CN114128767A - Milk protein partially hydrolyzed hypoallergenic infant formula containing breast milk oligosaccharides - Google Patents

Milk protein partially hydrolyzed hypoallergenic infant formula containing breast milk oligosaccharides Download PDF

Info

Publication number
CN114128767A
CN114128767A CN202111450898.9A CN202111450898A CN114128767A CN 114128767 A CN114128767 A CN 114128767A CN 202111450898 A CN202111450898 A CN 202111450898A CN 114128767 A CN114128767 A CN 114128767A
Authority
CN
China
Prior art keywords
infant formula
powder
protein
milk
intestinal
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
CN202111450898.9A
Other languages
Chinese (zh)
Other versions
CN114128767B (en
Inventor
李艳杰
刘彪
李威
孔小宇
吴春梅
王雯丹
吉塞拉·阿德里安娜·怀斯
司徒文佑
王燕霞
周名桥
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Inner Mongolia Yili Industrial Group Co Ltd
Original Assignee
Inner Mongolia Yili Industrial Group Co Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Inner Mongolia Yili Industrial Group Co Ltd filed Critical Inner Mongolia Yili Industrial Group Co Ltd
Priority to CN202111450898.9A priority Critical patent/CN114128767B/en
Publication of CN114128767A publication Critical patent/CN114128767A/en
Application granted granted Critical
Publication of CN114128767B publication Critical patent/CN114128767B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C9/00—Milk preparations; Milk powder or milk powder preparations
    • A23C9/20—Dietetic milk products not covered by groups A23C9/12 - A23C9/18
    • A23C9/203—Dietetic milk products not covered by groups A23C9/12 - A23C9/18 containing bifidus-active substances, e.g. lactulose; containing oligosaccharides
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C9/00—Milk preparations; Milk powder or milk powder preparations
    • A23C9/152—Milk preparations; Milk powder or milk powder preparations containing additives
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C9/00—Milk preparations; Milk powder or milk powder preparations
    • A23C9/152—Milk preparations; Milk powder or milk powder preparations containing additives
    • A23C9/1522—Inorganic additives, e.g. minerals, trace elements; Chlorination or fluoridation of milk; Organic salts or complexes of metals other than natrium or kalium; Calcium enrichment of milk
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C9/00—Milk preparations; Milk powder or milk powder preparations
    • A23C9/152—Milk preparations; Milk powder or milk powder preparations containing additives
    • A23C9/1526—Amino acids; Peptides; Protein hydrolysates; Nucleic acids; Derivatives thereof
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C9/00—Milk preparations; Milk powder or milk powder preparations
    • A23C9/152—Milk preparations; Milk powder or milk powder preparations containing additives
    • A23C9/1528—Fatty acids; Mono- or diglycerides; Petroleum jelly; Paraffine; Phospholipids; Derivatives thereof
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C9/00—Milk preparations; Milk powder or milk powder preparations
    • A23C9/152—Milk preparations; Milk powder or milk powder preparations containing additives
    • A23C9/156—Flavoured milk preparations ; Addition of fruits, vegetables, sugars, sugar alcohols or sweeteners
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C9/00—Milk preparations; Milk powder or milk powder preparations
    • A23C9/152—Milk preparations; Milk powder or milk powder preparations containing additives
    • A23C9/158—Milk preparations; Milk powder or milk powder preparations containing additives containing vitamins or antibiotics
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/51—Bifidobacterium

Landscapes

  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Food Science & Technology (AREA)
  • Polymers & Plastics (AREA)
  • Health & Medical Sciences (AREA)
  • Nutrition Science (AREA)
  • Oil, Petroleum & Natural Gas (AREA)
  • Biophysics (AREA)
  • Inorganic Chemistry (AREA)
  • Biochemistry (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Dairy Products (AREA)

Abstract

The invention provides a milk protein partial hydrolysis hypoallergenic infant formula containing breast milk oligosaccharides. Specifically, the invention provides an infant formula comprising breast milk oligosaccharides, wherein the breast milk oligosaccharides comprise lacto-N-tetraose, and the total content of the lacto-N-tetraose in the infant formula is 14.2-2273.0mg/100g of powder based on the total mass of the infant formula, or 0.02-3.0g/L based on the converted milk; and the total protein content in the infant formula is 9-20 g/100g based on the total mass of the infant formula, the total protein comprises hydrolyzed milk protein, and the protein with the hydrolysis degree of 8-23 and the molecular weight distribution of less than 3000dal accounts for more than 80% of the total protein. The invention also provides a preparation method and related application of the infant formula.

