Nutrient with large lipid globules comprising milk phospholipid coated vegetable fat for lipid digestion

Abstract

The present invention relates to a nutritional composition comprising lipid globules larger than conventional lipid globules and comprising a coating comprising a milk-derived phospholipid, which nutritional composition has beneficial effects on lipid digestion, in particular for improving lipid digestion kinetics.

Description

[0001] This application is a divisional application of patent application No. 201680033608.1 entitled "Nutrients with large lipid globules of plant fats coated with milk phospholipids for lipid digestion". The original application corresponds to international application PCT / NL2016 / 050242, filed on April 8, 2016, with a priority date of April 10, 2015. Technical Field

[0002] This invention relates to the field of nutritional compositions, particularly infant formula or follow-on formulas, which have beneficial effects on lipid digestion, especially for improving lipid digestion kinetics. Background Technology

[0003] Human milk is the primary energy source for many infants in the early stages of their lives and is the preferred method of feeding. Human milk is rich in lipids, which are a major energy source and essential nutrient for infants. Lipid globules have a mean mode diameter of approximately 4 μm by volume. In human or cow's milk, the lipid globules are covered by a triple membrane, namely the milk lipid globule membrane (MFGM), composed of phospholipids and other polar lipids, as well as membrane proteins.

[0004] Breastfeeding is the preferred method of feeding infants. However, there are situations that make breastfeeding unfeasible or less than ideal. In these cases, infant formula is a good alternative. The composition of modern infant formula has been modified to meet the many specific nutritional needs of rapidly growing and developing infants. It still appears that the composition of infant formula can be improved. A comparison between lipids in human milk and those in existing infant formula reveals differences in physical structure. Lipid droplets produced during IMF processing have a modal diameter of approximately 0.3 to 0.5 µm by volume and are primarily coated with proteins such as casein.

[0005] Garcia et al., Food Hydrocolloids, 2014, 35:494-504, disclosed that lipid composition can modulate the bioavailability of fatty acids, leading to health problems. They digested natural (milk) fat globules of various sizes, either covered by biomembranes or homogenized and heat-treated, under simulated human physiological conditions in the stomach and duodenum. Smaller fat globules were hydrolyzed more efficiently by gastric, gastric+pancreatic, and pancreatic lipases than larger ones due to their larger lipid interfacial area. However, homogenization, which significantly reduced the size of fat globules and thus increased the lipid / water interfacial area, failed to improve lipolysis in the stomach or duodenum, attributed to changes in the globule surface composition (particularly proteins relative to phospholipids).

[0006] On the other hand, Boullieu et al., Food Chemistry, 2015, 182:224-235, used semi-dynamic in vitro gastric digestion to compare minimally processed emulsions containing natural (cow's milk) milk fat globules with infant formula processed under two different conditions (homogenization or homogenization / pasteurization), the latter containing newly formed, predominantly protein-coated small lipid globules. It was found that the minimally processed emulsion underwent lipolysis more slowly than the processed formula, and the surface area of ​​the droplets was a key parameter controlling gastric lipolysis kinetics, the pattern of fatty acid release, and proteolysis by faster hydrolysis of adsorbed proteins. Of all structural parameters, the specific surface area of ​​the droplets in the infant formula was a key parameter controlling the rate of gastric lipolysis, the overall extent of gastric lipolysis, and the pattern of fatty acid release. No effect was observed on the globular surface composition.

[0007] However, applying the methods described above for enriching components of milk fat or obtaining large natural milk fat globules to infant formula is not commercially viable because the fatty acid composition of milk fat contains too few essential fatty acids to meet regulatory requirements for infant formula. Furthermore, such methods are economically infeasible for large-scale production. Additionally, for food safety reasons, heat treatment is required during the production of infant formula, and further heat is applied in the spray drying step to produce powdered formulas—preferred forms of infant cultures and stage 2 formulas.

[0008] In WO 2011 / 108934, the postprandial digestive kinetics of standard soybean oil were measured after 2 weeks of consuming a diet containing lipid globules coated with milk phospholipids. The study found altered fat metabolism, which, relative to the standard fat composition, dulled the peak of postprandial fat absorption and / or plasma triglyceride levels, prolonged postprandial fat absorption and / or plasma triglyceride levels, and increased postprandial fat bioavailability.

[0009] Fondaco et al., Food Biophysics, 12 / 2014; DOI: 10.1007 / s11483-014-9388-6, disclosed the in vitro digestion of several different infant formulas. They found a positive correlation between the lipolysis rate of infant formula and the surface area of ​​lipid droplets per gram of infant formula. However, they found that breast milk did not follow this trend.

[0010] US 2014 / 0255538 discloses a nutritional composition comprising a carbohydrate source, a protein source, and a lipid source, the lipid source including (bovine) milk fat globules formed from enriched lipids, the composition being used as an infant formula to promote lipid digestion in infants. Increased lipolysis is indicated because the milk fat globules are more accessible to lipases than fat globules in standard infant formula. Summary of the Invention

[0011] Infant formula, stage 2 formula, or growing-up formula is required that i) contains lipid globules containing plant fats rich in essential fatty acids, ii) is newly formed during the homogenization step of the production process, and iii) preferably undergoes heat treatment during the production of such formulas, wherein postprandial fat digestion kinetics are slowed and more similar to those of human milk.

[0012] Lipid digestion kinetics were evaluated in an in vitro model closely mimicking infant digestive conditions. The inventors of this invention discovered that nutritional compositions using large lipid globules exhibit a reduced lipolysis rate compared to smaller lipid globules, thus demonstrating lipid digestion kinetics more closely resembling those of human milk. These large lipid globules comprise plant fats rich in essential fatty acids, neoformed during production in mild homogenization steps in both aqueous and fatty phases, and subjected to heat treatment during production, having a mode diameter of 2 to 6 μm by volume, and / or 0.5 and 15 μm. 2 / g lipid specific surface area, and is coated with phospholipids.

[0013] Surprisingly, specific surface area is not the only determining factor. The inventors of this invention discovered that the lipolysis rate is also affected by the coating lipid globules. Large lipid globules coated with milk-derived phospholipids exhibit slower lipolysis than large lipid globules coated with the same amount of lecithin or soybean phospholipids. Transmission electron microscopy images revealed differences in surface composition.

[0014] Furthermore, compared to large lipid globules that primarily contain plant fats, large lipid globules containing a mixture of milk fat and plant fats exhibit a consistently slightly lower rate of lipolysis, which is more similar to human milk.

[0015] Therefore, the present invention relates to a nutritional composition having a reduced postprandial or gastric lipid digestion kinetic rate compared to that of a standard infant formula, and more similar to the lipolysis kinetics observed when feeding human milk, due to the presence of lipid globules that are larger or have a smaller specific surface area than usual, and contain a coating containing milk-derived phospholipids. Attached Figure Description

[0016] Figure 1 Microscopic (20x) images of IMFs with different lipid globules during in vitro gastric digestion are shown.

[0017] Figure 2 This represents a portion of a typical human milk lipid globule.

