Assessment of Regulatory Compliance Testing for Vitamin D in Infant Formula-Impact of Delegated Regulation (EU) 2019/828.

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.

Nutritional content, protein quantity, protein quality and carbon footprint of plant-based drinks and semi-skimmed milk in the Netherlands and Europe.

OBJECTIVE: To compare the nutritional composition of bovine milk and several plant-based drinks with a focus on protein and essential amino acid content and to determine the ratio of essential amino acids to greenhouse gas emission. DESIGN: Nutritional information on the label was extracted for semi-skimmed milk, soy, oat, almond, coconut and rice drink from the Innova database between January 2017 and March 2020 for the Netherlands, Belgium, Germany, Spain, Italy, and Sweden. Protein and amino acids were measured and carbon footprint was calculated for a selection of Dutch products. Protein quality was determined by calculating the contribution to the WHO essential amino acids requirements. SETTING: The bovine milk and plant-based drinks market in Netherlands, Belgium, Germany, Spain, Italy, and Sweden. PARTICIPATING PRODUCTS: Semi-skimmed bovine milk and soy-, oat-, almond-, coconut- and rice drink. RESULTS: Nutritional label information was collected for 399 products. Milk naturally contains many micronutrients, e.g. vitamin B2, B12, and calcium. Approximately 50% of the regular plant-based drinks was fortified with calcium, whereas the organic plant-based drinks were mostly unfortified. Protein quantity and quality were highest in milk. Soy drink had the best protein quality to carbon footprint ratio and milk came second. CONCLUSIONS: The nutrition - climate change balance presented in this study, is in line with previous literature, which shows that semi-skimmed bovine milk and fortified soy drink deserve a place in a sustainable diet.

Lysine blockage of milk proteins in infant formula impairs overall protein digestibility and peptide release.

During heat processing of milk and dairy products, for example infant formula, the Maillard reaction occurs. In vitro and animal studies suggest that Maillard reaction induced lysine blockage impairs protein digestibility. Most studies that investigate the effect of glycation on protein digestion use a mixture of isolated milk protein with reducing sugars. In this study, infant formulas with 6.5%, 8.4%, 11.2%, 14.8%, 20.8%, and 44.5% of blocked lysine (BL) were digested in an in vitro infant digestion model and tested for protein hydrolysis and peptide release. OPA (o-phthalaldehyde) assay was used to assess the degree of protein hydrolysis. SDS-PAGE was conducted to monitor the hydrolysis of specific proteins. Peptides formed after gastric and intestinal digestion were identified by LC/MS. Protein hydrolysis of the 6.5% BL sample was significantly higher after 10 minutes of intestinal digestion compared to all other samples. Most differences were observed after intestinal digestion. A significant change in peptide patterns was observed for the 45% BL sample resulting in a relatively higher number of peptides with more than 14 amino acids. Mainly casein-derived peptides were affected. Overall, the average peptide length was significantly increased for the 44.5% BL glycated product (on average 10.2 amino acids for 6-21% BL vs. 11.4 amino acids for 45% BL; p < 0.001). In conclusion, glycation of milk proteins in an infant formula product can impair overall protein digestibility. These findings emphasize the importance of mild processing and having low BL levels in infant formula to ensure optimal digestion of proteins.

Identification of a TLR2 Inhibiting Wheat Hydrolysate.