Description

Milk protein partially hydrolyzed hypoallergenic infant formula containing breast milk oligosaccharides
Technical Field
The invention relates to hypoallergenic infant formula food, in particular to milk protein partial hydrolysis hypoallergenic infant formula food containing breast milk oligosaccharides, a preparation method and related applications thereof, and belongs to the technical field of special medical hypoallergenic infant food.
Background
In recent years, the incidence of food allergy (FH/FA) has been on the rise year by year and has become a focus of research. Among the food allergies among infants, milk and egg allergies are the most common. Milk proteins are the most common source of protein for common infant formulas. CMPA refers to the adverse reaction of the body to the milk protein, which is mediated by an immune mechanism and can be mediated by IgE (immunoglobulin E) mediation, non-IgE mediation or a mixture of the two. Symptomatically, non-IgE-mediated mild to moderate CMPA is more prone to gastrointestinal symptoms, most commonly irritability, i.e., intestinal spasm; in addition, emesis, diarrhea, etc.; skin symptoms include itching, erythema, nonspecific rash, significant atopic eczema, and the like. The IgE-mediated CMPA has more prominent skin symptoms, such as acute pruritus, erythema, urticaria, angioedema and acute diffuse atopic eczema; gastrointestinal symptoms include vomiting, diarrhea, abdominal pain/intestinal cramps; respiratory symptoms are acute rhinitis and/or conjunctivitis, whereas non-IgE-mediated CMPA rarely shows respiratory symptoms within 1 year of age.
Avoidance therapy is currently used as a guideline opinion for milk protein allergic children. In the process, a milk protein deep hydrolysis formula (eHF) is used for diagnosis and treatment, and a milk protein partial hydrolysis formula (pHF) and a whole protein formula are used for a family reintroduction test. However, clinical studies show that long-term consumption of the deeply hydrolyzed formula avoids the need for a whole protein diet, which can lead to a certain degree of growth and development slowness in infants. Moreover, the long-term use of hydrolyzed formulas imposes an economic burden on the infant's family. Clinical data indicate that the milk protein allergic population, when subjected to the reintroduction experiment, has a higher proportion of tolerance than the whole protein formulation, thereby switching from the extensively hydrolyzed formulation to the partially hydrolyzed formulation earlier. It is significant that the infant improves the symptoms as soon as possible by the milk protein deep-hydrolyzed formula and then switches from the milk protein partial-hydrolyzed formula to the whole protein formula as soon as possible.
On the other hand, intestinal leakage (leaky gut), refers to a phenomenon in which intestinal permeability is increased to cause harmful substances such as bacteria and toxins in the intestines to pass through the intestinal mucosa into other tissues, organs and blood circulation in the human body. Gut permeability is associated with gut barrier function, and normal gut permeability depends on the integrity of the gut mucosal barrier. The intestinal mucosal barrier is a complex multi-layered system including physical, chemical, biological and immunological barriers. The intestinal physical barrier, which is the first line of defense against the external environment, occupies a central position in the intestinal barrier structure and is composed of intestinal epithelial cells and intercellular junctions. Intestinal cell permeability is largely divided into transepithelial or transcellular permeability and paracellular permeability. Cell bypass permeability is dependent on transport through interstitial spaces between cells. The intercellular junction includes tight junction, adhesion junction, desmosomes, etc., and most importantly, the tight junction is located at the apical end of the intestinal epithelial cell outer membrane and has a long and narrow band-like structure to block the intercellular space, thereby preventing macromolecular substances such as bacteria and toxins, etc. in the intestinal lumen from entering the blood circulation through the intercellular space. The intercellular tight junction structure has high dynamic stability, and its permeability determines the barrier function of the whole intestinal epithelial cell, and is regulated by intracellular and extracellular signals, and can be affected by diet, diseases, stress, etc. The intestinal chemical barrier is mainly composed of the mucus layer, which can alter the sites of intestinal microorganisms, preventing them from coming into direct contact with the host intestinal tissue cells. In addition, some substances produced in the intestinal tract, such as bile salts, mucopolysaccharides, lysozyme and glycoproteins, can also play a role in chemical barriers. The intestinal biological barrier is a micro-ecosystem with dynamic stability formed by intestinal symbiotic bacteria and a host, the intestinal symbiotic bacteria are attached to a mucous membrane layer on the surface of the intestinal tract of the