteases prior to in vitro digestion, being highest in D. Peptide numbers increased following pepsin and further during pancreatin digestion (β-casein > β-Lg > β-La), and released β-Lg peptides, typically 5-8 amino acids in length, contained several bioactivities, e.g. , dipeptidyl-peptidase IV (DPP-IV) and angiotensin converting enzyme (ACE) inhibition.

Exposure to naturally derived estrogen receptor activators, such as the phytoestrogen genistein, can occur at physiologically relevant concentrations in the human diet. Soy-based infant formulas are of particular concern because infants consuming these products have serum genistein levels almost 20 times greater than those seen in vegetarian adults. Comparable exposures in animal studies have adverse physiologic effects. The timing of exposure is particularly concerning because infants undergo a steroid hormone-sensitive period termed "minipuberty" during which estrogenic chemical exposure may alter normal reproductive tissue patterning and function. The delay between genistein exposure and reproductive outcomes poses a unique challenge to collecting epidemiological data. In 2010, the U.S. National Toxicology Program monograph on the safety of the use of soy formula stated that the use of soy-based infant formula posed minimal concern and emphasized a lack of data from human subjects. Since then, several new human and animal studies have advanced our epidemiological and mechanistic understanding of the risks and benefits of phytoestrogen exposure. Here we aim to identify clinically relevant findings regarding phytoestrogen exposure and female reproductive outcomes from the past 10 years, with a focus on the phytoestrogen genistein, and explore the implications of these findings for soy infant formula recommendations. Research presented in this review will inform clinical practice and dietary recommendations for infants based on evidence from both clinical epidemiology and basic research advances in endocrinology and developmental biology from mechanistic in vitro and animal studies., (Published by Elsevier Inc.)

Human milk oligosaccharides (HMO) have attracted great interest in recent years due to their role in boosting infants and adults health. According to several in vitro , in vivo and clinical studies, gastrointestinal and immune physiological systems benefit the most from HMO intake. Other organ systems, such as the respiratory, central nervous, circulatory, locomotor, and urinary systems have also been found to be affected by the HMO consumption in the recent decade. Due to their positive impact on human health, the incorporation of HMO into the infant formula or other functional foods has become highly desirable. Currently, their large-scale production is limited to 2'-fucosyllactose (2'FL) and lacto- N -neotetraose (LNnT) that are obtained through fermentation and added to the infant formula as fortifiers. Fewer advances have been made for other HMO to reach the industrial scale synthesis. The present paper summarizes the latest research on HMO in terms of their health benefits and synthetic methodologies, with the overall aim to establish the current status and trends in both fields.

Necrotizing enterocolitis (NEC) is a significant cause of mortality and morbidity in preterm infants. The pathogenesis of NEC is not completely understood; however, intestinal immaturity and excessive immunoreactivity of intestinal mucosa to intraluminal microbes and nutrients appear to have critical roles. Dietary fats are not only the main source of energy for preterm infants, but also exert potent effects on intestinal development, intestinal microbial colonization, immune function, and inflammatory response. Preterm infants have a relatively low capacity to digest and absorb triglyceride fat. Fat may thereby accumulate in the ileum and contribute to the development of NEC by inducing oxidative stress and inflammation. Some fat components, such as long-chain polyunsaturated fatty acids (LC-PUFAs), also exert immunomodulatory roles during the early postnatal period when the immune system is rapidly developing. LC-PUFAs may have the ability to modulate the inflammatory process of NEC, particularly when the balance between n3 and n6 LC-PUFAs derivatives is maintained. Supplementation with n3 LC-PUFAs alone may have limited effect on NEC prevention. In this review, we describe how various fatty acids play different roles in the pathogenesis of NEC in preterm infants.

Cronobacter sakazakii is a foodborne, emerging opportunistic pathogen that causes severe bacteremia, necrotizing enterocolitis, and sepsis with a mortality rate of up to 80%. In this study, we developed a simple and sensitive fluorescent turn-off aptasensor with rolling circle amplification assay for viable C. sakazakii detection in powdered infant formula. The results showed that the proposed aptasensor has good performance and specificity for detecting viable C. sakazakii in pure culture and powdered infant formula samples within 3 h. Under the optimal reaction conditions, there is a linear relationship between fluorescent intensity at 490 nm and logarithmic concentration of C. sakazakii in the range of 2.7 × 10 5 to 2.7 × 10 2 cfu/mL, with a limit of detection of 2.7 × 10 2 cfu/mL in pure culture. The proposed aptasensor achieved a recovery of 104 to 111% in pure culture, and 96 to 107% in spiked powdered infant formula samples. The proposed aptasensor does not require complicated DNA extraction steps or antibodies, and can be performed at 37°C, making it a convenient and sensitive strategy for C. sakazakii detection., (Copyright © 2021 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Children with medical complexities, such as multi-system disorders and/or neurological impairments, often experience feeding difficulties and need enteral nutrition. They frequently have impaired motility and digestive-absorbing functions related to their underlying condition. If a cow's milk allergy (CMA) occurs as a comorbidity, it is often misdiagnosed, due to the symptoms' overlap. Many of the commercialized mixtures intended for enteral nutrition are composed of partially hydrolyzed cow's milk proteins, which are not suitable for the treatment of CMA; thus, the exclusion of a concomitant CMA is mandatory in these patients for obtaining symptoms relief. In this review, we focus on the use of elemental and semi-elemental formulas in children with neurological diseases and in preterm infants as clinical "models" of medical complexity. In children with neurodisabilities, when gastrointestinal symptoms persist despite the use of specific enteral formula, or in cases of respiratory and/or dermatological symptoms, CMA should always be considered. If diagnosis is confirmed, only an extensively hydrolyzed or amino-acid based formula, or, as an alternative, extensively hydrolyzed nutritionally adequate formulas derived from rice or soy, should be used. Currently, enteral formulas tailored to the specific needs of preterm infants and children with neurological impairment presenting concomitant CMA have not been marketed yet. For the proper monitoring of the health status of patients with medical complexity, multidisciplinary evaluation and involvement of the nutritional team should be promoted.

