Analysis of Taurine in Infant Formulas and Adult Nutritionals by Hydrophilic Interaction Liquid Chromatography-Mass Spectrometry: First Action 2022.03.

BACKGROUND: Taurine is recognized as an essential growth factor and as being critical in the maintenance of functional tissue regulation. OBJECTIVE: To evaluate the analytical performance of a hydrophilic interaction liquid chromatography-tandem mass spectrometry (HILIC-MS/MS) method for compliance with AOAC Standard Method Performance Requirements (SMPR®) for taurine analysis described in SMPR 2014.013. METHOD: Following protein precipitation with Carrez solutions, taurine is extracted and separated by HILIC with detection by triple quadrupole MS using multiple reaction monitoring (MRM). Stable isotope labeled (SIL) taurine internal standard is used for quantification to correct for losses in extraction and variations in ionization in the ion source. RESULTS: The method was shown to meet the requirements specified in the SMPR with a linear range of 0.27-2700 mg/hg RTF (ready-to-feed), a limit of detection of 0.14 mg/hg RTF, acceptable recovery of 97.2-100.1%, and acceptable repeatability of 1.6-6.4% relative standard deviation. Additionally, the method was found to have no statistically significant bias compared with reference values for National Institute of Standards and Technology (NIST) 1849a certified reference material (CRM) (P-value = 0.95) and 1869 CRM (P-value = 0.31), and with results from AOAC 997.05 (P-value = 0.10). CONCLUSIONS: A recent review of the method and validation data by the Stakeholder Program on Infant Formula and Adult Nutritionals (SPIFAN) Expert Review Panel (ERP) found that this method met all the criteria for analysis of taurine specified in SMPR 2014.013 and voted to adopt this method as First Action AOAC Official MethodSM2022.03. HIGHLIGHTS: A method for the analysis of taurine in infant formulas and adult nutritionals by HILIC-MS/MS is described. A single-laboratory validation (SLV) study demonstrated the applicability of the method to meet requirements of SMPR 2014.013. In December 2022, the SPIFAN ERP voted to adopt this method as First Action AOAC Official Method 2022.03.

Designing blenderized tube feeding diets for children and investigating their physicochemical and microbial properties and Dietary Inflammatory Index.

BACKGROUND: Due to the benefits of blenderized tube feeding (BTF) diets, the interest in using them is increasing. This study aimed to design BTFs for children and investigate their physicochemical and microbial properties, as well as Dietary Inflammatory Index (DII). METHODS: Five BTF diets were formulated mainly with fresh foods; their DII, physical (viscosity), and chemical (moisture, ash, protein, fat, energy, and micronutrients) characteristics were assessed. Also, the Hazard Analysis and Critical Control Points (HACCP) system was implemented for quality assurance of preparation, storage, and delivery of BTFs to patients in hospital. The microbial contamination (total count, Salmonella, Escherichia coli, Bacillus cereus, Listeria monocytogenes, coliforms, Staphylococcus aureus coagulase positive, mold, and yeast) was analyzed. RESULTS: Energy and percentages of protein, fat, and carbohydrate in BTFs were in the range of 103-112 kcal/100 ml, 16%-22%, 28%-34%, and 48%-52%, respectively. The viscosity of the five developed BTFs was between 29 and 64 centipoises, which allows the formulas to flow without syringe pressure. The DII of all BTFs was between -0.73 and -2.24. Due to the implementation of HACCP, monitoring the production line of BTFs, and performance of corrective measures, no microbial contamination was observed by indicator pathogenic microorganisms. CONCLUSION: A planned BTF diet can be an excellent selection for children using enteral nutrition with tube feeding especially when they are made from fresh and anti-inflammatory foods such as recipes prepared in this study.

Microbiological quality of enteral formulations handled at home: A systematic review.

BACKGROUND: Studies have shown an increase in the number of patients who use enteral nutrition at home and the benefits of this type of nutrition care. However, little is known about the risk of bacterial contamination of enteral formulations prepared at home. Therefore, the aim of the study was to assess the microbiological quality of home-prepared enteral formulations. METHODS: This study is a systematic review, registered in PROSPERO. The search for articles was carried out in databases and gray literature. Eligible studies that microbiologically analyzed homemade enteral preparations (HEPs), blended enteral preparations (BEPs), and commercial enteral formulas (CEFs) that were prepared at home were selected. The types and quantities of microorganisms, sources of contamination in the handling area, and the consequences of contamination (signs and symptoms) were the subjects extracted from the studies. RESULTS: Five studies evaluated 217 enteral formulations. It was found that 72.81% of the enteral formulations exceeded the acceptable bacterial count in the case of at least one of the analyzed microorganisms. This result corresponded to 93.58% (n = 73) of the HEPs, 81.96% (n = 50) of the BEPs, and 44.87% (n = 35) of the CEFs. The presence of 10 different microorganisms was identified in the enteral formulations, and total coliforms and mesophilic aerobics were the microorganisms found in the greatest quantity in the samples. CONCLUSION: The three types of home-prepared enteral formulations showed unsatisfactory microbiological quality, indicating poor hygiene conditions during food handling.