SCOPE: Wheat hydrolysates are used in medical nutrition to provide undernourished patients a readily digestible protein source, for instance to recover from chemotherapy-induced intestinal mucosal inflammation. Since many hydrolysates of different sources can modulate the immune system, likely via Toll-like receptors (TLRs), it is hypothesized that also wheat hydrolysates might interact with TLR signaling, which could be a way to prevent intestinal inflammation and damage. METHODS AND RESULTS: The capacity of three wheat hydrolysates to modulate immunity by interfering with TLR signaling is determined. All wheat hydrolysates have TLR modulating effects but only one has strong TLR2 inhibiting effects, attenuating both TLR2/1 and TLR2/6 signaling in a reporter cell system. This is likely induced by direct TLR2-ectodomain binding, as confirmed by ELISA. Furthermore, this TLR2 blocking hydrolysate reduces IL-6 production in human dendritic cells. Application of reversed-phase-ultra HPLC combined with MS reveals that the presence of peptide WQIPEQSR is associated with the observed TLR2 inhibiting capacity. CONCLUSION: The study demonstrates TLR2-inhibiting capacities of a wheat hydrolysate. The findings provide a good start for further research to investigate whether this hydrolysate might contribute to the management of intestinal mucosal inflammation in cancer patients receiving chemotherapy.

Biochemical ripening of dredged sediments. Part 1. Kinetics of biological organic matter mineralization and chemical sulfur oxidation.

After dredged sediments have settled in a temporary upland disposal site, ripening starts, which turns waterlogged sediment into aerated soil. Aerobic biological mineralization of organic matter (OM) and chemical oxidation of reduced sulfur compounds are the major biochemical ripening processes. Quantitative data describing these processes are scarce. Therefore, aerobic oxidation and mineralization of five previously anaerobic dredged sediments were studied during a 160-d laboratory incubation experiment at 30 degrees C. A double exponential decay model could adequately describe sulfur oxidation and OM mineralization kinetics. During the first 7 d of incubation, 23 to 80% of the total sulfur was oxidized, after which no further sulfur oxidation was observed. Oxygen used for sulfur oxidation amounted up to 95% of the total oxygen uptake in the first 7 d and up to 45% of the oxygen uptake during the entire incubation period. Mineralization rates of the rapidly mineralizable OM fractions that degraded during the first 14 to 28 d of incubation were 10(2) to 10(3) times higher than the mineralization rates of the slowly mineralizable OM during the remaining period. First-order mineralization rates of the slowly mineralizable OM were 0.22 x 10(-3) to 0.54 x 10(-3) d(-1) and can be compared with those of terrestrial soils. Yields of biomass on substrate ranged from 0.08 to 0.45 g C(biomass)/g C(OM) and appeared to be higher for rapidly mineralizing OM than for slowly mineralizing OM. The results of this study can be used to optimize conditions during temporary disposal of sediments, to estimate the potential decrease in OM, and for future studies on the possible link between OM mineralization and degradation of hydrophobic organic contaminants.

Biochemical ripening of dredged sediments. Part 2. Degradation of polycyclic aromatic hydrocarbons and total petroleum hydrocarbons in slurried and consolidated sediments.

Ripening of polycyclic aromatic hydrocarbons (PAH) and total petroleum hydrocarbons (TPH) polluted dredged sediment can be considered as a bioremediation technique. Aerobic biodegradation of PAH and TPH was studied in five previously anaerobic-slurried sediments during a 350-d laboratory incubation experiment. In addition, oxygen penetration and degradation of PAH and TPH were studied in three consolidated (physically ripened) sediments. All experiments were conducted in the laboratory at 30 degrees C. A double exponential decay model could adequately describe PAH and TPH degradation kinetics in the slurried sediments. First-order degradation rate constants for the rapidly degradable fractions (12-58%) were approximately 0.13 and 0.058 d(-1) for PAH and TPH, respectively, whereas the rate constants for the slowly degradable fractions were approximately 0.36 x 10(-3) (PAH) and 0.66 x 10(-3) d(-1) (TPH). Rate constants for the rapidly and slowly degrading fractions have the same order of magnitude as the mineralization rate constants of the rapidly and slowly mineralizing organic matter (OM) fractions in the sediments. Oxygen uptake by degradation of PAH and TPH was negligible compared to the oxygen uptake by sulfur oxidation and OM mineralization. In consolidated sediments, PAH and TPH degradation was limited to the oxygenated part. Amounts of PAH and TPH that degraded in the oxygenated parts of the consolidated sediments during 21 d of incubation were similar to the amounts that degraded during 21 d in the slurried sediments.