Formula milk powder with immunomodulatory functional activity and its preparation method and application

Abstract

This invention provides a formula milk powder with immunomodulatory activity, its preparation method, and its application. The formula milk powder of this invention contains sphingomyelin, cephalin, and zinc, with a weight ratio of sphingomyelin / zinc = A, cephalin / zinc = B, and the A:B ratio being (0.45-3.45):1. Furthermore, based on the total mass of the formula milk powder, the total protein content is 9-27 g / 100g, whey protein accounts for 20%-100% of the total protein, the fat content is 10-35 g / 100g, the dietary fiber content is 0.1-8.0 g / 100g, and the carbohydrate content is 40-60 g / 100g. This invention also provides a method for preparing the formula milk powder. The formula milk powder of this invention helps to simultaneously increase intestinal flora diversity, reduce harmful bacteria in the intestinal flora, and increase the number of intestinal IgA secretory cells, thus exhibiting immunomodulatory activity.

Description

Technical Field

[0001] This invention relates to a formula milk powder with immunomodulatory activity, its preparation method and application, specifically to a formula milk powder containing sphingomyelin, cephalin and zinc that has immunomodulatory activity and is particularly suitable for infants and young children, its preparation method and related applications, belonging to the field of formula milk powder technology. Background Technology

[0002] The gut microbiota is closely related to the health of newborn mammals and humans. The colonization and development of the gut microbiota begin in early life, influenced by various factors before and after birth, and changes dynamically over time. For example, in human infants, the structure and composition gradually resemble that of adults around age 3, and then remain stable. Infancy is a critical period for gut microbiota development; changes in the gut microbiota during infancy can have a profound impact on the growth and development, energy metabolism, and the maturation of the nervous and immune systems. For instance, after birth, the majority of gut microbiota in human infants are Proteobacteria, Firmicutes, Bacteroidetes, Actinobacteria, and Fusobacteria. The gut microbiota gradually transitions from a developmental stage with low diversity and predominantly Bifidobacteria to a transitional stage with increased diversity and Proteobacteria and Clostridium difficile as dominant bacteria, ultimately evolving into a stable stage that approximates an adult-type gut microbiota structure dominated by Firmicutes and supplemented by Bacteroidetes. Higher levels of gut microbiota diversity contribute to the development and maturation of the infant's gut. Studies have shown that offspring of mothers experiencing perinatal anxiety and depression have lower alpha diversity in their gut microbiota; gut microbiota dysbiosis in early life can cause abnormal activation of the infant's immune system, disordered secretion of pro-inflammatory factors, and affect the normal immune development process.

[0003] B cells are a major type of immune cell in the gut. They mature in Pell's nodes, where activated B cells continuously convert to immunoglobulin A (IgA) secretory cells in both T-cell-dependent and non-T-cell-dependent ways. Upon antigen stimulation, B cells enter the lamina propria of the mucosa to form IgA secretory cells. Mature IgA secretory cells can further differentiate into sIgA. As the most abundant immunoglobulin in the mucosal layer, sIgA has functions such as defending against pathogens and toxins, and immune regulation. sIgA traps bacteria in intestinal mucus, preventing direct contact between intestinal symbionts and the gut, and can also neutralize toxins and pathogens in the intestinal lumen, thus avoiding harmful stimulation of the gut by intestinal contents. The recognition of pathogens and symbionts by sIgA guides immune stress and immune tolerance responses in immune cells of Pell's nodes, lymphoid follicles, or the lamina propria.

[0004] Therefore, an increase in gut microbiota diversity and the number of gut IgA-secreting cells, as well as a decrease in harmful bacteria in the gut microbiota, are beneficial to the health of newborn mammals or humans.

[0005] Previously, researchers focused on osteopontin and Bifidobacterium longum to increase gut microbiota diversity. Probiotics were previously the focus for reducing harmful bacteria in the gut microbiota and increasing the number of IgA-secreting cells in the gut.

[0006] After research, the inventors believe that osteopontin, Bifidobacterium longum, and probiotics have limited effects on increasing the diversity of intestinal flora, and also have limited effects on reducing the number of harmful bacteria and intestinal IgA secretory cells in the intestinal flora.

[0007] Therefore, there has been a lack of nutritional compositions that can effectively increase the diversity of gut microbiota in newborn mammals or humans, and reduce the number of harmful bacteria and IgA-secreting cells in the gut microbiota.

[0008] Furthermore, research on the efficacy of sphingomyelin and cephalin has largely focused on their neurogenic activity and bone development. For example, CN104883909A discloses a nutritional composition that lists sphingomyelin and cephalin as ingredients, which has an impact on the overall health of the brain and nervous system. CN115590195A discloses a nutritional composition that lists sphingomyelin and cephalin as ingredients, which has non-therapeutic uses in promoting bone growth, development, and health in humans or animals, such as improving bone density. However, these documents do not disclose the inclusion of zinc in their nutritional compositions, nor the effects of sphingomyelin and cephalin on the gut microbiota and intestinal IgA-secreting cells.

[0009] The effects of the combination of sphingomyelin, cephalin, and zinc on immunomodulatory activity, particularly on the regulatory activity of the immune system and gut microbiota (especially gut microbiota diversity, harmful bacteria in the gut microbiota, and the number of gut IgA-secreting cells), have not been reported in detail. Summary of the Invention

[0010] One object of the present invention is to provide a formula milk powder with immunomodulatory activity.

[0011] Another object of the present invention is to provide a method for preparing the formula milk powder.

[0012] Another object of the present invention is to provide the application of the aforementioned formula milk powder.

[0013] The inventors of this case discovered in experimental research that a nutritional composition containing sphingomyelin, cephalin, and zinc in a specific ratio can effectively increase the diversity of gut microbiota in newborn mammals (such as human infants and piglets), reduce the number of harmful bacteria and intestinal IgA secretory cells in the gut microbiota, and has immunomodulatory activity. Based on this, they provided a formula milk powder with immunomodulatory activity.