Analysis of Bovine Lactoferrin in Infant Formula and Adult Nutritional Products by Optical Biosensor Immunoassay: First Action 2021.07.

BACKGROUND: Bovine lactoferrin is increasingly being used as an ingredient in infant formula manufacture to enhance nutritional efficacy through the provision of immunoprotective, growth, and antimicrobial factors to the neonate. OBJECTIVE: To evaluate the analytical performance of an optical biosensor immunoassay for compliance with the method performance requirements described in SMPR 2020.005. METHOD: Following dilution of the sample in buffer, an automated, label-free, real-time optical biosensor immunoassay was used in a direct assay format to quantitate bovine lactoferrin by its interaction with an immobilized anti-lactoferrin antibody. Quantitation was accomplished by the external standard technique with interpolation from a 4-parameter calibration regression. RESULTS: The analytical range (0-200 mg/hg), method detection limit (0.8 mg/hg), recovery (96.1-109.2%), and repeatability (1.0-5.3%) complied with the requirements given in the lactoferrin SMPR. The method was shown to be specific for native, intact lactoferrin; thermally denatured lactoferrin generated no measurable binding response. CONCLUSION: The method described is suitable for the quantification of intact, undenatured lactoferrin in milk products, infant formulas (bovine milk protein-based, soy protein-based, and amino acid-based), and adult nutritionals and has been demonstrated to meet the performance requirements defined in SMPR 2020.005. HIGHLIGHTS: A single-laboratory validation (SLV) of an automated biosensor immunoassay for the determination of intact, undenatured lactoferrin is described.

Determination of Total Amino Acids in Infant Formulas, Adult Nutritionals, Dairy, and Cereal Matrixes by UHPLC-UV: Interlaboratory Validation Study, Final Action 2018.06.

BACKGROUND: A method for the quantification of total amino acids (including taurine and excluding tryptophan) using ultra- HPLC separation coupled to UV detection (UHPLC-UV) was granted First Action status (AOAC 2018.06) by the AOAC INTERNATIONAL Stakeholder Program for Infant Formula and Adult Nutritionals (SPIFAN) in 2018. OBJECTIVE: An interlaboratory study was conducted to further assess method performance against the AOAC Standard Method Performance Requirements (AOAC SMPR® 2014.013). Dairy and cereal matrixes were added to expand the scope of the method in collaboration with IDF (International Dairy Federation), ISO (International Organization for Standardization), and AACCI (American Association of Cereal Chemists International, now Cereals & Grains Association). METHODS: Sixteen different matrixes were chosen to cover the requirements of AOAC, IDF/ISO, and AACCI. Blind duplicate samples were organized into specific series to ensure that each pair was analyzed on the same day. Fifteen laboratories returned results. Data from four laboratories were considered invalid and removed from the dataset. Remaining data were assessed according to the Appendix D of the AOAC Official Methods of AnalysisSM (guidelines for collaborative study procedures). RESULTS: This method generally met the requirements listed in the SMPR for infant formulas and adult nutritionals, except for taurine. Method performance was comparable in dairy and cereal matrixes. Five different UHPLC instruments were used with either commercial or in-house reagents, demonstrating that the method is not limited to a single supplier. CONCLUSION: This method was recommended for Final Action in infant and adult/pediatric nutritional formulas by the AOAC SPIFAN Nutrients Expert Review Panel in April 2021, with the exception of taurine. The corresponding IDF/ISO Draft International Standard (DIS) was approved by national bodies in May 2022, and comments collected during the ballot were incorporated into this manuscript. HIGHLIGHTS: AOAC Official Method 2018.06 for the determination of total amino acids in infant formulas, adult nutritionals, dairy, and cereal matrixes was successfully validated in an interlaboratory study.

Lactose Concentration in Low-Lactose and Lactose-Free Milk, Milk Products, and Products Containing Dairy Ingredients by High Sensitivity Enzymatic Method (K-LOLAC), Collaborative Study: Final Action 2020.08.