[0018] Figure 3a and 3bThis refers to lipid spheres of IMF 1 with a thin film (5-10 nm) and a small number of protein aggregates.

[0019] Figure 4 The lipid droplets representing the control IMF 3 have a thicker (20-100 nm) and more densely stained protein membrane than the thin film surrounding the concept IMF 3 lipid spheres. Detailed Implementation

[0020] Therefore, the present invention relates to a method for reducing the rate of gastric lipolysis and / or the rate of postprandial lipolysis in a subject, comprising administering the subject a nutritional composition comprising lipid globules, said lipid globules comprising triglycerides derived from plant fats and phospholipids derived from non-human mammalian milk, wherein said lipid globules have: - The mode diameter is 2 to 6 µm, or - 0.5 to 15 m 2 Specific surface area per g of lipid, or - Mode diameter of 2 to 6 µm and 0.5 to 15 m 2 Specific surface area per g of lipid Furthermore, the lipid spheres contain a coating containing phospholipids.

[0021] In one implementation, the method for reducing the rate of gastric lipolysis and / or the rate of postprandial lipolysis in subjects is a non-medical approach.

[0022] In other words, the present invention relates to a nutritional composition comprising lipid globules, said lipid globules comprising triglycerides derived from plant fats and phospholipids derived from non-human mammalian milk, wherein said lipid globules have: - The mode diameter is 2 to 6 µm, or - 0.5 to 15 m 2 Specific surface area per g of lipid, or - Mode diameter of 2 to 6 µm and 0.5 to 15 m 2 Specific surface area per g of lipid Furthermore, the lipid globules contain a phospholipid-containing coating. The nutritional composition is used for - Reduce the rate of gastric lipolysis, and / or - Reduce the rate of postprandial lipolysis.

[0023] This invention can also be described as the use of a composition comprising lipid globules for preparing a nutritional composition, said lipid globules comprising triglycerides derived from plant fats and phospholipids derived from non-human mammalian milk, wherein said lipid globules have: - The mode diameter is 2 to 6 µm, or - 0.5 to 15 m2 Specific surface area per g of lipid, or - Mode diameter of 2 to 6 µm and 0.5 to 15 m 2 Specific surface area per g of lipid Furthermore, the lipid globules contain a phospholipid-containing coating. The nutritional composition is used for - Reduce the rate of gastric lipolysis, and / or - Reduce the rate of postprandial lipolysis.

[0024] The present invention can also be described as having the following uses for a nutritional composition comprising lipid globules: - Reduces the rate of gastric lipolysis, and / or - Reduces the rate of postprandial lipolysis. The lipid globules comprise triglycerides derived from plant fats and phospholipids derived from non-human mammalian milk, wherein the lipid globules have: - The mode diameter is 2 to 6 µm, or - 0.5 to 15 m 2 Specific surface area per g of lipid, or - Mode diameter of 2 to 6 µm and 0.5 to 15 m 2 Specific surface area per g of lipid Furthermore, the lipid spheres contain a coating containing phospholipids.

[0025] Throughout the instruction manual, the terms “fat” and “lipids” or “multiple lipids” are used interchangeably.

[0026] lipid components The composition provided by the method or use according to the invention comprises lipids. The composition comprises at least triglycerides derived from vegetable fats and phospholipids derived from the milk of non-human mammals. Preferably, the composition comprises at least 70% by weight, more preferably at least 80% by weight, more preferably at least 85% by weight, and even more preferably at least 90% by weight of triglycerides based on total lipids. The lipids may also comprise one or more free fatty acids, monoglycerides, and diglycerides.

[0027] The lipids preferably provide 30 to 60% of the total calories of the composition. More preferably, the compositions of the present invention contain lipids providing 35 to 55% of the total calories, and even more preferably, the compositions of the present invention contain lipids providing 40 to 50% of the total calories. When the composition is in liquid form, such as an instant liquid, it preferably contains 2.1 to 6.5 g lipids / 100 ml, more preferably 3.0 to 4.0 g / 100 ml. In one embodiment, the composition given according to the method or use of the present invention contains at least 15% by weight of lipids based on the dry weight of the composition. Based on dry weight, the composition preferably contains 10 to 50% by weight, more preferably 12.5 to 40% by weight of lipids, even more preferably 15 to 35% by weight of lipids, and even more preferably 19 to 30% by weight of lipids.

[0028] The composition provided by the method or use according to the invention comprises triglycerides derived from vegetable fats. In one embodiment, "derived from vegetable fats" means derived from plant fats. Preferably, the composition comprises plant lipids. The present plant lipids advantageously achieve an optimal fatty acid profile, with a high content of polyunsaturated fatty acids and more similar to human milk fat. Using lipids from ruminant milk (particularly cow's milk or other domestic ruminant milk) does not provide an optimal fatty acid profile. This less-than-ideal fatty acid profile (e.g., a high content of saturated fatty acids) is known to be disadvantageous. Preferably, the composition of the present invention comprises at least one, preferably at least two, lipid sources selected from: linseed oil, rapeseed oil (e.g., colza oil, low-erucic acid rapeseed oil, and canola oil), sage oil, perilla oil, purslane oil, blueberry oil, sea buckthorn oil, hemp oil, sunflower seed oil, high-oleic sunflower seed oil, safflower oil, high-oleic safflower oil, olive oil, blackcurrant seed oil, blue thistle oil, coconut oil, palm oil, and palm kernel oil. Preferably, the composition of the present invention comprises at least one, preferably at least two, lipid sources selected from: linseed oil, canola oil, coconut oil, sunflower seed oil, and high-oleic sunflower seed oil. Preferably, the composition comprises 30 to 99.5% by weight of plant lipids based on total lipids, more preferably 35 to 99% by weight, and even more preferably 40 to 95% by weight based on total lipids.

[0029] Phospholipids The compositions provided according to the method or use of the invention comprise phospholipids derived from non-human mammalian milk. In one embodiment, "derived from non-human mammalian milk" means originating from non-human mammalian milk. Phospholipids derived from non-human mammalian milk include glycerophospholipids and sphingomyelin. Phospholipids derived from non-human mammalian milk are preferably present in a coating on the surface of lipid globules. "Coating" means that the outer surface layer of the lipid globules contains phospholipids, which are not actually present in the core of the lipid globules. It has been found that non-human mammalian milk phospholipids present as a coating or outer layer of lipid globules in the diet modulate lipid digestion rates to tend towards human milk lipid digestion kinetics.

[0030] Not all phospholipids present in the composition need to be included in the coating, but it is preferable that most phospholipids are included in the coating. Preferably, more than 30% by weight, more preferably more than 50% by weight, more preferably more than 70% by weight, even more preferably more than 85% by weight, and most preferably more than 95% by weight of the phospholipids present in the composition are included in the coating of the lipid spheres.

