CN114568526A - Mother emulsified infant formula powder for improving intestinal microenvironment health and application thereof - Google Patents

Abstract

The invention provides a maternal emulsified infant formula powder for improving intestinal microenvironment health and application thereof. Specifically, the invention provides an application of breast milk oligosaccharide in preparing infant formula powder for improving intestinal microenvironment health, and also provides infant formula powder with the effect of improving intestinal microenvironment health, wherein the breast milk oligosaccharide is fucosyl oligosaccharide, sialyl oligosaccharide or lacto-N-tetrasaccharide; the improving the intestinal microenvironment health comprises: regulate production of butyric acid in the gut system, increase overall production of short chain fatty acids, be utilized by the gut flora as prebiotics in the gut system and produce gas, reduce production of isobutyric acid and/or isovaleric acid, and/or lower pH to maintain intestinal microenvironment health. According to the technology provided by the invention, the breast milk oligosaccharide is added into the infant formula powder, so that the intestinal microenvironment health can be improved, and the generation of butyric acid can be regulated and controlled.

Description

Breastmilk infant formula powder for improving intestinal microenvironment health and its application

technical field

The present invention relates to a new application of breast milk oligosaccharides, specifically, to the application of breast milk oligosaccharides in the preparation of infant formula powder with the effect of improving intestinal microenvironment health (especially reducing intestinal branched-chain fatty acids). , and the prepared infant formula powder and related preparation method.

Background technique

Human Milk Oligosaccharides (HMOs) are the third most abundant substances in breast milk except lactose and fat. Its total content varies during the various stages of lactation and is approximately 12-14 g/L in mature milk and approximately 20-24 g/L in colostrum. The structure of each breast milk oligosaccharide has a lactose at the reducing end, most of which have polylactosamine as the structural backbone, and contain fucose, sialic acid or both at the chain end. Breast milk oligosaccharides are mainly composed of three categories: fucosyl oligosaccharides, represented by 2'-fucosyl oligosaccharides and 3'-fucosyl oligosaccharides; sialic acid-based oligosaccharides; Oligosaccharides, represented by 3'-sialyllactose and 6'-sialyllactose; oligosaccharides formed by the core sugar chain structure without fucosyl or sialic acid N-tetrasaccharides and lactose-N-neotetrasaccharides are representative substances. The presence and content of HMOs vary among individuals and are related to the Lewis-secretory composition of nursing mothers. Since the raw material of infant formula is usually cow's milk, and cow's milk usually contains no or very little of these oligosaccharides, HMOs have become a gap that infant formula powder must bridge to be closer to the composition of breast milk.

Intestinal flora is an important constituent of the human intestinal micro-ecosystem and plays an important role in human health. The anaerobic bacilli, bifidobacteria, eubacteria, streptococci and lactobacilli in the intestinal flora can release metabolites short chain fatty acids (SCFA) by fermenting carbohydrates, proteins and lipids. It mainly includes acetic acid, propionic acid and butyric acid. SCFA can regulate various physiological functions of the body and play an important role in regulating the health of the intestinal microenvironment. For example, SCFA can provide energy and regulate electrolytes, acetic acid is an important source of host energy, propionic acid can participate in the process of reversing pyruvate into glucose, and butyric acid is taken up by epithelial cells and is the main energy source for epithelial cells. SCFAs also have anti-inflammatory, intestinal barrier function and antibacterial properties. The SCFA released by the fermentation of intestinal flora can reduce the intestinal pH, thereby increasing the growth of beneficial bacteria in the intestine and reducing the proliferation of harmful bacteria. Other physiological functions of SCFA include reducing the risk of early-onset type 1 diabetes, reducing anxiety, and promoting bone growth. The production of butyrate was significantly associated with a lower incidence of neonatal skin surface allergy, food allergy, and respiratory allergy.

At present, infant formula powder needs a solution that can generate butyric acid and regulate the micro-ecological health of infants and young children. According to the content and composition of breast milk oligosaccharides, the inventors have developed a carbohydrate formula close to that of breast milk carbohydrates, which provides a technical basis for breast emulsification of infant formula powder in the future. Combined with the existing breast milk research direction, continue to retain the α+β protein combination of protein breast milk, and develop new infant formula powder, which not only improves the baby's digestion and absorption of protein, enhances the baby's resistance, but also removes oligosaccharides from breast milk. The angle gives the baby double care.

SUMMARY OF THE INVENTION

One object of the present invention is to provide a new application of breast milk oligosaccharide, particularly its application in the preparation of breast milk infant formula powder with the effect of improving the health of intestinal microenvironment.

Another object of the present invention is to provide a breast milk formula powder for infants and young children.

Another object of the present invention is to provide an application of breast milk formula powder for infants and young children.

The present invention finds that some breast milk oligosaccharides have the effect of significantly improving the health of the intestinal microenvironment, which is embodied in regulating the production of butyric acid in the intestinal system, increasing the overall production of short-chain fatty acids, and acting as prebiotics in the intestinal system. The gut microbiota utilizes and produces gas, reduces the production of isobutyric acid, isovaleric acid and/or lowers pH to maintain a healthy gut microenvironment, thus providing a new application of breast milk oligosaccharides.

Specifically, the present invention provides the application of breast milk oligosaccharides in preparing food for improving the health of intestinal microenvironment, wherein the breast milk oligosaccharides are fucosyl oligosaccharides, sialic acid-based oligosaccharides Oligosaccharides or lactose-N-tetrasaccharides.

The invention also provides an infant formula powder with the effect of improving the health of the intestinal microenvironment, which contains breast milk oligosaccharides, and the breast milk oligosaccharides are fucosyl oligosaccharides, sialic acid-based oligosaccharides sugar or lactose-N-tetrasaccharide. Wherein, the improving the health of the intestinal microenvironment includes: the improving the health of the intestinal microenvironment includes: regulating the production of butyric acid in the intestinal system, increasing the overall production of short-chain fatty acids, and being absorbed by the intestines as a prebiotic in the intestinal system. The gut flora utilizes and produces gas, reduces isobutyric acid and/or isovaleric acid production, and/or lowers pH to maintain a healthy gut microenvironment.

Known breast milk oligosaccharides include fucosyllactose, sialyllactose, and basic sugar chain structures of breast milk oligosaccharides without fucosyl or sialic acid groups (typical representative substances include lactose-N-tetrasaccharide and its isomer lactose-N-neotetraose).

Among them, 2'-fucosyllactose (2'-fucosyllactose, 2'-FL or 2-FL or 2FL) is a trisaccharide structure formed by fucose and lactose, and is a representative of fucosyl oligosaccharides. sexual substances. Commercially available substances are usually prepared by microbial fermentation and have the same structure as oligosaccharides found in human milk.

3-fucosyllactose (3-fucosyllactose, 3'-FL or 3-FL or 3FL) is a trisaccharide structure formed by fucose and lactose, and is identical to 2'-fucosyllactose Construct. It is a representative substance of fucosyl oligosaccharides. This substance is prepared by microbial fermentation and has the same structure as oligosaccharides found in human milk.

Lactose-N-tetraose (LNT) is a hexasaccharide structure formed by lactose and tetrasaccharide. It is an oligomer based on a core sugar chain and does not contain fucosyl or sialic acid groups. A representative substance of sugar. This substance is prepared by microbial fermentation and has the same structure as oligosaccharides found in human milk.

3'-sialyllactose (3'-sialyllactose, 3'-SL or 3-SL or 3SL), a trisaccharide structure formed by sialic acid and lactose, is a representative substance of sialic acid-based oligosaccharides. This substance is prepared by microbial fermentation and has the same structure as oligosaccharides found in human milk.

6'-sialyllactose (6'-sialyllactose, 6'-SL or 6-SL or 6SL), a trisaccharide structure formed by sialic acid and lactose, is a representative substance of sialic acid-based oligosaccharides. This substance is prepared by microbial fermentation and has the same structure as oligosaccharides found in human milk.

According to a specific embodiment of the present invention, the infant formula powder with the effect of improving intestinal microenvironment health of the present invention contains 14.2-1515.3 mg/100g of breast milk oligosaccharides, or 0.02 mg of breast milk oligosaccharides in terms of milk. -2.0g/L.