host to form a microbial barrier formed by bacteria, and the intestinal biological barrier inhibits the colonization and the propagation of pathogenic bacteria through mechanisms such as competitive adhesion. In the intestinal immunity barrier, intestinal tract-related lymphoid tissues play a role in juggling, wherein lymphocytes, macrophages and the like play a role in resisting pathogen invasion, and in intestinal tract immunity effector molecules, secretory immunoglobulin A (sIgA) plays a key role, is produced in an intestinal tract lamina propria and is secreted into an intestinal cavity after being processed by intestinal tract epithelial cells, can block adhesion of antigens such as bacteria, toxins and viruses on mucous membranes, and plays a role in clearing the antigens. The interaction of these barriers allows the intestinal tract to maintain a permeability balance, preventing the loss of water and electrolytes and the entry of antigens and microorganisms into the body, while allowing molecular exchange between the body and the environment and the absorption of nutrients in the food.
Leakage of gut due to abnormal barrier function in the gut is a major potential cause of many health problems, particularly microbial toxins produced by the metabolism of the intestinal flora or other toxic substances that enter with food, through the intestinal wall with increased permeability ("intestinal leakage") into the blood circulation, causing various autoimmune symptoms by stimulating the autoimmune system or poisoning internal organs, thus causing various diseases. Compared with the infants fed by breast milk, the infants not fed by breast milk have poor intestinal development and are easy to have intestinal leakage.
In most of the milk protein hydrolysis formula milk powder for infants in the market at present, the prebiotics are fructo-oligosaccharide or galacto-oligosaccharide, or no prebiotics are added, so that the problem of improving the integrity of the intestinal epithelium cannot be fundamentally solved, intestinal leakage cannot be prevented or reduced, and allergic infants cannot be promoted to tolerate whole protein formula as soon as possible.
Therefore, there is a need for solutions to improve the intestinal health of infants.
Disclosure of Invention
It is an object of the present invention to provide an infant formula that improves the intestinal health of an infant.
It is another object of the present invention to provide a method for preparing said infant formula.
It is another object of the present invention to provide the use of said infant formula.
The inventor finds in experimental research that the breast milk oligosaccharide lactose-N-tetraose (LNT) can be used alone or in combination with other breast milk oligosaccharides to prevent or improve intestinal leakage or intestinal barrier damage, and is beneficial to improving intercellular intestinal permeability and intestinal barrier, improving intercellular gaps, and particularly beneficial to maintaining intestinal health of infants. Thus, the present invention provides an infant formula that prevents intestinal leakage and promotes intestinal health by adding breast milk oligosaccharides including lacto-N-tetraose to the infant formula.
In particular, in one aspect, the present invention provides an infant formula comprising breast milk oligosaccharides including lacto-N-tetraose, wherein the total content of lacto-N-tetraose in the infant formula is from 14.2 to 2273.0mg/100g powder, based on the total mass of the infant formula, or from 0.02 to 3.0g/L, based on milk;
and the total protein content in the infant formula is 9-20 g/100g based on the total mass of the infant formula, the total protein comprises hydrolyzed milk protein, and the protein with the hydrolysis degree of 8-23 and the molecular weight distribution of less than 3000dal accounts for more than 80% of the total protein.
lacto-N-tetraose, a hexasaccharide structure formed by lactose and tetraose, is a representative substance of oligosaccharides having a core sugar chain as a basic structure and not containing fucosyl or sialyl groups, which is generally commercially available through a microbial fermentation process and has the same structure as lacto-N-tetraose found in human milk.
According to a particular embodiment of the invention, the amount of lacto-N-tetraose in the infant formula of the invention is 70.9-1515.3mg/100g powder, or 0.1-2.0g/L on a milk basis; preferably 70.9-757.7mg/100g powder, or 0.1-1.0g/L in terms of milk.
According to a particular embodiment of the invention, the breast milk oligosaccharides further comprise 3 '-sialyllactose (3' -sialyllactose, 3 '-SL or 3SL), the amount of 3' -sialyllactose used in the infant formula being 14.2-1515.3mg/100g powder, based on the total mass of the infant formula, or 0.02-2.0g/L, calculated as milk. Preferably, the amount of 3' -sialyllactose in the infant formula is from 70.9 to 454.6mg/100g powder, or from 0.1 to 0.6g/L on a converted to milk basis; more preferably 70.9 to 227.3mg/100g of the powder, or 0.1 to 0.3g/L in terms of milk.