[0014] This invention includes the following aspects:

[0015] <First Aspect>

[0016] The first aspect of the present invention is to provide a formula milk powder containing sphingomyelin, cephalin, and zinc, wherein, by weight ratio, sphingomyelin / zinc = A, cephalin / zinc = B, and the ratio of A:B is (0.45-3.45):1; and, by total mass of the formula milk powder, the total protein content is 9-27g / 100g, the whey protein accounts for 20%-100% of the total protein, the fat content is 10-35g / 100g, the dietary fiber content is 0.1-8.0g / 100g, and the carbohydrate content is 40-60g / 100g.

[0017] In some specific embodiments, in the formula milk powder, sphingomyelin / zinc = A, cephalin / zinc = B, and the ratio of A:B is (0.5-3.45):1, preferably (0.6-3.45):1, more preferably (0.7-2.5):1, and even more preferably (1-1.5):1.

[0018] In some specific embodiments, based on solid components, in the formula milk powder, relative to the total amount of sphingomyelin and cephalin 100% by mass, the content of sphingomyelin is 30%-75% by mass, preferably 40%-70% by mass, and the content of cephalin is 30%-70% by mass, preferably 35%-65% by mass.

[0019] In some specific embodiments, based on solid components, the zinc content in the formula milk powder is 1%-10% by mass, preferably 3%-6% by mass, relative to 100% by mass of the total amount of sphingomyelin and cephalin.

[0020] In some specific embodiments, based on solid components, the total amount of sphingomyelin, cephalin, and zinc relative to 100g of formula milk powder is 50mg-200mg, preferably 80mg-160mg, more preferably 90mg-150mg, and especially preferably 100mg-140mg or 120mg-140mg.

[0021] In some specific embodiments, the protein-providing raw materials in the formula milk powder may include any one or more of raw milk, whole milk powder, skim milk powder, demineralized whey powder, casein powder, and whey protein powder; preferably, the whey protein powder includes α-lactalbumin powder and milk fat globule membrane powder. In some specific embodiments, based on 1000 parts by weight of formula milk powder, the amount of raw materials added includes any one or more of the following: 800-3500 parts by weight of raw milk, 0-450 parts by weight of whole milk powder, 0-500 parts by weight of skim milk powder, 0-500 parts by weight of whey protein powder, 0-500 parts by weight of demineralized whey powder, 0-80 parts by weight of α-lactalbumin powder, 0-50 parts by weight of β-casein powder, and 0-50 parts by weight of milk fat globule membrane powder.

[0022] In some specific embodiments, the raw materials providing fat in the formula milk powder include any one or more of milk fat raw materials and vegetable oils; the milk fat raw materials include one or more of raw milk, whole milk powder, and anhydrous milk; the vegetable oils include one or more of sunflower seed oil, corn oil, soybean oil, low-erucic acid rapeseed oil, coconut oil, flaxseed oil, palm oil, perilla oil, walnut oil, and OPO structured lipids. In some specific embodiments, based on 1000 parts by weight of formula milk powder, the raw materials include: 0-90 parts by weight of sunflower seed oil, 0-20 parts by weight of flaxseed oil, and 0-150 parts by weight of OPO structured lipids.

[0023] In some specific embodiments, the raw materials providing dietary fiber in the formula milk powder include any one or more of fructooligosaccharides, galactooligosaccharides, polyfructose, raffinose, and human milk oligosaccharides. In some specific embodiments, based on 1000 parts by weight of formula milk powder, the raw materials include: 0-50 parts by weight of galactooligosaccharides, 0-50 parts by weight of fructooligosaccharides, and 0-30 parts by weight of human milk oligosaccharides.

[0024] In some specific embodiments, the raw material providing carbohydrates in the formula milk powder includes lactose (which may be a base raw material containing lactose, or raw lactose). In some specific embodiments, based on 1000 parts by weight of formula milk powder, the raw material includes: 0-500 parts by weight of raw lactose.

[0025] The sources of sphingomyelin and cephalin in this invention are not limited; they can be derived from commercially available food-grade sphingomyelin and cephalin, or from food-grade sphingomyelin and cephalin prepared using conventional techniques such as column chromatography, membrane filtration, and supercritical fluid extraction. In some specific embodiments, the raw materials providing sphingomyelin and cephalin in the formula milk powder include: raw materials containing milk fat, whey protein powder, milk fat globule membrane powder, phospholipids, and any one or more of these. In some specific embodiments of this invention, based on 1000 parts by weight of formula milk powder, the raw materials include: 800-3500 parts by weight of raw milk, 0-50 parts by weight of α-lactalbumin powder, 0-20 parts by weight of milk fat globule membrane powder, and 0-10 parts by weight of phospholipids.

[0026] The source of zinc in this invention is not limited, as long as it is a zinc-containing compound that can be used in food. In some specific embodiments, the raw materials providing zinc in the formula milk powder include: zinc sulfate, zinc gluconate, zinc glycinate, zinc oxide, zinc lactate, zinc citrate, zinc chloride, zinc acetate, zinc carbonate, and / or zinc citrate. In some specific embodiments of this invention, based on 1000 parts by weight of formula milk powder, the zinc content is 1-10 mg / 100g, and the source is zinc sulfate.

[0027] In some specific embodiments, the formula milk powder of the present invention further includes compound vitamins and compound minerals. These compound nutrients are a combination of nutritional components that meet national standards, and are added in different amounts according to different formulas. Preferably, based on 1000 parts by weight of formula milk powder, the raw materials include: 7-35 parts by weight of compound nutrients containing calcium powder, vitamins, and minerals. More preferably, the compound nutrients are added at least in the form of compound vitamin packets, calcium powder packets, and mineral nutrient packets. Specifically, the compound nutrients are preferably added in the form of the following nutrient packets:

[0028] Minerals, per gram of mineral packet: iron 40-120mg, zinc 20-55mg, copper 2600-5000μg, selenium 0-180μg, iodine 400-1800μg, manganese 0-750μg, total weight 0.5-2 parts by weight;

[0029] Calcium powder, each gram of calcium powder packet contains: calcium 100-420mg, phosphorus 0-100mg, sodium 0-150mg, potassium 0-50mg, total weight 5-30 parts by weight;

[0030] Magnesium chloride nutritional packs contain 10%-16% effective magnesium and weigh 0-3kg.

