Nutritional composition with large lipid globules and native protein

Abstract

The present invention relates to nutritional compositions such as an infant formula, a follow-on formula, a young child formula or a medical nutritional composition that comprises lipid globules having a volume weighted mode diameter of at least 1.0 µm, and / or wherein at least 40 vol.%, based on total volume, has a diameter between 2 µm and 12 µm, and wherein the whey protein has a nativity of 40 to 95%. The invention further relates to health benefits linked to this nutritional composition.

Description

[0001] NUTRITIONAL COMPOSITION WITH LARGE LIPID GLOBULES AND NATIVE PROTEIN

[0002] TECHNICAL FIELD OF THE INVENTION

[0003] The present invention relates to nutritional compositions, such as an infant formula, a follow-on formula, a young child formula or a medical nutritional composition, that comprises lipid globules having a volume weighted mode diameter of at least 1.0 pm, and / or wherein at least 40 vol.%, based on lipid volume, has a diameter between 2 pm and 12 pm, and wherein 40 to 95 wt.% of the whey protein is native whey protein.

[0004] BACKGROUND OF THE INVENTION

[0005] Infant or follow-on formulae are commonly used when breastfeeding is inadequate or unsuccessful for medical reasons, or because of a choice not to breastfeed. Commercial infant formulae are commonly used to provide supplemental or sole source of nutrition in early life. These formulae comprise a range of nutrients to meet the nutritional needs of the growing infant, and typically include fat, carbohydrate, protein, vitamins, minerals, and other nutrients helpful for optimal infant growth and development.

[0006] Nutritional compositions for infants and young children are often sold as powders to be reconstituted with water or in some instances as ready to drink or concentrated liquid compositions. Those compositions are intended to cover most or all the nutritional needs of the infants or young children.

[0007] Human milk lipids have a distinct physical structure composed of large lipid globules with an average volume-weighted mode diameter of about 4 pm existing of a triglyceride core coated by a tri-layer of membranes, the milk fat globule membrane (MFGM). The volume-weighted mode diameter of lipid droplets in standard infant formula is about 0.3 to 0.5 pm due to the industrial processing procedures to achieve stable and reproducible end products and is not surrounded by MFGM but mostly by proteins such as casein. Standard commercially available formulae also mostly contain vegetable oils and have small lipid droplets with proteins adhering to the surface.

[0008] Infant formula with lipid globules with an architecture more similar to the lipid globules in human milk and a process to prepare those have been described in for example WO2010027258 or WO2010027259.

[0009] Following these and several further disclosures numerous health effects are associated with the consumption of nutritional composition having larger lipid globules which are more similar to the lipid globules in human milk.

[0010] It is also known that native whey protein has beneficial health effects. For example, native whey is associated with effects on reducing allergy, see W02019160416, and with improving intestinal maturation, see W02020159373, treating and / or preventing intestinal infection, see W02020159372 and improving gastro-intestinal tolerance, see W02022090269. The presence of native proteins in nutritional compositions is known to lead to severe foaming. As confirmed in Dissanayake, M. et al., ‘Functional properties of whey proteins affected by heat treatment and hydrodynamic high-pressure shearing’, Journal of Dairy science | 92 (4): 1387-1397, 2009.

[0011] While this is desirable when trying to make, e.g. a coffee creamer, foaming is disadvantageous when preparing nutritional compositions for infants, young children or subjects with health challenges, or when reconstituting these nutritional compositions in water when they are provided as a powder. Foaming is also a disadvantage when measured amounts of a product need to be fed to a subject. When foam is present it is a challenge to achieve dosing accuracy when feeding liquid tube feeding nutrition using a feed pump for example. Not only for reconstitutable powders, but also for ready to drink (RTD) liquid products excessive foaming is a disadvantage as it is often recommended to shake RTD liquids before use and with excessive foaming transferring the liquid to a bottle, cup or any other format is inconvenient. Furthermore, foaming means that air is entrapped in a formulation which may provide intestinal discomfort to an infant or a subject being fed with the nutritional composition.

[0012] SUMMARY OF THE INVENTION

[0013] It has now been found that when including native proteins into a nutritional composition wherein the lipid is significantly present in the form of large lipid globules, the excessive foaming is severely suppressed.

[0014] Quite unexpectedly, the nutritional compositions of the present invention with lipid globules with an architecture more similar to the lipid globules in human milk, viz which contain the large lipid globules with a volume weighted mode diameter of more than 1 .0 pm and / or wherein a substantial part of the lipid globules have a diameter between 2 pm and 12 pm and are optionally coated with phospholipids as in the disclosures referred to above, in which the whey protein is has a nativity of between 40 and 95% provides for compositions that have limited tendency to foam, which has advantages for their production process but also makes preparing bottles for babies or patients much easier, i.e. leads to substantial advantages for parents or consumers.

[0015] The inventors of the present invention have found that the foaming characteristics of highly native whey protein is suppressed in nutritional compositions containing large lipid globules with a volume weighted mode diameter of more than 1 .0 pm and / or wherein a substantial part, at least 40 vol.%, based on lipid volume, of the lipid globules have a diameter between 2 pm and 12 pm.

[0016] Accordingly, the present invention provides a nutritional composition comprising lipid and protein, and optionally carbohydrates, wherein the lipid is in the form of lipid globules and wherein the lipid comprises vegetable lipid; the lipid globules have a mode diameter, based on volume, of at least 1 .0 pm and / or at least 40 vol.% of the lipid globules, based on lipid volume, have a diameter of 2 to 12 pm; the protein comprises whey protein, the nutritional composition comprises 3-10 wt.% whey protein by dry weight of the nutritional composition; and

[0017] 40 to 95 wt.% of the whey protein is native whey protein; wherein the composition is an infant formula, a follow on formula, a young child formula, or a medical nutritional composition.

[0018] The present invention further provides a nutritional composition, as described herein above, for use in prevention of development of obesity later in life, preventing infections, improving the recovery from infections such as intestinal infections, reducing / preventing intestinal permeability, reducing gastrointestinal intolerance, reducing the risk of gut health problems, improving recovery of gut health problems, and / or reducing / preventing allergic response.

[0019] The present invention also relates to the non-therapeutic use of the nutritional composition, as described herein above, for promotion of metabolic health, promotion of development of good body composition, promotion of balanced growth, promotion of lean growth, promotion of cognitive development, improving brain health, improving gut health, and / or providing beneficial prebiotic effects.

[0020] DETAILED DESCRIPTION OF THE INVENTION

[0021] As used herein, the following terms have the following meanings.

[0022] The term “infant” as used herein refers to a child under the age of 12 months.

[0023] “Infant formula” or “follow-on formula” or “young child formula” means that it concerns a composition that is artificially made or in other words that it is a synthetic composition (i.e., the synthetic composition is not breast milk). Hence the nutritional composition is preferably an artificial infant formula or an artificial follow-on formula or an artificial young child formula or a synthetic infant formula or a synthetic follow- on formula or a synthetic young child formula.

[0024] Infant formula refers to nutritional compositions, artificially made, intended for infants of 0 months to about 4 months to 6 months of age and are intended as a substitute for human milk. Typically, infant formulae are suitable to be used as sole source of nutrition. Such infant formulae are also known as starter formula. Follow-on formulae are for infants starting with at 4 months to 6 months of life to 12 months of life and are intended to be supplementary feedings for infants that start weaning on other foods. Infant formulae and follow-on formulae are subject to strict regulations, for example for the EU regulations no. 609 / 2013 and no. 2016 / 127.

[0025] The term “young child” as used herein refers to a child aged between one and three years, also called a toddler. Young child formula refers to nutritional compositions, artificially made, intended for infants of 12 months to 36 months, which are intended to be supplementary feedings for infants. In the context of the present invention, young child formula can also be named growing-up milk. A medical nutritional product is a product that provides nutrition and strengthens or improves health. Such products are for subjects that have a disease or disorder to treat, delay or prevent the disorder or disease, or, for healthy subjects to prevent or delay (the development of) a disease or disorder.

[0026] “Base powders” for an infant formula, follow on formula, or young child formula are particulate nutritional compositions to which in a further blending step one or more further ingredients are added to make a complete formula meeting the regulations referred to above. In preferred embodiments the base powder contains at least a lipid and protein ingredient, in more preferred embodiments the base powder contains at least a lipid, a protein and a carbohydrate ingredient. Ingredients that are blended with the base powder are in preferred embodiments one or more from the group of digestible carbohydrates, non- digestible oligosaccharides, probiotics, vitamins and minerals. Most preferred this blending step is a dry blending step and the one or more further ingredients are selected from non-digestible oligosaccharides, probiotics, vitamins and minerals are blended in their powder form.