Infant formula contains thermal processing contaminants, such as dietary advanced glycation end-products (dAGEs), glycidyl esters (GEs), 2-monochloropropane-1,3-diol esters and 3-monochloropropane-1,2-diol esters (MCPDEs). This systematic review aimed to gain insights into the occurrence of these contaminants in different types of infant formula, to understand potential effects of the formulation and processing of infant formulas on these contaminants, as well as into possible mitigation strategies. The occurrence of dAGEs in infant formula depends on the recipes and processing conditions. Hydrolyzed protein formulations promote dAGEs formation in infant formula since peptides are more prone to glycation than intact proteins, which is reflected in high dAGEs concentration in hypoallergenic infant formula. Different carbohydrates in recipes result into different glycation extents of infant formula: maltodextrin containing formulas contained less dAGEs than those with lactose. Concerning mitigation strategies, applying ultra-high-temperature (UHT) treatment during milk processing leads to less dAGEs formation than using in-bottle sterilization. Although data are limited, evidence showed that encapsulation of raw ingredients or the use of antioxidants or enzymes in recipes is promising. The occurrence of MCPDEs and GEs in infant formula fully depends on the vegetable oils used in the recipe. High levels of these contaminants can be found when relatively high amounts of palm oils or fats are used. The mitigation of MCPDEs and GEs should therefore be performed on fats and oils before their application to infant formula recipes. Data and knowledge gaps identified in this review can be useful to guide future studies., (© 2021 The Authors. Comprehensive Reviews in Food Science and Food Safety published by Wiley Periodicals LLC on behalf of Institute of Food Technologists.)

Objective: A gas chromatography-mass spectrometry(GC-MS) method for the determination of 16 European priority polycyclic aromatic hydrocarbons(16 EUPAHs) in infant formula milk powder was established, and the characterization and investigation of 16 EUPAHs in 70 milk formula powders were carried out in 2020., Methods: After hydrolysis, extraction, saponification and solid phase extraction, infant formula milk powder was detected by GC-MS using DB-EUPAH capillary column(20 m×0.18 mm, 0.14 μm)and quantified by internal standard method., Results: The average recoveries ranged from 67.8% to 116.2% and the relative standard deviations ranged from 2.0% to 15.1%(n=6). The limits of quantification and detection of the method were 0.5 and 0.2 μg/kg, respectively. The content of 16 EUPAHs was <0.2-0.48 μg/kg, including PAH4 content in the range of <0.2-0.91 μg/kg, the characterization and investigation of infant formula powder in 16 EUPAHs mainly chrysene, cyclopenta[c, d]pyrene, benz[a]anthracene, benzo[b]fluoranthene, benzo[g, h, i]perylene., Conclusion: The method is simple, accurate and suitable for the determination of 16 EUPAHs in infant formula milk powder. The result showed that the content of 16 EUPAHs in commercially available infant formula milk powder in Hangzhou was low and all of them met the limit requirement of European Union.

Adequate protein intake by very-low-birth-weight preterm infants (≤1,500 g at birth) is essential to optimize growth and development. The estimated needs for this population are the highest of all humans, however, the recommended intake has varied greatly over the past several years. A literature search was conducted in PubMed, Embase, CINAHL (Cumulative Index to Nursing and Allied Health Literature), and Cochrane Central databases to identify randomized controlled trials evaluating the effect of prescribed protein intake and identified outcomes. Articles were screened by 2 reviewers, risk of bias was assessed, data were synthesized quantitatively and narratively, and each outcome was separately graded for certainty of evidence. The literature search retrieved 25,384 articles and 2 trials were included in final analysis. No trials were identified that evaluated effect of protein amount on morbidities or mortality. Moderate certainty evidence found a significant difference in weight gain when protein intake of greater than 3.5 g/kg/day from preterm infant formula was compared with lower intakes. Low-certainty evidence found no evidence of effect of protein intake of 2.6 vs 3.1 vs 3.8 g/kg/day on length, head circumference, skinfold measurements, or mid-arm circumference. Low-certainty evidence found some improvement in development measures when higher protein intake of 3.8 vs 3.1 vs 2.6 g/kg/day were compared. Low-certainty evidence found no significant difference in bone mineral content when these protein intakes were compared. No studies were identified that compared protein intake greater than 4.0 g/kg/day. This systematic review found that protein intake between 3.5 and 4.0 g/kg/day promotes weight gain and improved development., (Copyright © 2021 Academy of Nutrition and Dietetics. Published by Elsevier Inc. All rights reserved.)

The presence of 3-monochloropropane-1,2-diol (3-MCPD) esters and 2-MCPD esters in infant formulas have raised a number of food safety concerns. Here, a dietary exposure assessment was conducted for 3-and 2-MCPD esters in infant formulas available for consumption in Chinese infant and toddlers aged 0-3 years old. This work presents the occurrence data for 3-and 2-MCPD ester in 874 infant formulas purchased in China between 2015 and 2017. The concentrations of 3-MCPD esters ranged from ND to 1.469 mg/kg, with concentrations of 2-MCPD esters ranging from ND to 0.218 mg/kg. The LODs of 3-and 2-MCPD esters were 0.027-0.074 mg/kg. The mean exposures of infants and toddlers to 3-MCPD esters from formulas were lower than the tolerable daily intake (TDI, 2 μg/kg bw/day, established by EFSA), while high exposures (95th percentile) to 3-MCPD esters ranged from 0.907 to 2.520 μg/kg bw/day. On the whole, the health risk of Chinese infant and toddlers exposed to 3-MCPD esters was low, but the health risk of some infants aged 0-6 months with high formula consumption (95th percentile) raises some concern.