[0031] Potassium chloride nutritional pack, with an effective potassium content of 49%-53%, and a total weight of 0-7kg;

[0032] Choline chloride nutritional pack, with an effective choline content of 72%-76%, and a total weight of 0.5-4kg;

[0033] Each gram of this vitamin pack may contain: Vitamin A 1200-2900μg RE; Vitamin D 20-80μg; Vitamin E 10-50mg α-TE; Vitamin K1 100-400μg; Vitamin B1 1000-3600μg; Vitamin B2 600-4500μg; Vitamin B6 1000-2500μg; Vitamin B12 3-10μg; Nicotinamide 11000-21000μg; Folic acid 180-550μg; Pantothenic acid 5400-12500μg; Biotin 20-120μg; Taurine 130-300mg; Vitamin C 20-500mg; Inositol 0-250mg; L-carnitine 0-100mg; Total weight 2-5kg.

[0034] In some specific embodiments, the raw materials of the formula milk powder of the present invention may further include one or more combinations of appropriate DHA, ARA, nucleotides, lactoferrin, probiotics, etc. Preferably, based on 1000 parts by weight of the formula milk powder of the present invention, the raw materials include: 0-25 parts by weight of DHA, 0-35 parts by weight of ARA, 0-3 parts by weight of lactoferrin, 0-2 parts by weight of nucleotides, and 0-1 parts by weight of probiotics.

[0035] In some specific embodiments, the formula milk powder of the present invention is an infant formula milk powder. Specifically, the formula milk powder may be a formula powder for infants aged 0-6 months, a formula powder for older infants aged 6-12 months, or a formula powder for toddlers aged 12-36 months.

[0036] In some specific embodiments, the formula milk powder of the present invention is a special medical purpose formula food.

[0037] <Second aspect>

[0038] A second aspect of the present invention provides a method for preparing the aforementioned formula milk powder, the method comprising:

[0039] The content of sphingomyelin, cephalin, and zinc in food ingredients is adjusted so that, by weight ratio, sphingomyelin / zinc = A, cephalin / zinc = B, and the ratio of A:B is (0.45-3.45):1, to prepare the formula milk powder.

[0040] In some specific embodiments, the process flow of the formula milk powder preparation method of the present invention mainly includes: ingredient mixing, homogenization, concentration and sterilization, spray drying, and dry mixing to obtain the finished product. Specific preparation methods include:

[0041] Mix milk that has been coarsely filtered, homogenized and sterilized, powdered raw materials and melted oil raw materials, add galactooligosaccharide syrup to the small hopper, and add compound nutrient fortifier, magnesium chloride nutrient pack, potassium chloride nutrient pack and choline chloride nutrient pack to the nutrient tank to obtain a mixed liquid.

[0042] The mixed liquid is filtered, homogenized, cooled, concentrated and sterilized, spray dried, and fluidized bed dried and cooled to obtain dried milk powder, which is then mixed with DHA, ARA, lactoferrin, nucleotides, bifidobacteria, etc., selectively included in the formula, and screened to obtain the formula milk powder.

[0043] In some specific embodiments, in the method for preparing formula milk powder of the present invention, the preferred primary pressure for homogenization of the mixed liquid is 105±5 bar, and the secondary pressure is 32±3 bar.

[0044] In some specific embodiments, in the preparation method of the formula milk powder of the present invention, the concentration and sterilization preferably adopts dual-effect concentration, more preferably, the sterilization temperature is ≥83℃ and the sterilization time is 25 seconds; even more preferably, the output concentration is 48%-52% dry matter.

[0045] In some specific embodiments, in the preparation method of the formula milk powder of the present invention, the inlet air temperature of the spray drying is preferably 165-180°C, the exhaust air temperature is 75-90°C, the high pressure pump pressure is 160-210 bar, and the tower negative pressure is -4 mbar to -2 mbar.

[0046] In some specific embodiments, the preparation method of the formula milk powder of the present invention preferably includes two drying and cooling processes, with the temperature of the milk powder after the second drying and cooling being 25-30°C; at the same time, the phospholipids are mixed with the carrier and heated to 60-65°C, and then uniformly dispersed on the surface of the milk powder under the action of compressed air.

[0047] In one specific embodiment of the present invention, an infant formula milk powder is prepared, and the preparation process may include the following specific steps:

[0048] 1) Milk coarse filtration: After coarse filtration and degassing in the balance tank, the milk is preheated by a plate heat exchanger and then separated by a separator to remove impurities.

[0049] 2) Milk homogenization and sterilization: After removing impurities, part of the milk is homogenized in a homogenizer, while the other part is not homogenized. The homogenized milk and the unhomogenized milk are mixed and then enter the sterilization system for sterilization. After sterilization, the milk enters the mixing tank.

[0050] 3) Powder addition: Various powder raw materials are metered according to the formula and then added to the powder mixing tank for storage through the air conveying system.

[0051] 4) Vacuum powder suction: Various powder raw materials in the powder mixing tank are sucked into the mixing tank through a vacuum system.

[0052] 5) Melting and blending oil: Put the oil specified in the formula into the melting room according to the formula requirements. The temperature of the melting room should be maintained at 50-90℃. After the oil is melted, pump it into the mixed oil storage tank through the oil pump and flow meter according to the formula ratio requirements.

[0053] 6) Storage of mixed oils: Mixed oils are stored in oil storage tanks at a temperature of 40-50℃ for less than 12 hours to prevent fat oxidation.

[0054] 7) Weighing: Pump the mixed oil into the mixing tank according to the formula requirements.

[0055] 8) Adding compound nutrient fortifiers: Add compound nutrient fortifier I, compound nutrient fortifier II, compound nutrient fortifier III, compound magnesium chloride nutrient pack, compound potassium chloride nutrient pack and choline chloride separately, dissolve them in 100-200kg of purified water, and then pump them into the mixing tank. After each type is pumped, rinse the addition tank and pipeline with 100kg of purified water.

[0056] 9) Adding small ingredients: Add galacto-oligosaccharide syrup to the small hopper and then transfer it to the mixing tank.