BACKGROUND: The AOAC Stakeholder Panel on Strategic Food Analytical Methods issued a call for methods in 2018 for the measurement of lactose in low-lactose and lactose-free products under Standard Method Performance Requirement (SMPR®) 2018.009. Megazyme's Lactose Assay Kit (K-LOLAC) was reviewed and accepted as a First Action Official MethodSM in 2020 (2020.08). OBJECTIVE: A collaborative study was conducted to evaluate the to evaluate the reproducibility of AOAC Official MethodSM2020.08 for the measurement of lactose concentration in low-lactose and lactose-free milk, milk products, and products containing dairy ingredients. METHOD: Samples are deproteinated and clarified by treatment with Carrez reagents, and then free glucose is removed using a glucose oxidase and catalase treatment system. Quantification of lactose is based on the hydrolytic activity of ß-galactosidase, which hydrolyses lactose to glucose and galactose. Any remaining free D-glucose is first measured using a hexokinase (HK)/glucose 6-phosphate dehydrogenase (G-6PDH)/6-phosphogluconate dehydrogenase (6-PGDH) based assay procedure, and then ß-galactosidase is added to hydrolyze the lactose in the same reaction vessel with concurrent measurement of the released D-glucose. The samples analyzed included a number of lactose-free and low-lactose milk samples, lactose-free infant formula, lactose-free milkshake, lactose-free adult nutritional drink, lactose-free cream, and lactose-free cheese. RESULTS: All materials had repeatability relative standard deviations (RSDr) <7%. The reproducibility relative standard deviation (RSDR) varied from 3.8 to 14.9% with seven of the 10 test samples having an RSDR of <10%. CONCLUSIONS: The Lactose Assay Kit (K-LOLAC) meets the requirements for reproducibility set out under SMPR 2018.009. HIGHLIGHTS: The Lactose Assay (K-LOLAC) is a robust, simple, and reproducible method for analysis of lactose in foodstuffs and beverages.

A mathematical description for the nutrition therapy ecosystem.

Introduction: Background: nutrition therapy is a complex area of healthcare systems that encompasses patient characteristics, medical decision making, nutritional formula characteristics, and costs, composing a complex ecosystem. The integration of these different domains is actualized in medical prescription in a heuristic and iterative way, taking into account patient characteristics and formulas, with a limited capacity for in-scale calculations and inclusion of factors involved in the prescription of nutritional formulas and other ecosystem elements. From a practical standpoint, depicting the four areas as equalities could provide the necessary equivalence to study dependence and consequence from left- and right-side terms. Objectives: the objective of this theoretical study is to provide a mathematical model that describes and integrates different aspects of nutrition therapy. Methods: in this theoretical study, we deducted a mathematical representation for nutrition therapy using first-grade equations and simple calculus techniques. Results: a formula that coordinates four elements of the nutrition therapy ecosystem was found: cashflow, compliance adherence, patient, and macronutrient mass, formula density and unitary cost. Conclusion: factors involving decision-making in nutrition therapy can be unified in a mathematical model.

Introducción: Antecedentes: la terapia nutricional es un área compleja de los sistemas de salud que abarca las características del paciente, la toma de decisiones médicas, las características de la fórmula nutricional y los costos, componiendo un ecosistema complejo. La integración de estos diferentes dominios se actualiza en la prescripción médica de forma heurística e iterativa, teniendo en cuenta las características y fórmulas del paciente, con una capacidad limitada para cálculos a escala e inclusión de otros factores relacionados con la prescripción de la terapia nutricional y los elementos del ecosistema. Desde un punto de vista práctico, describir las cuatro áreas como igualdades podría proporcionar la equivalencia necesaria para estudiar la dependencia y la consecuencia de los términos del lado izquierdo y derecho. Objetivos: el objetivo de este estudio teórico es brindar un modelo matemático que describa e integre diferentes aspectos de la terapia nutricional. Métodos: en este estudio teórico se dedujo una representación matemática para la terapia nutricional utilizando ecuaciones de primer grado y técnicas de cálculo simple. Resultados: se encontró una fórmula que coordina cuatro elementos del ecosistema de la terapia nutricional: flujo de caja, adherencia al cumplimiento, masa del paciente y macronutrientes, densidad de la fórmula y costo unitario. Conclusión: los factores que involucran la toma de decisiones en la terapia nutricional se pueden unificar en un modelo matemático.

Effect of Artificial Diet on Immature Stage of the Great Eggfly, Hypolimnas bolina (Lepidoptera: Nymphalidae).