According to a specific embodiment of the present invention, the infant formula powder with the effect of improving the health of the intestinal microenvironment of the present invention is an infant formula powder, which contains 14.2-1515.3 mg/100g of breast milk oligosaccharides, and the infant formula The total protein content of the powder is 10~19g/100g, the proportion of whey protein to total protein is 38~70%, the content of α-whey protein is 1.25~2.5g/100g, and the content of β-casein is 2.2~4.0g /100g, the fat content is 17～29g/100g, the linoleic acid content is 2500～4500mg/100g, the α-linolenic acid content is 330～450mg/100g, the dietary fiber is 0.95～6.3g/100g, and the dietary fiber includes low Polygalactose, fructooligosaccharides and/or breast milk oligosaccharides, the carbohydrate content is 50-57g/100g.

The breast milk oligosaccharide-containing breast milk formula powder for infants and young children of the present invention has components, especially the composition and proportion of milk protein, fat and oligosaccharide, which are closer to breast milk, and the feeding effect is also closer to breast milk, which can improve the immunity of infants and young children. force.

The inventor of this case collected colostrum and mature breast milk, and collected bovine colostrum and mature milk samples in key areas of the country, and conducted a large number of research and analysis, focusing on the adjustment of the microstructure of the formula powder. Breast milk is designed, and specific raw materials are selected to prepare infant formula powder, so that the formula powder is closer to breast milk in terms of subdivision composition and feeding effect.

The inventor of this case has conducted an analysis and research on the nutrients required for the growth and development of infants aged 0-3 years, and investigated the corresponding feeding situations of various infant milk powders on the market, focusing on the adjustment of the microstructure of the formula powder, and the protein , fat, dietary fiber (prebiotics) and other aspects have been adapted to provide an infant formula powder containing breast milk oligosaccharides suitable for infants aged 0-3 years old. The formula powder can improve the intestinal bacteria of infants and young children. group, boost immunity.

According to a specific embodiment of the present invention, in the breast milk oligosaccharide-containing breast milk infant formula powder of the present invention, the total protein content is 10-19 g/100 g, and the total protein mainly includes milk protein. In addition, the proportion of whey protein in the total protein is usually controlled to be 38% to 70%. Specifically, the raw material for providing milk protein includes one or more of basic raw milk, whole milk powder, skimmed milk powder, whey protein powder, and demineralized whey powder; preferably, based on 1000 parts by weight of the milk containing breast milk The oligosaccharide breast milk formula powder for infants and young children, its raw materials include: 850-1750 parts by weight of raw cow milk, 0-275 parts by weight of skimmed milk powder, the raw cow milk and skim milk powder can be partially or completely used in an equivalent amount of whole milk powder, Skim milk substitute. Further, in whey protein powder (such as whey protein powder WPC 80%, whey protein powder WPC 34%, etc.), demineralized whey powder (such as demineralized whey powder D70, D90, etc.) added to fortify whey protein One or more, preferably including desalinated whey powder, and whey protein powder (eg whey protein powder WPC 80% and/or whey protein powder WPC 34%); and alpha-lactalbumin in fortified products The raw material α-whey protein powder is further added, and the raw material β-casein powder is further added to strengthen the β-casein in the product; preferably, based on 1000 parts by weight of the one containing breast milk oligosaccharides The breast milk formula powder for infants and young children comprises: 0-170 parts by weight of whey protein powder (preferably including 0-170 parts by weight of whey protein powder WPC34%); 25-225 parts by weight of demineralized whey powder; 3-40 parts by weight of albumin powder; 0.5-25 parts by weight of β-casein powder.

In the breast milk oligosaccharide-containing breast milk oligosaccharide-containing breast milk formula for infants and young children, in addition to the basic raw materials containing milk fat (such as the aforementioned raw milk, skimmed milk powder), the raw material for providing fat may also include vegetable oil, and the vegetable oil may include sunflower. One or more of seed oil, corn oil, soybean oil, canola oil, coconut oil, palm oil, walnut oil, preferably including sunflower oil, corn oil and soybean oil, the addition of these vegetable oils is one aspect To provide the product with fat components, on the other hand, linoleic acid and α-linolenic acid (preferably, the content of α-linolenic acid in the milk powder of the present invention is 310-450 mg/100 g). In addition, the raw material for providing the fat may optionally include the raw material OPO structural lipid added to provide 1,3-dioleate-2-palmitic acid triglyceride. Due to the different purities of OPO structural lipid raw materials currently on the market, that is, the content of the active ingredient 1,3-dioleic acid-2-palmitic acid triglyceride varies, usually about 40% to 70%. In order to distinguish the active ingredient 1,3-dioleic acid-2-palmitic acid triglyceride and its raw materials, the term "1,3-dioleic acid-2-palmitic acid triglyceride" is used when describing the active ingredient, When describing food raw materials that provide the active ingredient 1,3-dioleic acid-2-palmitic acid triglyceride, the commonly known "OPO structural lipid" is used. The specific addition amount of the OPO structural fat can be converted according to the content requirements of the 1,3-dioleic acid-2-palmitic acid triglyceride in the milk powder product of the present invention and the raw material purity of the OPO structural fat. More preferably, based on 1000 parts by weight of the breast milk oligosaccharide-containing breast milk oligosaccharide-containing infant formula powder, the raw materials include: 0-80 parts by weight of sunflower oil; 0-40 parts by weight of corn oil; 0-80 parts by weight of soybean oil parts by weight; OPO structural grease 0-140 parts by weight.

Preferably, the content of linoleic acid and α-linolenic acid in the raw materials sunflower oil, corn oil, soybean oil and OPO structural fat used in the present invention are 7.6-8.9%, 0.25-0.38%, 53.0-56.20%, 0.9-1.6%, 50.0-53.5%, 7.6-9.6%, 5.9-6.3%, 0.4-0.62%, the content of canola oil linoleic acid and α-linolenic acid used are 16-19%, 8.0-10.6%, and the contents of linoleic acid and alpha-linolenic acid in coconut oil are 1-3% and 0-1% respectively. The effective content of 1,3-dioleic acid-2-palmitic acid triglyceride in the OPO structural lipid raw material is 40% to 70%.

According to a specific embodiment of the present invention, in the infant formula powder with the effect of improving the health of the intestinal microenvironment of the present invention, the raw materials for providing breast milk oligosaccharides are directly derived from commercial breast milk oligosaccharides.

According to a specific embodiment of the present invention, the breast milk oligosaccharide-containing breastmilk infant formula powder of the present invention may further contain probiotics, and the probiotics are preferably bifidobacteria. Preferably, based on 1000 parts by weight of the breast milk oligosaccharide-containing breastmilk infant formula powder, the amount of bifidobacteria added is 0.1-0.2 parts by weight; more preferably 0.18-0.2 parts by weight. More preferably, the amount of bifidobacteria contained in each part by weight of bifidobacteria powder is 3×10 ¹⁰ CFU or more.

According to a specific embodiment of the present invention, in the breast milk oligosaccharide-containing breast milk infant formula powder of the present invention, a part of carbohydrates comes from lactose-containing basic raw materials such as milk, whole milk powder and/or skimmed milk powder, etc., and additionally added Lactose raw material to provide carbohydrates. That is, in the infant formula powder of the present invention, the raw material for providing carbohydrates includes raw lactose in addition to the basic raw material containing lactose. Preferably, based on 1000 parts by weight of the breast milk oligosaccharide-containing breast milk infant formula powder, the raw materials include: 125-325 parts by weight of lactose. The specific addition amount of lactose can be adjusted within the range so that the carbohydrate content of the breast milk oligosaccharide-containing breastmilk infant formula powder of the present invention is preferably 52-56 g/100 g.

According to a specific embodiment of the present invention, the raw material of the breast milk oligosaccharide-containing breast milk infant formula powder of the present invention further includes one or more of appropriate DHA, ARA, lactoferrin, etc., and may also include Nutrient complex containing calcium powder, vitamins and minerals, in addition to the carrier anhydrous cream and phospholipids used in the spray drying process for milk powder preparation. Preferably, based on 1000 parts by weight of the breast milk oligosaccharide-containing breast milk infant formula powder, the raw materials include: 2 to 12 parts by weight of DHA; 3 to 15 parts by weight of ARA; a compound comprising calcium powder, vitamins and minerals 8-16 parts by weight of nutrients; 1-4 parts by weight of phospholipids; 0-2 parts by weight of anhydrous cream.