Human clinical trials have shown that a specific partially hydrolyzed 100% whey-based infant formula (pHF-W) reduces AD risk in the first yeast of life. Meta-analyses with a specific pHF-W (pHF-W1) confirm a protective effect while other meta-analyses pooling different pHF-W show conflicting results. Here we investigated the molecular composition and functional properties of the specific pHF-W1 as well as the stability of its manufacturing process over time. This specific pHF-W1 was compared with other pHF-Ws. We used size exclusion chromatography to characterize the peptide molecular weight (MW), a rat basophil degranulation assay to assess the relative level of beta-lactoglobulin (BLG) allergenicity and a preclinical model of oral tolerance induction to test prevention of allergic sensitization. To analyze the exact peptide sequences before and after an HLA binding assay, a mass cytometry approach was used. Peptide size allergenicity and oral tolerance induction were conserved across pHF-W1 batches of production and time. The median MW of the 37 samples of pHF-W1 tested was 800 ± 400 Da. Further oral tolerance induction was observed using 10 different batches of the pHF-W1 with a mean reduction of BLG-specific IgE levels of 0.76 log (95% CI = -0.95; -0.57). When comparing pHF-W1 with three other formulas (pHF-W2 3 and 4), peptide size was not necessarily associated with allergenicity reduction in vitro nor oral tolerance induction in vivo as measured by specific IgE level ( p < 0.05 for pHF-W1 and 2 and p = 0.271 and p = 0.189 for pHF-W3 and 4 respectively). Peptide composition showed a limited overlap between the formulas tested ranging from 11.7% to 24.2%. Furthermore nine regions in the BLG sequence were identified as binding HLA-DR. In conclusion, not all pHF-Ws tested have the same peptide size distribution decreased allergenicity and ability to induce oral tolerance. Specific peptides are released during the different processes used by different infant formula producers.

Triacylglycerol (TAG) components in human milk during different lactation periods, infant formulas with different fat sources, other mammalian milk (cow, goat, donkey, and yak milk), and plant oil (sunflower, rapeseed, corn, soybean, palm, palm kernel, and coconut oil) were analyzed and compared using ultraperformance supercritical fluid chromatography and quadrupole time-of-flight mass spectrometry (UPSFC-Q-TOF-MS). We identified 191 TAGs (86, 102, 101, and 54 TAGs in human milk, infant formula, mammalian milk, and plant oil, respectively). TAGs esterified with palmitic acid (16:0) were major TAG structures in human milk (59.08% of total TAGs) and contained 30 TAG types. The sn -O/P/O regioisomer constituted more than 80% of the O/P/O content of human milk, whereas the sn -O/O/P levels were higher in other samples. The carbon number (CN) 52 content was higher than the CN 54 content in human milk, with the opposite observed in infant formula. TAGs with CN < 40 content were abundant in cow, goat, and yak milk; donkey milk was rich in CN 52 content. TAGs composed of medium-chain fatty acids (MCFAs) and long-chain fatty acids (LCFAs) were rich in human milk, while TAGs with three MCFAs were rich in infant formula. The TAG characteristics of infant formula were directly related to its fat resource. TAGs with fewer double bonds were abundant in the plant oil formula; however, highly unsaturated TAGs were prominent in the cow and goat milk formulas, similar to plant oil and mammalian milk. Significant differences in the TAG distribution were observed among the different species.

Introduction: A decrease in weight velocity and feeding difficulties in infants may be caused by an inadequate caloric intake and underlying medical conditions., Case Description: By focusing on four clinical cases, this article illustrates the temporary use of a special infant formula in orally-fed and enterally-fed infants with unsatisfactory weight gain and special medical conditions such as gastrointestinal and neurological disorders. The formula was a nutritionally complete hypercaloric infant formula containing partially hydrolyzed whey protein. It was used after full consideration of all feeding options including breastfeeding., Conclusion: Implementing appropriate feeding behaviors, adapted to age and potential comorbidities, is an essential prerequisite for therapeutic management. The use of a nutritionally complete hypercaloric infant formula can be helpful to manage unsatisfactory weight gain and feeding difficulties in infants.

This study determined the intakes of complementary foods (CFs) and milk-based formulas (MFs) by a total of 119 subjects aged 6-23.9 months from urban day care centers. Dietary intakes were assessed using two-day weighed food records. Intake adequacy of energy and nutrients was compared to the Recommended Nutrient Intakes (RNI) for Malaysia. The most commonly consumed CFs were cereals (rice, noodles, bread). The subjects derived approximately half of their energy requirements (kcals) from CFs (57 ± 35%) and MFs (56 ± 31%). Protein intake was in excess of their RNI requirements, from both CFs (145 ± 72%) and MFs (133 ± 88%). Main sources of protein included meat, dairy products, and western fast food. Intake of CFs provided less than the RNI requirements for vitamin A, thiamine, riboflavin, folate, vitamin C, calcium, iron, and zinc. Neither CF nor MF intake met the Adequate Intake (AI) requirements for essential fatty acids. These findings indicate imbalances in the dietary intake of the subjects that may have adverse health implications, including increased risk of rapid weight gain from excess protein intake, and linear growth faltering and intellectual impairment from multiple micronutrient deficiencies. Interventions are needed to improve child feeding knowledge and practices among parents and child care providers.

Breast milk has the most suitable composition for the proper development in the first year of a child's life. However, it is often replaced with artificial milk. The aim of the study was to analyze the composition of essential elements: Na, K, Ca, P, Mg, Fe, Zn, Cu, and Mn as well as toxic elements: Ni, Pb, Sr, Li, and In in 18 formulas available in Poland. The daily supply was also estimated. The study was performed by Inductively Coupled Plasma Optical Emission Spectrometry method. The results showed the presence of all essential elements tested, but the content of P and Mn significantly differed from the concentrations declared. Such discrepancies can have significant impact on the daily dose of the bioelements taken. However, the content of elements was within the reference standards established by the EU Directive with exception of P, the amount of which exceeded the norms 5.23-18.80-times. Daily supply of P in tested milk as well as Fe and Mn provided with first and hypoallergenic formula exceeded the adequate intake. Analysis revealed the contamination with harmful elements-Pb, Sr, Li, and In were detected in almost all products. The study confirms the data concerning some discrepancies in composition and the contamination of food and may provide information on the feeding quality of children and estimation of health risk associated with exposure to toxic elements.