[0027] The term “whey protein” as used herein refers to whey protein such as found in mammal milk. Whey protein include proteins such as betalactoglobuline, alpha-lactalbumin, serum albumin, and immunoglobulins.

[0028] The term “native (or undenatured) protein” as used herein refers to protein with an intact primary structure, secondary structure, tertiary structure and (if applicable) quaternary structure.

[0029] The term “denatured protein” as used herein refers to protein with only an intact primary structure (i.e. the amino acid sequence is not disrupted). The secondary, tertiary and quaternary structure of the protein are absent.

[0030] The term “intact protein” as used herein refers to protein with at least an intact primary structure. The secondary, tertiary and quaternary structure of the protein may be intact or absent. In other words, intact protein covers both native protein and denatured intact protein.

[0031] Nativity in this specification is determined following the below method:

[0032] The protein concentrations were measured using the method of Dumas (Flash 4000 N / protein analyzer, Thermo Fischer Scientific). The nitrogen concentrations were converted into a protein concentration using a conversion factor of 6.25. The soluble protein fraction was determined by dissolving 20 gram nutritional composition powder into 80 gram water and acidifying the sample to pH 4.6 using 7M HCI solution. At pH 4.6 only the native whey protein remains soluble and denatured whey protein and casein proteins precipitate. The solution was centrifuged for 30 min at 4500g (Megafuge 16, Thermo Fisher Scientific) after which the protein concentration of the supernatant was determined. The nativity (%) was calculated as follows: wt. % soluble proteinpH 46

[0033] Nativity (%) = xlOO wt. % total whey protein

[0034] The term “native whey protein” as used herein refers to whey protein that is soluble in an aqueous solution at 20 °C having a pH of 4.6.

[0035] In this document and in its claims, the verb "to comprise" and its conjugations is used in its non-limiting sense to mean that items following the word are included, but items not specifically mentioned are not excluded. In addition, reference to an element by the indefinite article "a" or "an" does not exclude the possibility that more than one of the elements is present, unless the context clearly requires that there be one and only one of the elements. The indefinite article "a" or "an" thus usually means "at least one".

[0036] Formula

[0037] In a preferred embodiment, the invention pertains to a nutritional composition comprising lipid and protein, and optionally carbohydrates, wherein the lipid is in the form of lipid globules and wherein a. the lipid globules have a mode diameter based on volume of at least 1 .0 pm and / or at least 40 vol.% of the lipid globules have a diameter of between 2 and 12 pm; and b. the protein contains whey protein, wherein the nativity of the whey protein is 40 to 95%.

[0038] A first aspect of the invention pertains to a nutritional composition comprising lipid and protein, and optionally carbohydrates, wherein the lipid is in the form of lipid globules and wherein the lipid comprises vegetable lipid; the lipid globules have a mode diameter, based on volume, of at least 1 .0 pm and / or at least 40 vol.% of the lipid globules, based on lipid volume, have a diameter of 2 to 12 pm; the protein comprises whey protein, the nutritional composition comprises 3-10 wt.% whey protein by dry weight of the nutritional composition; and

[0039] 40 to 95 wt.% of the whey protein is native whey protein; wherein the composition is an infant formula, a follow on formula, a young child formula, or a medical nutritional composition.

[0040] The nutritional composition is preferably an infant formula, a follow-on formula, or a young child formula. The nutritional composition is preferably an infant formula or a follow-on formula. More preferably the nutritional composition is an infant formula. This means that the composition is not human milk. It also means that the nutritional composition is not native cow’s milk or native milk from another mammal. It further means that the nutritional composition is a synthetic nutritional composition.

[0041] In some embodiments the nutritional composition is preferably a base powder for any of the above products to which some ingredients are still to be added in a (dry) blending step. The nutritional composition preferably comprises protein, lipid, and digestible carbohydrates, wherein the lipid provides 30 to 60% of the total calories, the protein provides 5% to 20% of the total calories and the digestible carbohydrates provide 25% to 75% of the total calories.

[0042] The nutritional composition is preferably an infant formula or an follow-on formula and preferably comprises 3 g to 7 g lipid / 100 kcal, more preferably 4 g to 6 g lipid / 100 kcal, even more preferably 4.5 g to 5.5 g lipid / 100 kcal, preferably comprises 1 .7 g to 3.5 g protein / 100 kcal, more preferably 1 .8 g to 2.1 g protein / 100 kcal, even more preferably 1 .8 g to 2.0 g protein / 100 kcal and preferably comprises 5 g to 20 g digestible carbohydrate / 100 kcal, more preferably 6 g to 16 g digestible carbohydrate / 100 kcal, even more preferably 10 g to 15 g digestible carbohydrate / 100 kcal.

[0043] Preferably the nutritional composition is an infant formula or follow-on formula, and preferably has an energy density of 60 kcal / 100 ml to 75 kcal / 100 ml, more preferably 60 kcal / 100 ml to 70 kcal / 100 ml, when in a ready-to-drink form. This caloric density ensures an optimal balance between hydration and caloric intake.

[0044] Suitably, the nutritional composition is a powdered (i.e. particulate) nutritional composition, which can be reconstituted with water or other food grade aqueous liquid, to form a reconstituted ready-to-drink liquid. It was found that lipid globules maintained their size and coating when reconstituted.

[0045] Alternatively preferably, the nutritional composition is ready-to-drink liquid nutritional composition.

[0046] When in (reconstituted) ready-to-drink liquid form, the dry matter content in the nutritional composition is between 8-20 g / 100 ml, more preferably between 10-15 g / 100 ml.

[0047] The powdered nutritional composition preferably has a dry matter content of at least 90 wt.%, more preferably 92.5-100 wt.%, even more preferably 95-99.9 wt.%.

[0048] Preferably, the powdered nutritional composition comprises 80-99.5 wt.% of base powder particles, more preferably 85-99 wt.% of base powder particles. Further particles that may be added are components that can be added in a dry blending step, such as digestible carbohydrates, non-digestible oligosaccharides, LCPUFA’s, probiotics, minerals and vitamins.

[0049] The weight ratio between the base powder particles and the further particles in the powdered nutritional composition is preferably at least 10:1 , more preferably 25:1-1500:1 , and most preferably 50:1-1000:1 .

[0050] Process

[0051] A preferred process for producing the nutritional composition, as described herein, comprises the steps of a. Providing an aqueous phase with a dry matter content of 5-75 wt.%, based on total weight of the aqueous phase, wherein the aqueous phase comprises whey protein with a nativity of 40 to 100%, preferably 50 to 100%, even more preferably 60 to 100%; b. Providing a lipid phase which comprises at least one lipid; c. Mixing the lipid phase with the aqueous phase to provide an oil in water emulsion; d. Spraying the oil in water emulsion to provide particles by employing an atomization step; e. Optionally further drying the obtained sprayed product; f. Optionally dry blending further particles with the particles obtained in step e.

[0052] It should be understood that the process is preferably done at conditions under which whey protein denaturation is limited as much as possible. Conditions at which whey protein do not denature are well known to a person skilled in the art. By employing the measurement method to establish nativity, as described in this document, it is possible to check if whey protein denaturing takes place and it is possible to adjust the process conditions to better preserve nativity.

[0053] Nevertheless, when all the steps of above process are performed under conditions to avoid loss of nativity, a loss of nativity of a few percent may be unavoidable. For example, spray drying whey protein may result in a small loss of nativity of one or a few percent.

[0054] As the protein is subjected to a spraying step in which it may lose some of its nativity, it is within the scope of the present invention to employ whey protein with a higher nativity in step a. and to account for some loss of nativity during the process.

[0055] Also, in the process of the invention in some preferred embodiments a heat treatment is included to sterilize, pasteurize or better dissolve, homogenize or mix the separate components. Such a heat treatment could lead to some loss of nativity. It is within the normal skills of a person in the field of milk proteins to know the conditions that can be used to limit the loss of nativity of milk proteins during heat treatment.

[0056] Heat treatment conditions at which whey protein do not - or only minimally - denature can preferably be: keeping the temperature below 75 °C; if a temperature of 75-95 °C is employed, ensuring that this does not last longer than 60 seconds; if a temperature of 95-125 °C is employed, ensuring that this does not last longer than 5 seconds; or if a temperature of above 125 °C is employed, ensuring this is done instantaneous (e.g. DSI). Minimizing the time for heating is recommended to preserve nativity as much as possible.