<b>Background and Objective:</b> One of the Nymphalidae butterfly species found in West Sumatra in <i>Hypolimnas bolina</i>. Currently, research on the artificial diet for the Nymphalidae butterfly is relatively rare in Padang, West Sumatra. The objectives of this study were to analyze the preferences of <i>H. bolina</i> larvae, duration of the immature stage and mortality of <i>H. bolina</i> in artificial diet treatment. <b>Materials and Methods:</b> Some biological aspects of <i>H. bolina</i> in corresponding to artificial diet and its effect were investigated in the laboratory. <b>Results:</b> The result showed that there was no significant difference in the frequency of visits of the larvae in the two diet treatments namely natural (<i>Laportea interrupta</i> leaves) and artificial diets (Sig = 0.289, p>0.05) but the duration of the visit of <i>H. bolina</i> larvae was significantly different (Sig = 0.000, p<0.05). The visit duration of the immature stage of <i>H. bolina </i>was significantly different, except the prepupa and pupal stage. There was no mortality of instar larvae and prepupa stage observed in both of the two-diet treatments. However, the mortality of pupae in an artificial diet was 4%. Of the total of 24 individual larvae fed with artificial diet, all of them successfully emerged, consisted of 12 males and 12 females but there was one male with abnormal wings. The average living period in the artificial diet of imago was 14.82 days for males and 16.77 days for a female. The average larval weight was no significant difference (Sig = 0.981, p>0.05) but the average pupal weight of the natural diet was slightly higher than the artificial diet. <b>Conclusion:</b> The formulation of an artificial diet is suitable for <i>H. bolina</i> larvae based on the results of immature mortality and adult emergences. Therefore, the formulation of an artificial diet is suitable for <i>H. bolina</i> with its composition almost similar to <i>L. interrupta</i> leaves (natural diet).

Serum cytokine patterns are modulated in infants fed formula with probiotics or milk fat globule membranes: A randomized controlled trial.

BACKGROUND: Proteins and lipids of milk fat globule membrane (MFGM) and probiotics are immunomodulatory. We hypothesized that Lactobacillus paracasei ssp. paracasei strain F19 (F19) would augment vaccine antibody and T helper 1 type immune responses whereas MFGM would produce an immune response closer to that of breastfed (BF) infants. OBJECTIVE: To compare the effects of supplementing formula with F19 or bovine MFGM on serum cytokine and vaccine responses of formula-fed (FF) and BF infants. DESIGN: FF infants were randomized to formula with F19 (n = 195) or MFGM (n = 192), or standard formula (SF) (n = 194) from age 21±7 days until 4 months. A BF group served as reference (n = 208). We analyzed seven cytokines (n = 398) in serum at age 4 months using magnetic bead-based multiplex technology. Using ELISA, we analyzed anti-diphtheria IgG (n = 258) and anti-poliovirus IgG (n = 309) concentrations in serum before and after the second and third immunization, respectively. RESULTS: Compared with SF, the F19 group had greater IL-2 and lower IFN-γ concentrations (p<0.05, average effect size 0.14 and 0.39). Compared with BF, the F19 group had greater IL-2, IL-4 and IL-17A concentrations (p<0.05, average effect size 0.42, 0.34 and 0.26, respectively). The MFGM group had lower IL-2 and IL-17A concentrations compared with SF (p<0.05, average effect size 0.34 and 0.31). Cytokine concentrations were comparable among the MFGM and BF groups. Vaccine responses were comparable among the formula groups. CONCLUSIONS: Contrary to previous studies F19 increased IL-2 and lowered IFN-γ production, suggesting that the response to probiotics differs across populations. The cytokine profile of the MFGM group approached that of BF infants, and may be associated with the previous finding that infectious outcomes for the MFGM group in this cohort were closer to those of BF infants, as opposed to the SF group. These immunomodulatory effects support future clinical evaluation of infant formula with F19 or MFGM.

Effectiveness and cost-effectiveness of 4 supplementary foods for treating moderate acute malnutrition: results from a cluster-randomized intervention trial in Sierra Leone.