[0057] 10) Filtration: Mix the raw materials in a mixing tank, and filter the resulting liquid through a filter screen to remove any physical impurities that may have been introduced from the raw materials.

[0058] 11) Homogenization: The mixed liquid is homogenized by a homogenizer. The first pressure is 105±5 bar and the second pressure is 32±3 bar. The fat globules are mechanically processed and dispersed into uniform fat globules.

[0059] 12) Cooling and storage: The homogenized liquid enters the plate heat exchanger for cooling. It is cooled to below 20°C and temporarily stored in the pre-storage tank. It will enter the next process within 6 hours. The agitator is turned on according to the set requirements.

[0060] 13) Concentration and sterilization: Double-effect concentration is used during production, with a sterilization temperature of ≥83℃ and a sterilization time of 25 seconds; the output concentration is 48%-52% dry matter.

[0061] 14) Concentrated milk storage, preheating and filtration, spray drying: The concentrated milk is temporarily stored in a concentrated milk balance tank; it is preheated to 60-70℃ by a scraper preheater, and after preheating, the material is filtered through a 1mm pore size filter and then spray-dried in a drying tower by a high-pressure pump. Fine powder is agglomerated at the top of the tower or in a fluidized bed as required; the inlet air temperature is 165-180℃, the outlet air temperature is 75-90℃, the high-pressure pump pressure is 160-210 bar, and the tower negative pressure is -4mbar to -2mbar.

[0062] 15) Fluidized bed drying and cooling: After the milk powder from the drying tower is dried twice in a fluidized bed (first stage), it is cooled to 25-30℃ in a fluidized bed (second stage); at the same time, the phospholipids are mixed with the carrier and heated to 60-65℃. Under the action of compressed air, the phospholipids are evenly dispersed on the surface of the milk powder, causing the powder particles to agglomerate and increase their particle size and solubility.

[0063] 16) Packaging: Weigh and seal DHA, ARA, lactoferrin, nucleotides, and bifidobacteria according to the formula requirements.

[0064] 17) Dry mixing: Mix the weighed DHA, ARA, lactoferrin, nucleotides, bifidobacteria and milk powder in a dry mixer.

[0065] 18) Sieving: The milk powder is sieved to make the particle size uniform and the powder residue is disposed of as waste.

[0066] 19) Powder discharge: Collect powder in a sterilized powder collection box and transport it from the powder discharge room to the powder loading room.

[0067] 20) Powdering: Pour the milk powder into the powder storage tank on the large and small packaging machines according to the packaging requirements.

[0068] 21) Packaging: Nitrogen-filled packaging using automatic packaging machines of different specifications; oxygen content is below 1-5% during nitrogen filling.

[0069] 22) Packing: Put the packaged small bags into the carton and add the powder scoop at the same time, and seal the carton with a sealing machine.

[0070] 23) Finished product inspection: Sampling inspection of packaged products according to the inspection plan.

[0071] 24) Warehousing and storage: Products that have passed inspection shall be stored in the warehouse at room temperature with a humidity of ≤65%.

[0072] <Third aspect>

[0073] A third aspect of the present invention provides the use of sphingomyelin, cephalin, and zinc in the preparation of products that promote growth and / or have immunomodulatory activity, wherein the products are the formula milk powder described in the first aspect of the present invention.

[0074] In some specific implementations, the growth promotion includes increasing the length of the small intestine per unit weight of infants.

[0075] In some specific implementations, the immunomodulatory activity includes: increasing gut microbiota diversity, reducing harmful bacteria in the gut microbiota, and / or increasing the number of gut IgA-secreting cells.

[0076] In some specific implementations, the gut microbiota diversity is expressed as diversity using the chao index and the PD whole tree index.

[0077] In some specific embodiments, the harmful bacteria are Corynebacterium, Staphylococcus, and / or Allobaculum.

[0078] A fourth aspect of the invention further comprises providing the use of the nutritional composition of the first aspect of the invention in the preparation of products that increase the length of the small intestine per unit weight of infants.

[0079] The formula milk powder of the present invention can simultaneously increase the diversity of intestinal flora in infants (especially increase the chao index and PD whole tree index), reduce the number of harmful bacteria in the intestinal flora (especially reduce the number of Corynebacterium, Staphylococcus and / or Allobaculum) and increase the number of intestinal IgA secretory cells, but will not affect the normal growth and development, food intake and blood routine indicators of newborns.

[0080] The formula milk powder of the present invention can regulate the intestinal flora, mainly in two aspects: [1] regulating the composition of the intestinal flora: reducing the abundance of harmful bacteria Corynebacterium, Staphylococcus and Allobaculum; [2] reducing Allobaculum. Studies have shown that the content of Allobaculum is high in high-fat diets; Allobaculum isolated from IBD patients aggravates colitis in newborn mice and causes systemic antibody response of antigen-specific mucosa; it is negatively correlated with Akk bacteria.

[0081] In summary, the present invention provides a formula milk powder, its preparation method and application, wherein the formula milk powder helps to simultaneously increase the diversity of intestinal flora, reduce harmful bacteria in the intestinal flora and increase the number of intestinal IgA secretory cells, and has immunomodulatory activity. Attached Figure Description

[0082] Figure 1 The changes in piglet weight in each experimental group are shown.

[0083] Figure 2 The study shows the weight gain of piglets in each experimental group.

[0084] Figure 3 The feed intake of piglets in each experimental group is shown.

[0085] Figure 4 The effects of different nutritional compositions on the red blood cell count (A), white blood cell count (B), neutrophil count (C), and lymphocyte count (D) of piglets are shown.

[0086] Figure 5 The effects of different nutritional compositions on the CHAO index (A), GOODS Coverage index (B), Simpson index (C), and PD Whole Tree index (D) of the gut microbiota of piglets are shown.

[0087] Figure 6 The effects of different groups of nutrient compositions (Example Group 3M and Control Group 5M) on the gut microbiota species of piglets were shown. Figure 6 The horizontal axis represents different bacterial genera: Genera 1: Lachnoclostridium; Genera 2: Corynebacterium; Genera 3: Allobaculum; Genera 4: Jeotgalicoccus; Genera 5: Dubosiella; Genera 6: Staphylococcus.