[0057] For example, very short term (seconds) of high temperature treatments such as DSI (direct steam injection), UHT, or steam infusion can be performed as they lead to no or very limited loss of nativity. Heat treatments, for example up to 85 °C, can also be performed but also here it is important to limit them in time as indicated above.

[0058] It is also possible and known to the person skilled in the art to employ filtration steps to remove bacteria and maintain nativity of the protein. Also to end up with a nativity in the desired range the nativity of the starting material is of importance.

[0059] In a particularly preferred embodiment, step c. and step d. of the process are as follows: c. Mixing the lipid phase and the aqueous phase to provide an oil in water emulsion, wherein at least 10 vol.% of the lipid globules in the oil in water emulsion have a diameter of >12 pm and / or wherein the lipid globules in the oil in water emulsion have a volume-weighted mode diameter from 5 to 25 pm, preferably 7 to 15 pm; d. Spraying the oil in water emulsion to provide particles by employing an atomization step, wherein the size of the lipid globules is reduced so as to obtain particles comprising lipid globules, wherein less than 10 vol.% of the lipid globules have a diameter of >12 pm and / or wherein the lipid globules have a volume-weighted mode diameter from 2.5 to 7 pm, preferably 3 to 6 pm.

[0060] Further details about performing the step c. and d. in this manner can be found in WO2016146496. In this process the gist is to control the shear during step c. and to ensure that step c. provides relatively large lipid globules. Advantageously during the atomization step it is quite convenient to end up with lipid globules in the desired mode diameter range as claimed.

[0061] Preferably, the process comprises dry blending of further particles with the particles obtained in step e. These further particles preferably comprise one or more of digestible carbohydrates, non-digestible oligosaccharides, LCPUFAs, probiotics, minerals and vitamins.

[0062] Preferably, in step a. an aqueous phase is provided with a dry matter content of 10 to 60 wt.% (based on total weight of the aqueous phase), which comprises at least one protein component, and optionally carbohydrate, even more preferably 20 to 50 wt.%

[0063] Preferably, in step c. the lipid phase is mixed with the aqueous phase in a ratio of 5 wt.% to 50 wt.% using a mixer to provide an oil in water emulsion. Preferably, the mixer in step c. is an inline mixer, rotor stator mixer, or a static mixer; more preferably an inline mixer.

[0064] The oil in water emulsion preferably has a dry matter content of 30 to 70 wt.%, more preferably it is higher than 40 wt.%.

[0065] The atomizers employed in step d. of the process can be any atomizer known in the art, preferably a rotary atomizer or a pneumatic atomizer is employed, such as a 2 fluid nozzle. Protein

[0066] The protein concentration in the nutritional composition is determined by the sum of protein, peptides, and free amino acids.

[0067] The nutritional composition comprises whey protein, wherein 40 to 95 wt.% of the whey protein is native whey protein. In a preferred embodiment, 50 to 90 wt.%, even more preferably 60 to 85 wt.% of the whey protein is native whey protein.

[0068] The source of the protein is preferably selected in such a way that the minimum requirements for essential amino acid content are met, and satisfactory growth is ensured. Hence preferable protein sources are derived from mammals’ milk, such as goat, sheep or cow’s milk, more preferably derived from cow's milk.

[0069] Suitable sources of whey protein are preferably selected from acid whey, sweet whey, whey protein isolate, whey protein concentrate, or mixtures thereof. More preferably the source of whey protein is whey protein concentrate.

[0070] Preferably the nutritional composition further comprises casein. Preferably, the nutritional composition comprises whey protein and casein in a wt. ratio of 40:60 to 80:10.

[0071] The protein preferably provides 5% to 20% of the total calories. More preferably the nutritional composition comprises protein that provides 6% to 12% of the total calories. Preferably the nutritional composition comprises less than 3.5 g protein per 100 kcal, more preferably the nutritional composition comprises between 1 .5 g and 2.1 g protein per 100 kcal, even more preferably between 1 .6 g and 2.0 g protein per 100 kcal. A low protein concentration advantageously is closer to human milk as human milk comprises a lower amount of protein based on total calories compared to cow’s milk. Based on a (reconstituted) ready-to-drink liquid product the nutritional composition preferably comprises less than 1.5 g protein per 100 ml, more preferably between 1.2 g and 1.5 g per 100ml, even more preferably between 1 .25 g and 1 .35 g per 100ml.

[0072] Based on dry weight the nutritional composition preferably comprises less than 12 wt.% protein, more preferably between 9 wt.% and 12 wt.%, even more preferably between 10 wt.% and 11 wt.%. Worded alternatively, when the nutritional composition is in powder form, the proteins are preferably present in an amount of 9-12 g / 100 g dry weight, more preferably 10-11 g / 100 g dry weight of the composition.

[0073] The nutritional composition comprises 3-10 wt.% of whey protein, based on dry weight of the nutritional composition, more preferably 4-8 wt.% of whey protein, based on dry weight of the nutritional composition. Preferably, the protein-to-fat ratio in the nutritional composition is between 1 :20 and 1 :1.5, more preferably between 1 :10 and 1 :2, and most preferably between 1 :5 and 1 .2.2.

[0074] Lipid globule size

[0075] The lipid is present in the nutritional composition in the form of lipid globules. When the nutritional composition is in liquid form, these lipid globules are emulsified in the aqueous phase. Alternatively, when the nutritional composition is in powder form, the lipid globules are present in the powder and the powder is suitable for reconstitution with water or another food grade aqueous phase. The lipid globules comprise a core and a surface.

[0076] The lipid globules in the nutritional composition preferably have mode diameter, based on volume, of at least 1 .0 pm, more preferably at least 2.0 pm, even more preferably at least 3.0 pm, and most preferably at least 4.0 pm. Preferably, the lipid globules have a mode diameter, based on volume, between 1 .0 and 10 pm, more preferably between 2.0 and 8.0 pm, even more preferably between 3.0 and 7.0 pm, and most preferably between 4.0 pm and 6.0 pm.

[0077] Alternatively, or preferably in addition, the size distribution of the lipid globules is preferably in such a way that at least 45 volume % (vol.%), more preferably at least 55 vol.%, even more preferably at least 65 vol.%, and most preferably at least 75 vol.% of the lipid globules have a diameter between 2 and 12 pm. In a more preferred embodiment, at least 45 vol.%, preferably at least 55 vol.%, more preferably at least 65 vol.%, and most preferably at least 75 vol.% of the lipid globules have a diameter between 2 and 10 pm. In an even more preferred embodiment, at least 45 vol.%, more preferably at least 55 vol.%, yet even more preferably at least 65 vol.%, and most preferably at least 75 vol.% of the lipid globules have a diameter between 4 and 10 pm. Preferably less than 5 vol.% of the lipid globules have a diameter above 12 pm.

[0078] The volume percentage of lipid globules is based on volume of total lipid. The mode diameter relates to the diameter which is the most present based on volume of total lipid, or the peak value in a graphic representation, having on the X-axis the diameter and on the Y-axis the volume (%).

[0079] The volume of the lipid globules and its size distribution can suitably be determined using a particle size analyzer such as a Mastersizer 2000 (Malvern Instruments, Malvern, UK), for example by the method described in Michalski et al, 2001 , Lait 81 : 787-796.

[0080] Phospholipid

[0081] The lipid in the nutritional composition preferably comprises 0.5 to 20 wt.% phospholipids based on total lipid and the lipid globules preferably have a coating on the surface comprising said phospholipids. Preferably, the nutritional composition comprises 0.6 to 10 wt.%, more preferably 0.7 to 8 wt.%, even more preferably 0.8 to 6 wt.%, and most preferably 1 to 5 wt.% phospholipids based on total lipid. Phospholipids are amphipathic of nature and include glycerophospholipids and sphingomyelin. By ‘coating’ is meant that the outer surface layer of the lipid globules comprises phospholipid, whereas phospholipid is virtually absent in the core of the lipid globule. A suitable way to determine whether phospholipid is located on the surface of lipid globules is confocal laser scanning microscopy or transmission electron microscopy; see for instance Gallier et al. (A novel infant milk formula concept: Mimicking the human milk fat globule structure, Colloids and Surfaces B: Biointerfaces, 136 (2015), 329- 339).