3' -sialyllactose is a trisaccharide structure formed by sialic acid and lactose, and is a representative substance of sialyl oligosaccharides. The commercial products of 3 '-sialyllactose in the prior art are mostly prepared by microbial fermentation methods and have the same structure as 3' -sialyllactose found in human milk.
According to a specific embodiment of the present invention, the infant formula of the present invention comprises the following human milk oligosaccharides, wherein the mass ratio of lacto-N-tetraose to 3' -sialyllactose is (3-5): 1. the research of the invention shows that the breast milk oligosaccharide composition can more obviously prevent or improve intestinal leakage or intestinal barrier damage, is favorable for improving intercellular intestinal permeability and intestinal barrier, improves intercellular gaps, and is particularly favorable for maintaining the intestinal health of infants.
According to a particular embodiment of the invention, the breast milk oligosaccharides further comprise 2 '-fucosyllactose (2' -fucosyllactise, 2 '-FL or 2FL), and the amount of 2' -fucosyllactose used in the infant formula is 14.2-3182.2mg/100g powder, based on the total mass of the infant formula, or 0.02-4.2g/L, based on milk. Preferably, the amount of 2' -fucosyllactose in the infant formula is 70.9-1818.4mg/100g powder, or 0.1-2.4g/L in terms of milk; more preferably, it is 70.9 to 1515.3mg/100g of the powder, or 0.1 to 2.0g/L in terms of milk.
2' -fucosyllactose is a trisaccharide structure formed by fucose and lactose, and is a representative substance of fucosyl oligosaccharide. Commercially available materials are usually prepared by microbial fermentation and have the same structure as oligosaccharides found in human milk.
According to a particular embodiment of the invention, the total content of the breast milk oligosaccharides in the infant formula of the present invention is below 4000mg/100g powder.
In some embodiments of the invention, the infant formula of the invention, wherein the breast milk oligosaccharides are prepared from (1-4): 1 of 2' -fucosyllactose and lacto-N-tetraose. Such breast milk oligosaccharide compositions significantly inhibit or reduce the production of intestinal branched-chain fatty acids, particularly in infants not receiving breast feeding (formula fed infants), significantly reduce the production of intestinal branched-chain fatty acids such as isovaleric acid, and further reduce the production of distal colonic isobutyric acid, lower intestinal pH, be utilized by the intestinal flora and produce gas in the intestinal system as prebiotics, and/or regulate the production of beneficial short-chain fatty acids including formic, acetic, propionic, butyric and/or lactic acids in the intestinal system.
In some embodiments of the invention, the infant formula of the invention, wherein the breast milk oligosaccharides are prepared from (3-5): (1.5-2.5): 1 of 2 '-fucosyllactose, lacto-N-tetraose and 3' -sialyllactose. The present study shows that such breast milk oligosaccharide compositions are useful for improving the intestinal immune response, and in particular for improving the immune response activity of intestinal monocytes differentiating into macrophages. In some embodiments of the present invention, the improvement of the immune response activity of monocytes to differentiate into macrophages is to increase the immune response activity of monocytes to respond to stimulation after differentiation into macrophages or to prevent the decline of the immune response activity of monocytes after training to differentiate into macrophages. In particular, flora metabolic disturbance can generate abnormal concentrations of toxins such as lipopolysaccharide in human intestinal tracts, and the breast milk oligosaccharide composition can improve the immune response activity of macrophages for the stimulation of the toxins. Furthermore, if monocytes are stimulated by some enterotoxins (i.e. trained conditions), their immune response activity is reduced after re-differentiation into macrophages, whereas the breast milk oligosaccharide composition of the present invention prevents a reduction in immune response activity, even further enhances immune response activity. According to some embodiments of the invention, the immune response activity is characterized in the present invention by measuring the amount of secreted TNF- α following differentiation of monocytes into macrophages. Experiments prove that the breast milk oligosaccharide composition can improve the secretion of TNF-alpha when monocytes are differentiated into macrophages and then respond to stimulation (such as lipopolysaccharide stimulation).
According to a specific embodiment of the present invention, the infant formula of the present invention provides a raw material for human milk oligosaccharides using a commercially available food grade human milk oligosaccharide raw material.
According to a particular embodiment of the invention, the infant formula of the invention is intended for reducing the high risk of milk protein allergy in infants, whereby it is required that the major component (more than 80%) of the total protein is hydrolysed milk protein, and that the degree of hydrolysis is between 8 and 23, and that proteins with a molecular weight distribution below 3000dal account for more than 80% of the total protein.