[0088] Figure 7 The effects of different nutrient compositions on the weight and length of the small intestine in piglets are shown.

[0089] Figure 8 The effects of different nutritional compositions on the number of IgA-secreting cells in the piglet intestine were shown.

[0090] Figure 9 This image shows a pathological diagram of characteristic IgA-secreting cells in the small intestine. Figure 9 (A) is the control group 5M. Figure 9 (B) is Example Group 3M. Detailed Implementation

[0091] To provide a clearer understanding of the technical features, objectives, and beneficial effects of this invention, the technical solution of this invention will now be described in detail with reference to specific examples and accompanying drawings. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of this invention. Unless otherwise specifically defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art.

[0092] Example 1: Infant formula milk powder for 0-6 months of age

[0093] This embodiment provides an infant formula milk powder for infants aged 0-6 months, with a raw material composition of approximately 1000 kg:

[0094] Milk 1000kg, demineralized whey powder 300kg, blended vegetable oil (containing OPO) 236kg, lactose 210kg, galactooligosaccharides 40.5kg, α-lactalbumin powder 27kg, fructooligosaccharides 17.5kg, casein 22kg, milk fat globule membrane 5kg, phospholipids 4kg. Compound vitamins 3.5kg, compound calcium powder 12kg, compound mineral powder 1kg, potassium chloride 4kg, magnesium chloride 1.7kg, choline chloride 1.5kg. DHA 12kg, ARA 14kg, bifidobacteria 0.18kg, nucleotides 0.65kg, lactoferrin 0.65kg.

[0095] in:

[0096] (1) The total weight of the compound vitamins is 3.5 kg, and each gram of compound vitamins contains:

[0097]

[0098]

[0099] (2) The total weight of the calcium powder packets is 12kg, and each gram of calcium powder packet contains:

[0100]

[0101] (3) The total weight of the mineral pack is 1kg, and each gram of mineral pack contains:

[0102] Copper, μg Copper sulfate 3800 Iron, mg Ferrous sulfate 50 Zinc, mg Zinc sulfate 35 Manganese, μg manganese sulfate 500 Iodine, μg Potassium iodide 1000 Selenium, μg Sodium selenite 185 base material lactose —

[0103] The preparation process of the formula milk powder in this embodiment is as follows:

[0104] 1) Milk coarse filtration: After coarse filtration and degassing in the balance tank, the milk is preheated by a plate heat exchanger and then separated by a separator to remove impurities.

[0105] 2) Milk homogenization and sterilization: After removing impurities, part of the milk is homogenized in a homogenizer, while the other part is not homogenized. The homogenized milk and the unhomogenized milk are mixed and then enter the sterilization system for sterilization. After sterilization, the milk enters the mixing tank.

[0106] 3) Powder addition: Various powder raw materials are metered according to the formula and then added to the powder mixing tank for storage through the air conveying system.

[0107] 4) Vacuum powder suction: Various powder raw materials in the powder mixing tank are sucked into the mixing tank through a vacuum system.

[0108] 5) Melting and blending oil: Put the oil specified in the formula into the melting room according to the formula requirements. The temperature of the melting room should be maintained at 50-90℃. After the oil is melted, pump it into the mixed oil storage tank through the oil pump and flow meter according to the formula ratio requirements.

[0109] 6) Storage of mixed oils: Mixed oils are stored in oil storage tanks at a temperature of 40-50℃ for less than 12 hours to prevent fat oxidation.

[0110] 7) Weighing: Pump the mixed oil into the mixing tank according to the formula requirements.

[0111] 8) Adding compound nutritional fortifiers: Add vitamin packets, calcium powder packets, mineral packets, magnesium chloride packets, potassium chloride packets, and choline chloride packets separately. Dissolve each packet separately in 100-200 kg of purified water and then add them to the mixing tank. After each packet is added, rinse the addition tank and pipeline with 100 kg of purified water.

[0112] 9) Adding small ingredients: Add galacto-oligosaccharide syrup to the small hopper and then transfer it to the mixing tank.

[0113] 10) Filtration: Mix the raw materials in a mixing tank, and filter the resulting liquid through a filter screen to remove any physical impurities that may have been introduced from the raw materials.

[0114] 11) Homogenization: The mixed liquid is homogenized by a homogenizer. The first pressure is 105±5 bar and the second pressure is 32±3 bar. The fat globules are mechanically processed and dispersed into uniform fat globules.

[0115] 12) Cooling and storage: The homogenized liquid enters the plate heat exchanger for cooling. It is cooled to below 20°C and temporarily stored in the pre-storage tank. It will enter the next process within 6 hours. The agitator is turned on according to the set requirements.

[0116] 13) Concentration and sterilization: Double-effect concentration is used during production, with a sterilization temperature of ≥83℃ and a sterilization time of 25 seconds; the output concentration is 48%-52% dry matter.

[0117] 14) Concentrated milk storage, preheating and filtration, spray drying: The concentrated milk is temporarily stored in a concentrated milk balance tank; it is preheated to 60-70℃ by a scraper preheater, and after preheating, the material is filtered through a 1mm pore size filter and then spray-dried in a drying tower by a high-pressure pump. Fine powder is agglomerated at the top of the tower or in a fluidized bed as required; the inlet air temperature is 165-180℃, the outlet air temperature is 75-90℃, the high-pressure pump pressure is 160-210 bar, and the tower negative pressure is -4mbar to -2mbar.

[0118] 15) Fluidized bed drying and cooling: After the milk powder from the drying tower is dried twice in a fluidized bed (first stage), it is cooled to 25-30℃ in a fluidized bed (second stage); at the same time, the phospholipids are mixed with the carrier and heated to 60-65℃. Under the action of compressed air, the phospholipids are evenly dispersed on the surface of the milk powder, causing the powder particles to agglomerate and increase their particle size and solubility.

[0119] 16) Packaging: Weigh and seal DHA, ARA, lactoferrin, nucleotides, and bifidobacteria according to the formula requirements.

[0120] 17) Dry mixing: Mix the weighed DHA, ARA, lactoferrin, nucleotides, bifidobacteria and milk powder in a dry mixer.