Breast feeding and human milk are the standards for infant feeding and nutrition. Human milk oligosaccharides (HMOs) are the third most abundant solid component in human milk. To date, more than 200 structural different HMOs have been identified and some can be synthesized by the food industry. HMOs are one of the major differences between human milk and formula milk, and current evidence demonstrates their various beneficial effects toward infants' health: acting as anti-adhesive antimicrobials, immune modulators, and intestinal cell response modulators, as well as providing prebiotics effect and neurodevelopment and cognition effects. HMOs compositions vary among mothers, influenced by the stage of lactation, duration of pregnancy and maternal genetic factors. However, there are still some unknown factors affecting the compositions of HMOs and requiring further research for clarification. A combination of preclinical and clinical cohort studies may help to identify whether an individual HMO contributes to disease protection. In recent years, 2'-fucosyllactose (2'-FL) and lacto-N-neotetraose (LNnT) have been approved as food ingredients by official authorities. Infant formulae supplemented with these HMOs are well-tolerated. However, more prospective clinical studies are warranted to elucidate HMOs' significance in infant nutrition. Breast milk feeding remains the best option for infants nutrition and development. Whenever breast milk is not adequate or unavailable, infant formula supplemented with HMOs might be considered as an alternative., Competing Interests: Declaration of competing interest The authors did not receive any financial or non-financial benefits from any commercial entity in support of this article., (Copyright © 2021. Published by Elsevier B.V.)

An efficient and simple method for determining vanillin, methyl vanillin and ethyl vanillin in milk and dairy products was developed using a liquid-liquid extraction (LLE) procedure coupled to high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS). Different extraction procedures were tested and optimised by spiking three vanillin compounds into a blank matrix in which none of any food additives were detected, and the extraction with acetonitrile solution and n -hexane as cleaning sorbent allowed an efficient recovery of 87.6-101.7% with RSDs less than 5%. The limit of detection (LOD) ranged from 6.2 to 20.1 μg/kg. High sensitivity, accuracy and selectivity were found for the in-house validated method, which can eliminate the interferences from complicated matrices effectively, and fulfil the quality criteria for routine laboratory application for real samples. The developed method was then finally applied to screen the three analytes in 65 milk and dairy products including infant formula milk powders from local markets to check for compliance with Chinese Regulation. Concentrations of the total vanillin and ethyl vanillin ranged from 0.0323 to 246.3 mg/kg, which is within the limits of Chinese regulations.

Protein modifications in liquid infant formula (IF) have been widely studied, but distinguishing between heat- and storage-induced structural changes remains challenging. A generic liquid IF was subjected to direct or indirect UHT treatment and stored at 40 °C up to 180 days. Colour and pH were monitored and structural changes were characterised by dynamic light scattering, SDS-PAGE and centrifugal field-flow fractionation (FFF) coupled with multi-angle light scattering (MALS) and UV detectors to evaluate whether heat-induced differences would level out during storage. Both direct- and indirect UHT treatment led to structural changes, where the higher heat load of the indirect UHT treatment caused more pronounced changes. Indications were that storage-induced changes in pH, browning and non-reducible cross-links were not dependent on UHT treatment. However, FFF-MALS-UV analysis allowed characterisation of complex aggregates, where structural changes continued to be most pronounced in indirect UHT treated samples, and different storage-induced aggregation behaviour was observed., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Infant Milk Formula (IMF) is designed as a breastmilk substitute to satisfy the nutritional requirements during the first months of life. This study investigates the effects of two IMF processing technologies on cow milk protein digestion using an infant static in vitro gastrointestinal model. The degree of protein hydrolysis at the end of the gastric phase was 3.7-fold higher for IMF produced by high temperature (IMF-HT), compared to IMF produced by cascade membrane filtration (IMF-CMF), as assessed by free N-terminal group analysis. The processing type also influenced the panel of bioavailable peptides detected in basolateral compartments of Caco-2 monolayers exposed to gastrointestinal digested IMFs. In addition, IMF-CMF significantly increased tight junction protein, claudin 1, whilst IMF-HT significantly reduced tight junction integrity. In conclusion, producing IMF by CMF may preserve intestinal barrier integrity and can deliver its own unique inventory of bioavailable peptides with potential bioactivity., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Gut microbiota development in formula-fed and breast-fed infants is known to differ. This could relate to the usage of unmodified vegetable oil instead of mammalian fat in infant formula (IF), causing the enhanced formation of the poorly soluble soap calcium palmitate (CP) in the infant's gut. Here we investigate in vitro the possible influence of CP on the infant gut bacteria. The growth of several bacterial species dominant in the infant's gut was analyzed by culturing in media with CP. Faecalibacterium prausnitzii as a sensitive representative was analyzed in detail by scanning transmission electron microscopy, membrane staining, gas chromatography, and microbial fuel cell experiments. Of all bacteria tested, the growth of several bifidobacteria and F. prausnitzii was reduced at 0.01 mg/ml CP, Bifidobacterium infantis stopped growing completely. CP reduced the cell envelope thickness of F. prausnitzii, disturbed the cell membrane fatty acids and function of membrane proteins involved in electron transport. CP inhibited the growth of bifidobacteria and faecalibacteria. This suggests that modification of fat in IF may benefit the development of the gut microbiota in formula-fed infants by supporting the colonization of important beneficial bacteria in early life. Future clinical studies are needed to confirm this., (© 2021 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.)