In the breast milk oligosaccharide-containing breast milk oligosaccharide-containing formula powder for infants and young children, the compound nutrients are combinations of nutritional components that meet national standards, and different addition amounts are used according to different formulas. The formula powder of the present invention can selectively adopt any one or any combination of the following compound nutrients if nutrients are added as needed. Preferably, the compounded nutrients at least include compounded vitamins, calcium powder, and mineral nutrition packs, and the dosage of each component is:

1) Compound vitamins, in each gram of compound vitamins:

Vitamin A: 1700～5800μgRE

Vitamin D: 30～70μg

Vitamin B1: 3600～6800μg

Vitamin B2: 3500～6900μg

Vitamin B6: 2400～4000μg

Vitamin B12: 8～20μg

Vitamin K1: 400～700μg

Vitamin C: 330～700mg

Vitamin E: 27～70mgα-TE

Niacinamide: 26000～41550μg

Folic acid: 700～920μg

Biotin: 100～245μg

Pantothenic acid: 12000～25230μg

2) Mineral II, per gram of Mineral II:

Calcium: 300～455mg

Phosphorus: 75～150mg

3) Minerals, per gram of minerals:

Iron: 40～85mg

Zinc: 23～48mg

Copper: 2600～4180μg

Iodine: 500～995μg

Selenium: 0～200μg

Manganese: 0～579μg

4) Magnesium chloride, per 1000 kg of milk powder:

Magnesium: 80～170g

5) Potassium chloride, per 1000 kg of milk powder:

Potassium: 450～1000g.

In addition, the compounded nutrients may optionally include choline chloride (300-950 g of choline per 1000 kilograms of milk powder), and the compounded nutrients may optionally include magnesium chloride (per 1000 kilograms of milk powder). Magnesium 80-170g), potassium chloride (450-1000g potassium per 1,000 kg of milk powder) can optionally be included in the compounded nutrients, and taurine (180-1000g potassium per 1000 kg of milk powder) can be optionally included in the compounded vitamins 340mg/g). The base material of the compound nutrients is preferably lactose. Preferably, based on 1000 parts by weight of the breast milk oligosaccharide-containing breast milk infant formula powder, the amount of compound nutrients added is 7-16 parts by weight, wherein the compound vitamin nutrition package is preferably 2-3 parts by weight In parts by weight, the calcium powder nutrition package is preferably 2-6 weight parts, the mineral nutrition package is preferably 0.5-3 weight parts, and the base material of each nutrition package is preferably lactose. The content of each component of the above-mentioned compound nutrients refers to the addition amount of the nutrient substance for strengthening, excluding the content of nutrient components in other raw materials of milk powder, for example, calcium powder (calcium carbonate), "calcium" in every 1000 kg of milk powder. : 1300 ~ 1600g" means to strengthen the calcium element in the product, based on 1000 kg of milk powder, adding calcium powder (eg calcium carbonate) in which the weight of the calcium element is 1300 ~ 1600g.

In the breast milk oligosaccharide-containing breast milk oligosaccharide-containing infant formula powder, the raw material phospholipids and anhydrous cream are mainly used for forming powder particles in the spray drying process, and the phospholipids can be soybean lecithin and/or lecithin. Phospholipids and anhydrous cream are used less, but also contribute to the fat content of milk powder products.

According to a preferred specific embodiment of the present invention, a breast milk oligosaccharide-containing breast milk infant formula powder of the present invention, its raw materials include:

7 to 16 parts by weight of compound nutrients containing calcium powder, vitamins and minerals;

2 to 12 parts by weight of DHA;

3-15 parts by weight of ARA;

Bifidobacterium 0.1 to 0.2 parts by weight.

It can be understood that, in the breast milk oligosaccharide-containing breastmilk infant formula powder of the present invention, the specific dosage of each raw material should be adjusted and determined on the premise that the product index requirements of the formula powder are met. In the breast milk oligosaccharide-containing breast milk infant formula powder of the present invention, the product performance indicators not specified or listed should be implemented in accordance with the national standards for infant formula food and relevant standards and regulations.

In the breast milk oligosaccharide-containing breast milk oligosaccharide formula powder for infants and young children of the present invention, each raw material can be obtained commercially, and the selection of each raw material should meet the requirements of relevant standards, wherein the breast milk oligosaccharide should also meet the requirements of the present invention. . In addition, the compounded nutrients can also be compounded by themselves. In the present invention, "compound" is only used for convenience of expression, and it does not mean that each component in the compound must be mixed together before application. All raw materials should be added and used under the premise of meeting relevant regulations.

On the other hand, the present invention also provides a method for preparing the breast milk oligosaccharide-containing breast milk formula powder for infants and young children. Other raw materials are mixed to prepare the infant formula powder. The technical process of its preparation mainly includes: batching, homogenization, concentration and sterilization, spray drying, and dry mixing to obtain a finished product. Specifically, the method for preparing the breast milk oligosaccharide-containing breast milk infant formula powder of the present invention comprises:

Ingredients: Mix the raw milk and formula powder with other raw materials except the spray carrier (phospholipid and anhydrous cream) and post-mixtures (such as DHA, ARA, lactoferrin, bifidobacteria, etc.) to obtain a mixed liquid ;

Homogenization: The mixed liquid is homogenized;

Concentration and sterilization: the homogenized feed liquid is concentrated and sterilized. The conditions of concentration and sterilization are: double-effect concentration, sterilization temperature ≥83°C, sterilization time 20-30 seconds, and control the discharge concentration to be 48%-52% dry matter;

Spray drying: The thick milk is preheated to 60~70℃ by a scraper preheater. After preheating, the material is filtered by a filter with a pore size of 0.8~1.2mm, and then sprayed into a drying tower for spray drying. The spray drying conditions are controlled as follows: The air temperature is 165-180℃, the exhaust air temperature is 75-90℃, the pressure of the high-pressure pump is 160-210bar, and the negative pressure of the tower is -4--2mbar;

Fluidized bed drying and cooling: the powder from the drying tower is then dried by the primary fluidized bed, and then cooled to 25-30°C by the secondary fluidized bed, and heated to 60-60°C under the action of compressed air. The mixture of phospholipid (lecithin and/or soybean phospholipid) and carrier (anhydrous cream) at ℃ is uniformly dispersed on the surface of the powder to obtain powder particles;

Post-mixing: dry-mixing the post-mixing (DHA, ARA, lactoferrin, bifidobacteria) with the powder particles after drying and cooling in the fluidized bed, and packaging to obtain the finished milk powder.

According to a specific embodiment of the present invention, in the preparation method of the breast milk oligosaccharide-containing breastmilk infant formula powder of the present invention, the order of adding nutrients and stirring and dissolving time are also very important. , Minerals are added to the other powder and oil mixture in sequence, and after adding calcium powder, stir and dissolve for 15-25 minutes, add vitamins and stir and dissolve for 15-20 minutes, add minerals and stir and dissolve for 15-20 minutes. minute. The inventor found in the research that calcium can be combined with casein in milk protein to form colloidal calcium by adding calcium powder first, so the precipitation of calcium can be avoided or weakened. However, this combining process requires sufficient time. At the same time, if calcium and minerals are added at the same time, the calcium powder can adsorb some minerals such as iron, which can cause it to change from ferrous iron to ferric iron, and ferric iron is red, so it is easy to cause yellow to red precipitation. Secondly, adding vitamins, the purpose is that the vitamins contain ingredients that are easily oxidized by metal ions, such as VC. If minerals are added first, the loss of vitamins will be caused due to the high local vitamin concentration. And if the local concentration of VC is too high, it will accelerate the oxidation of ferrous iron. Minerals are added at the end because minerals are dispersed in water slower than vitamins. At the same time, in the process of mineral dispersion, a water film must be formed on the surface of its particles to stabilize it and not cause damage to milk proteins. This process requires sufficient in between. Adding minerals after the vitamins are fully mixed with milk and diluted can reduce the loss of minerals to vitamins.

According to a specific embodiment of the present invention, in the preparation method of the breast milk oligosaccharide-containing breast milk infant formula powder of the present invention, the batching process includes:

Coarse filtration of milk: After the milk is coarsely filtered and degassed in the balance tank, it is preheated by the plate heat exchanger, and then the impurities are separated by the separator.

Milk homogenization and sterilization: Part of the raw milk after removing impurities enters the homogenizer for homogenization, and the other part is not homogenized. After homogenization, the two are mixed and entered into the sterilization system for sterilization.