[0031] In one embodiment, the composition given according to the method or use of the invention comprises at least 0.5% by weight of phospholipids based on total lipids. Preferably, the composition given according to the method or use of the invention comprises 0.5 to 20% by weight of phospholipids based on total lipids, more preferably 0.5 to 10% by weight, more preferably 1 to 10% by weight, even more preferably 1.0 to 5% by weight, even more preferably 1.0 to 2.0% by weight of phospholipids. Preferably, at least 80% by weight of the phospholipids are derived from non-human mammalian milk, more preferably at least 90% by weight, even more preferably at least 95% by weight or 99% by weight, or preferably all of the phospholipids are derived from non-human mammalian milk.

[0032] Preferably, the composition given according to the method or use of the invention comprises sphingomyelin. Sphingomyelin has phosphoric acid choline and phosphoroethanolamine molecules esterified with the 1-hydroxyl group of ceramide. Preferably, the composition given according to the method or use of the invention comprises 0.05 to 10% by weight of sphingomyelin based on total lipids, more preferably 0.1 to 5% by weight, even more preferably 0.2 to 2% by weight. Preferably, the nutritional composition comprises at least 15% by weight of sphingomyelin based on total phospholipids, more preferably at least 20% by weight. Preferably, the amount of sphingomyelin is less than 50% by weight based on total phospholipids.

[0033] Preferably, the composition provided according to the method or use of the invention comprises glycerophospholipids. Glycerophospholipids are a class of lipids formed by: fatty acids esterified at hydroxyl groups on carbon-1 and carbon-2 of the glycerol moiety in the main chain and a negatively charged phosphate group linked to carbon-3 of glycerol via an ester bond; and optionally linked to the phosphate group a choline group (in the case of phosphatidylcholine (PC), a serine group (in the case of phosphatidylserine (PS), an ethanolamine group (in the case of phosphatidylethanolamine (PE), an inositol group (in the case of phosphatidylinositol (PI), or a glycerol group (in the case of phosphatidylglycerol (PG)). Preferably, the composition contains PC, PS, PI and / or PE, more preferably at least PS.

[0034] Preferably, the composition given according to the method or use of the invention comprises at least 1% by weight, preferably at least 2% by weight, of phosphatidylserine based on total phospholipids. Preferably, the amount of phosphatidylserine is less than 10% by weight based on total phospholipids.

[0035] Phospholipids derived from non-human mammalian milk include phospholipids isolated from milk lipids, cream lipids, cream whey lipids, butter whey lipids, β-whey lipids, whey lipids, cheese lipids, and / or buttermilk lipids. Buttermilk lipids are typically obtained during the preparation of buttermilk. Butter whey lipids or β-whey lipids are typically obtained during the preparation of anhydrous milk fat from cream or butter. Preferably, the phospholipids are obtained from cream. Phospholipids are preferably derived from the milk of cows, donkeys, sheep, goats, buffalo, horses, and camels, and most preferably from cow's milk. Lipid extracts isolated from cow's milk are most preferably used. A suitable source of phospholipids derived from non-human mammalian milk is a fraction of milk that can be isolated from a membrane called the milk fat globule membrane (MFGM). Therefore, in one embodiment, the phospholipid provided for use in the method of the present invention is MFGM.

[0036] Phospholipids derived from milk fat are more advantageous in reducing the rate of gastric lipolysis and / or postprandial lipolysis than polar lipids from other sources. The phospholipids are located on the surface of lipid globules, i.e., contained within a coating or outer layer. In one embodiment, the lipid globules comprise a monolayer containing phospholipids derived from milk fat. A suitable method for determining whether polar lipids are located on the surface of lipid globules is laser scanning microscopy or transmission electron microscopy. Therefore, the simultaneous use of polar lipids (especially phospholipids) derived from domestic animal milk and triglycerides derived from plant lipids enables the preparation of coated lipid globules with a coating more similar to human milk, while providing optimal fatty acid distribution.

[0037] milk fat Preferably, the composition given according to the method or use of the invention comprises fat or lipids from mammalian milk. Preferably, the composition comprises lipids from ruminant animal milk, preferably cow's milk, goat's milk, sheep's milk, buffalo milk, yak milk, reindeer milk, and camel milk, most preferably cow's milk. Preferably, the mammalian milk is not human milk. Therefore, in one embodiment, the composition given according to the method or use of the invention comprises non-human mammalian milk fat. Preferably, the mammalian milk component comprises at least 70% by weight of triglycerides, more preferably at least 90% by weight, and even more preferably at least 97% by weight.

[0038] Preferably, the mammalian milk lipids are derived from butter, butter fat, butteroil, and anhydrous milk fat, more preferably anhydrous milk fat and butter. These types of milk fat lipid sources have a high triglyceride content. Furthermore, these lipid sources are in continuous fat phase form or water-in-oil emulsion form. Using these milk fat sources in the preparation of the nutritional compositions of the present invention enables the formation of lipid globules, each globul containing a mixture of vegetable fat and milk fat.

[0039] The milk fat of this invention refers to the total lipid components of milk produced by mammals (e.g., cattle) and is present in commercial dairy products and milk-derived products.

[0040] As defined in this article, butter is a water-in-oil emulsion containing more than 80% by weight of milk fat.

[0041] As defined in this article, milk fat refers to all the fat components in milk that can be separated by churning; in other words, all the fat components present in butter.

[0042] Anhydrous milk fat (AMF) is a term known in the art and refers to extracted milk fat. AMF typically contains more than 99% by weight of lipids based on total weight. It can be prepared by extracting milk fat from cream or butter. Anhydrous butter, as defined herein, is synonymous with AMF.

[0043] Cream is also a term known in the art. It generally refers to a milk lipid extract having more than 98% by weight of lipids, and is often a precursor in the process of preparing anhydrous milk fat or anhydrous butter.

[0044] Preferably, the composition comprises 5 to 70% by weight of non-human mammalian milk lipids based on total lipids. In one embodiment, the composition comprises 10 to 65% by weight, more preferably 15 to 60% by weight, and even more preferably 25 to 55% by weight of non-human mammalian milk lipids based on total lipids. Preferably, these milk lipids are selected from butter, milk fat, cream, and anhydrous milk fat.

[0045] Preferably, the ratio of vegetable fat to milk fat is 3 / 7 to 20 / 1.

[0046] The composition may further comprise non-vegetable lipids and non-dairy fats, such as animal fats other than dairy fats, such as fish oil and ovoid lipids, as well as microbial oils, algal oils, fungal oils, or single-cell oils. Preferably, the non-vegetable fats and non-dairy fats are present in an amount of up to 10% by weight, more preferably up to 5% by weight, based on total lipids. Preferably, the lipids in the nutritional composition of the present invention comprise fat sources containing long-chain polyunsaturated fatty acids (LC-PUFAs), selected from fish oil, marine oil, algal oil, microbial oil, single-cell oil, and ovoid lipids, in an amount of 0.25 to 10% by weight, preferably 0.5 to 10% by weight, based on total lipids.