Background: A method has been developed and validated for selective, accurate, and precise determination of total proteinogenic amino acids and taurine from infant formula and adult/pediatric nutritional formulas (powders, ready-to-feed liquids, and liquid concentrates). The method was reviewed by the AOAC INTERNATIONAL SPIFAN Expert Review Panel (ERP) during the 133rd AOAC Annual Meeting and Expo on September 7, 2019 in Denver, CO, USA, and was recommended to First Action Official MethodsSM status., Objective: The method involves protein hydrolysis to amino acids, a simple pre-column derivatization of amino acids and separation of derivatized amino acids by UHPLC. The quantification of amino acids is performed by multi-point calibration using norvaline as the internal standard. The analytical method is capable of quantitative determination for 22 proteinogenic amino acids, but cannot be used to quantitate tryptophan, which is destroyed during the acid hydrolysis step. Asparagine is determined as aspartic acid and glutamine as glutamic acid. Cystine and cysteine are converted to S-2-carboxyethylthiocysteine (CYSx) and the derivative is separated from the other amino acids. Citrulline, which is present in some matrixes, is separated from other amino acids and is not included in the method performance evaluation in the single-laboratory validation (SLV)., Method: The proposed method met all the performance requirement limits set in Standard Method Performance Requirement (SMPR®) 2014.013 for total proteinogenic amino acids and taurine. The correlation coefficient of multi-point calibration was not less than 0.999 for any amino acids at any point in the SLV study, confirming the validity of linear dynamic range (LDR) and linearity of the method. The individual amino acids in the chromatogram were identified by absolute retention time and relative retention time (RRT) with respect to the internal standard norvaline. There were no significant (S/N <10) interferences from the reagents or byproducts of derivatization and targeted matrixes. The method demonstrated high selectivity., Result: Accuracy of the method was validated using standard reference materials (SRMs; NIST SRM 1869 and 1849a) and spike recovery studies. The amino acid results in the SRMs were within the ranges of Reference Mass Fraction Values. The accuracy of the method was corroboratively validated by spike recovery studies. The average spike recovery ranged between 93 and 107% ensuring the accuracy of the method for amino acids and compliance with AOAC SMPR 2014.013., Conclusions: Precision data of the method demonstrate that it meets the stakeholder requirements as per the SMPR. The mean RSDr for all the amino acids for 17 matrixes selected for the SLV were not more than 4%. The method is very sensitive and the LOQ can go down to approximately ten times lower than the SMPR requirements. The sensitivity of method is a direct reflection of its signal-to-noise ratio which ensures guaranteed method performance at the lower levels of amino acids in these matrixes., Highlights: Taurine (aminoethane sulfonic acid), unlike the other amino acids, is a beta-sulfonic amino acid that is not used in protein synthesis but is found as a free amino acid in tissues. The acidic functional group (-COOH) in common amino acids is replaced with a sulfonic acid (-SO3H) group in taurine. The method offers baseline separation of citrulline which is an alpha amino acid generally present in infant formula and adult/pediatric nutritional products. The separation of citrulline eliminates the risk of interference of this compound with other amino acids. The method can also separate and quantitate hydroxyproline, an important component of collagen that is often used to quantitate collagen. The method is simple and does not include any proprietary chemicals or instruments, and it can be performed on any basic reverse phase UHPLC system with UV detection (1-6)., (© AOAC INTERNATIONAL 2020. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Background: Recent research has uncovered the potential for excess manganese (Mn) intakes causing significant neurotoxic effects for early brain development., Methods: We identified the Mn tolerable intakes (TI) published by the U.S. Institute of Medicine (IOM), World Health Organization (WHO), Agence nationale de sécurité sanitaire (ANSES), and U.S. Environmental Protection Agency (US EPA) and examined the primary studies on which regulatory TIs are based. We converted the TIs to μg of Mn/kg/day using standard assumptions specific to each agency. We estimated μg of Mn/kg/day intakes due to formulas. Using our estimates for formula intakes, weights, and kcal content, we converted regulatory maxima and minima from μg of Mn/100 kcals to estimates of μg of Mn/kg/day., Results: Except for the proposed ANSES TI for drinking water, none of the primary studies on which Mn intake guidelines and regulations are based measured health outcomes. Some infant formulas may exceed the regulatory TIs, especially if prepared with water containing considerable concentrations of Mn (e.g. 250 μg/L), even while meeting national and international regulatory standards or guidelines., Conclusions: Infant formula regulations must be revised to reduce the potential for excess manganese intakes and the practice of manganese supplementation of infant formulas should be ceased., (Copyright © 2020 The Author(s). Published by Elsevier GmbH.. All rights reserved.)

Background: High protein intake in early life is associated with an increased risk of childhood obesity. Dietary protein intake may be a key mechanistic modulator through alterations in endocrine and metabolic responses., Objective: We aimed to determine the impact of different protein intake of infants on blood metabolic and hormonal markers at the age of four months. We further aimed to investigate the association between these markers and anthropometric parameters and body composition until the age of two years., Design: Term infants received a modified low-protein formula (mLP) (1.7 g protein/100 kcal) or a specifically designed control formula (CTRL) (2.1 g protein/100 kcal) until 6 months of age in a double blinded RCT. The outcomes were compared with a breast-fed (BF) group. Glucose, insulin, leptin, IGF-1, IGF-BP1, -BP2, and -BP3 levels were measured at the age of 4 months. Anthropometric parameters and body composition were assessed until the age of 2 years. Groups were compared using linear regression analysis., Results: No significant differences were observed in any of the blood parameters between the formula groups ( n = 53 mLP; n = 44 CTRL) despite a significant difference in protein intake. Insulin and HOMA-IR were higher in both formula groups compared to the BF group ( n = 36) ( p < 0.001). IGF-BP1 was lower in both formula groups compared to the BF group ( p < 0.01). We found a lower IGF-BP2 level in the CTRL group compared to the BF group ( p < 0.01) and a higher IGF-BP3 level in the mLP group compared to the BF group ( p = 0.03). There were no significant differences in glucose, leptin, and IGF-1 between the three feeding groups. We found specific associations of all early-life metabolic and hormonal blood parameters with long-term growth and body composition except for IGF-1., Conclusions: Reducing protein intake by 20% did not result in a different metabolic profile in formula-fed infants at 4 months of age. Formula-fed infants had a lower insulin sensitivity compared to breast-fed infants. We found associations between all metabolic and hormonal markers (except for IGF-1) determined at age 4 months and growth and body composition up to two years of age.