Powder addition: All kinds of powder raw materials are uniformly added to the powder mixing tank through the air delivery system after being metered according to the formula, and sucked into the vacuum mixing tank through the vacuum system;

Dissolving oil: put the oil specified in the formula into the carburetor room according to the formula requirements, the temperature in the carburetor room is kept at 50~90℃, after the oil is dissolved, put it into the mixed oil storage tank, and mix the oil according to the formula requirements. The oil is pumped into the mixing tank by the oil pump;

Nutrient dissolution and addition: calcium powder, vitamins and minerals are dissolved in pure water respectively, and then added to the mixing tank in sequence to obtain a mixed material liquid.

In a specific embodiment of the present invention, the preparation method of the breast milk oligosaccharide-containing breast milk infant formula powder of the present invention is carried out according to the following operations:

1) Addition of powder: All kinds of powder raw materials are metered according to the formula and added to the powder mixing tank uniformly through the air delivery system for storage.

2) Vacuum powder suction: various powder raw materials in the powder mixing tank are sucked into the vacuum mixing tank through the vacuum system.

3) Dissolving oil distribution: Put the grease specified in the formula into the carburetor room according to the formula requirements, the temperature in the carburetor room should be kept at 50~90℃, and after the oil is melted, inject it through the oil pump and flowmeter according to the formula requirements in the mixed oil storage tank.

4) Storage of mixed oil: The mixed oil is stored in an oil storage tank at a temperature of 40 to 50 °C, and the storage time is less than 12 hours to prevent fat oxidation.

5) Weighing: According to the formula requirements, the mixed oil is pumped into the mixing tank through the oil pump.

6) Nutrient dissolution and addition: calcium powder, minerals, vitamins, etc. are added separately, and 100-200kg of pure water is used to dissolve them separately, and then put into a wet mixing tank. After each type of filling, rinse the addition tank and pipeline with 100kg of pure water.

7) Filtration: The mixed feed liquid is filtered through a filter screen to remove the physical impurities that may be brought into the raw material.

8) Homogenization: the mixed feed liquid is homogenized by a homogenizer, the first-stage pressure of homogenization is 105±5bar, and the first-stage pressure of homogenization is 32±3bar, and the fat globules are mechanically processed to disperse them into uniform Consistent fat globules.

9) Cooling and storage: The homogenized feed liquid enters the plate heat exchanger for cooling: it is cooled to below 20°C, temporarily stored in the pre-storage tank, and enters the next process within 6 hours, and the agitator is turned on according to the set requirements.

10) Concentration and sterilization: double-effect concentration is used in production, the sterilization temperature is ≥83°C, and the sterilization time is 25 seconds. The discharge concentrations are all 48% to 52% dry matter.

11) Thick milk storage, preheating filtration, spray drying: the concentrated milk is temporarily stored in the thick milk balance tank. It is preheated to 60~70℃ by a scraper preheater. After preheating, the material is filtered by a filter with a 1mm aperture, and then pumped into a drying tower for spray drying with a high-pressure pump. The fine powder is agglomerated at the top of the tower or in a fluidized bed as required. . Inlet air temperature: 165～180℃, exhaust air temperature 75～90℃, high pressure pump pressure 160～210bar, tower negative pressure -4～-2mbar.

12) Fluidized bed drying and cooling: the powder from the drying tower is then dried by the fluidized bed (first stage) and then cooled to 25-30°C through the fluidized bed (secondary). At the same time, the phospholipid is mixed with the carrier and heated to 60-65°C. Under the action of compressed air, it is uniformly dispersed on the surface of the powder to make the powder particles agglomerate and increase its particle size and instant solubility.

13) Packing: According to the formula requirements, the personnel in the milling workshop will weigh and pack DHA, ARA lactoferrin and Bifidobacterium into sealed bags.

14) Dry mixing: Mix the weighed post-mix (DHA, ARA, Lactoferrin, Bifidobacterium) and milk powder in a dry mixer.

15) Sieve powder: The powdered milk powder is uniform in particle size through a vibrating sieve, and the powder residue is discarded.

16) Powder output: use a sterilized powder collecting box to collect powder, and transport it from the powder output room to the powder room.

17) Powdering: Pour the milk powder into the powder storage tank on the large and small packaging machine according to the packaging requirements.

18) Packing: 400g automatic packing machine filled with nitrogen. The oxygen content during nitrogen filling is less than 1%. The oxygen content of 900 grams of iron cans with automatic nitrogen filling is less than 5%.

19) Packing: put the packaged sachet into the carton and add the powder scoop at the same time, and seal it with a carton sealing machine.

20) Finished product inspection: Sampling inspection of the packaged products according to the inspection plan.

21) Storage in storage: The products that have passed the inspection are stored in storage, and are required to be stored at room temperature with humidity ≤65%.

In the present invention, the breast milk oligosaccharides can be mixed together during the batching, or can be added during the post-mixing process.

On the other hand, the present invention also provides the application of the infant formula powder in the preparation of food with the effect of improving the health of the intestinal microenvironment. Among them, the improvement of the health of the intestinal microenvironment includes: regulating the production of butyric acid in the intestinal system, increasing the overall production of short-chain fatty acids, and being used as a prebiotic in the intestinal system by the intestinal flora and producing gas, reducing the abnormal Production of butyric acid and/or isovaleric acid, and/or lowering of pH to maintain a healthy gut microenvironment.

According to a specific embodiment of the present invention, in the infant formula powder with the effect of improving intestinal microenvironment health of the present invention, the breast milk oligosaccharide is a fucosyl-based oligosaccharide or a sialic acid-based oligosaccharide or Lactose-N-tetrasaccharide. Preferably, the fucosyl oligosaccharide is 2'-FL and/or 3-FL, and the sialic acid oligosaccharide is 3-SL and/or 6-SL.

According to some specific embodiments of the present invention, the breast milk oligosaccharide in the infant formula powder of the present invention is used for regulating the production of butyric acid in the proximal colon, and the breast milk oligosaccharide is 6-SL. Under this application, the application amount of 6-SL in milk powder is 14.2-1515.3mg/100g powder, or 0.02-2.0g/L in terms of milk liquid; preferably 70.9-606.1mg/100g powder, or 0.1-0.8 g/L in terms of milk liquid; more preferably 70.9-454.6 mg/100g powder, or 0.1-0.6 g/L in terms of milk liquid conversion.

According to some specific embodiments of the present invention, the breast milk oligosaccharide in the infant formula powder of the present invention is for regulating the production of butyric acid in the distal colon, and the breast milk oligosaccharide is 3-SL and/or 6-SL SL. Preferably, the breast milk oligosaccharide is further used to reduce the production of isobutyric acid and/or isovaleric acid in the distal colon, and in this application, the breast milk oligosaccharide is further preferably 3-SL. The application amount of 3-SL in milk powder is 14.2-1515.3mg/100g powder, or 0.02-2.0g/L in terms of conversion to milk; preferably 70.9-454.6mg/100g powder, or in terms of conversion to milk Calculated as 0.1-0.6g/L; more preferably 70.9-227.3mg/100g powder, or 0.1-0.3g/L in terms of milk. The application amount of 6-SL in milk powder is 14.2-1515.3mg/100g powder, or 0.02-2.0g/L in terms of conversion to milk; preferably 70.9-606.1mg/100g powder, or in terms of conversion to milk Calculated as 0.1-0.8g/L; more preferably 70.9-454.6mg/100g powder, or 0.1-0.6g/L in terms of milk.

According to some specific embodiments of the present invention, the breast milk oligosaccharides in the infant formula powder of the present invention are used as prebiotics in the proximal colon to be utilized by intestinal flora and produce gas, and the breast milk oligosaccharides are 3 -SL and/or 6-SL. The application amount of 3-SL in milk powder is 14.2-1515.3mg/100g powder, or 0.02-2.0g/L in terms of conversion to milk; preferably 70.9-454.6mg/100g powder, or in terms of conversion to milk Calculated as 0.1-0.6g/L; more preferably 70.9-227.3mg/100g powder, or 0.1-0.3g/L in terms of milk. The application amount of 6-SL in milk powder is 14.2-1515.3mg/100g powder, or 0.02-2.0g/L in terms of conversion to milk; preferably 70.9-606.1mg/100g powder, or in terms of conversion to milk Calculated as 0.1-0.8g/L; more preferably 70.9-454.6mg/100g powder, or 0.1-0.6g/L in terms of milk.