[0121] 18) Sieving: The milk powder is sieved to make the particle size uniform and the powder residue is disposed of as waste.

[0122] 19) Powder discharge: Collect powder in a sterilized powder collection box and transport it from the powder discharge room to the powder loading room.

[0123] 20) Powdering: Pour the milk powder into the powder storage tank on the large and small packaging machines according to the packaging requirements.

[0124] 21) Packaging: Nitrogen-filled packaging using automatic packaging machines of different specifications; oxygen content is below 1-5% during nitrogen filling.

[0125] 22) Packing: Put the packaged small bags into the carton and add the powder scoop at the same time, and seal the carton with a sealing machine.

[0126] 23) Finished product inspection: Sampling inspection of packaged products according to the inspection plan.

[0127] 24) Warehousing and storage: Products that have passed inspection shall be stored in the warehouse at room temperature with a humidity of ≤65%.

[0128] In this embodiment of the 0-6 month infant formula, cerebrosides and sphingomyelins are mainly introduced by milk fat globule membrane powder, with the final product containing 44 mg / 100g of cerebrosides and 87 mg / 100g of sphingomyelins; zinc is mainly introduced by the mineral package, with the final product containing 4.5 mg / 100g. Based on the total mass of the formula, the total protein content is 10.50g / 100g, with whey protein accounting for 60% of the total protein; the fat content is 27.00g / 100g; the dietary fiber content is 3.0g / 100g; and the carbohydrate content is 54.6g / 100g.

[0129] Example 2: Formula milk powder for older infants aged 6-12 months

[0130] This embodiment provides a formula milk powder for infants aged 6-12 months, with a raw material composition of approximately 1000 kg:

[0131] Milk 1000kg, demineralized whey powder 250kg, blended vegetable oil (containing OPO) 183kg, lactose 250kg, galactooligosaccharides 12.5kg, α-lactalbumin powder 10kg, fructooligosaccharides 5kg, casein 10kg, milk fat globule membrane 5kg, phospholipids 4kg. Compound vitamins 3.5kg, compound calcium powder 12kg, compound mineral powder 1kg, potassium chloride 2kg, magnesium chloride 0.85kg, choline chloride 1.5kg. DHA 12kg, ARA 14kg, bifidobacteria 0.18kg, nucleotides 0.65kg, lactoferrin 0.65kg.

[0132] in:

[0133] (1) The total weight of the compound vitamins is 3.5 kg, and each gram of compound vitamins contains:

[0134]

[0135]

[0136] (2) The total weight of the calcium powder packets is 12kg, and each gram of calcium powder packet contains:

[0137]

[0138] (3) The total weight of the mineral pack is 1kg, and each gram of mineral pack contains:

[0139] Copper, μg Copper sulfate 3100 Iron, mg Ferrous sulfate 60 Zinc, mg Zinc sulfate 29 Manganese, μg manganese sulfate 810 Iodine, μg Potassium iodide 500 Selenium, μg Sodium selenite 130 base material lactose —

[0140] The preparation process of the formula milk powder in this embodiment is basically the same as that in Example 1.

[0141] In this embodiment, the infant formula for 6-12 month old infants contains cerebrosides and sphingomyelin primarily introduced by milk fat globule membrane powder, with the final product containing 43 mg / 100g of cerebrosides and 80 mg / 100g of sphingomyelin. Zinc is introduced by the mineral package, with the final product containing 4.5 mg / 100g. Based on the total mass of the formula, the total protein content is 12.00g / 100g, with whey protein accounting for 40% of the total protein; the fat content is 23.00g / 100g; the dietary fiber content is 1.91g / 100g; and the carbohydrate content is 57.1g / 100g.

[0142] Example 3: Infant formula milk powder for 12-36 month olds

[0143] This embodiment provides a formula milk powder for infants aged 12-36 months, with a raw material composition of approximately 1000 kg:

[0144] Milk 1000kg, demineralized whey powder 250kg, blended vegetable oil (containing OPO) 181kg, lactose 170kg, galactooligosaccharides 13.5kg, α-lactalbumin powder 4kg, fructooligosaccharides 6kg, casein 1kg, milk fat globule membrane 5kg, phospholipids 4kg. Compound vitamins 3.5kg, compound calcium powder 10kg, compound mineral powder 1kg, potassium chloride 2kg, magnesium chloride 0.85kg, choline chloride 1.5kg. DHA 12kg, ARA 14kg, bifidobacteria 0.18kg, nucleotides 0.65kg, lactoferrin 0.65kg.

[0145] in:

[0146] (1) The total weight of the compound vitamins is 3.5 kg, and each gram of compound vitamins contains:

[0147] Vitamin A, μg RE Retinyl acetate 1800 Vitamin D, μg Cholecalciferol 42 Vitamin E, mg α-TE dl-α-tocopherol acetate 8 <![CDATA[Vitamin K1, μg]]> Plant-based menaquinone 156 <![CDATA[Vitamin B1, μg]]> Thiamine nitrate 1620 <![CDATA[Vitamin B2, μg]]> Riboflavin 2100 <![CDATA[Vitamin B6, μg]]> Pyridoxine hydrochloride 1200 <![CDATA[Vitamin B 12 , μg]]> Cyanocobalamin 4 Niacin, μg Niacinamide 14500 Folic acid, μg folic acid 320 Pantothenic acid, μg D-Calcium pantothenate 7800 Biotin, μg D-Biotin 40 Taurine, mg Taurine 80 Inositol, mg Inositol 22 L-carnitine, mg L-carnitine tartrate 140 Vitamin C, mg Sodium L-ascorbate 125 base material lactose /

[0148] (2) The total weight of the calcium powder packets is 10kg, and each gram of calcium powder packet contains:

[0149]

[0150] (3) The total weight of the mineral pack is 1kg, and each gram of mineral pack contains:

[0151] Copper, μg Copper sulfate 3200 Iron, mg Ferrous sulfate 67 Zinc, mg Zinc sulfate 25 Iodine, μg Potassium iodide 450 base material lactose —

[0152] The preparation process of the formula milk powder in this embodiment is basically the same as that in Example 1.