BACKGROUND: Moderate acute malnutrition (MAM) affects 33 million children annually. Investments in formulations of corn-soy blended flours and lipid-based nutrient supplements have effectively improved MAM recovery rates. Information costs and cost-effectiveness differences are still needed. OBJECTIVES: We assessed recovery and sustained recovery rates of MAM children receiving a supplementary food: ready-to-use supplementary food (RUSF), corn soy whey blend with fortified vegetable oil (CSWB w/oil), or Super Cereal Plus with amylase (SC + A) compared to Corn Soy Blend Plus with fortified vegetable oil (CSB+ w/oil). We also estimated differences in costs and cost effectiveness of each supplement. METHODS: In Sierra Leone, we randomly assigned 29 health centers to provide a supplement containing 550 kcal/d for ∼12 wk to 2691 children with MAM aged 6-59 mo. We calculated cost per enrollee, cost per child who recovered, and cost per child who sustained recovery each from 2 perspectives: program perspective and caregiver perspective, combined. RESULTS: Of 2653 MAM children (98.6%) with complete data, 1676 children (63%) recovered. There were no significant differences in the odds of recovery compared to CSB+ w/oil [0.83 (95% CI: 0.64-1.08) for CSWB w/oil, 1.01 (95% CI: 0.78-1.3) for SC + A, 1.05 (95% CI: 0.82-1.34) for RUSF]. The odds of sustaining recovery were significantly lower for RUSF (0.7; 95% CI 0.49-0.99) but not CSWB w/oil or SC + A [1.08 (95% CI: 0.73-1.6) and 0.96 (95% CI: 0.67-1.4), respectively] when compared to CSB+ w/oil. Costs per enrollee [US dollars (USD)/child] ranged from $105/child in RUSF to $112/child in SC + A and costs per recovered child (USD/child) ranged from $163/child in RUSF to $179/child in CSWB w/oil, with overlapping uncertainty ranges. Costs were highest per sustained recovery (USD/child), ranging from $214/child with the CSB+ w/oil to $226/child with the SC + A, with overlapping uncertainty ranges. CONCLUSIONS: The 4 supplements performed similarly across recovery (but not sustained recovery) and costed measures. Analyses of posttreatment outcomes are necessary to estimate the full cost of MAM treatment. This trial was registered at clinicaltrials.gov as NCT03146897.

Development of weaning food with prebiotic effects based on roasted or extruded quinoa and millet flour.

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.

The effect of cooling and rehydration methods in high moisture meat analogs with pulse proteins-peas, lentils, and faba beans.

Pulse proteins (PLP) can be ideal alternative-sources that produce a meat-like textured product, known as a high moisture meat analog (HMMA). In this research, each commercial PLP: pea (16%), lentil (16%), and faba-bean (20%) was mixed with pea isolate (63%, 63%, and 59%, respectively) and constant ingredients which are canola oil (6%) and wheat gluten (15%) and texturized to produce HMMA using a twin-screw extruder (TX-52) with a cooling die. Soy concentrate and soy isolate were mixed with the constant ingredients and texturized into an HMMA and used as a control. Before freezing for storage, each sample was cooled by air, water, or a brine solution (2% or 4%) for 10 min. Frozen samples were thawed at room temperature (25 °C) for 3 hr and rehydrated by soaking at 25 °C for 2 hr, warm-soaking at 50 °C for 12 hr, or boiling for 2 min. Color, moisture content (MC), specific density (SD), water absorption index (WAI), water solubility index (WSI), and texture were measured. Compared to the control, samples with PLP had less lightness and texture and greater redness, yellowness, MC and WSI. The 2% brine solution used for cooling reduced WSI without textural change compared to other cooling methods. Boiling for rehydration increased lightness while warm-soaking decreased lightness and increased yellowness. In addition, boiling resulted in the least MC, SD, WSI, and WAI following soaking and warm-soaking. Therefore, these PLP can be used as alternative meat sources to soy proteins and a 2% brine solution for cooling and rehydration by boiling are recommended to reduce the WSI. PRACTICAL APPLICATION: Pulses are an excellent food ingredient because they are rich in protein and have an exceptional nutritional profile. In this study, high moisture meat analogs containing pea proteins, lentil proteins, faba bean proteins, and pea isolate instead of soy concentrate and soy isolate were produced. According to the results, pulse proteins can be an alternative source to soy proteins. Since they formed relatively well-defined orientation. Further research can be conducted using modified processing conditions for texturization to improve its quality. In addition, this research can help researchers and product developers understand proper handling methods for HMMA products after production such as cooling before freezing for storage and thawing and rehydrating after freezing.

Different Dietary N-3 Polyunsaturated Fatty Acid Formulations Distinctively Modify Tissue Fatty Acid and N-Acylethanolamine Profiles.

We investigated the influence of different dietary formulation of n-3 polyunsaturated fatty acids (PUFA) on rat tissue fatty acid (FA) incorporation and consequent modulation of their bioactive metabolite N-acylethanolamines (NAE). For 10 weeks, rats were fed diets with 12% of fat from milk + 4% soybean oil and 4% of oils with different n-3 PUFA species: soybean oil as control, linseed oil rich in α-linolenic (ALA), Buglossoides arvensis oil rich in ALA and stearidonic acid (SDA), fish oil rich in eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), Nannochloropsis microalga oil rich in EPA or Schizochytrium microalga oil rich in DHA. FA and NAE profiles were determined in plasma, liver, brain and adipose tissues. Different dietary n-3 PUFA distinctively influenced tissue FA profiles and consequently NAE tissue concentrations. Interestingly, in visceral adipose tissue the levels of N-arachidonoylethanolamide (AEA) and N-docosahexaenoylethanolamide (DHEA), NAE derived from arachidonic acid (AA) and DHA, respectively, significantly correlated with NAE in plasma, and circulating DHEA levels were also correlated with those in liver and brain. Circulating NAE derived from stearic acid, stearoylethanolamide (SEA), palmitic acid and palmitoylethanolamide (PEA) correlated with their liver concentrations. Our data indicate that dietary n-3 PUFA are not all the same in terms of altering tissue FA and NAE concentrations. In addition, correlation analyses suggest that NAE levels in plasma may reflect their concentration in specific tissues. Given the receptor-mediated tissue specific metabolic role of each NAE, a personalized formulation of dietary n-3 PUFA might potentially produce tailored metabolic effects in different pathophysiological conditions.