[0047] lipid globule size According to the invention, lipids are present in the composition in the form of lipid spheres. When in liquid form, these lipid spheres are emulsified in an aqueous phase. Alternatively, the lipid spheres are present in powder form, and the powder is suitable for reconstitution with water or another food-grade aqueous phase. Typically and preferably, the lipid spheres comprise a core and a surface. The core preferably comprises vegetable fats and milk fats, and preferably comprises at least 80% by weight, more preferably at least 90% by weight, of triglycerides, and more preferably consists substantially of triglycerides. Not all triglyceride lipids present in the composition must be included in the core of the lipid spheres, but preferably most of them are, preferably more than 50% by weight, more preferably more than 70% by weight, even more preferably more than 85% by weight, even more preferably more than 95% by weight, and most preferably more than 98% by weight of triglyceride lipids present in the composition are included in the core of the lipid spheres.

[0048] The lipid microspheres provided according to the method or use of the present invention have a mode diameter of greater than 2.0 µm, preferably greater than 2.5 µm, and more preferably greater than 3.0 µm based on volume. The lipid microspheres provided according to the method or use of the present invention have a mode diameter of less than 6.0 µm, preferably less than 5.5 µm, and more preferably less than 5.0 µm based on volume. Preferably, the lipid microspheres have a mode diameter of 2 to 6 µm, preferably 2.0 to 6.0 µm, more preferably 2.5 to 6.0 µm, more preferably 3.0 to 6.0 µm, even more preferably 3.0 to 5.5 µm, and even more preferably 3.0 to 5.0 µm based on volume.

[0049] Standard infant formula or growing milk contains lipid globules with a mode diameter much smaller than 1.0 µm, typically around 0.5 µm. Larger lipid globules have been found to have improved lipid digestion kinetics.

[0050] The percentage of lipid globules is based on the volume of total lipids. The mode diameter refers to the diameter most commonly found based on the volume of total lipids, or the peak value in a graph where X is the diameter and Y is the volume (%). A suitable method for determining the volume of lipid globules and their size distribution is, for example, the method described by Michalski et al., 2001, Lait 81:787-796, using a Mastersizer particle size analyzer (Malvern Instruments, Malvern, UK). The specific surface area of ​​lipid globules is the surface area per unit weight of lipid, which decreases with increasing globule size. Therefore, the specific surface area of ​​lipid globules can be calculated from the particle size distribution of lipid globules and the concentration and density of lipids. The specific surface area of ​​lipid globules given by the method or use according to the invention is preferably 0.5 to 15 m². 2 / g lipid, preferably 1.0 to 10.0 m 2 / g, more preferably 1.5 to 8.0 m 2 / g, or even more preferably 2.0 to 7.0 m 2 / g lipid. D[3,2] is the surface moment average or Sotter mean diameter, and can be determined using Mastersizer particle size analyzer software. D[4,3] is the volume moment average diameter or DeBrouckere mean diameter.

[0051] Methods for obtaining lipid microspheres with increased size and / or coated with phospholipids are disclosed in WO 2010 / 0027258 and WO2010 / 0027259.

[0052] Fatty acid composition In this article, LA refers to linoleic acid and / or an acyl chain (18:2 n6); ALA refers to α-linolenic acid and / or an acyl chain (18:3 n3); LC-PUFA refers to long-chain polyunsaturated fatty acids and / or acyl chains containing at least 20 carbon atoms in the fatty acyl chain and having more than two unsaturated bonds; DHA refers to docosahexaenoic acid and / or an acyl chain (22:6, n3); EPA refers to eicosapentaenoic acid and / or an acyl chain (20:5 n3); ARA refers to arachidonic acid and / or an acyl chain (20:4 n6); DPA refers to docosapentaenoic acid and / or an acyl chain (22:5 n3). PUFA refers to polyunsaturated fatty acids and / or acyl chains containing more than two unsaturated bonds; MUFA refers to fatty acids and / or acyl chains containing one unsaturated bond; SFA refers to saturated fatty acids and / or acyl chains. Medium-chain fatty acids (MCFAs) are fatty acids and / or acyl chains with a chain length of 8 to 12 carbon atoms. Butyric acid (BA) is a fatty acid and / or acyl chain containing 4 carbon atoms.

[0053] Preferably, the composition given according to the method or use of the invention comprises 10 to 25% by weight of PUFA based on total fatty acids. Amounts greater than 25% by weight would be much higher than those present in human milk and would present technical problems (e.g., stability of the nutritional composition).

[0054] LA is preferably present in sufficient quantities to promote healthy growth and development. Therefore, the composition preferably contains less than 20% by weight of LA based on total fatty acids, more preferably 5 to 15% by weight. Preferably, the composition contains more than 5% by weight of LA based on total fatty acids, more preferably at least 10% by weight. Preferably, ALA is present in sufficient quantities to promote healthy growth and development in infants. Therefore, the composition of the present invention preferably contains at least 0.5% by weight of ALA based on total fatty acids, more preferably at least 1.0% by weight of ALA based on total fatty acids. Preferably, the composition contains at least 1.4% by weight of ALA based on total fatty acids, more preferably at least 1.5% by weight. Preferably, the composition contains less than 10% by weight of ALA based on total fatty acids, more preferably less than 5.0% by weight. In one embodiment, the composition given according to the method or use of the present invention contains at least 10% by weight of LA and at least 1% by weight of ALA, preferably 10 to 20% by weight of LA and 1 to 5% by weight of ALA.

[0055] In one embodiment, the composition given according to the method or use of the invention comprises at least 0.3% by weight of BA based on total fatty acids. In one embodiment, the nutritional composition comprises 0.3 to 4.0% by weight of BA based on total fatty acids. For triglycerides derived from ruminant milk (e.g., cow's milk), a relatively high amount of BA is characteristic. BA is not present in vegetable fats or MCT-rich fats such as coconut oil, nor is it found in polar lipids derived from milk fat. Therefore, an alternative method of describing the presence of milk fat triglycerides in the composition is to define the fatty acid distribution by a BA content of 0.3 to 4.0% by weight based on total fatty acids. Based on the total weight of fatty acids, the composition preferably comprises at least 0.3% by weight of BA, preferably at least 0.5% by weight, more preferably at least 0.6% by weight, more preferably at least 0.8% by weight. Preferably, the composition has less than 4% by weight of BA based on total fatty acids, more preferably less than 3% by weight, more preferably less than 2.5% by weight. It is not intended to be limited to theory, but the presence of BA can provide beneficial effects.

[0056] In one embodiment, the composition given according to the method or use of the invention comprises at least 10% by weight of LA, at least 1% by weight of ALA and at least 0.3% by weight of BA based on the total weight of fatty acids, preferably 10 to 20% by weight of LA, 1 to 5% by weight of ALA and 0.3 to 3% by weight of BA based on the total weight of fatty acids.

[0057] Preferably, the composition given according to the method or use of the invention comprises at least 5% by weight of MCFA based on total fatty acids, more preferably at least 7% by weight. The composition advantageously comprises less than 15% by weight of MCFA based on total fatty acids, more preferably less than 10% by weight.