[0082] The nutritional composition preferably comprises glycerophospholipids. Examples of glycerophospholipids are phosphatidylcholine (PC), phosphatidylserine (PS), phosphatidylethanolamine (PE), phosphatidylinositol (PI) and phosphatidylglycerol (PG). Preferably the nutritional composition comprises one or more of PC, PS, PI and PE, more preferably the nutritional composition comprises at least PC.

[0083] The nutritional composition preferably comprises sphingomyelin. Sphingomyelins have a phosphorylcholine or phosphorylethanolamine molecule esterified to the 1 -hydroxy group of a ceramide. They are classified as phospholipid as well as sphingolipid, but are not classified as a glycerophospholipid nor as a glycosphingolipid. Preferably the nutritional composition comprises 0.05 to 10 wt.% sphingomyelin based on total lipid, more preferably 0.1 to 5 wt.%, even more preferably 0.2 to 2 wt.%. Preferably the nutritional composition comprises at least 5 wt.%, more preferably 5 to 40 wt.% sphingomyelin based on total phospholipid, more preferably 10 to 35 wt.%, even more preferably 15 to 35 wt.% sphingomyelin, based on total phospholipid.

[0084] The nutritional composition preferably comprises glycosphingolipids. The term glycosphingolipids in the present context particularly refers to glycolipids with an amino alcohol sphingosine. The sphingosine backbone is O-linked to a charged head-group such as ethanolamine, serine or choline backbone. The backbone is also amide linked to a fatty acyl group. Glycosphingolipids are ceramides with one or more sugar residues joined in a beta-glycosidic linkage at the 1-hydroxyl position, and include gangliosides. Preferably the nutritional composition contains gangliosides, more preferably at least one ganglioside selected from the group consisting of GM3 and GD3. Preferably the nutritional composition comprises 0.1 to 10 wt.% glycosphingolipids based on total lipid, more preferably 0.5 to 5 wt.%, even more preferably 2 to 4 wt.% glycosphingolipids, based on total lipid.

[0085] The nutritional composition preferably comprises cholesterol. The nutritional composition preferably comprises at least 0.005 wt.% cholesterol based on total lipid, more preferably at least 0.02 wt.%, even more preferably at least 0.05 wt.%, and most preferably at least 0.1 wt.% cholesterol based on total lipid. Preferably the amount of cholesterol does not exceed 10 wt.% based on total lipid, more preferably does not exceed 5 wt.%, most preferably does not exceed 1 wt.% of cholesterol based on total lipid in the nutritional composition. Preferred sources for providing the phospholipid, glycosphingolipid and / or cholesterol are egg lipids, milk fat, buttermilk fat and butter serum fat (such as beta serum fat). Another preferred source for phospholipid, particularly PC, is soy lecithin and / or sunflower lecithin.

[0086] The nutritional composition preferably comprises phospholipid derived from mammalian milk. Preferably the nutritional composition comprises phospholipid and glycosphingolipid derived from mammalian milk. Preferably also cholesterol is derived from mammalian milk. The nutritional composition preferably comprises phospholipid, glycosphingolipid and / or cholesterol derived from mammalian milk of cows, mares, sheep, goats, buffalos, horses and camels. More preferably the nutritional composition comprises phospholipid, glycosphingolipid and / or cholesterol derived from cow’s milk.

[0087] Phospholipid derived from mammalian milk includes preferably phospholipid that is derived from milk lipid, cream lipid, cream serum lipid, butter serum lipid (beta serum lipid), whey lipid, cheese lipid and / or buttermilk lipid. Buttermilk lipid is typically obtained during the manufacture of buttermilk. Butter serum lipid or beta serum lipid is typically obtained during the manufacture of anhydrous milk fat from butter. More preferably the phospholipid, glycosphingolipid and / or cholesterol is derived from whey, e.g., a whey protein concentrate. Suitable commercially available sources for phospholipid from milk are BAEF, SM2, SM3 and SM4 powder of Corman, Salibra of Glanbia, Vivinal MFGM of FrieslandCampina and LacProdan MFGM-10 or PL20 from Aria.

[0088] The use of phospholipid from mammalian milk fat advantageously comprises the use of milk fat globule membranes, which are more reminiscent to the situation in human milk. The concomitant use of phospholipid derived from mammalian milk and triglycerides derived from vegetable lipids therefore enables the manufacture of coated lipid globules with a coating more similar to human milk, while at the same time providing an optimal fatty acid profile.

[0089] Preferably the phospholipid is derived from mammalian milk, more preferably derived from or forms part of milk fat globule membrane (MFGM). Preferably the phospholipid is derived from cow’s milk, more preferably derived from or forms part of cow’s MFGM.

[0090] Preferably the nutritional composition comprises phospholipid and glycosphingolipid and more preferably the weight ratio of phospholipid : glycosphingolipid is from 2:1 to 12:1 , more preferably from 2:1 to 10:1 and even more preferably 2:1 to 5:1.

[0091] Methods for obtaining lipid globules with an increased size and / or coating with phospholipid are for example described in WO 2010 / 027258 and WO 2010 / 027259.

[0092] Lipid The nutritional composition comprises lipid. The term “lipid” as used herein refers to one or more selected from the group consisting of triglycerides, polar lipids (such as phospholipids, cholesterol, glycolipids, sphingomyelin), free fatty acids, monoglycerides and diglycerides.

[0093] The lipid provides preferably 30 to 60% of the total calories of the nutritional composition. More preferably the nutritional composition comprises lipid providing 35 to 55% of the total calories, even more preferably the nutritional composition comprises lipids providing 40 to 50% of the total calories. The lipids are preferably present in an amount of 4 to 6 g per 100 kcal. When in liquid form, e.g., as a (reconstituted) ready-to-feed liquid, the nutritional composition preferably comprises 2.1 to 6.5 g lipids per 100 ml, more preferably 3.0 to 4.0 g per 100 ml. Based on dry weight, the nutritional composition preferably comprises 10 to 50 wt.%, more preferably 12.5 to 40 wt.% lipids, even more preferably 19 to 30 wt.% lipids.

[0094] The lipid comprises vegetable lipid. The presence of vegetable lipids advantageously enables an optimal fatty acid profile, high in polyunsaturated fatty acids and / or more reminiscent to human milk fat. Lipids from mammalian milk alone, e.g., cow’s milk, do not provide an optimal fatty acid profile. The amount of essential fatty acids is too low in mammalian milk.

[0095] Preferably the nutritional composition comprises at least one, preferably at least two vegetable lipid sources selected from the group consisting of linseed oil (flaxseed oil), rape seed oil (such as colza oil, low erucic acid rape seed oil and canola oil), sunflower oil, high oleic sunflower oil, safflower oil, high oleic safflower oil, olive oil, coconut oil, palm oil and palm kernel oil.

[0096] In one preferred embodiment, the nutritional composition comprises 5 to 98 wt.% vegetable lipids based on total lipids, more preferably 10 to 95 wt.%, more preferably 20 to 80 wt.%, even more preferably 25 to 75 wt.%, most preferably 40 to 60 wt.% of vegetable lipids based on total lipids. Preferably, the nutritional composition also comprises non-vegetable lipids. Preferably, said non-vegetable lipids are one or more non-vegetable lipids selected from mammalian milk fat, mammalian milk derived lipid as a preferred source of phospholipid, and fish, marine and / or microbial oils as source of LC-PUFA.

[0097] Faty acid composition

[0098] SFA relates to saturated fatty acids and / or acyl chains, MUFA relates to mono-unsaturated fatty acid and / or acyl chains, PUFA refers to polyunsaturated fatty acids and / or acyl chains with 2 or more unsaturated bonds; LC-PUFA refers to long chain polyunsaturated fatty acids and / or acyl chains comprising at least 20 carbon atoms in the fatty acyl chain and with 2 or more unsaturated bonds; DHA refers to docosahexaenoic acid and / or acyl chain (22:6, n3); EPA refers to eicosapentaenoic acid and / or acyl chain (20:5 n3); ARA refers to arachidonic acid and / or acyl chain (20:4 n6); DPA refers to docosapentaenoic acid and / or acyl chain (22:5 n3). n3 or omega 3 PUFA refers to polyunsaturated fatty acids and / or acyl chains with 2 or more unsaturated bonds and with an unsaturated bond at the third carbon atom from the methyl end of the fatty acyl chain, n6 or omega 6 PUFA refers to polyunsaturated fatty acids and / or acyl chains with 2 or more unsaturated bonds and with an unsaturated bond at the sixth carbon atom from the methyl end of the fatty acyl chain.