A collagen hydrolysate/milk protein-blend stimulates muscle anabolism equivalently to an isoenergetic milk protein-blend containing a greater quantity of essential amino acids in older men.

BACKGROUND & AIMS: Nutritional composition is key for skeletal muscle maintenance into older age. Yet the acute effects of collagen protein blended with other protein sources, in relation to skeletal muscle anabolism, are ill-defined. We investigated human muscle protein synthesis (MPS) responses to a 20 g blend of collagen protein hydrolysate + milk protein (CP+MP, 125 ml) oral nutritional supplement (ONS) vs. 20 g non-blended milk protein source (MP, 200 ml) ONS, in older adults. METHODS: Healthy older men (N = 8, 71±1 y, BMI: 27±1 kg·m-2) underwent a randomized trial of 20 g protein, from either a CP+MP blend (Fresubin®3.2 kcal DRINK), or a kcal-matched (higher in essential amino acids (EAA) ONS of MP alone. Vastus lateralis (VL) MPS and plasma AA were determined using stable isotope-tracer mass spectrometry; anabolic signaling was quantified via immuno-blotting in VL biopsies taken at baseline and 2/4 h after ONS feeding. Plasma insulin was measured via enzyme-linked immunosorbent assay (ELISA). Measures were taken at rest, after the feed (FED) and after the feed + exercise (FED-EX) conditions (unilateral leg exercise, 6 × 8, 75% 1-RM). RESULTS: MP resulted in a greater increase in plasma leucine (MP mean: 152 ± 6 µM, CP+MP mean: 113 ± 4 µM (Feed P < 0.001) and EAA (MP mean: 917 ± 25 µM, CP+MP mean: 786 ± 15 µM (Feed P < 0.01) than CP+MP. CP + MP increased plasma glycine (peak 385 ± 57 µM (P < 0.05)), proline (peak 323 ± 29 µM (P < 0.01)) and non-essential amino acids (NEAA) (peak 1621 ± 107 µM (P < 0.01)) with MP showing no increase. Plasma insulin increased in both trials (CP+MP: 58 ± 10 mU/mL (P < 0.01), MP: 42 ± 6 mU/mL (P < 0.01), with peak insulin greater with CP+MP vs. MP (P < 0.01). MPS demonstrated equivalent increases in response to CP+MP and MP under both FED (MP: 0.039 ± 0.005%/h to 0.081 ± 0.014%/h (P < 0.05), CP+MP: 0.042 ± 0.004%/h to 0.085 ± 0.007%/h (P < 0.05)) and FED-EX (MP: 0.039 ± 0.005%/h to 0.093 ± 0.013%/h (P < 0.01), CP+MP: 0.042 ± 0.004%/h to 0.105 ± 0.015%/h, (P < 0.01)) conditions. FED muscle p-mTOR fold-change from baseline increased to a greater extent with CP+MP vs. MP (P < 0.05), whilst FED-EX muscle p-eEF2 fold-change from baseline decreased to a greater extent with CP+MP vs. MP (P < 0.05); otherwise anabolic signaling responses were indistinguishable. CONCLUSION: Fresubin®3.2 kcal DRINK, which contains a 20 g mixed blend of CP+MP, resulted in equivalent MPS responses to MP alone. Fresubin® 3.2 Kcal DRINK may provide a suitable alternative to MP for use in older adults and a convenient way to supplement calories and protein to improve patient adherence and mitigate muscle mass loss.