[0153] In this embodiment, the infant formula milk powder for children aged 12-36 months contains cerebrosides and sphingomyelin primarily introduced through milk fat globule membrane powder, with the final product containing 47 mg / 100g of cerebrosides and 75 mg / 100g of sphingomyelin. Zinc is mainly introduced through the mineral package, with the final product containing 4.5 mg / 100g. Based on the total mass of the formula milk powder, the total protein content is 13.80g / 100g, with whey protein accounting for 20% of the total protein; the fat content is 22.00g / 100g; the dietary fiber content is 2.01g / 100g; and the carbohydrate content is 55.2g / 100g.

[0154] Efficacy experiment of phospholipid, sphingomyelin and zinc nutritional composition

[0155] [Experimental Materials]

[0156] <Experimental Base Powder>

[0157] The source information for the base powder is as follows:

[0158] Product Name: Piglet Milk Replacer

[0159] Brand: Precision

[0160] Manufacturer: Experimental Base of Chinese Academy of Agricultural Sciences, Changping District, Beijing

[0161] No significant amounts of sphingomyelin, cephalin, and zinc were detected in the base powder.

[0162] <Sphingomyelin and Cephalin>

[0163] Sphingomyelin and cephalin were commercially available and are food grade.

[0164] Zinc source

[0165] The zinc source is food-grade zinc sulfate.

[0166] [Preparation of the experimental formula powder]

[0167] <Formula Powder 1 to 6>

[0168] Formula powders 1 to 6 were prepared by adding 104 mg of each nutrient composition to 100 g of base powder. The proportions and amounts of sphingomyelin (SM), cephalin (PE), and zinc sulfate in each nutrient composition are shown in Tables 1 and 2. The proportion of zinc (Zn) is calculated based on the zinc content in zinc sulfate.

[0169] Table 1. Addition ratios of SM, PE, and Zn in each experimental group

[0170]

[0171] Table 2. Dosage of SM, PE, and Zn added to each experimental group (mg / 100g)

[0172]

[0173] (The data in Table 2 allow for an error of ±5% due to random factors such as the detection method.)

[0174] [Detection methods for SM, PE and zinc content]

[0175] To accurately determine the actual feeding amounts of the three components, the addition amounts in Table 2 were obtained by detecting the content of the three components in the actual fed formula. The Zn content in each experimental group sample was detected according to the method of GB 5009.14-2017; the SM and PE content in each experimental group sample was detected based on the LC-MS detection method.

[0176] [Evaluation of Experimental Preparation]

[0177] <Preliminary Preparations>

[0178] The experimental animals were ordinary male three-way crossbred piglets aged 2 days. The piglets weighed between 1.3 and 2.0 kg, and 10 piglets were included in each experimental group (example group 1M to 4M, control group 5M and 6M).

[0179] Animal husbandry conditions

[0180] The conditions for raising piglets shall be in accordance with GB14925-2010, specifically as follows:

[0181] Temperature: Room temperature 16-26℃;

[0182] Humidity: Relative humidity 40-70%;

[0183] Lighting: Artificial lighting, with 12 hours of alternating light and dark;

[0184] Air exchange rate: ≥8 times / hour, 100% fresh air;

[0185] <Milk preparation and feeding>

[0186] Milk preparation: Based on the nutritional needs of piglets, mix the formula powder with warm water (37-40℃) to prepare a 20% milk solution. Prepare the milk solution again every 4-5 hours.

[0187] Feeding amount: Daily feed amount for piglets = 285mL / Kg body weight on the day + 50mL.

[0188] For example, when feeding a piglet that weighs 2kg on the same day, the feeding amount is 620ml (285×2+50) of milk.

[0189] Feeding method: Divide the daily feed amount into 5 portions and feed at 9am, 1pm, 5pm, 9pm and 1am.

[0190] Animal Admission and Feeding

[0191] Select 2-day-old male piglets, weighing 1.3-2.0 kg, and introduce 5-10 piglets per batch, depending on the sow's farrowing status. Adjust the daily feed amount according to the piglets' weight on the day of farrowing. Raise the piglets until 30 days postpartum.

[0192] [Evaluation of experimental design, implementation, and results]

[0193] <Physiological and intestinal indicators of piglets>

[0194] During the experiment, the fasting weight and feed intake of piglets were recorded every morning. On the last day of the experiment (day 30), blood and small intestine samples were collected from piglets for routine blood tests, intestinal tissue analysis (intestinal weight, number of IgA secretory cells), and intestinal flora analysis.

[0195] <Fasting body weight and feed intake of piglets>

[0196] The initial weight of piglets was recorded when they were admitted to the group, and the weight of piglets was recorded at a fixed time every day during the testing phase to track changes in piglet weight.

[0197] like Figure 1 As shown, there were no significant differences in the weight changes of piglets in each group, indicating that the tested nutritional composition can meet the normal growth needs of piglets.

[0198] In addition, such as Figure 2 As shown, there were no significant differences in daily weight changes and weight gain among the groups of animals during the experiment, indicating that the tested nutritional composition can meet the normal growth needs of piglets.

[0199] like Figure 3 As shown, the daily feed intake of piglets is positively correlated with their body weight, and the trends of the two are consistent. Moreover, there are no significant differences among the groups, indicating that the tested formula does not affect the daily feed intake of piglets.

[0200] Evaluation results of complete blood total white blood cell count, white blood cell differential analysis (neutrophils and lymphocytes), and routine blood tests.

[0201] Blood cells can be divided into three categories: red blood cells (RBCs), white blood cells (WBCs), and platelets. White blood cells can be further divided into five types: neutrophils, eosinophils, basophils, monocytes, and lymphocytes. Neutrophils are the most abundant type of white blood cell, accounting for 50-70%, while lymphocytes account for 20%-40%. All types of white blood cells participate in the body's defense functions, primarily by phagocytosis to clear invading bacteria and viruses, and by forming antibodies or sensitizing lymphocytes to destroy or eliminate invading pathogens.

[0202] Whole blood was collected from the jugular vein of piglets during dissection and analyzed using a fully automated blood biochemistry analyzer.