According to some specific embodiments of the present invention, the breast milk oligosaccharide in the infant formula of the present invention is for increasing the overall production of short-chain fatty acids in the proximal colon and/or the distal colon, and the breast milk oligosaccharide is 3-SL and/or 6-SL. Under this application, the application amount of 3-SL in milk powder is 14.2-1515.3mg/100g powder, or 0.02-2.0g/L in terms of milk liquid; preferably 70.9-454.6mg/100g powder, or 0.1-0.6 g/L in terms of milk liquid; more preferably 70.9-227.3 mg/100g powder, or 0.1-0.3 g/L in terms of milk liquid conversion. The application amount of 6-SL in milk powder is 14.2-1515.3mg/100g powder, or 0.02-2.0g/L in terms of conversion to milk; preferably 70.9-606.1mg/100g powder, or in terms of conversion to milk Calculated as 0.1-0.8g/L; more preferably 70.9-454.6mg/100g powder, or 0.1-0.6g/L in terms of milk.

According to some specific embodiments of the present invention, the breast milk oligosaccharides in the infant formula of the present invention are used to lower the pH of the proximal colon to maintain a healthy intestinal microenvironment. The breast milk oligosaccharides are 3-SL and/or 6-SL. Under this application, the application amount of 3-SL in milk powder is 14.2-1515.3mg/100g powder, or 0.02-2.0g/L in terms of milk liquid; preferably 70.9-454.6mg/100g powder, or 0.1-0.6 g/L in terms of milk liquid; more preferably 70.9-227.3 mg/100g powder, or 0.1-0.3 g/L in terms of milk liquid conversion. The application amount of 6-SL in milk powder is 14.2-1515.3mg/100g powder, or 0.02-2.0g/L in terms of conversion to milk; preferably 70.9-606.1mg/100g powder, or in terms of conversion to milk Calculated as 0.1-0.8g/L; more preferably 70.9-454.6mg/100g powder, or 0.1-0.6g/L in terms of milk.

According to some specific embodiments of the present invention, the breast milk oligosaccharides in the infant formula powder of the present invention are also beneficial to promote the production of short-chain fatty acids such as formic acid, propionic acid, and acetic acid that are beneficial to the human body in the intestinal system.

To sum up, the present invention finds that breast milk oligosaccharides can significantly improve the health of intestinal microenvironment, and it is used to add it to infant formula powder, which is closer to breast milk, can regulate the production of butyric acid, and improve intestinal The microenvironment is healthy and has broad application prospects.

Description of drawings

FIG. 1A is a schematic diagram of fecal inoculation culture in the SHIME device of the present invention.

Figure 1B shows a schematic diagram of the SHIME fermentation grouping of the present invention.

Figure 2 shows the microbiota of the simulated proximal colon (left) and distal colon (right) after two weeks of culture in the SHIME device mimicking the infant colon.

Figure 3A shows the detection results of pH changes of the proximal colon in the small batch fermentation experiment of each HMO in the present invention.

FIG. 3B shows the detection results of pH changes of the distal colon in the small batch fermentation experiment of each HMO in the present invention.

FIG. 4A shows the detection results of the air pressure caused by the proximal colon over time in the small batch fermentation experiment of each HMO in the present invention.

FIG. 4B shows the detection results of the air pressure caused by the distal colon over time in the small batch fermentation experiment of each HMO in the present invention.

Figure 5 shows the overall detection results of short-chain fatty acids produced by small batch fermentation of each HMO in a simulated proximal colon environment in the present invention.

Fig. 6 shows the detection results of butyric acid produced in the fecal batch fermentation experiment of the breast milk oligosaccharide in the present invention to simulate the proximal colon of infants.

Figure 7 shows the detection results of formic acid produced in the fecal batch fermentation experiment of the breast milk oligosaccharide monomers of the present invention to simulate the proximal colon of infants.

Figure 8 shows the detection results of acetic acid produced in the fecal batch fermentation experiment of the breast milk oligosaccharide monomers of the present invention to simulate the proximal colon of infants.

Figure 9 shows the detection results of propionic acid produced in the fecal batch fermentation experiment of the breast milk oligosaccharide monomers of the present invention to simulate the proximal colon of infants.

Figure 10 shows the overall detection results of short-chain fatty acids produced in the fecal batch fermentation experiment of the breast milk oligosaccharide monomers of the present invention mimicking the distal colon of infants.

Figure 11 shows the detection results of butyric acid produced in the fecal batch fermentation experiment of the breast milk oligosaccharide monomers of the present invention to simulate the distal colon of infants.

Figure 12 shows the detection results of formic acid produced in the fecal batch fermentation experiment of the breast milk oligosaccharide monomers of the present invention to simulate the distal colon of infants.

Figure 13 shows the detection results of acetic acid produced in the fecal batch fermentation experiment of the breast milk oligosaccharide monomers of the present invention to simulate the distal colon of infants.

Figure 14 shows the detection results of propionic acid produced in the fecal batch fermentation experiment of the breast milk oligosaccharide monomers of the present invention to simulate the distal colon of infants.

Figure 15A and Figure 15B show the detection results of isobutyric acid and isovaleric acid produced in the fecal batch fermentation experiment of the breast milk oligosaccharide monomers of the present invention to simulate the distal colon of infants.

Detailed ways

In order to have a clearer understanding of the technical features, purposes and beneficial effects of the present invention, the following detailed description is now given in conjunction with specific embodiments and the technical solutions of the present invention. It should be understood that these examples are only used to illustrate the present invention and not to limit the scope.

In the examples, each original reagent material can be obtained commercially, and the experimental methods without specific conditions are conventional methods and conventional conditions well known in the art, or according to the conditions suggested by the instrument manufacturer.

Example 1

Breastmilk infant formula containing breast milk oligosaccharides (preparation of 1000 kg):

Milk 1000, lactose 300kg, whey protein powder WPC80% 40kg, desalted whey powder D90 D90 175kg, sunflower oil 30kg, corn oil 20kg, soybean oil 65kg, OPO structural fat 110kg, α- Whey protein powder 27kg, beta-casein powder 9kg, lecithin 2kg, anhydrous cream 1kg, fructooligosaccharide powder 17kg, galactooligosaccharide syrup 40kg, human milk oligosaccharide (HMO) 1kg , Compound nutrients 14.35 kg, DHA 8 kg, ARA 8 kg, Bifidobacterium 0.2 kg.

One kilogram of human milk oligosaccharide (HMO) is 3-SL or 6-SL.

Among them, compound nutrients include about 2.5 kg of compound vitamin nutrition package, about 0.75 kg of choline chloride nutrition package, about 6 kg of calcium powder nutrition package, about 1 kg of mineral nutrition package, magnesium chloride and potassium chloride nutrition package. The base material of the package is lactose, and the specific main nutrient composition is as follows: 1) Compound vitamins, in each gram of compound vitamins, taurine: 180 mg; vitamin A (retinyl acetate): 2300 μg RE; vitamin D (cholecalciferol) : 34 μg; Vitamin B1 (thiamine nitrate): 4000 μg; Vitamin B2 (riboflavin): 1200 μg; Vitamin B6 (pyridoxine hydrochloride): 2050 μg; Vitamin B12 (cyanocobalamin): 11.2 μg; Menadione): 400 μg; Vitamin C (L-Ascorbic Acid): 495 mg; Vitamin E (dl-α-Tocopheryl Acetate): 38 mgα-TE; Nicotinamide: 23050 μg; Folic Acid: 530 μg; Biotin (D-Biotin) : 88μg; pantothenic acid (D-calcium pantothenate): 15000μg; 2) calcium powder, per gram of calcium powder, calcium (calcium carbonate, tricalcium phosphate): 230mg; phosphorus (tricalcium phosphate): 90mg; 3) minerals, Per gram of minerals, iron (ferrous sulfate): 80 mg; zinc (zinc sulfate): 45 mg; copper (copper sulfate): 4000 μg; iodine (potassium iodide): 980 μg; selenium (sodium selenite): 170 μg; manganese ( manganese sulfate): 550 μg; 4) magnesium chloride, per 1000 kg of milk powder, magnesium (magnesium chloride): 150 g; 5) potassium chloride, per 1000 kg of milk powder, potassium (potassium chloride): 1050 g; 6) choline chloride , per 1000 kg of milk powder, choline (choline chloride): 700g.

The content of linoleic acid and α-linolenic acid in high oleic sunflower oil, corn oil, soybean oil and structured oil (OPO) used as raw materials were 8.20%, 0.3%, 55.50%, 1.2%, 52.10%, 8.8%, 6.18%, 0.47%.