Estudio en vida real de efectividad de una fórmula hipercalórica hiperproteica en el mantenimiento y mejora del estado nutricional en pacientes con indicación de nutrición enteral a largo plazo / Real-life study of the effectiveness of a high calorie and high protein formula in the maintenance and improvement of nutritional state in patients with an indication of long-term enteral nutrition

La selección de la fórmula más adecuada en nutrición enteral domiciliaria a largo plazo es un tema controvertido. Nuestro objetivo fue estudiar una fórmula hipercalórica hiperproteica en pacientes con alimentación exclusivamente con sonda a largo plazo (180 días). MÉTODOS: Estudio multicéntrico observacional prospectivo en vida real con fórmula hipercalórica hiperproteica (2kcal/ml y 20% de proteínas). Se recogieron datos generales, antropométricos, analíticos y de calidad de vida mediante escala analógica visual del European Quality of Life-5 Dimensions al inicio, 60, 120 y 180 días. La tolerancia gastrointestinal se evaluó con una escala analógica visual y escala de heces de Bristol y la valoración del riesgo de desnutrición mediante NRS-2002. RESULTADOS: Un total de 51 pacientes (88,2% varones, edad media de 62,0 años), con patología oncológica en el 72,5%. No hubo diferencias en datos antropométricos, aunque sí se redujo el porcentaje de pacientes con riesgo de desnutrición del 75 al 8,3% (p < 0,0001). No se observaron diferencias en albúmina, prealbúmina, transferrina, linfocitos o hematocrito. La calidad de vida mejoró de 3,84 (1,27) a 5,37 (1,12) en la escala analógica visual (p < 0,0001). Se observó una reducción de la sintomatología gastrointestinal a lo largo del seguimiento. Tanto el número como el porcentaje de deposiciones consideradas normales según la escala de Bristol se mantuvieron estables. CONCLUSIÓN: Nuestro estudio apoya que el empleo de fórmulas hipercalóricas hiperproteicas durante un tratamiento nutricional a 6 meses permite una adecuada evolución nutricional sin riesgo de deshidratación y con una buena tolerancia, incluso mejoría de sintomatología gastrointestinal, y puede contribuir a una mejora en la calidad de vida

The selection of the most appropriate formula in long-term home enteral nutrition is a controversial issue. Our objective was to study a high protein hypercaloric enteral nutrition formula in patients with long-term feeding (180 days). METHODS: Prospective observational multicenter real-life study with high-protein hypercaloric formula (2kcal/ml and 20% protein). General, anthropometric, analytical and quality of life data were collected by visual analog scale of the European Quality of Life-5 Dimensions at the beginning, 60, 120 and 180 days. Gastrointestinal tolerance was assessed with a visual analog scale and Bristol Stool Scale and the risk of malnutrition was assessed using NRS-2002. RESULTS: 51 patients (88.2% men, mean age 62.0 years), with oncological diseases in 72.5%. No differences in anthropometric data were observed, although the percentage of patients at risk of malnutrition according to NRS 2002 was reduced from 75% to 8.3% (p < 0.0001). No differences were observed in albumin, prealbumin, transferrin, lymphocytes or hematocrit. The quality of life improved from 3.84 (1.27) to 5.37 (1.12) on the visual analog scale (p < 0.0001). A reduction in gastrointestinal symptoms was observed throughout the period of enteral nutrition. Both the number and percentage of stools considered normal according to the Bristol scale remained stable. CONCLUSION: Our study supports that the use of high-protein hypercaloric formulas during a 6-month nutritional treatment allows an adequate nutritional evolution without risk of dehydration and with a good tolerance, even improvement of gastrointestinal symptoms, and can contribute to an improvement in the quality of lifetime

Simultaneous Determination of Total Vitamins B1, B2, B3, and B6 in Infant Formula and Related Nutritionals by Enzymatic Digestion and LC-MS/MS-A Multi-Laboratory Testing Study Final Action: AOAC Method 2015.14.

BACKGROUND/OBJECTIVE: A multi-laboratory study was completed for AOAC First Action Method 2015.14. Ten laboratories from eight countries participated. Each laboratory analyzed (in duplicate) a placebo and 14 fortified nutritionals. Product matrices analyzed included milk, soy, partially hydrolyzed milk, partially hydrolyzed soy, and elemental-based infant formula powders, milk based infant formula ready-to-feed liquids, adult low-fat powders, and adult high fat and high protein ready-to-drink nutritionals. Data was then compared to standard method performance requirements (SMPR). METHOD: Samples were prepared by enzymatic digestion with papain, α-amylase, and phosphatase to hydrolyze protein and complex carbohydrate and to free phosphorylated vitamin forms respectively. Stable-isotope labeled internal standards were incorporated into the sample preparation to correct for variability in both the sample preparation and instrument response. Prepared samples and working standard solutions were injected onto an ultra-high-pressure liquid chromatograph, interfaced to a triple-quadrupole mass spectrometer (MS/MS). Reverse phase gradient chromatography, using mobile phases of methanol and 20 mM ammonium formate in water on a C-18 column, were used for the analysis. The MS/MS was configured to monitor precursor-fragment ion pairs for each analyte and internal standard. Vitamins B1 (thiamine), B2 (riboflavin), B3 (niacin), and B6 (pyridoxine) were quantified by least squares regression using the response ratio of the analyte to its internal standard. RESULTS/CONCLUSIONS: Total vitamins B1, B2, B3, and B6 had average repeatability of 2.3%, 3.9%, 2.7%, and 2.2% RSD, and reproducibility of 8.2%, 6.9%, 6.7%, and 5.8% RSD, respectively. Repeatability and reproducibility SMPR were met for 53/56 and 50/56 fortified-product/analyte pairs analyzed, respectively.