[0099] The nutritional composition preferably comprises LA, which refers to linoleic acid and / or acyl chain (18:2 n6). LA is an n6 PUFA and the precursor of n6 LC-PUFA and is an essential fatty acid as it cannot be synthesized by the human body. LA preferably is present in a sufficient amount to promote a healthy growth and development, yet in an amount as low as possible to prevent negative, competitive, effects on the formation of n3 PUFA and a too high n6 / n3 ratio. The nutritional composition therefore preferably comprises less than 25 wt.%, more preferably less than 20 wt.%, more preferably less than 15 wt.% LA based on total fatty acids. The nutritional composition preferably comprises at least 5 wt.% LA based on fatty acids, preferably at least 7.5 wt.%, more preferably at least 10 wt.% based on total fatty acids.

[0100] The nutritional composition preferably comprises ALA, which refers to alpha-linolenic acid and / or acyl chain (18:3 n3). ALA is a n3 PUFA and the precursor of n3 LC-PUFA and is an essential fatty acid as it cannot be synthesized by the human body. Preferably ALA is present in a sufficient amount to promote a healthy growth and development of the infant. The nutritional composition therefore preferably comprises at least 0.5 wt.%, more preferably at least 1.0 wt.%, more preferably the nutritional composition comprises at least 1.5 wt.%, even more preferably at least 2.0 wt.% ALA based on total fatty acids. Preferably the nutritional composition comprises less than 10 wt.% ALA, more preferably less than 5.0 wt.% ALA based on total fatty acids.

[0101] The weight ratio LA / ALA preferably is well balanced to ensure an optimal n6 / n3 PUFA, n6 / n3 LC PUFA and DHA / ARA ratio in the cellular membranes. Therefore, the nutritional composition preferably comprises a weight ratio of LA / ALA from 2 to 20, more preferably from 3 to 15, more preferably from 5 to 12, more preferably from 5 to 10. Preferably the n6 PUFA / n3 PUFA weight ratio is from 3 to 20, more preferably from 3 to 15, more preferably from 5 to 12, more preferably from 5 to 10.

[0102] Preferably, the nutritional composition comprises n3 LC-PUFA, such as EPA, DPA and / or DHA, more preferably DHA. As the conversion of ALA to DHA may be less efficient in infants, preferably both ALA and DHA are present in the nutritional composition. Preferably the nutritional composition comprises at least 0.05 wt.%, preferably at least 0.1 wt.%, more preferably at least 0.2 wt.%, of DHA based on total fatty acids. Preferably the nutritional composition comprises not more than 2.0, preferably not more than 1 .0 wt.%, of DHA based on total fatty acids.

[0103] The nutritional composition preferably comprises ARA. Preferably the nutritional composition comprises at least 0.05 wt.%, preferably at least 0.1 wt.%, more preferably at least 0.2 wt.%, of ARA based on total fatty acids. As the group of n6 fatty acids, especially ARA counteracts the group of n3 fatty acids, especially DHA, the nutritional composition preferably comprises relatively low amounts of ARA. Preferably the nutritional composition comprises not more than 2.0 wt.%, preferably not more than 1 .0 wt.%, of ARA based on total fatty acids. Preferably the weight ratio between DHA and ARA is between % to 4 / 1 , more preferably between % to 2 / 1 , more preferably between 0.6 and 1 .5.

[0104] Palmitic acid (PA) at sn-2 position of triglyceride

[0105] Triglycerides are preferably the major fraction of the lipids in the nutritional composition. Triglycerides comprise a glycerol moiety to which, via ester bonds, three fatty acid residues are attached, which may be the same or different, and which are generally chosen from saturated and unsaturated fatty acids containing 4 to 26 carbon atoms. Such triglycerides may differ in the fatty acid residues that are present and / or may differ in the respective position(s) of the fatty acid residues to the glycerol backbone (e.g. in the sn-1 , sn-2 and / or sn-3 position).

[0106] Preferably, the nutritional composition comprises at least 70 wt.%, more preferably at least 80 wt.%, even more preferably at least 90 wt.% and most preferably at least 95 wt.% triglycerides based on total lipids.

[0107] Preferably, the lipid in the nutritional composition has an increased amount of palmitic acid (PA) located at the sn-2 position in a triglyceride, based on total PA. PA relates to palmitic acid and / or acyl chains (C16:0).

[0108] Lipids that can be used to enhance the amount of PA located at the sn-2 position in triglycerides based on total PA are commercially available-e.g. from Loders Croklaan under the name Betapol™ and / or can be prepared in a manner known per se, for instance as described in EP 0698078 and / or EP 0758846. Another suitable source is InFat™ of Enzymotec. In case these lipids are obtained by trans- or interesterification of vegetable triglycerides, these sources are in the context of the present invention regarded as vegetable lipids.

[0109] A preferred source for triglycerides to enhance PA at the sn-2 or beta position in a triglyceride is mammalian milk fat, preferably non-human mammalian milk fat, more preferably cow’s milk fat. Preferably mammalian milk fat, in particular cow’s milk fat, is used in the form of anhydrous milk fat, butter oil, butter fat or butter.

[0110] In a preferred embodiment the lipid comprises vegetable lipid and mammalian milk fat. In a particularly preferred embodiment, the lipid in the nutritional composition comprises: a. 30 to 90 wt.% vegetable fat based on total lipid, and b. 10 to 70 wt.% mammalian milk fat based on total lipid.

[0111] More preferably, the lipid in the nutritional composition comprises: a. 35 to 75 wt.% vegetable fat based on total lipid, and b. 25 to 65 wt.% mammalian milk fat based on total lipid.

[0112] Most preferably, the lipid in the nutritional composition comprises: a. 40 to 60 wt.% vegetable fat based on total lipid, and b. 40 to 60 wt.% mammalian milk fat based on total lipid.

[0113] The amount of palmitic acid (PA) that is present in the total lipid of the nutritional composition is preferably at least 10 wt.% based on total fatty acids, more preferably 12-30 wt.%, even more preferably 14 to 24 wt.% and most preferably from 16 to 19 wt.% of PA by weight of total fatty acids.

[0114] Preferably, at least 15 wt.%, more preferably at least 20 wt.%, even more preferably at least 25 wt.%, and most preferably at least 30 wt.% of PA is in the sn-2 or beta position in a triglyceride, based on total PA. Preferably the amount of PA in the sn-2 position in a triglyceride is not more than 45 wt.%, preferably not more than 40 wt.% based on total PA. Preferably the amount of PA in the sn-2 position in a triglyceride is from 25 to 40 wt.% based on total PA present in the total lipid.

[0115] In the context of the present invention, a weight percentage of fatty acids based on total fatty acids is calculated as if all fatty acids are free fatty acids, hence it is not taken into account whether a fatty acid is attached to a glycerol backbone or not.

[0116] Digestible carbohydrates

[0117] The nutritional composition preferably comprises digestible carbohydrates. The digestible carbohydrates preferably provide 30 to 80% of the total calories of the nutritional composition. Preferably the digestible carbohydrates provide 40 to 60% of the total calories. Based on calories the nutritional composition preferably comprises of 5 to 20 g of digestible carbohydrates per 100 kcal, more preferably 7.5 to 15 g. When in liquid form, e.g. as a (reconstituted) ready-to-feed liquid, the nutritional composition preferably comprises 3 to 30 g digestible carbohydrate per 100 ml, more preferably 6 to 20 g, even more preferably 7 to 10 g per 100 ml. Based on dry weight, the nutritional composition preferably comprises 20 to 80 wt.%, more preferably 40 to 65 wt.% digestible carbohydrates.

[0118] Preferred digestible carbohydrate sources are lactose, glucose, sucrose, fructose, galactose, maltose, starch, and maltodextrin. Lactose is the main digestible carbohydrate present in human milk. Lactose advantageously has a low glycemic index. The nutritional composition preferably comprises lactose. The nutritional composition preferably comprises digestible carbohydrate, wherein at least 35 wt.%, more preferably at least 50 wt.%, more preferably at least 75 wt.%, and most preferably at least 95 wt.% of the digestible carbohydrate is lactose. Based on dry weight the nutritional composition preferably comprises at least 25 wt.% lactose, preferably at least 40 wt.%.

[0119] Human milk oligosaccharides

[0120] The nutritional composition preferably comprises human milk oligosaccharides (HMO). The term “human milk oligosaccharides” or “HMO” as used herein refers to non-digestible oligosaccharides which are present in human breast milk. Human breast milk comprises two types of carbohydrates: lactose and HMO. HMO are the third most abundant component of human breast milk, after lactose and lipids. Human breast milk contains three major HMO types: fucosylated HMO, sialylated HMO and N- acetylated HMO.