The specific preparation process of breast-milk infant formula milk powder containing breast-milk oligosaccharide is as follows:

1) Addition of powder: All kinds of powder raw materials are metered according to the formula and added to the powder mixing tank uniformly through the air delivery system for storage.

2) Vacuum powder suction: various powder raw materials in the powder mixing tank are sucked into the vacuum mixing tank through the vacuum system.

3) Dissolving oil: put the grease specified in the formula into the carburetor room according to the formula requirements, and the temperature in the carburetor room should be kept at 60 °C. After the oil is dissolved, the mixed oil is pumped through the oil pump and flowmeter according to the formula requirements. in storage tank.

4) Weighing: According to the formula requirements, the mixed oil is pumped into the mixing tank through the oil pump.

5) Nutrient dissolution and addition: calcium powder, minerals, vitamins, etc. are added separately, dissolved separately with 150kg of pure water, and then poured into the mixing tank. After each type of mixing, 100kg of pure water is used to rinse the addition tank and pipeline.

6) Filtration: The mixed feed liquid is filtered through a filter screen to remove physical impurities that may be brought into the raw material.

7) Homogenization: The mixed feed liquid is homogenized by a homogenizer, and the fat globules are mechanically processed to disperse them into uniform fat globules.

8) Cooling and storage: The homogenized feed liquid enters the plate heat exchanger for cooling: it is cooled to below 20°C, temporarily stored in the pre-storage tank, and enters the next process within 6 hours, and the agitator is turned on according to the set requirements.

9) Concentration and sterilization: double-effect concentration is used in production, the sterilization temperature is ≥83 °C, and the sterilization time is 25 seconds. The discharge concentrations were all 50% dry matter.

10) Thick milk storage, preheating filtration, spray drying: the concentrated milk is temporarily stored in the thick milk balance tank. It is preheated to 60 ℃ by a scraper preheater. After preheating, the material is filtered by a filter with a 1mm aperture, and then pumped into a drying tower for spray drying with a high-pressure pump. The fine powder is agglomerated at the top of the tower or in a fluidized bed as required. Inlet air temperature: 180℃, exhaust air temperature 86℃, high pressure pump pressure 200bar, tower negative pressure -4mba.

11) Fluidized bed drying and cooling: the powder coming out of the drying tower is subjected to secondary drying in the fluidized bed (first stage), and then cooled to 30°C through the fluidized bed (secondary stage). At the same time, the lecithin and the carrier are mixed and heated to 60°C. Under the action of compressed air, the lecithin is uniformly dispersed on the surface of the powder, so that the powder particles are agglomerated to increase their particle size and instant solubility.

12) Sub-packaging: According to the formula requirements, the personnel in the milling workshop will weigh and pack the DHA, ARA or Bifidobacterium into sealed bags.

13) Dry mixing: Mix the weighed DHA, ARA or Bifidobacterium with milk powder in a dry mixing machine.

14) Sifting powder: The particle size of the milk powder is uniform through a vibrating sieve, and the powder residue is discarded.

15) Powder output: use a sterilized powder collecting box to collect powder, and transport it from the powder output room to the powder room.

16) Powdering: Pour the milk powder into the powder storage tank on the large and small packaging machine according to the packaging requirements.

17) Packing: 400g automatic packing machine packed with nitrogen. The oxygen content during nitrogen filling is less than 1%. The oxygen content of 900 grams of iron cans with automatic nitrogen filling is less than 5%.

18) Packing: Put the packaged pouch into the carton and add the powder scoop at the same time, and seal it with a carton sealing machine.

19) Finished product inspection: Sampling inspection of the packaged products according to the inspection plan.

20) Storage in storage: The products that have passed the inspection are stored in storage, and are required to be stored at room temperature with humidity ≤65%.

Example 2

Breastmilk infant formula containing breast milk oligosaccharides (preparation of 1000 kg):

Milk 1200, lactose 140kg, whole milk powder 100kg, skimmed milk powder 260kg, whey protein powder WPC34% 100kg, demineralized whey powder D90 210kg, sunflower oil 22kg, corn oil 15kg, soybean oil 48kg , OPO structural fat 85kg, α-whey protein powder 4kg, β-casein powder 1kg, soybean lecithin 2kg, anhydrous cream 1kg, fructooligosaccharide powder 5kg, galactooligosaccharide syrup 13kg, Human milk oligosaccharide (HMO) 1.2 kg, compound nutrients 10.3 kg, DHA 7 kg, ARA 8 kg, Bifidobacterium 0.18 kg.

1.2 kg of human milk oligosaccharides (HMO) were 3-SL or 6-SL.

Among them, the compound nutrients include about 2.6 kilograms of compound vitamin nutrition packs, about 0.35 kilograms of choline chloride nutrition packs, about 3.7 kilograms of calcium powder nutrition packs, about 1.3 kilograms of mineral nutrition packs, and magnesium chloride and potassium chloride nutrition packs. The base material of the package is lactose, and the specific main nutrient composition is as follows: 1) Compound vitamins, in each gram of compound vitamins, taurine: 190 mg; vitamin A (retinyl acetate): 2000 μg RE; vitamin D (cholecalciferol) : 40 μg; Vitamin B1 (thiamine nitrate): 3900 μg; Vitamin B2 (riboflavin): 1000 μg; Vitamin B6 (pyridoxine hydrochloride): 1500 μg; Vitamin B12 (cyanocobalamin): 5 μg; naphthoquinone): 350 μg; vitamin C (L-ascorbic acid): 400 mg; vitamin E (dl-α-tocopheryl acetate): 30 mgα-TE; niacinamide: 23000 μg; folic acid: 420 μg; biotin (D-biotin): 70μg; pantothenic acid (D-calcium pantothenate): 8000μg; 2) calcium powder, per gram of calcium powder, calcium (calcium carbonate, tricalcium phosphate): 250mg; phosphorus (tricalcium phosphate): 75mg; 3) minerals, each In gram minerals, iron (ferrous sulfate): 76mg; zinc (zinc sulfate): 26mg; copper (copper sulfate): 2700μg; iodine (potassium iodide): 600μg; selenium (sodium selenite): 160μg; 4) magnesium chloride , per 1000 kg of milk powder, magnesium (magnesium chloride): 85g; 5) potassium chloride, per 1000 kg of milk powder, potassium (potassium chloride): 1000g; 6) choline chloride, per 1000 kg of milk powder, choline (choline chloride): 700 g.

The content of linoleic acid and α-linolenic acid in the raw materials sunflower oil, corn oil, soybean oil and structural oil (OPO) used are 8.20%, 0.3%, 55.50%, 1.2%, 52.10%, 8.8%, respectively. 6.18%, 0.47%.

The specific preparation process of breast milk oligosaccharide (HMO)-containing infant formula milk powder is as follows:

1) Addition of powder: All kinds of powder raw materials are metered according to the formula and added to the powder mixing tank uniformly through the air delivery system for storage.

2) Vacuum powder suction: various powder raw materials in the powder mixing tank are sucked into the vacuum mixing tank through the vacuum system.

3) Dissolving oil: put the grease specified in the formula into the carburetor room according to the formula requirements, and the temperature in the carburetor room should be kept at 60 °C. After the oil is dissolved, the mixed oil is pumped through the oil pump and flowmeter according to the formula requirements. in storage tank.

4) Weighing: According to the formula requirements, the mixed oil is pumped into the mixing tank through the oil pump.

5) Nutrient dissolution and addition: calcium powder, minerals, vitamins, etc. are added separately, dissolved separately with 150kg of pure water, and then poured into the mixing tank. After each type of mixing, 100kg of pure water is used to rinse the addition tank and pipeline.

6) Filtration: The mixed feed liquid is filtered through a filter screen to remove physical impurities that may be brought into the raw material.

7) Homogenization: The mixed feed liquid is homogenized by a homogenizer, and the fat globules are mechanically processed to disperse them into uniform fat globules.

8) Cooling and storage: The homogenized feed liquid enters the plate heat exchanger for cooling: it is cooled to below 20°C, temporarily stored in the pre-storage tank, and enters the next process within 6 hours, and the agitator is turned on according to the set requirements.

9) Concentration and sterilization: double-effect concentration is used in production, the sterilization temperature is ≥83 °C, and the sterilization time is 25 seconds. The discharge concentrations were all 50% dry matter.