Background: High oral exposure and biological vulnerabilities may put formula-fed infants at risk for manganese-induced neurotoxicity., Objectives: We sought to characterize manganese concentrations in public drinking water and prepared infant formulas commonly purchased in the United States, integrate information from these sources into a health risk assessment specific to formula-fed infants, and examine whether households that receive water with elevated manganese concentrations avoid or treat the water, which has implications for formula preparation., Methods: Manganese was measured in 27 infant formulas and nearly all Minnesota community public water systems (CPWS). The risk assessment produced central tendency and upper-end exposure estimates that were compared to a neonatal animal-based health reference dose (RfD) and considered possible differences in bioavailability. A survey study assessed esthetic concerns, treatment, and use of water in a Twin Cities community with various levels of manganese in drinking water., Results: Ten percent of CPWSs were estimated to exceed the EPA health advisory level of 300   μ g / L . Manganese concentrations in formula ranged from 69.8 to 741   μ g / L , with amino acid > soy > cow ' s   milk formula concentrations. Central tendency estimates of soy and amino acid formula reconstituted with water at the CPWS 95th percentile manganese concentration exceeded the neonatal-based RfD. Upper-end estimates of manganese intake from formula alone, independent of any water contribution, equaled or exceeded the neonatal-based RfD. In the survey study, we observed increased awareness of esthetic issues and water avoidance at higher manganese concentrations, but these concentrations were not a reliable consumption deterrent, as the majority of households with inside tap drinking water results above 300   μ g / L reported drinking the water., Discussion: Excessive exposure to manganese early in life can have long-lasting neurological impacts. This assessment underscores the potential for manganese overexposure in formula-fed infants. U.S. agencies that regulate formula and drinking water must work collaboratively to assess and mitigate potential risks. https://doi.org/10.1289/EHP7901.

Background: The use of hypoallergenic infant formulas and the need for reliable tests to determine the presence of residual antigens have increased in parallel., Objective: An LC-MS method for quantitation of casein was validated using incurred samples and a matrix-matched external standard curve., Method: Powdered infant formula samples were extracted in a buffer of sodium deoxycholate and ammonium bicarbonate at 60°C and filtered through 7 kDa desalting columns. Samples were digested overnight with trypsin and precipitated with acid prior to analysis of marker peptides by tandem mass spectrometry., Results: Based on three marker peptides, the linear range for casein was 1.8-42 μg/g of powdered infant formula with an LOQ of 1.8 μg/g. The determination coefficients (R2) for each curve were ≥0.99 for casein peptides. Method repeatability was ≤22% RSD and intermediate precision was ≤23% RSD; recovery of casein from incurred material (2-20 µg/g) ranged from 78% to 118%., Conclusions: An LC-MS/MS method was developed and validated for confirmation of casein allergens in hypoallergenic infant formula., Highlights: A method was developed to accurately and reliably quantify casein allergens in extensively hydrolyzed casein infant formula by LC-MS without the need for custom peptide standards., (© AOAC INTERNATIONAL 2020.)

Weaning is the gradual process of introducing solids or semisolid foods into an infant's diet, in order to ensure their healthy growth. This study developed two kinds of formula weaning food based on roasted or extruded quinoa and millet flour, and evaluated their quality. A fructo-oligosaccharide (FOS)/galacto-oligosaccharide (GOS) mix was added to provide the prebiotic potential. The protein contents of the roasted quinoa-millet complementary food (RQMCF) and extruded quinoa-millet complementary food (EQMCF) were 16.7% and 17.74% higher, respectively, than that of commercial millet complementary food (CMCF). Both RQMCF and EQMCF provided sufficient levels of energy and minerals. Extrusion provided the foods with a lower viscosity, and higher solubility and water absorption ability than roasting. In vitro digestion results showed that EQMCF exhibited the highest starch and protein digestibility (89.76% and 88.72%, respectively) followed by RQMCF (87.75% and 86.63%) and CMCF (83.35% and 81.54%). The digestas of RQMCF and EQMCF after in vitro digestion exhibited prebiotic effects by promoting the growth of the probiotics (Lactobacillus plantarum and Lactobacillus delbrueckii). These results will contribute to developing complementary weaning foods for infants. PRACTICAL APPLICATION: This study has shown that extrusion is an efficient and stable processing method for producing infant complementary foods with low density, balanced nutrition, and high levels of starch and protein digestibility. Extruded quinoa-millet prebiotic complementary food can also promote the proliferation of probiotics. This will provide a new direction for developing novel infant formula weaning foods., (© 2021 Institute of Food Technologists®.)