[0203] like Figure 4 As shown, there were no significant differences in red blood cell count and white blood cell count among the experimental groups (all within the normal range), and there were no significant differences in neutrophil count and lymphocyte count among the experimental groups (all within the normal range).

[0204] The results indicate that none of the tested nutritional compositions had any effect on the blood routine indicators of piglets.

[0205] [Gut microbiota analysis]

[0206] <Analysis of Gut Microbiota Alpha Diversity>

[0207] During dissection, rectal contents were taken for 16S microbial diversity analysis. Primer region: V3V4. Forward primer: 343F - 5’-TACGGRAGGCAGCAG-3’; Reverse primer: 798R - 5'-AGGGTATCTAATCCT-3'.

[0208] As Figure 5 shown, the α-diversity of Example groups 2M, 3M, and 4M showed that the chao index and PD whole tree index were significantly higher than those of Control groups 5M and 6M.

[0209] <Analysis of differential microbial communities (species differences at the genus level)>

[0210] Compared with Control groups 5M and 6M, Example groups 1M - 4M were able to regulate the intestinal flora composition, especially significantly reducing the abundances of harmful bacteria such as Corynebacterium, Staphylococcus, and Allobaculum. Figure 6 An intuitive comparison between Example group 3M and Control group 5M is illustrated.

[0211] <Evaluation of total small intestine weight and length>

[0212] During dissection, the entire small intestine of piglets was taken and its length was measured with a tape measure. The weight of the small intestine was measured using a platform scale. Since the length and weight of the small intestine vary greatly among animals with different body weights, the small intestine length (cm) / body weight (kg) and small intestine weight (g) / body weight (kg) were used for statistical analysis. As Figure 7 shown, compared with Control groups 5M and 6M, there was no significant difference in the small intestine weight of each experimental group; but compared with Control groups 5M and 6M, Example groups 1M - 4M could increase the small intestine length per unit body weight of piglets. Especially, Example groups 1M and 2M could significantly increase the small intestine length per unit body weight of piglets, and Example group 2M had the best effect.

[0213] <Evaluation of the number of intestinal IgA-secreting cells in piglets>

[0214] For each experimental group of piglets, the number of intestinal IgA-secreting cells was analyzed using immunohistochemistry (IHC) method, and optical density analysis was used instead of cell counting. Specifically, tissue specimens fixed with 4% paraformaldehyde were paraffin-embedded, sectioned, stained, and then the target positive expression was observed under a microscope. As Figure 8 shown, compared with Control groups 5M and 6M, Example groups 1M - 4M could all significantly increase the number of intestinal IgA-secreting cells in piglets.

[0215] In addition, Figure 9(A) and (B) respectively illustrate the pathological features of small intestinal IgA secretory cells in the control group and the example group. Figure 9 In the image, the stained dots represent sIgA-secreting cells. Figure 9 The dots after staining (B)(3M) are larger than those after staining. Figure 9 The number of dots after staining with (A)(5M) was significantly greater, meaning that the number of sIgA secreting cells was significantly increased in the 3M group compared to the 5M group.

[0216] The pathological images of small intestinal IgA secretory cells provide direct evidence that the nutritional composition of the present invention can increase the number of small intestinal IgA secretory cells in piglets.

[0217] The above results indicate that the formula milk powder with the added nutritional composition of the present invention can simultaneously increase the diversity of intestinal flora in newborn mammals, reduce harmful bacteria in the intestinal flora, and increase the number of intestinal IgA secretory cells, without affecting the growth, development, food intake, and blood routine indicators of newborn mammals.

[0218] In addition, the formula milk powder of the present invention also has the effect of increasing the length of the small intestine in newborn mammals.

[0219] The above embodiments are only used to illustrate the technical solutions of the present invention, and are not intended to limit them. It is obvious to those skilled in the art that modifications can be made to the technical solutions described in the above embodiments, or equivalent substitutions can be made to some of the technical features, without departing from the general scope of the present invention.

Claims

1. Use of a composition consisting of sphingomyelin, cephalin and zinc for the manufacture of a product having a reduced abundance of harmful bacteria in the intestinal flora, wherein, The harmful bacteria are Corynebacterium and Heterobacterium. The product is a formula milk powder containing sphingomyelin, cephalin, and zinc. By weight ratio, sphingomyelin / zinc = A, cephalin / zinc = B, and the ratio of A:B is (0.6-2.25):

1. Relative to 100% mass of the total amount of sphingomyelin and cephalin, the zinc content is 3% to 6% by mass. Furthermore, based on the total mass of the formula milk powder, the total protein content is 9-27g / 100g, the whey protein accounts for 20%-100% of the total protein, the fat content is 10-35g / 100g, the dietary fiber content is 0.1-8.0g / 100g, and the carbohydrate content is 40-60g / 100g. Relative to 100g of formula milk powder, the total amount of sphingomyelin, cephalin, and zinc is 100mg-140mg.

2. The application according to claim 1, wherein: The raw materials that provide protein include any one or more of the following: raw milk, whole milk powder, skim milk powder, demineralized whey powder, casein powder, and whey protein powder; The raw materials providing fat include any one or more of milk fat raw materials and vegetable oils; the milk fat raw materials include one or more of raw milk, whole milk powder, and anhydrous milk; the vegetable oils include one or more of sunflower seed oil, corn oil, soybean oil, low erucic acid rapeseed oil, coconut oil, flaxseed oil, palm oil, perilla oil, walnut oil, and OPO structured oils. Raw materials that provide dietary fiber include any one or more of fructooligosaccharides, galactooligosaccharides, polyfructooligosaccharides, raffinose, and human milk oligosaccharides; The raw materials that provide carbohydrates include lactose.

3. Use according to claim 2, wherein, The whey protein powder includes α-lactalbumin powder and milk fat globule membrane powder.

4. The use according to any one of claims 1 to 3, wherein, The formula milk powder mentioned is infant formula milk powder.

5. The application according to any one of claims 1-3, wherein, The formula milk powder is for infants aged 0-6 months, for older infants aged 6-12 months, or for toddlers aged 12-36 months.

6. The application according to any one of claims 1-3, wherein, The formula milk powder is a special medical purpose formula food.

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