[0058] Preferably, the composition provided by the method or use according to the invention comprises LC-PUFA, more preferably n-3 LC-PUFA. More preferably, the composition of the invention comprises EPA, DPA and / or DHA, even more preferably DHA. Because low concentrations of DHA, DPA and / or EPA are already effective, and normal growth and development are important, the content of n-3 LC-PUFA (more preferably DHA) in the composition of the invention preferably does not exceed 5% by weight of the total fatty acid content. Preferably, the composition comprises at least 0.15% by weight of the total fatty acid content, preferably at least 0.35% by weight, more preferably at least 0.75% by weight of n-3 LC-PUFA, more preferably DHA. The composition preferably comprises at least 0.25% by weight of LC-PUFA based on the total fatty acid content. Preferably, the lipids in the composition comprise a fat source comprising 0.25% to 5% by weight of LC-PUFA based on total fatty acids, including at least 0.15% by weight of n-3 LC-PUFA based on total fatty acids, wherein the n-3 LC-PUFA is selected from DHA, EPA and DPA, more preferably DHA.

[0059] Since the n-6 fatty acid group (especially arachidonic acid (ARA) and LA as its precursor) offsets the n-3 fatty acid group (especially DHA and EPA and ALA as their precursor), the composition preferably contains a relatively small amount of ARA. Based on total fatty acids, the content of n-6 LC-PUFA (preferably ARA) is preferably no more than 5% by weight, more preferably no more than 2.0% by weight, more preferably no more than 0.75% by weight, and even more preferably no more than 0.5% by weight. Because ARA is important for optimally functional membranes (especially membranes of neural tissue) in infants, the amount of n-6 LC-PUFA (preferably ARA) is preferably at least 0.02% by weight, more preferably at least 0.05% by weight, more preferably at least 0.1% by weight, and more preferably at least 0.2% by weight, based on total fatty acids. The presence of a preferably small amount of ARA in nutrients given to infants under 6 months of age is beneficial, as infant formula is often the sole source of nutrition for these infants. Preferably, the weight ratio of n-6 LC-PUFA to n-3 LC-PUFA is less than 3, more preferably less than 2, and even more preferably less than 1.

[0060] Digestible carbohydrates Preferably, the composition given according to the method or use of the invention comprises digestible carbohydrates. The digestible carbohydrates preferably provide 30 to 80% of the total calories of the composition. Preferably, the digestible carbohydrates provide 40 to 60% of the total calories. When in liquid form (e.g., as an instant liquid), each 100 ml of the composition preferably contains 3.0 to 30 g of digestible carbohydrates, more preferably 6.0 to 20 g per 100 ml, and even more preferably 7.0 to 10.0 g per 100 ml. Based on dry weight, the compositions of the invention preferably contain 20 to 80% by weight, more preferably 40 to 65% by weight of digestible carbohydrates.

[0061] Preferred sources of digestible carbohydrates include lactose, glucose, sucrose, fructose, galactose, maltose, starch, and maltodextrin. Lactose is the main digestible carbohydrate found in human milk. Lactose advantageously has a low glycemic index. The compositions of the present invention preferably contain lactose. The compositions of the present invention preferably contain digestible carbohydrates, wherein at least 35% by weight, more preferably at least 50% by weight, more preferably at least 75% by weight, even more preferably at least 90% by weight, and most preferably at least 95% by weight of the digestible carbohydrates are lactose. Based on dry weight, the compositions of the present invention preferably contain at least 25% by weight of lactose, preferably at least 40% by weight.

[0062] Indigestible carbohydrates In one embodiment, preferably, the composition given according to the method or use of the invention comprises an indigestible oligosaccharide. Preferably, the composition of the invention comprises an indigestible oligosaccharide with a degree of polymerization (DP) of 2 to 250, more preferably 3 to 60.

[0063] Preferably, the composition of the present invention comprises fructo-oligosaccharide, galacto-oligosaccharide, and / or galacturonic acid oligosaccharide, more preferably galacto-oligosaccharide, and most preferably trans-galacto-oligosaccharide. In a preferred embodiment, the composition comprises a mixture of trans-galacto-oligosaccharide and fructo-oligosaccharide. Suitable indigestible oligosaccharides are, for example, Vivinal GOS (FrieslandcampinaDOMO), Raftilin HP, or Raftilose (Orafti).

[0064] Preferably, each 100 ml of the composition contains 80 mg to 2 g of indigestible oligosaccharides, more preferably 150 mg to 1.50 g per 100 ml, and even more preferably 300 mg to 1 g per 100 ml. On a dry weight basis, the composition preferably contains 0.25% to 20% by weight, more preferably 0.5% to 10% by weight, and even more preferably 1.5% to 7.5% by weight.

[0065] protein Preferably, the composition given according to the method or use of the invention comprises protein. The protein preferably provides 5 to 15% of the total calories. Preferably, the composition comprises protein providing 6 to 12% of the total calories. More preferably, the protein present in the composition is less than 9% based on a calorie count. Human milk contains less protein than cow's milk based on a calorie count. The protein concentration in the nutritional composition is determined by the sum of protein, peptides, and free amino acids. Based on dry weight, the composition preferably contains less than 12% by weight of protein, more preferably 9.6 to 12% by weight, and even more preferably 10 to 11% by weight. Based on ready-to-drink liquid products, the composition preferably contains less than 1.5 g of protein per 100 ml, more preferably 1.2 to 1.5 g, and even more preferably 1.25 to 1.35 g.

[0066] Protein sources should be selected in a manner that meets the minimum requirements for essential amino acid content and ensures satisfactory growth. Therefore, protein sources based on bovine milk proteins, such as whey, casein, and mixtures thereof, and proteins based on soybeans, potatoes, or peas are preferred. Preferably, the composition provided according to the method or use of the invention comprises casein and whey protein. If whey protein is used, the protein source is preferably based on acidic or sweet whey, whey protein isolate, or mixtures thereof, and may contain α-lactalbumin and β-lactoglobulin. More preferably, the protein source is acidic or sweet whey from which casein glycomacropeptide (CGMP) has been removed. Preferably, the composition comprises casein, preferably containing at least 3% by weight of casein based on dry weight. Preferably, the casein is intact and / or unhydrolyzed. The proteins of the present invention comprise peptides and free amino acids.

[0067] other Preferably, the composition administered according to the method or use of the invention is suitable for providing daily nutritional needs for humans under 36 months of age, particularly under 24 months of age, even more preferably under 18 months of age, most preferably infants under 12 months of age, and most preferably infants aged 0 to 6 months. Therefore, the nutritional composition is intended for feeding or for feeding human subjects, preferably infants under 36 months of age, preferably under 24 months of age, even more preferably infants under 18 months of age. Most preferably, the composition in the method or use of the invention is intended for administration or for feeding infants under 12 months of age.