The presence of Cronobacter sakazakii must be controlled in infant powder plants, because it may cause infectious disease in infants, with high mortality. Testing for C. sakazakii in powdered infant formula should be performed before delivery, and it requires rapid and specific detection methods. In this study, we established a surface-enhanced Raman scattering (SERS) immunochromatographic test strip for the quantitative determination of C. sakazakii in powdered infant formula. Monoclonal antibodies for C. sakazakii were labeled with p-aminothiophenol-bound colloidal gold nanoparticles. Color change in the test line indicated the presence of C. sakazakii. A highly sensitive and quantitative test method was developed based on the Raman signal produced by the p-aminothiophenol bonding on gold nanoparticles. The SERS immunochromatographic test strip assay required a short analysis time (12 min) and exhibited a linearity range from 10 2 to 10 7 cfu/mL. The limit of detection was 201 cfu/mL without preculture. The SERS immunochromatographic test strip assay is a promising tool for the simple and rapid quantitative analysis of C. sakazakii and other pathogenic bacteria., (Copyright © 2021 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Exclusive breastfeeding ideally up to 6 months of life is the feed of choice for infants and should be promoted by healthcare professionals. However, when human milk is not sufficient or not available, infant formula, generally cow's milk-based, meeting strictly regulated nutritional and safety requirements, are recommended. Human breastmilk feeding has a positive health impact for both mother and child, but there is limited evidence that it has a long-term protective effect on the development of allergic disease. Some studies have found an association of an increased risk to develop cow's milk allergy with early exposure to cow's milk protein in formula milk. As a result, over the last 30 years, partially hydrolyzed formulas (pHF) have gained popularity and, more recently, become embroiled in a debate about their role in the primary prevention of allergic outcomes. Similar debates exist in regards to the potential preventative effects of pre-, pro- and synbiotics as well as nutritional factors, notably vitamin D and omega-3 fatty acids. This paper aims to critically address these aspects, drawing information from published data interpreted by an international expert group in paediatrics, allergy, gastro-intestinal diseases and nutrition. This group of experts emphasize that human milk is the optimal source of infant nutrition. With regards to pHFs, whilst no harm has been shown with their use and some studies have suggested potential benefit preventing atopic dermatitis in at risk infants, there is insufficient evidence for or against their routine recommendation for primary allergy prevention. The method of hydrolysation differs for every formula. There is insufficient evidence to recommend supplementation with vitamin D, omega-3 LCPUFA, specific prebiotic oligosaccharides or specific probiotic strains during pregnancy, lactation and early life to prevent the development of allergic disease in children. There remains a need for well-designed trials with the currently commercialised pHFs and supplements to allow for better clarity and evidence-based recommendations., Competing Interests: Conflict of interest YVs has participated as a clinical investigator, and/or advisory board member, and/or consultant, and/or speaker for Abbott Nutrition, Biocodex, Biogaia, CHR Hansen, Danone, Nestle Health Science, Nestle Nutrition Institute, Nutricia, Mead Johnson Nutrition, United Pharmaceuticals, Wyeth. RM has participated as a speaker and/or advisory board member for Nestle, Danone, Abbott and Mead Johnson. JPC has participated as a clinical investigator, and/or consultant and/or a speaker for Biostime, Mead-Johnson, Nestlé and Nutricia. CD has participated as advisory board member, and/or consultant, and/or speaker for Nutricia, Nestlé Health Science, United Pharmaceuticals, Nestlé Waters. AF currently sponsored research by Danone, Sanofi, Hipp, Ferrero. Advisory boards of Danone, Stallergenes, Menarini, Abbott, DBV, Novartis, and Hipp. SS has participated as advisory board member, and/or consultant, and/or speaker for Bioproject, Danone, Mellin, Nestlé, Novalac. RS has participated as a clinical investigator, and/or advisory board member, and/or consultant, and/or speaker for Abbott, Danone, Douxmatok, Else, Frutarum, Nestlé Nutrition Institute, Nestle Health Science, NGS, Teva, Ukko, YMINI. HS has participated as a clinical investigator, and/or advisory board member, and/or consultant, and/or speaker for Arla, Danone, Nestlé, Nestlé Nutrition Institute, Nutricia. NT has participated as a speaker and/or advisory board member, and/or consultant for Danone and Takeda. CV has provided educational material and review of educational material for Reckitt Benckiser, Abbott Nutrition, Danone, DBV technologies and NNI. She has received research funding from Reckitt Benckiser and the National Peanut Board. VV has participated as a consultant, and/or speaker for Nestlé Nutrition Institute, Nestle Health Science, Nutricia, Biostime., (Copyright © 2020 Elsevier Ltd and European Society for Clinical Nutrition and Metabolism. All rights reserved.)

Previous research shows that factors such as time, temperature, nutritional make-up of a liquid, and type of thickening agent can significantly alter the resulting thickness of liquids. This study sought to determine the effect of three distinct mixing methods on the resulting thickness of ready to feed infant formulas mixed to Mildly and Moderately Thick (International Dysphagia Diet Standardization Initiative [IDDSI] Levels 2 and 3) with three different thickening agents. Eight commercially available infant formulas were mixed with three different thickening agents by three different mixing methods. The IDDSI Syringe Flow Test was used to categorize the thickened formulas. Chi square analyses were completed to determine the impact of mixing method on the thickened formulas. The majority (94%) of thickened formula combinations prepared to a target Mildly Thick consistency produced thickened formula that was thinner or thicker than the target. In contrast, the majority (76%) of thickened formula combinations prepared to a target Moderately Thick (IDDSI Level 3) consistency produced thickened formula that was equivalent to the target consistency. A statistically significant relationship was found between mixing methods and resulting IDDSI category for samples mixed to a target of Moderately Thick. The thickening agent and method of mixing must be considered carefully when preparing infant formulas to Mildly and Moderately Thick target IDDSI categories. Based on results of this study, it is recommended that providers utilize a clinical testing method, such as the IDDSI Syringe Flow Test, when attempting to create a Mildly Thick formula consistency., (© 2020 Wiley Periodicals LLC.)

This study aimed to evaluate the risk concerning child population's health because of the occurrence of AFM 1 in UHT milk, powdered milk (PM) and infant formulae (IF). Determination of AFM 1 was performed in 60 samples and evaluation of the mycotoxin exposure was carried out through the determination of the estimated daily intake (EDI), whereas risk characterization was evaluated with the calculation of the risk of Hepatocellular Carcinoma (HCC) and the Margin of Exposure (MOE). AFM 1 ranged from 150 to 1020 ng/kg, and all the positive samples exceeded the limits stablished by European Community. The EDI for AFM 1 ranged according to the age group of the population studied (0-5 years old) from 0.828 to 2.523, 0-2.113 and 0.029-0.833 ng/kg b. w./day in UHT, PM and IF, respectively. The number of HCC cases associated with AFM 1 exposure (0.0015 a 0.0045) was higher than the limit of 0.001 case/100,000. MOE values for AFM 1 were 728 to 239, considerably below the security margin of 10,000. These results point to a potential risk to the health of Brazilian child population exposed to AFM 1 in dairy products., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Human milk is a source of nutrients and contains many distinct bioactive components. Among these, human milk oligosaccharides (HMOs) have attracted considerable attention and are being investigated as a "novel foods". Human milk is unique in its oligosaccharide composition. Recent research has focused on the complexity of HMOs by highlighting their diversity, structural variability, concentration variance, and structure-function relationships. In vitro and in vivo studies have demonstrated that HMOs drive infant gut microbiota, improve intestinal barrier functions, and modulate cell receptor signaling, thereby contributing to the development of infant immunity. These studies, combined with epidemiological data, indicate that some HMO may confer health benefits by preventing infections and diseases such as necrotizing enterocolitis and allergies. However, randomized controlled trials are restricted to structurally simple compounds such as 2' fucosyllactose and lacto-N-neotetraose. More controlled clinical trials are needed to justify routine supplementation of formula. It is felt that a better understanding of the role of HMOs leading to the development of inexpensive methods for large-scale HMO production is needed.