10) Thick milk storage, preheating filtration, spray drying: the concentrated milk is temporarily stored in the thick milk balance tank. It is preheated to 60 ℃ by a scraper preheater. After preheating, the material is filtered by a filter with a 1mm aperture, and then pumped into a drying tower for spray drying with a high-pressure pump. The fine powder is agglomerated at the top of the tower or in a fluidized bed as required. Inlet air temperature: 180℃, exhaust air temperature 86℃, high pressure pump pressure 200bar, tower negative pressure -4mba.

11) Fluidized bed drying and cooling: the powder coming out of the drying tower is subjected to secondary drying in the fluidized bed (first stage), and then cooled to 30°C through the fluidized bed (secondary stage). At the same time, the soybean lecithin is mixed with the carrier and heated to 60°C. Under the action of compressed air, it is uniformly dispersed on the surface of the powder, so that the powder particles are agglomerated to increase their particle size and instant solubility.

12) Sub-packaging: According to the formula requirements, the personnel in the milling workshop will weigh and pack the DHA, ARA or Bifidobacterium into sealed bags.

13) Dry mixing: Mix the weighed DHA, ARA or Bifidobacterium with milk powder in a dry mixing machine.

14) Sifting powder: The particle size of the milk powder is uniform through a vibrating sieve, and the powder residue is discarded.

15) Powder output: use a sterilized powder collecting box to collect powder, and transport it from the powder output room to the powder room.

16) Powdering: Pour the milk powder into the powder storage tank on the large and small packaging machine according to the packaging requirements.

17) Packing: 400g automatic packing machine packed with nitrogen. The oxygen content during nitrogen filling is less than 1%. The oxygen content of 900 grams of iron cans with automatic nitrogen filling is less than 5%.

18) Packing: Put the packaged pouch into the carton and add the powder scoop at the same time, and seal it with a carton sealing machine.

19) Finished product inspection: Sampling inspection of the packaged products according to the inspection plan.

20) Storage in storage: The products that have passed the inspection are stored in storage, and are required to be stored at room temperature with humidity ≤65%.

Breast milk oligosaccharide efficacy test

Fecal inoculation culture in SHIME device

蛋白胨(2g/L)，黏蛋白(6g/L)，乳糖(2.1g/L)，酪蛋白(0.2g/L)，乳清蛋白-乳白蛋白(2.7g/L)，L-半胱氨酸盐酸盐(0.2g/L)。本发明各实验中的食物料参考Le Blay等人(2010)调整了乳糖、酪蛋白和乳清蛋白的比例约为12:1:15并保证营养素稳定均衡，来模拟常规的母乳或婴配粉喂养情况下，婴儿的肠道微生态可能接触的食物组成。婴儿粪便菌群在SHIME模型中稳定了两周后，近端和远端结肠会被取样，溶于甘油中形成原液，并在厌氧条件下存放于-80℃。Using the SHIME device (see diagram in Figure 1A), fresh fecal samples containing microbiota were obtained from a healthy 5-month-old spontaneously delivered and exclusively breastfed infant and inoculated into containers corresponding to the proximal and distal colons . Food rations were fed to the gastric/small intestine end of the device three times a day for two weeks to support the growth and colonization of flora in the proximal and distal colons. Among them, the food that is digested by the small intestine and enters the colon is prepared by adjusting the ratio of lactose, casein and whey protein on the basis of the standard food provided by ProDigest, the manufacturer of the SHIME device. The standard food is composed of the following ingredients: Gum (1g/L), Glucose (1g/L), Starch (1g/L), Cellobiose (1g/L),

Peptone (2g/L), Mucin (6g/L), Lactose (2.1g/L), Casein (0.2g/L), Whey Protein-Lactalbumin (2.7g/L), L-cysteine Hydrochloride (0.2g/L). The food materials in each experiment of the present invention refer to Le Blay et al. (2010) to adjust the ratio of lactose, casein and whey protein to about 12:1:15 and ensure stable and balanced nutrients to simulate conventional breast milk or infant formula The composition of foods that the infant's gut microbiome may be exposed to under feeding conditions. After the infant fecal flora was stabilized in the SHIME model for two weeks, the proximal and distal colons were sampled, dissolved in glycerol to form a stock solution, and stored at -80°C under anaerobic conditions.

Analytical testing of the composition of the microbiota will focus on specific strains: Lactobacillus, Bifidobacterium, Rosetella, Eubacterium and Faecalibacterium, as they are known to be associated with (prebiotic) health benefits. Detection analysis is based on qPCR.

Small batch fermentation

After the infant flora was inoculated into the SHIME model for 2 weeks of stable growth (as described in the aforementioned "fecal inoculation culture in the SHIME device"), 10 mL of the flora of the proximal and distal colons were taken and transferred to fermentation flasks under anaerobic conditions. Bulk fermentation. On the basis of 43mL basal buffer (for adjusting pH and simulating the corresponding colonic environment), each fermentation flask also contains 20mL PBS buffer (for dissolving and bringing in HMO test substance) supplemented with different contents of HMO , so that the final concentrations of various HMOs were 0.02 g/L, 0.2 g/L, 2 g/L, the pH of the proximal colon was set to 5.6, and the pH of the distal colon was set to 6.5. The vials were incubated at 37°C with shaking. During incubation, air pressure was checked at 0, 6 hours, 24 hours and 48 hours, followed by samples for pH and short chain fatty acids. The assay was repeated three times.

During the HMO intervention, the gas production of the groups was compared by measuring changes in air pressure. Analysis of short chain fatty acids included isobutyric acid, isovaleric acid, butyric acid, propionic acid, and acetic acid were also analyzed, and formic acid was also analyzed, each by HPLC.

SHIME fermentation

The fecal flora of infants sampled and stored in "Stool Inoculation Culture in SHIME Device" was used to inoculate the SHIME model to investigate the fermentation of HMO in the SHIME device. Two batches of experiments were performed before and after, and three sample groups (or controls) were simultaneously performed in each batch of experiments. In each of the three experimental devices, the devices mimicking the extreme and distal colons were seeded separately (see Figure 1B for an illustration). The chow was fed three times a day, and after 4 days of culture, HMO was added to the chow (the control group did not have HMO). 280 mg of HMO (concentration of 2 g/L) and 60 mL of pancreatic juice were added to 140 mL of feed solution for each food material. Only one experiment was performed for each group of HMOs and controls, so the biological replicate was 1. SHIME fermentation continued until the 14th day after HMO intervention, and samples were taken at different time periods during the fermentation period.

data analysis

A two-tailed, paired t-test was performed on the data results. If there is a significant difference between the two groups, and p<0.05, it is indicated with an asterisk *. Two asterisks** indicate p<0.01. Three asterisks *** indicate p<0.001.

Simulates the microbiota in the proximal and distal colonic environment

Figure 2 shows the identification of fecal flora after inoculation and culture in the SHIME device for two weeks. After stabilizing in the SHIME model for two weeks, after the microflora determination, it was found that the content of Bifidobacterium and Lactobacillus was very low and almost undetectable, which was consistent with previous literature reports (Laforest-Lapointe, 2017). It was demonstrated that in a simulated infant gut environment, after feeding with a standard diet containing lactose/casein/whey, the corresponding colonic microbiota was more towards formula-fed infants than breast-fed infants.

Variation of pH with time for each HMO in small batch fermentation experiments

See Figure 3A for the detection results of pH changes in the proximal colon of each HMO in small batch fermentation experiments. See Figure 3B for the detection results of pH changes in the distal colon of each HMO in the small batch fermentation experiment. As can be seen, in the proximal colon, the pH reduction at 6 hours was more pronounced than at other time points. It can be seen that with the prolongation of fermentation time, HMO will be utilized by the flora in infant feces to produce short-chain fatty acids, thereby lowering the pH.

Variation of air pressure with time induced by each HMO in small batch fermentation experiments

Figure 4A shows the time-dependent changes in air pressure induced by the proximal colon of each HMO in the small batch fermentation experiment. Figure 4B shows the time-dependent changes in air pressure induced by the distal colon of each HMO in the small batch fermentation experiment. It can be seen that the two sialic acid oligosaccharides, 3-SL and 6-SL, have higher air pressure in the proximal colon under the condition of 2g/L, which proves that under this condition, the two oligosaccharides Can be better utilized by fecal flora. In the simulated distal colon, all HMOs caused an increase in air pressure at 2 g/L.