[0068] Preferably, the composition provided according to the method or use of the invention comprises lipids, proteins, and digestible carbohydrates, wherein lipids preferably provide 30 to 60% of the total calories, proteins preferably provide 5 to 20% of the total calories, more preferably 5 to 15% of the total calories, and digestible carbohydrates preferably provide 25 to 75% of the total calories. Preferably, the composition comprises lipids providing 35 to 50% of the total calories, proteins providing 6 to 12% of the total calories, and digestible carbohydrates providing 40 to 60% of the total calories. In one embodiment, proteins provide 5 to 9% of the total calories. The total calorie amount is determined by the sum of calories obtained from proteins, lipids, and carbohydrates.

[0069] According to international guidelines for infant formula, the composition preferably contains other ingredients, such as vitamins and minerals.

[0070] In one embodiment, the composition given according to the method or use of the invention is an infant formula, a stage 2 formula, or a growing milk.

[0071] To meet the calorie requirements of infants, the composition preferably contains 45 to 200 kcal / 100 ml of liquid, more preferably 60 to 90 kcal / 100 ml of liquid, and even more preferably 60 to 75 kcal / 100 ml of liquid. This calorie density ensures an optimal ratio between water and calorie consumption. The molar osmolarity of the composition of the present invention is preferably 150 to 420 mOsmol / L, more preferably 260 to 320 mOsmol / L. A low molar osmolarity is intended to reduce gastrointestinal pressure.

[0072] Preferably, the composition is in liquid form and viscometered at 20°C for 100 s⁻¹. -1 The viscosity, as measured by shear rate, is less than 35 mPa·s, more preferably less than 6 mPa·s. In one embodiment, the composition is administered according to the method or use of the invention. The composition is suitably in powder form, which can be reconstituted with water or other food-grade aqueous liquids to form a liquid, or in the form of a liquid concentrate that should be diluted with water. It has been found that the lipid globules retain their size and encapsulation when reconstituted. In one embodiment, the composition administered according to the method or use of the invention is in powder form. In one embodiment, the powder is obtained after heat treatment of the composition in which lipid globules are formed. When the composition is in liquid form, the preferred daily volume is about 80 to 2500 ml, more preferably about 450 to 1000 ml per day.

[0073] application In the first six months of a exclusively breastfed infant's life, human milk lipids contribute approximately 45-55% of the milk's energy content, equivalent to about 5.5 kg of total fat intake. Lipids are primarily in the form of triglycerides, which are digested by lipases in the gastrointestinal tract. In adults, acidic lipases (tongue lipase and gastric lipase) account for 30% of lipolysis during digestion, with gastric lipase contributing the most. These acidic lipases do not require bile acids or colipases to achieve optimal enzymatic activity. In infants, acidic lipases are much more important, providing up to 50% of total lipolytic activity. Pancreatic lipases are secreted from the pancreas. Bile salts secreted from the liver and stored in the gallbladder are released into the duodenum, where they coat and emulsify large lipid droplets into smaller microdroplets, increasing the total surface area of ​​fat and allowing lipases to break it down more efficiently. The resulting monomers (two free fatty acids and one 2-monoacylglycerol) then move along the small intestine via peristalsis to be absorbed into the lymphatic system. In infants, pancreatic lipase and bile salt levels are low, so the products of gastric lipolysis play an important role in milk lipid digestion by compensating for the low levels of pancreatic lipase and emulsification.

[0074] It has been found that nutritional compositions containing larger lipid globules than conventional lipid globules, compared to standard infant formula, exhibit lower lipolysis rates and are more similar to human milk. These lipid globules comprise plant fats with a modal diameter of 2 to 6 μm based on volume, and / or a specific surface area of ​​0.5 and 15 m². 2 / g lipids, and coated with phospholipids derived from non-human mammalian milk. Furthermore, even lower lipolysis rates were observed when large lipid globules contained a combination of plant and milk fats. The slow lipolysis observed using human milk and the nutritional composition of this invention ensures lipid absorption throughout the gastrointestinal tract, thereby activating feedback mechanisms (e.g., ileal immobilization) to a lesser extent. For conventional IMF, rapid lipolysis overloads the small intestine with free fatty acids and induces feedback mechanisms to a greater extent. The total amount of fat ingested by the body is the same, therefore there is no effect on energy absorption. A slower rate of lipolysis regulates fat release in the body and can affect hormone signaling transduction, and is considered to have generally beneficial health effects on infants.

[0075] Therefore, the present invention aims to reduce the rate of gastric lipolysis and / or the rate of postprandial lipolysis in subjects. Preferably, the subjects are human infants under 36 months of age. Preferably, the composition according to the method or use of the present invention is administered orally to the subjects, preferably human infants.

[0076] Example Example 1: In vitro digestion kinetics of lipids in infant formula with different lipid globules Three infant formulas were tested. Infant formula 1 is the infant formula of this invention, and contains 3.4 g of lipids per 100 ml (24.7% by weight on a dry weight basis), of which 3.2 g are plant lipids, including 0.1% by weight of butyric acid, 18.4% by weight of palmitic acid, 13.5% by weight of linoleic acid, 2.5% by weight of alpha-linolenic acid, 0.36% by weight of arachidonic acid, and 0.2% by weight of docosahexaenoic acid, based on total fatty acids. The amount of phospholipids is 54.4 mg / 100 ml, which is 1.6% by weight on a total lipid basis, of which 1.5% by weight is derived from bovine fat globule membranes. As determined using the Malvern Mastersizer method, the mode diameter is approximately 4.4 µm on a volume basis, and approximately 60% by volume of the lipid globules have a size of 2 to 12 µm. A suitable method for determining the volume and size distribution of lipid globules is, for example, the method described by Michalski et al., 2001, Laid 81:787-796, using a Mastersizer particle size analyzer (Malvern Instruments, Malvern, UK). The specific surface area is 7 m². 2 / g lipids.

[0077] Infant formula 2 is also an infant formula of the present invention and is similar to infant formula 1, but it contains about 48% by weight of milk lipids and at least 45% by weight of plant lipids. It contains 1.39% by weight of butyric acid, 17.7% by weight of palmitic acid (of which at least 20% of the residues are at the sn2 position), 14.0% by weight of linolenic acid, 2.6% by weight of α-linolenic acid, 0.31% by weight of arachidonic acid, and 0.2% by weight of docosahexaenoic acid.

[0078] Infant formula 3 is a control formula with a similar composition to infant formula 1, but without milk-derived phospholipids. The mode diameter, measured by volume, is approximately 0.4 µm, as determined using the Malvern Mastersizer method, and the size of lipid globules less than 15% by volume ranges from 2 to 12 µm. The specific surface area is approximately 20 m². 2 / g lipids.