[0121] Suitable HMO for the preparation of the nutritional composition are commercially available, for example from Kyowa Hakko Bio, Japan; Friesland Campina, The Netherlands; Glycom DSM, Denmark and Chr. Hansen, Denmark. Otherwise, it is well within the reach of the skilled person to obtain HMO by isolation from suitable sources or by chemical synthesis using methods known in the art.

[0122] Preferably, the wt. ratio of HMO to mammalian milk derived phospholipids in the nutritional composition is between 1 :10 to 30:1 , more preferably between 1 :5 to 20:1 , and most preferably between 1 :1 to 10:1 .

[0123] When the nutritional composition is a (reconstituted) ready-to-drink liquid nutritional composition, the composition preferably comprises 20-400 mg HMO per 100 ml, more preferably 30-300 mg HMO per 100 ml and most preferably 40-250 mg HMO per 100 ml HMO.

[0124] When the nutritional composition is a powdered nutritional composition, the composition preferably comprises 300-4000 mg HMO per 100 g dry weight, more preferably 450-2000 mg HMO per 100 g dry weight.

[0125] When expressed in amounts based on calories, preferably the nutritional composition comprises 30- 600 mg HMO per 100 kcal, more preferably 45-450 mg HMO per 100 kcal and most preferably 60-375 mg HMO per 100 kcal.

[0126] The HMO in the nutritional composition is preferably selected from 2’-fucosyllactose (2’FL), 3- fucosyllactose (3-FL), lacto-N-tetraose (LNT), lacto-N-neotetraose (LNnT), para-lacto-N-neohexaose (para-LNnH), sialic acid, 3' sialyllactose (3’SL), 6' sialyllactose (6’SL), difucosyllactose (DFL), lacto-N- fucopentaose, lacto-N-fucohexaose, lacto-N-difucohexaose, sialyl-lacto-N-tetraose (LSTa), sialyl-lacto- N-tetraose b (LSTb), sialyl-lacto-N-tetraose c (LSTc), disialyllacto-N-tetraose (DSLNT), lacto-N- neodifucohexaose (LNnDFH I), fucosyllacto-N-hexaose, fucosyllacto-N-neohexaose, difucosyllacto-N- hexaose I, difuco-lacto-N-neohexaose, difucosyllacto-N-neohexaose I, difucosyllacto-N-neohexaose II, fucosyl-para-Lacto-N-hexaose, and tri-fuco-para-Lacto-N-hexaose I and combinations thereof.

[0127] The HMO preferably comprises at least 2 types of HMO, more preferably at least 3 types of HMO, even more preferably at least 4 types of HMO, and most preferably at least 5 types of HMO. Preferably, these types of HMO are selected from 2’-fucosyllactose (2’FL), 3-fucosyllactose (3-FL), lacto-N-tetraose (LNT), lacto-N-neotetraose (LNnT), para-lacto-N-neohexaose (para-LNnH), sialic acid, 3' sialyllactose (3’SL), 6' sialyllactose (6’SL), difucosyllactose (DFL), lacto-N-fucopentaose, lacto-N-fucohexaose, lacto- N-difucohexaose, sialyl-lacto-N-tetraose (LSTa), sialyl-lacto-N-tetraose b (LSTb), sialyl-lacto-N-tetraose c (LSTc), disialyllacto-N-tetraose (DSLNT), lacto-N-neodifucohexaose (LNnDFH I), fucosyllacto-N- hexaose, fucosyllacto-N-neohexaose, difucosyllacto-N-hexaose I, difuco-lacto-N-neohexaose, difucosyllacto-N-neohexaose I, difucosyllacto-N-neohexaose II, fucosyl-para-Lacto-N-hexaose, and tri- fuco-para-Lacto-N-hexaose I and combinations thereof.

[0128] In a preferred embodiment, the HMO is selected from 2’FL, 3-FL, DFL, LNT, LNnT, 3’SL, 6’SL, and combinations thereof. More preferably the HMO is selected from 2’FL, 3-FL, LNT, 3’SL, 6’SL and combinations thereof.

[0129] In a particularly preferred embodiment, the nutritional composition comprises 5 types of HMO, said 5 types of HMO being 2’FL, 3-FL, LNT, 3’SL, and 6’SL. More preferably, the HMO comprises 40-60 wt.% 2’FL, 10-20 wt.% 3-FL, 20-30 wt.% LNT, 2-7 wt.% 3’SL, and 4-8 wt.% 6’SL based on total HMO weight.

[0130] In alternative preferred embodiment, the HMO comprise at least 40 wt.%, more preferably 45-100 wt.%, and most preferably 50-90 wt.% of 2’FL based on total HMO weight.

[0131] In yet another alternative preferred embodiment, the HMO comprises a combination of 2’FL and LNnT, more preferably the HMO consists of the combination of 2’FL and LNnT. Preferably, the HMO comprises 60-90 wt.% 2’FL and 10-40 wt.% LNnT based on total HMO weight, more preferably the HMO comprises 65-85 wt.% 2’FL and 15-35 wt.% LNnT and most preferably the HMO comprises 70-80 wt.% 2’FL and 20-30 wt.% LNnT.

[0132] GOS and FOS

[0133] The nutritional composition preferably comprises galacto-oligosaccharides (GOS) and / or fructooligosaccharides (FOS), more preferably the nutritional composition comprises GOS and FOS. GOS and FOS are both non-digestible oligosaccharides which act as a prebiotic.

[0134] The GOS are preferably transgalacto-oligosaccharides. A suitable GOS is commercially available, for example VivinalOGOS (FrieslandCampina DOMO). Preferably the GOS are short chain galactooligosaccharides (scGOS) with an average degree of polymerization (DP) in the range of 1 to 10, more preferably in the range of 3 to 7.

[0135] A suitable FOS is commercially available, for example RaftilinOHP or Raftilose® (Orafti). Preferably the FOS are long chain fructo-oligosaccharides (IcFOS) with an average DP in the range of 10-100, more preferably in the range of 20 to 60.

[0136] Preferably, the weight ratio of GOS to FOS ranges from 100:1 to 1 :10, more preferably from 20:1 to 1 :1 , even more preferably from 7:1 to 10:1 , and most preferably the weight ratio is 9:1. Preferably these weight ratio’s apply to scGOS and IcFOS.

[0137] Preferably, the weight ratio of GOS and / or FOS combined to HMO ranges from 20:1 to 1 :10, more preferably from 15:1 to 1 :5 and most preferably from 10:1 to 1 :1 . Preferably, the nutritional composition comprises 80 mg to 2 g of GOS and / or FOS per 100 ml, more preferably 150 mg to 1 .5 g, most preferably 300 mg to 1 g of GOS / FOS per 100 ml.

[0138] Based on dry weight, the nutritional composition preferably comprises 0.25-20 wt.%, more preferably 0.5-10 wt.%, and most preferably 1 .5-7.5 wt.% of GOS and / or FOS.

[0139] Probiotics

[0140] The nutritional composition preferably comprises probiotic bacteria. The term “probiotic bacteria” as used herein refers to live beneficial bacteria that provide health benefits when consumed, generally by improving or restoring the gut microbiota. The term “gut microbiota” as used herein refers to all microorganisms, including bacteria, archaea, virus, and fungi, that are found in the digestive tract of a human subject.

[0141] The probiotic bacteria are preferably selected from Lactobacillus, Bifidobacterium and combinations thereof, more preferably the probiotic bacteria comprise Bifidobacterium, even more preferably the probiotic bacteria are Bifidobacterium, yet even more preferably the probiotic bacteria are Bifidobacterium breve and most preferably the probiotic bacteria are the strain B. breve M-16V (Morinaga).

[0142] The probiotic bacteria are preferably provided in therapeutically effective amounts. The nutritional composition preferably contains between 104and 1010colony forming units (cfu) probiotic bacteria per gram dry weight of the present composition, more preferably between 105and 109, and most preferably between 106and 108CFU probiotic bacteria per gram dry weight.

[0143] When the nutritional composition is a liquid nutritional composition, preferably a (reconstituted) ready- to-drink liquid nutritional composition, the liquid nutritional composition preferably comprises 106and 101° colony forming units (cfu) probiotic bacteria per 100 ml, more preferably between 107and 109CFU per 100 ml.

[0144] Application of the nutritional composition

[0145] The nutritional composition according to the first aspect of the invention are suitable for achieving beneficial effects in subjects, preferably in human subjects, more preferably in human infants, toddlers, and children.