Simulating the production of short-chain fatty acids by small-batch fermentation of each HMO in a proximal colonic environment

Figure 5 shows the results of the total amount of short-chain fatty acids produced by small batch fermentation of each HMO in a simulated proximal colon environment. It can be seen that after 48 hours of fermentation for all HMOs, the short-chain fatty acids produced are higher than other time points, especially 3-SL and 6-SL under the condition of 2g/L, produce significantly more SCFA, among which 2g/L of 3-SL also produced more SCFA after 24 hours of fermentation.

Figure 6 shows the detection results of butyric acid produced by small batch fermentation of each HMO in a simulated proximal colon environment. It can be seen that in the simulated proximal colon environment, only 6-SL can significantly increase the production of butyrate after 48 hours of fermentation.

Figure 7 shows the detection results of formic acid produced by small batch fermentation of each HMO in a simulated proximal colon environment. As can be seen, only 3-SL and 6-SL significantly increased formate after 48 hours of simulated proximal colonic fermentation. Among them, the effect of 6-SL is more significant.

Figure 8 shows the detection results of acetic acid produced by small batch fermentation of each HMO in a simulated proximal colon environment. As can be seen, only 3-SL and 6-SL significantly increased acetate after 48 hours of simulated proximal colonic fermentation. Among them, the effect of 3-SL is more significant.

See Figure 9 for the detection results of small batch fermentation of each HMO to produce propionic acid in a simulated proximal colon environment. As can be seen, only 6-SL significantly increased propionate after 48 hours of simulated proximal colonic fermentation.

Simulating the production of short-chain fatty acids by small-batch fermentation of each HMO in a distal colonic environment

Figure 10 shows the detection results of the total amount of short-chain fatty acids produced by small batch fermentation of each HMO in a simulated distal colon environment. It can be seen that in the distal colon, the production of SCFA by each HMO is more active than that in the proximal colon, which also confirms the generally accepted scientific view that most of the fermentation of the intestinal flora occurs in the distal colon. The SCFA produced increased with time from 0, 6, 24, to 48 hours. At 6 hours, 0.02 g/L of 3-SL and 0.02 g/L of LNT produced relatively more SCFAs; at 24 hours, 2 g/L of 6-SL and 2 g/L of 3 -FL produced relatively more short-chain fatty acids; and at the time point of 48 hours, all HMOs could produce more short-chain fatty acids at 2 g/L.

Figure 11 shows the detection results of butyric acid produced by small batch fermentation of each HMO in a simulated distal colon environment. As can be seen, only 3-SL and 6-SL significantly increased butyrate after 48 hours of simulated distal colonic fermentation.

Figure 12 shows the detection results of formic acid produced by small batch fermentation of each HMO in a simulated distal colon environment. As can be seen, all HMOs significantly increased formate after 48 hours of simulated distal colonic fermentation. Among them, the effect of 3-SL, 6-SL, LNT and 3-FL is more significant.

Figure 13 shows the detection results of acetic acid produced by small batch fermentation of each HMO in a simulated distal colon environment. As can be seen, all HMOs significantly increased acetate after 48 hours of simulated distal colonic fermentation. Among them, the effect of 3-SL, 6-SL and LNT is more significant.

Figure 14 shows the detection results of small batch fermentation of each HMO to produce propionic acid in a simulated distal colon environment. As can be seen, all HMOs significantly increased propionate after 48 hours of simulated distal colonic fermentation. Among them, the effect of 3-SL and 3-FL is more significant.

The detection results of isobutyric acid and isovaleric acid produced by small batch fermentation of each HMO under the simulated distal colon environment are shown in Figure 15A and Figure 15B, respectively. It can be seen that in the distal colon, 2'-FL, 3-FL and 3-SL, LNT can significantly reduce the production of isobutyric acid. Both sialic acid-type oligosaccharides and two fucosyl-type oligosaccharides can reduce isovaleric acid production, respectively.

Simulation of the production of butyrate by HMO fermentation in the SHIME model in the distal colon environment

See Table 1 for the detection results of butyric acid produced by the fermentation of each HMO in the SHIME model under the simulated distal colon environment.

Table 1

In the case of SHIME model fermentation, after 14 days of fermentation, the butyric acid produced by various HMOs decreased compared with the initial day 1 of fermentation, but the control group (without HMO) had the highest decrease. Compared with the first day, the fold reduction on day 14 was significantly improved and improved. Among them, the fold reduction of 3-SL was the lowest, followed by 3-FL. Compared with the control, 6-SL, 2'-FL and LNT were all There is a better improvement effect. It can be seen that each HMO has certain advantages in the regulation of butyric acid in the fermentation product compared with the control group without HMO.

Claims (10)

1. The application of breast milk oligosaccharide in the preparation of infant formula powder for improving the health of intestinal microenvironment, wherein the breast milk oligosaccharide is fucosyl oligosaccharide, sialic acid-based oligosaccharide Or lactose-N-tetrasaccharide; the improvement of intestinal microenvironment health includes: regulating the production of butyric acid in the intestinal system, increasing the overall production of short-chain fatty acids, and being utilized by the intestinal flora as a prebiotic in the intestinal system And produce gas, reduce isobutyric acid and/or isovaleric acid production, and/or lower pH to maintain a healthy gut microenvironment. 2. An infant formula powder with the effect of improving intestinal microenvironment health, which contains breast milk oligosaccharides, wherein the breast milk oligosaccharides are fucosyl oligosaccharides, sialic acid-based oligosaccharides Or lactose-N-tetrasaccharide; the improvement of intestinal microenvironment health includes: regulating the production of butyric acid in the intestinal system, increasing the overall production of short-chain fatty acids, and being utilized by the intestinal flora as a prebiotic in the intestinal system And produce gas, reduce isobutyric acid and/or isovaleric acid production, and/or lower pH to maintain a healthy gut microenvironment. 3. The infant formula powder according to claim 2, which contains 14.2-1515.3 mg/100g of breast milk oligosaccharides, or 0.02-2.0 g/L of breast milk oligosaccharides in terms of milk. 4. The infant formula powder according to claim 2, which contains 14.2-1515.3mg/100g of breast milk oligosaccharides, and the total protein content of the infant formula powder is 10-19g/100g, and whey protein accounts for the total The proportion of protein is 38-70%, the content of α-whey protein is 1.25-2.5g/100g, the content of β-casein is 2.2-4.0g/100g, the content of fat is 17-29g/100g, and the content of linoleic acid is 2500～4500mg/100g, α-linolenic acid content is 330～450mg/100g, dietary fiber is 0.95～6.3g/100g, and the dietary fiber includes galactooligosaccharides, fructooligosaccharides and/or breast milk oligosaccharides, carbohydrates The content is 50～57g/100g. 5. The infant formula powder according to claim 2, 3 or 4, wherein the breast milk oligosaccharide is 3-SL and/or 6-SL. 6. The preparation method of the formula powder for infants and young children according to any one of claims 2 to 5, which adopts a wet or dry production process, and mixes breast milk oligosaccharides with other raw materials in the formula to prepare the infants and young children formula powder. 7. The application of the infant formula powder according to any one of claims 2 to 5 in the preparation of infant formula food for improving the health of the intestinal microenvironment, wherein the improving the health of the intestinal microenvironment comprises: regulating intestinal microenvironment Production of butyric acid in the gut system, increased overall production of short-chain fatty acids, utilized in the gut system as a prebiotic by gut flora and gas production, reduced production of isobutyric acid and/or isovaleric acid, and/or Lower pH to maintain a healthy gut microenvironment. 8. application according to claim 7, wherein, the breast milk oligosaccharide in infant formula powder is for regulating the production of butyric acid in distal colon, preferably further reducing the production of isobutyric acid and/or isovaleric acid , the breast milk oligosaccharide is 3-SL and/or 6-SL. 9. The use according to claim 7, wherein the breast milk oligosaccharides in the infant formula are used as prebiotics in the proximal colon to be utilized by intestinal flora and gas production, and/or reduce the proximal colon pH value, the breast milk oligosaccharides are 3-SL and/or 6-SL. 10. The use according to claim 7, wherein the breast milk oligosaccharides in the infant formula are used to increase the overall production of beneficial short chain fatty acids in the proximal colon and/or distal colon, the beneficial Short chain fatty acids include formic acid, acetic acid and/or propionic acid, and the breast milk oligosaccharides are 3-SL and/or 6-SL.

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