[0079] As described by Van den Braak et al. (ClinNutr. 2013 Oct;32(5):765-71), 150 ml of infant formula was subjected to gastric and intestinal conditions at 37°C using a pH and substrate pump-controlled device suitable for simulating the physiological conditions of human infants. In short, gastric digestion was simulated by gradually decreasing the pH (from pH 6.8 to pH 4.3) over 120 minutes after the addition of 1 M hydrochloric acid, 25 ml of 0.6 mg / ml α-amylase, 52.5 ml of 0.05 mg / ml porcine pepsin (SIGMA), and 0.125 mg / ml fungal lipase (Amano). Comparative experiments showed that the digestion of IMF 1 was very similar when using fungal lipase or rabbit gastric lipase (which is similar to human gastric lipase). Therefore, after adding 1 M sodium hydroxide / 1 M sodium carbonate, 0.75 ml of 2 mg / ml porcine trypsin, 135 ml of 0.015 g / ml trypsin, and 0.005 g / ml bile extract (SIGMA), enteric digestion was simulated by raising the pH to pH 7.2 over 120 minutes. Samples were taken at t=0-10-30-60-90-120 minutes (during gastric digestion) and t=130-150-180-210-240 minutes (equivalent to 10-30-60-90-120 minutes during enteric digestion). 1 ml sample volume was collected in glass tubes and frozen at -80°C for analysis of free fatty acids (FFA) by GC. Particle size distribution was measured using a Malvern Mastersizer.

[0080] The results are shown in Table 1.

[0081] Table 1: Free fatty acids formed after digestion of IMF with different lipid globule properties in an in vitro model for infant digestion (n=3).

[0082]

[0083] *: P < 0.05 vs IMF 3 For both IMFs of this invention, the concentration of FFA formed during simulated gastrointestinal digestion was significantly lower than that of the standard control IMF 3. Compared to conventional IMF 3, IMFs 1 and 2 showed slower average gastric lipolysis after 120 minutes of digestion: 37 ± 5 and 33 ± 9 vs. 62 ± 2 μg FFA / min (P=0.002). Lipolysis of IMF2 was slightly but consistently lower than that of IMF 1.

[0084] Compared to conventional IMF, the total gastrointestinal lipolysis rates (after 240 min) of IMFs 1 and 2 were slower: 55 ± 3 and 54 ± 6 vs. 72 ± 9 μg FFA / min. For all IMFs, the mean lipolysis rate after 120 min of digestion under small intestinal conditions was more comparable (approximately 60 µg FFA / min).

[0085] Example 2: Particle size characteristics of IMFs with different lipid globules during in vitro gastric digestion. In the experiment of Example 1, 10 ml samples of IMF 1, IMF 2, and IMF 3 were taken during gastric digestion and collected in 15 ml tubes placed on ice for structural analysis on the same day. A drop of sample was placed on a microscope slide and examined using an optical microscope. Particle size distribution was measured using a Malvern Mastersizer. The results are shown in Table 2 and... Figure 1 middle.

[0086] Figure 1 Microscopic (20x) images of IMFs with different lipid globules during in vitro gastric digestion are shown.

[0087] Table 2: Particle size characteristics of IMFs with different lipid globules during in vitro gastric digestion (n=3).

[0088]

[0089] During gastric digestion, the particle distribution and microscopic images of IMFs 1, 2, and 3 differed significantly. In conventional IMF 3, the specific surface area (SSA) was significantly reduced due to the formation of large protein aggregates. For the IMFs 1 and 2 of this invention, far fewer protein aggregates were formed, and large, intact lipid droplets remained, with less incremental change in particle size distribution (see [link]). Figure 1Up to 60 minutes, the SSA did not change significantly, and the SSA of IMF 3 was higher than that of other IMFs. However, the SSA decreased at t=90 and t=120, and was lowest in IMF 3. Meanwhile, in conventional IMF 3, the volume-weighted average diameter (D[4,3]) and surface-weighted average diameter (D[3,2]) of the particles were lower than those of other IMFs until 60 minutes. After 60 minutes, the particle size increased in all IMFs, but the increase was most significant in IMF 3, making the particles in IMF 3 significantly larger compared to other IMFs, due to the formation of large protein aggregates. For the IMFs of this invention, far fewer protein aggregates were formed, and large, intact lipid droplets remained, with less incremental change in particle size distribution.

[0090] Example 3: Effects of different phospholipid coatings on lipolysis Similar experiments were performed as described in Example 1, except that different phospholipid sources were tested. IMF 1 is similar to IMF 1 in Example 1.

[0091] IMF 2 is similar to IMF 1, except that it uses 1.5% by weight lecithin instead of milk-derived phospholipids. IMF 3 contains 1.5% by weight soybean phospholipids. IMF 4 is a standard control IMF, similar to IMF 3 in Example 1. In IMF 2, the volumetric modal diameter of the lipid globules is approximately 5.1 µm, while in IMF 3 it is approximately 4.6 µm.

[0092] The results are shown in Table 3.

[0093] Table 3: Free fatty acids formed after digestion of IMFs with different lipid globule properties in an in vitro model for infant digestion (n=3).

[0094]

[0095] For IMF 1 of the present invention, the concentration of FFA formed during simulated gastrointestinal digestion was lower than that of the standard control IMF 4, similar to Example 1. IMFs 2 and 3, coated with lecithin or soybean lecithin respectively, showed moderate effects, but were more similar to the control IMF 4, especially in terms of the initial rate of lipolysis in the gastric stage (see Table 3). The kinetics of FFA formation between IMF 2 and IMF 3 were very similar, showing higher FFA formation than IMF 1 and a slightly lower rate than IMF 4.

[0096] Compared to conventional IMF 4, IMF 1 showed slower average gastric lipolysis after 120 minutes of digestion: 0.13 vs 0.22 mM FFA / min. The average gastric lipolysis rates for IMF 2 and IMF 3 were 0.11 and 0.12 mM FFA / min, respectively.

[0097] Compared to conventional IMF 4, IMF 1 had a slower total gastrointestinal lipolysis rate (after 240 min): 0.29 vs. 0.35 mM FFA / min. The total gastrointestinal lipolysis rates for IMF 2 and IMF 3 were 0.31 and 0.32 mM FFA / min, respectively.

[0098] For IMF 1 and IMF 4, under small intestinal conditions, the average lipolysis was more comparable (approximately 0.35 mM FFA / min) after the subsequent 120 minutes of digestion, and lower than that of IMF 2 and IMF 3 (approximately 0.45 mM FFA / min).

[0099] Example 4: Dynamic Intestinal Model (TIM) As described by Abrahamse et al. (2012, Food Dig. 3: 63-77), a semi-dynamic intestinal model (TIM-1) was used in in vitro digestion experiments 1, IMF 1, IMF 2 and IMF 3, which were adapted to simulate the physiological conditions of human infants and specific settings were used to determine lipid digestion and absorption.

[0100] The results are shown in Table 4.

[0101] Compared to IMF 3, IMF 1 and IMF 2 exhibited lower and slower lipolysis between 90 and 120 minutes of gastric and intestinal digestion, similar to the results in Example 1. This difference in hydrolysis begins early in gastric digestion (within 20 minutes) and continues to play a role during intestinal digestion. For all IMFs, lipid absorption was optimal and similar at the end of digestion (approximately 93%).

[0102] Table 4: Lipolysis % (n=3) of IMFs with different lipid globule properties after digestion in a dynamic in vitro model for infant digestion.