[0146] In the next embodiments ‘nutritional composition, as described herein before’ refers to the nutritional composition according to the first aspect of the invention and all preferred embodiments equally apply to the second and the third aspect of the invention. In a preferred embodiment, the present invention provides a nutritional composition, as described herein above, for use in promotion of metabolic health, promotion of development of good body composition, prevention of development of obesity later in life, promotion of balanced growth, promotion of lean growth, promotion of cognitive development, improving brain health, improving gut health, providing beneficial prebiotic effects, increasing immune cell function and immune health, preventing infections, improving the recovery from infections such as intestinal infections, reducing / preventing intestinal permeability, stimulating / improving intestinal barrier functions I epithelial cell modulators, reducing gastrointestinal intolerance and so to improve gut health and reducing the risk of gut health problems and / or improving a recovery of a gut health problems, and / or reducing / preventing allergic response.

[0147] A second aspect of the invention pertains to a nutritional composition, as described herein before, for use in prevention of development of obesity later in life, preventing infections, improving the recovery from infections such as intestinal infections, reducing / preventing intestinal permeability, reducing gastrointestinal intolerance, reducing the risk of gut health problems, improving recovery of gut health problems, and / or reducing / preventing allergic response.

[0148] The invention also covers a method for prevention of development of obesity later in life, preventing infections, improving the recovery from infections such as intestinal infections, reducing / preventing intestinal permeability, reducing gastrointestinal intolerance, reducing the risk of gut health problems, improving recovery of gut health problems, and / or reducing / preventing allergic response, said method comprising administrating a nutritional composition as described herein before.

[0149] The invention may also be worded as the use of lipid globules and native whey protein in the manufacture of a nutritional composition for prevention of development of obesity later in life, preventing infections, improving the recovery from infections such as intestinal infections, reducing / preventing intestinal permeability, reducing gastrointestinal intolerance, reducing the risk of gut health problems, improving recovery of gut health problems, and / or reducing / preventing allergic response, wherein the nutritional composition is as described herein before.

[0150] Worded differently, the invention also pertains to the use of a nutritional composition, as described herein before, for prevention of development of obesity later in life, preventing infections, improving the recovery from infections such as intestinal infections, reducing / preventing intestinal permeability, reducing gastrointestinal intolerance, reducing the risk of gut health problems, improving recovery of gut health problems, and / or reducing / preventing allergic response.

[0151] Allergic response is in preferred embodiments a skin allergic response. Gut health includes improving the microbiota in the gut so as to treat and / or prevent gut dysbiosis, to decrease the abundance of opportunistic pathogens, and / or to increase the abundance of beneficial bacteria in the gut. Preferably the nutritional composition, as described herein before, is for use in prevention of development of obesity later in life, reducing the risk of gut health problems and / or preventing allergic response. More preferably, the nutritional composition, as described herein before, is for use in reducing the risk of gut health problems and / or preventing allergic response.

[0152] A third aspect of the invention pertains to the non-therapeutic use of the nutritional composition, as described herein above, for promotion of metabolic health, promotion of development of good body composition, promotion of balanced growth, promotion of lean growth, promotion of cognitive development, improving brain health, improving gut health, and / or providing beneficial prebiotic effects.

[0153] Preferably, the nutritional composition is for use in promotion of balanced growth, promotion of cognitive development, improving brain health and / or improving gut health, more preferably the nutritional composition is for use in promotion of cognitive development and / or improving gut health.

[0154] To support that the nutritional composition are effective for use as described above, reference is made to the following documents:

[0155] • Gallier S. et al., Natural and processed milk and oil body emulsions: Bioavailability, bioaccessibility and functionality. Food Structure 13, 2017,13-23.

[0156] • Breij L. et al., An infant formula with large, milk phospholipid-coated lipid droplets containing a mixture of dairy and vegetable lipids supports adequate growth and is well tolerated in healthy, term infants. Am J Clin Nutr, 2019;109(3): 586-596; and

[0157] • the following pre-published patent applications: o For formulas with large (phospholipid) coated lipid globules: WO2010027258, WO2010027259, W02011115490, WO2011115491 , WO2012173485,

[0158] WO2012173486, WO2013191542, WO2015065193, WO2016163881 ,

[0159] WO2017064304, WO 2018104512 and WO2024156893 o For native whey protein: W02019160416, W02020159373, W02020159372, W02022090269.

[0160] EXAMPLES

[0161] In all examples the below methods were employed to determine the below properties:

[0162] Volume based lipid globule diameter

[0163] The powders were reconstituted in water. The lipid globule and its size distribution was determined using a particle size analyzer (Mastersizer 2000) by the method described in Michalski et al., 2001 , Lait 81 : 787-796.

[0164] Nativity

[0165] The protein concentrations were measured using the method of Dumas (Flash 4000 N / protein analyzer, Thermo Fischer Scientific). The nitrogen concentrations were converted into a protein concentration using a conversion factor of 6.25. The soluble protein fraction was determined by dissolving 20 gram powder into 80 gram water and acidifying the sample to pH 4.6 using 7M HCI solution. The solution was centrifuged for 30 min at 4500g (Megafuge 16, Thermo Fisher Scientific) after which the protein concentration of the supernatant was determined. The nativity (%) was calculated as follows:

[0166] Foam height

[0167] The liquid foam height of the (reconstituted) powders was analyzed by adding 13 g powder into 90 ml water of 40 °C in a graded cylinder having an outer diameter 5 cm. The cylinder is horizontally shaken for 30 seconds after which the foam height is expressed as the total volume minus the liquid volume.

[0168] Example 1

[0169] A powdered infant formula was prepared comprising per kg final product about 5180 kcal, about 303 g lipid, about 484 g digestible carbohydrates, about 32 g non-digestible oligosaccharides and about 119 g protein (casein-whey wt. ratio of 40:60). The composition was prepared using whey protein concentrate, skimmed milk powder, lactose, a vegetable oil blend (fat) and non-digestible oligosaccharides. Also, vitamins, minerals, trace elements as known in the art were used.

[0170] An aqueous phase comprising whey protein with a nativity of more than 80%, digestible carbohydrates, and the other ingredients, except the fat and fat-soluble vitamins, was prepared by state-of-the-art techniques. The dry matter content of the aqueous phase was around 44 wt.%. The temperature of the aqueous phase was continuously below 60 °C.

[0171] A lipid phase was prepared by adding together the lipid and fat-soluble vitamins. The lipid phase was heated to 60°C and added to the aqueous phase and premixed with a Typhoon propeller mixer. The preemulsion was fed by a pump to a rotor stator machine (IKA process pilot 2000 / 04 generator 4M) operating at 4000 rpm and having a slit width of less than 1 mm. The resulting emulsion was collected in a tank. The collected composition was spray dried using pneumatic atomizers (2-fluid) at 2 bar air pressure and an inlet temperature of the drying gas being 195 °C resulting in powder 1 with relatively large lipid globules and the nativity as shown in Table 1 .

[0172] Comparative example C1

[0173] The procedure of Example 1 was repeated, but the pre-emulsion was fed to a high-pressure homogeniser (GEA Ariete homogeniser ns3006) with pressures of 100 bar (Stage 1) and 20 bar (Stage 2) instead of a rotor stator machine resulting in a comparative powder C1 with relatively small lipid globules and the nativity as shown in Table 1 .

[0174] Example 2 A powdered infant formula was prepared comprising per kg final product about 4600 kcal, about 210 g lipid, about 525 g carbohydrates, about 39 g non-digestible oligosaccharides and about 150 g protein (casein-whey wt. ratio of 50:50). The composition was prepared using whey protein concentrate, skimmed milk powder, lactose, a vegetable oil blend (fat) and non-digestible oligosaccharides. Also, vitamins, minerals, trace elements as known in the art were used.

[0175] An aqueous phase comprising whey protein with a nativity of more than 75%, digestible carbohydrates, and the other ingredients, except the fat and fat-soluble vitamins, was prepared by state-of-the-art techniques. The dry matter content of the aqueous phase was around 44 wt.%. The aqueous phase was pasteurized for 30 seconds at 85 °C.

[0176] A lipid phase was prepared by adding together the lipid and fat-soluble vitamins. The lipid phase was heated to 60°C and added to the aqueous phase and premixed with a Typhoon propeller mixer. The preemulsion was fed by a pump to a rotor stator machine (IKA process pilot 2000 / 04 generator 4M) operating at 4000 rpm and having a slit width of less than 1 mm. The resulting emulsion was collected in a tank. The collected composition was spray dried using pneumatic atomizers (2-fluid) at 2 bar air pressure and an inlet temperature of the drying gas being 195 °C resulting in powder 2 with relatively large lipid globules and the nativity as shown in Table 1 .