Quantification of the specific folate vitamers to estimate total folate in foods is not standardized. A collaborative study, including eight European laboratories, was conducted in order to determine the repeatability and reproducibility of the method for folate quantification in foods using the plant-origin γ-glutamyl hydrolase as part of the extraction procedure. The seven food samples analyzed represent the food groups; fruits, vegetables, dairy products, legumes, offal, fish, and fortified infant formula. The homogenization step was included, and six folate vitamers were analyzed using LC-MS/MS. Total folate content, expressed as folic acid equivalent, was 17-490 μg/100 g in all samples. Horwitz ratio values were within the acceptable range (0.60-1.94), except for fish. The results for fortified infant formula, a certified reference material (NIST 1869), confirmed the trueness of the method. The collaborative study is part of a standardization project within the Nordic Committee on Food Analysis (NMKL)., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Background: Although manganese (Mn) is an essential nutrient, recent research has revealed that excess Mn in early childhood may have adverse effects on neurodevelopment., Methods: We estimated daily total Mn intake due to breast milk at average body weights by reviewing reported concentrations of breast milk Mn and measurements of body weight and breast milk intake at 3 weeks, 4.25 months, 7 months, and 18 months. We compared these figures to the Mn content measured in 44 infant, follow-up, and toddler formulas purchased in the United States and France. We calculated Mn content of formula products made with ultra-trace elemental analysis grade water (0 μg Mn/L) and with water containing 250 μg Mn/L, a concentration which is relatively high but less than the World Health Organization Health-based value of 400 μg Mn/L or the United States Environmental Protection Agency Health Advisory of 350 μg Mn/L., Results: Estimated mean daily Mn intake from breast milk ranged from 1.2 μg Mn/kg/day (3 weeks) to 0.16 μg Mn/kg/day (18 months), with the highest intakes at the youngest age stage we considered, 3 weeks. Estimated daily Mn intake from formula products reconstituted with 0 μg Mn/L water ranged from 130 μg Mn/kg/day (3 weeks) to 4.8 μg Mn/kg/day (18 months) with the highest intakes at 3 weeks. Formula products provided 28-520 times greater than the mean daily intake of Mn from breast milk for the 4 age stages that we considered. Estimated daily Mn intake from formula products reconstituted with water containing 250 μg Mn/L ranged from 12 μg Mn/kg/day to 170 μg Mn/kg/day, which exceeds the United States Environmental Protection Agency Reference Dose of 140 μg Mn/kg/day for adults., Conclusions: Mn deficiency is highly unlikely with exclusive breast milk or infant formula feeding, but established tolerable daily intake levels for Mn may be surpassed by some of these products when following labeled instructions., (Copyright © 2020 The Author(s). Published by Elsevier GmbH.. All rights reserved.)

Palm oil/olein (PO/POL) is used in infant formulas to imitate the fatty acid profile of human milk (HM) and achieve similar levels of palmitic acid (PA). However, the positions of fatty acids on the triacylglyceride differ between PO/POL and HM, which affect fat absorption and produce unintended physiological consequences. Recent papers have reviewed evidence for physiological benefits of PO/POL and beta-palmitate (sn-2-palmitate) in infant formulas. The aim of the present review is to supplement the assessment of available clinical evidence on the physiological effects of PO/POL formulas in healthy infants. We intend to focus on PO/POL and not on sn-2-palmitate, since the latter was recently extensively reviewed. Clinical evidence supports that PO/POL in infant formulas leads to a lower fat, DHA, palmitate and calcium absorption, and bone mineralization; soft stools; and growth (weight accretion) compared to formulas without PO/POL. Consequently, it seems prudent to be considerate and cautious when adding PO/POL to infant formulas. While HM is the gold standard for infant nutrition, the development of infant formula should be based on achieving positive physiological outcomes, rather than just replicating HM nutrient composition.

Infant formula feeding, compared with human milk, has been associated with development of a distinct infant gut microbiome, but no previous study has examined effects of formula with added sugars. This work examined differences in gut microbiota among 91 Hispanic infants who consumed human milk [at breast (BB) vs. pumped in bottle (BP)] and 2 kinds of infant formula [(traditional lactose-based (TF) vs. lactose-reduced with added sugar (ASF)]. At 1 and 6 months, infant stool was collected to characterize gut microbiota. At 6 months, mothers completed 24-hour dietary recalls and questionnaires to determine infant consumption of human milk (BB vs. BP) or formula (TF vs. ASF). Linear regression models were used to determine associations of milk consumption type and microbial features at 6 months. Infants in the formula groups exhibited a significantly more 'mature' microbiome than infants in the human milk groups with the most pronounced differences observed between the ASF vs. BB groups. In the ASF group, we observed reduced log-normalized abundance of Bifidobacteriaceae (TF-BB Mean Difference = -0.71, ASF-BB Mean Difference = -1.10), and increased abundance of Lachnospiraceae (TF-BB Mean Difference = +0.89, ASF-BB Mean Difference = +1.20). We also observed a higher Community Phenotype Index of propionate, most likely produced by Lachnospiraceae , in the ASF group (TF-BB Mean Difference = +0.27, ASF-BB Mean Difference = +0.36). This study provides the first evidence that consumption of infant formula with added sugar may have a stronger association than birth delivery mode, infant caloric intake, and maternal BMI on the infant's microbiome at 6 months of age.