[0103]

[0104] Example 5: Evaluation of the surface properties of lipid microspheres by transmission electron microscopy (TEM) Samples prepared for TEM imaging were modified from those described by Gallier et al. (Food Chemistry 2013, 141:3215-3223). In 15 ml tubes, samples (human milk, IMF 1 and IMF 3 from Example 1) were mixed with 3% cryogenic gel agarose (stored at 50°C) at a 1:1 ratio. After cooling, the gelled samples were cut into 1 mm pieces. 3 The block was transferred to 3% glutaraldehyde in 0.1 M dimethylarstannate buffer (pH 7.2). After fixation, the block was rinsed with 0.1 M dimethylarstannate buffer (pH 7.2). Post-fixation was performed using 1% osmium tetroxide (OsO4). The block was rinsed with water. Whole-block staining was performed using 1% uranium acetate. The block was dehydrated in ethanol series (50-100%) and embedded in Epon resin. Sections (60 nm) were prepared using a Leica microtome (Wetzlar, Germany). Post-staining was performed on a grid (Cu 100M-H coated with Formvar membrane and carbon film) using 7% uranium acetate and Reynolds lead citrate. Imaging was performed using a Tecnai 10 / 12 transmission electron microscope (FEI, Hillsboro, OR, USA), equipped with a SISCCD camera MegaView II, a single tilting sample holder, and the dedicated software AnalySISPro.

[0105] Figure 2 This represents a portion of a typical human milk lipid globule. The thickness of the polysaccharide-coated MFGM trilayer ranges from 5 to 20 nm. Dashed arrows point to the MFGM bilayer; solid arrows point to the inner MFGM monolayer; dotted arrows point to the MFGM polysaccharide coating extending into the aqueous phase; the scale bar represents 50 nm.

[0106] Figures 3A and 3B show lipid spheres of IMF 1 with a thin film (5–10 nm) including a small number of protein aggregates. Some droplets have casein microspheres that interact with the interface. Furthermore, bovine MFGM fragments were detected in the aqueous phase, interacting at the lipid droplet interface. Dotted arrows point to casein microspheres interacting with the lipid sphere interface; solid arrows point to MFGM fragments in the aqueous phase located at the sphere interface; dashed arrows point to small interfacial protein aggregates; scale bars represent A: 1 μm; B: 200 nm.

[0107] Figure 4The lipid droplets represent control IMF 3, which have a thicker (20-100 nm) and more densely stained protein membrane than the film surrounding the lipid spheres of concept IMF 3. Protein aggregates were observed at the interface of the control IMF 3 lipid spheres, but these aggregates were larger than those at the interface of the IMF 1 lipid spheres of the present invention. The aggregation of lipid spheres may be due to protein bridging or protein-protein interactions between droplets. The MFGM fragment was not detected in control IMF 3. Dashed arrows point to interfacial protein aggregates; dotted arrows point to lipid spheres with a thick protein coating; the scale bar represents 500 nm.

[0108] The IMF has a whey protein:casein ratio of 60:40. β-lactoglobulin is the predominant whey protein in the IMF. During heat treatment above 65°C, β-lactoglobulin unfolds and interacts with itself or other proteins due to its free sulfhydryl groups, resulting in the formation of disulfide bonds between molecules. Aggregation of β-lactoglobulin with other proteins will produce a membrane thicker than the compact and thin (2-3 nm) layer formed by native β-lactoglobulin molecules. Since both IMFs of the present invention and the control IMF were heat-treated at temperatures above 65°C, β-lactoglobulin is more likely to exist as aggregated protein molecules. Unsaturated phospholipids form a very thin monolayer of about 2 nm, while saturated phospholipids such as sphingomyelin form monolayers up to 4 nm in length due to their more straight fatty acid chains. Therefore, the interface of the control IMF 3 lipid globules consists only of casein and whey protein, mainly in an aggregated state. The interface of the IMF lipid globules of this invention may consist of phospholipids, MFGM fragments or absorbed MFGM protein and lipids, as well as naturally occurring undenatured milk proteins and a small amount of casein microparticles linked to the interface. Transmission electron microscopy was used to characterize the HM lipid globules, the lipid globules of the IMF of this invention, and the currently standard IMF 3. The IMF of this invention contains lipid globules whose size and phospholipid coating are similar to those of lipid globules in human milk.

[0109] These examples demonstrate improved (i.e., reduced) gastric lipolysis rates and / or improved (i.e., reduced) postprandial lipolysis rates achieved by compositions comprising lipid globules that are larger than conventional lipid globules or have a lower specific surface area than conventional lipid globules, and that contain a coating of milk-derived phospholipids. It can be inferred that by feeding infants or young children formulations containing lipid globules, which are compositionally and in size and coating more similar to human milk lipid globules, there is a fat digestion mode and bioavailability of ingested fatty acids that are considered more similar to human milk. This effect exists even when the formulation contains newly formed lipid globules in a mild homogenization step and even when heat-treated in the presence of proteins.

Claims

1. A method for reducing the rate of gastric lipolysis and / or the rate of postprandial lipolysis in a subject, comprising administering the subject a nutritional composition comprising lipid globules, said lipid globules comprising triglycerides derived from plant fats and phospholipids derived from non-human mammalian milk, wherein said lipid globules have: - The mode diameter is 2 to 6 µm, or - 0.5 to 15 m 2 Specific surface area per g of lipid, or - Mode diameter of 2 to 6 µm and 0.5 to 15 m 2 Specific surface area per g of lipid Furthermore, the lipid spheres contain a coating containing phospholipids.

2. The method of claim 1, wherein the composition comprises non-human mammalian milk fat.

3. The method of claim 1 or 2, wherein the amount of lipid is at least 15% by weight based on the dry weight of the composition.

4. The method of any of the preceding claims, wherein the amount of phospholipids is at least 0.5% by weight based on total lipids.

5. The method of any of the preceding claims, wherein the phospholipid comprises at least 15% by weight of sphingomyelin.

6. The method of any of the preceding claims, wherein the phospholipids comprise at least 2% by weight of phosphatidylserine based on total phospholipids.

7. The method of any of the preceding claims, wherein the lipid comprises at least 10% by weight linoleic acid and at least 1% by weight α-linolenic acid.

8. The method of any of the preceding claims, wherein the lipid comprises at least 0.3% by weight of butyric acid based on total fatty acids.

9. The method of any of the preceding claims, wherein the nutritional composition comprises casein and whey protein.

10. The method of any of the preceding claims, wherein the nutritional composition is an infant formula, a stage 2 formula, and / or a growing milk.

11. The method of any of the preceding claims, the method being used to provide nutrition to human subjects under 36 months of age.

12. The method of any of the preceding claims, wherein the lipid globules comprise a monolayer containing phospholipids derived from milk fat.

13. The method of any of the preceding claims, wherein the phospholipid is provided in the form of a milky liposphere membrane.

14. The method of any of the preceding claims, wherein the specific surface area of ​​the lipid spheres is from 1.0 to 8.0 m². 2 / g lipids.

15. The method of any of the preceding claims, wherein the nutritional composition is in powder form.

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