[0177] Comparative Example C2

[0178] The procedure of Example 2 was repeated, but the pre-emulsion was fed to a high-pressure homogeniser (GEA Ariete homogeniser ns3006) with pressures of 100 bar (Stage 1) and 20 bar (Stage 2) instead of a rotor stator machine resulting in a comparative powder C2 with relatively small lipid globules and the nativity as shown in Table 1 .

[0179] For all compositions made in the Examples 1 , C1 , 2, and C2 the foam height was determined by the above method. The results are provided in below Table 1

[0180] Table 1

[0181] Example 3

[0182] A powdered infant formula was prepared comprising per kg final product about 5030 kcal, about 294 g lipid (35 wt.% palm oil, 25 wt.% coconut oil, 21 wt.% sunflower oil, 19 wt.% rapeseed oil), about 452 g digestible carbohydrates, about 66 g non-digestible oligosaccharides, about 5 g milk phospholipids and about 121 g protein (60 wt.% whey, 40 wt.% casein). Also, vitamins, minerals, trace elements as known in the art were used. The composition was prepared using whey protein, cream derived milk phospholipids (Lipamine M20 ex Lecico), skimmed milk powder, lactose, an oil blend (fat) and non- digestible oligosaccharides.

[0183] An aqueous phase comprising protein with a nativity of about 65%, milk phospholipid source, digestible carbohydrates, and the other ingredients, except the fat and fat-soluble vitamins, was prepared by state- of-the-art techniques. The dry matter content of the aqueous phase was around 44 wt.%. The aqueous phase was pasteurized for 30 seconds at 85 °C.

[0184] A lipid phase was prepared by adding together the lipid and fat-soluble vitamins. The lipid phase was heated to 60°C and added to the aqueous phase and premixed with a Typhoon propeller mixer. The preemulsion was fed by a pump to a rotor stator machine (IKA process pilot 2000 / 04 generator 4M) operating at 4000 rpm and having a slit width of less than 1 mm. The resulting emulsion was collected in a tank. The collected composition was spray dried using pneumatic atomizers (2-fluid) at 2 bar air pressure and an inlet temperature of the drying gas being 195 °C resulting in powder 3 with relatively large lipid globules and the nativity as shown in Table 2.

[0185] Example 4

[0186] A powdered infant formula was prepared using the same process as in Example 3, comprising per kg final product about 4830 kcal, about 261 g lipid (50 wt.% anhydrous milkfat, 25 wt.% sunflower oil, 17 wt.% rapeseed oil, 8 wt.% coconut oil), about 507 g digestible carbohydrates, about 66 g non-digestible oligosaccharides, about 4 g milk phospholipids and about 98 g protein (60 wt.% whey, 40 wt.% casein). The composition was prepared using whey protein, whey derived milk phospholipids (Lacprodan MFGM-10 ex Aria), skimmed milk powder, lactose, an oil blend (fat) and non-digestible oligosaccharides. The net nativity of the aqueous phase when initially mixed was about 55%. The net nativity after mixing the raw materials was lower than in Example 3 as the MFGM source contains a substantial amount of denatured whey protein. The resulting powder 4 has the lipid globules and nativity as shown in Table 2.

[0187] For the compositions prepared in the Examples 3 and 4 the foam height was determined following the same procedure as above. The results are provided in below Table 2

[0188] Table 2

[0189] Conclusion For powdered nutritional compositions containing small lipid globules, high nativity of whey protein is leading to more foam formation upon reconstitution. By making powdered nutritional compositions with larger lipid globules, with or without phospholipid coating, undesired foam formation is effectively suppressed.

[0190] Comparative Example 5

[0191] A powdered infant formula was prepared using the same process as comparative example C1 , comprising per kg final product about 4871 kcal, about 245 g lipid (mixture of vegetable oil and anhydrous milk fat), about 530 g digestible carbohydrates (lactose), about 58 g non-digestible oligosaccharides and about 96 g protein (60 wt.% whey, 40 wt.% casein). Further, vitamins, minerals, trace elements as known in the art were used. The characteristics of powdered infant formula Example 5 is shown in Table 3 below.

[0192] Whey protein denaturation

[0193] The effect of whey protein denaturation on foam formation was assessed by comparing two powdered infant formulas, i.e. Example 4 vs. Comparative Example 5.

[0194] Firstly, the foam height test as described herein above was applied for both powdered infant formulas. The outcome is described below in Table 3 under the heading ‘Before heat treatment’.

[0195] Further, for both powdered infant formulas, 13 g powder was reconstituted into 90 ml water of 40°C and subsequently heated at 90 °C for 30 minutes with the aim to denature whey protein. Next the heated suspension was allowed to cool down to 40°C and transferred into a graded cylinder having an outer diameter 5 cm. The cylinder was shaken horizontally for 30 seconds, after which the foam height was expressed as the total volume minus the liquid volume. The outcome is described below in Table 3 under the heading ‘After heat treatment’.

[0196] Table 3

Claims

CLAIMS1. A nutritional composition comprising lipid and protein, and optionally carbohydrates, wherein the lipid is in the form of lipid globules and wherein the lipid comprises vegetable lipid; the lipid globules have a mode diameter, based on volume, of at least 1.0 pm and / or at least 40 vol.% of the lipid globules, based on lipid volume, have a diameter of 2 to 12 pm; the protein comprises whey protein, the nutritional composition comprises 3-10 wt.% whey protein by dry weight of the nutritional composition; and40 to 95 wt.% of the whey protein is native whey protein; wherein the composition is an infant formula, a follow on formula, a young child formula, or a medical nutritional composition.

2. Composition of claim 1 , wherein the protein-to-fat ratio in the nutritional composition is between 1 :20 and 1 :1.5.

3. Composition of claim 1 or 2, wherein the nutritional composition comprises less than 1.5 g protein per 100 ml, when in (reconstituted) ready-to-drink liquid form.

4. Composition of any one of the preceding claims, wherein 50 to 90 wt.% of the whey protein is native whey protein.

5. Composition of any one of the preceding claims, wherein native whey protein refers to whey protein that is soluble in an aqueous solution at 20 °C having a pH of 4.6.

6. Composition of any one of the preceding claims, wherein the nutritional composition comprises 2.1 to 6.5 g lipid per 100 ml, when in (reconstituted) ready-to-drink liquid form and / or 10 to 50 wt.% of lipid, based on dry weight of the nutritional composition.

7. Composition of any one of the preceding claims, wherein the nutritional composition comprises at least 6 g digestible carbohydrate per 100 ml, when in (reconstituted) ready-to-drink liquid form.

8. Composition of any one of the preceding claims, wherein the nutritional composition comprises 9 to 12 wt.% of protein, based on dry weight of the nutritional composition.

9. Composition of any one of the preceding claims, comprising at least 0.5 wt.% of phospholipids based on total lipid and wherein preferably the lipid globules have a coating comprising the phospholipids.

10. Composition of claim 9, wherein the phospholipids are derived from milk fat globular membrane (MFGM).

11. Composition of any one of the preceding claims, wherein the protein comprises further protein besides whey protein, preferably the protein comprises casein.

12. Composition of any one of the preceding claims, wherein the lipid comprises vegetable lipid and mammalian milk fat.

13. Composition of any one of the preceding claims, that is an infant formula, a follow on formula, or a young child formula.

14. Composition of any one of the preceding claims, wherein the nutritional composition is a particulate nutritional composition suitable for reconstitution with water or a food grade aqueous liquid to form a reconstituted ready-to-drink liquid.

15. Composition of any one of claims 1-13, wherein the nutritional composition is a ready-to-drink liquid nutritional composition.

16. Nutritional composition of any one of the preceding claims, for use in prevention of development of obesity later in life, preventing infections, improving the recovery from infections such as intestinal infections, reducing / preventing intestinal permeability, reducing gastrointestinal intolerance, reducing the risk of gut health problems, improving recovery of gut health problems, and / or reducing / preventing allergic response.

17. Non-therapeutic use of the nutritional composition of any one of claims 1-15 for promotion of metabolic health, promotion of development of good body composition, promotion of balanced growth, promotion of lean growth, promotion of cognitive development, improving brain health, improving gut health, and / or providing beneficial prebiotic effects.

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