Determination of Fructans in Infant Formula and Adult/Pediatric Nutritional Formula by Anion-Exchange Chromatography with Pulsed Amperometric Detection after Enzymatic Treatment: Collaborative Study, Final Action 2016.14.

BACKGROUND: Fructans are added to infant formula and adult nutritionals for their prebiotic effect. A method (AOAC 2016.14) was developed for their analysis which has already demonstrated excellent performance during single laboratory validation. OBJECTIVE: To determine repeatability and reproducibility of the method through a collaborative study. METHODS: Fourteen laboratories from 11 different countries enrolled for the study. Participants analyzed a practice sample, then 8 formula or adult nutritionals in blind duplicate. Results and any method modifications were reported to the study director. RESULTS: Twelve laboratories provided results on time for reporting. Precision results for five samples met the requirements of the Standard Method Performance Requirements (SMPR 2014.002), with RSDr ranging from 3.60 to 4.25% and RSDR ranging from 5.90 to 11.7%. The practice sample also met the requirements of SMPR 2014.002, with RSDr and RSDR of 2.53% and 6.70% respectively. Precision results for three test samples did not fully meet the SMPR, with RSDr ranging from 2.27 to 7.65% and RSDR ranging from 12.8 to 15.1%. After review, the AOAC Stakeholder Panel for Infant Formula and Adult Nutritional Expert Review Panel (SPIFAN ERP) concluded that the data presented mostly met the SMPR and hence recommended that the method to be advanced for adoption as an AOAC Final Action method. CONCLUSIONS: The method described in AOAC 2016.14 is suitable for the determination of fructans in infant formula and adult nutritionals.

Determination of Free and Total Choline and Free and Total Carnitine in Infant Formula and Adult/Pediatric Nutritional Formula by Liquid Chromatography/Tandem Mass Spectrometry (HPLC-MS/MS): A Multi-laboratory Testing Study, Final Action 2015.10.

BACKGROUND: A multi-laboratory study was conducted on AOAC First Action Method 2015.10 "Determination of Free and Total Choline and Free and Total Carnitine in Infant Formula and Adult/Pediatric Nutritional Formula by Liquid Chromatography/Tandem Mass Spectrometry (HPLC-MS/MS)." OBJECTIVE: In this study, nine laboratories participated in the performance testing of the method using ten nutritional products tested as blind duplicates. METHOD: Both free and total carnitine and free and total choline content of the samples were determined using separate extractions for the free and total results. For free choline and carnitine analysis, samples are diluted in water. For total choline and carnitine analysis, samples are extracted using acid-assisted microwave hydrolysis with nitric acid. For both the free and total methods, samples are then diluted with acetonitrile and analyzed using strong cation exchange (SCX) liquid chromatography coupled to a triple quadrupole tandem mass spectrometer (LCMS). Stable isotope labeled internal standards were utilized in all analyses to compensate for extraction inefficiencies and ionization suppression.

Updated Food Composition Database for Cereal-Based Gluten Free Products in Spain: Is Reformulation Moving on?

We developed a comprehensive composition database of 629 cereal-based gluten free (GF) products available in Spain. Information on ingredients and nutritional composition was retrieved from food package labels. GF products were primarily composed of rice and/or corn flour, and 90% of them included added rice starch. The most common added fat was sunflower oil (present in one third of the products), followed by palm fat, olive oil, and cocoa. Only 24.5% of the products had the nutrition claim "no added sugar". Fifty-six percent of the GF products had sucrose in their formulation. Xanthan gum was the most frequently employed fiber, appearing in 34.2% of the GF products, followed by other commonly used such as hydroxypropyl methylcellulose (23.1%), guar gum (19.7%), and vegetable gums (19.6%). Macronutrient analysis revealed that 25.4% of the products could be labeled as a source of fiber. Many of the considered GF food products showed very high contents of energy (33.5%), fats (28.5%), saturated fatty acids (30.0%), sugars (21.6%), and salt (28.3%). There is a timid reformulation in fat composition and salt reduction, but a lesser usage of alternative flours and pseudocereals.