The raw materials of the infant formula in the invention for providing total protein can comprise one or more of hydrolyzed whey protein powder, hydrolyzed casein powder, hydrolyzed milk protein powder and hydrolyzed milk fat globule membrane protein.
According to a specific embodiment of the invention, the infant formula of the invention has a fat content of 15-29 g/100g, based on the total mass of the infant formula; the carbohydrate content is 50-58 g/100 g.
The infant formula of the present invention may comprise a base material containing milk fat as a raw material, and may further comprise vegetable oil, wherein the vegetable oil may comprise one or more of sunflower oil, corn oil, soybean oil, canola oil, coconut oil, palm oil and walnut oil, preferably sunflower oil, corn oil and soybean oil, and the vegetable oil is added to provide the product with fat components, linoleic acid and alpha-linolenic acid. In addition, the raw material for providing the fat may optionally include a raw material OPO structural fat added for providing the 1, 3-dioleoyl-2-palmitic acid triglyceride. More preferably, the raw materials comprise, based on 1000 parts by weight of the infant formula: 0-150 parts by weight of sunflower seed oil; 0-40 parts by weight of corn oil; 0-80 parts by weight of soybean oil; 0-140 parts of OPO structure grease.
According to a particular embodiment of the invention, the infant formula of the invention wherein the carbohydrates are partly derived from lactose and partly derived from non-lactose derived materials, such as pregelatinized starch, maltodextrin, corn syrup solids, glucose syrup. That is, the infant formula of the present invention, the carbohydrate-providing raw material includes, in addition to the lactose-containing base material, raw lactose and a starch-based material that is pre-hydrolyzed and gelatinized. Preferably, the raw materials comprise, based on 1000 parts by weight of the formula powder: 0-580 parts of lactose and 0-580 parts of non-lactose substances. The specific amount of lactose added can be adjusted within the stated range.
According to a particular embodiment of the invention, the infant formula of the invention further comprises one or more of nutrients, DHA, ARA, nucleotides, lactoferrin, probiotics. Preferably, the raw materials comprise, based on 1000 parts by weight of the infant formula: 8-15 parts of DHA and 14-28 parts of ARA; 0-0.7 parts by weight of lactoferrin; 7-50 parts by weight of compound nutrient containing calcium powder, vitamins and minerals.
According to a specific embodiment of the invention, in the infant formula of the invention, the compound nutrients are a combination of nutrient components meeting the national standard, and different addition amounts are used according to different formulas. According to the infant formula food, any one or any combination of the following compound nutrient components can be selectively adopted if the nutrient is added according to the needs. Preferably, the compound nutrient at least comprises compound vitamins, calcium powder and a mineral substance nutrient bag, and the dosage of each component is as follows:
1) compounding vitamins, wherein each gram of the compounding vitamins comprises the following components:
taurine: 140 to 340mg
Vitamin A: 1700 to 5800 mu gRE
Vitamin D: 25 to 70 μ g
Vitamin B1：2000～6800μg
Vitamin B2：3000～6900μg
Vitamin B6：1700～4000μg
Vitamin B12：8～20μg
Vitamin K1：200～700μg
Vitamin C: 0 to 700mg
Vitamin E: 10-70 mg of alpha-TE
Nicotinamide: 10000-41550 mu g
Folic acid: 350-920 mu g
Biotin: 70 to 245 mu g
Pantothenic acid: 7100-25230 μ g
Inositol: 0-250mg
L-carnitine: 0-60mg
2) Mineral two, per gram of mineral two:
sodium: 40 to 100mg
Potassium: 200 to 500mg
2) Mineral three, per gram mineral two:
calcium: 200 to 500mg
Phosphorus: 75-300 mg
3) Mineral one, per gram of mineral one:
iron: 20 to 110mg
Zinc: 23-90 mg
Copper: 2000-4180 μ g
Iodine: 500 to 995 mu g
Selenium: 0 to 200 μ g
Manganese: 0 to 579 μ g
4) Compounding magnesium chloride, wherein each gram of magnesium chloride is packaged:
magnesium: 80-170 mg
5) Choline chloride per gram of choline chloride
Choline: 300 to 950mg
The base material of the compound nutrient is preferably lactose, solid corn syrup or L-sodium ascorbate. Based on 1000 parts by weight of the milk protein partial hydrolysis formula powder containing breast milk oligosaccharides, the addition amount of compound nutrients is 7-52 parts by weight, wherein the preferable compound vitamin nutrition package is 2-4 parts by weight, the preferable mineral two nutrition package is 2-16 parts by weight, the preferable mineral three nutrition package is 0.5-20 parts by weight, the preferable mineral one nutrition package is 0.5-3 parts by weight, the preferable magnesium chloride is 0-3.5 parts by weight, the preferable choline chloride is 0-4.5 parts by weight, and the preferable base material of each nutrition package is lactose or sodium L-ascorbate.
In order to ensure the utilization efficiency of the nutrients, the invention selects a stable nutrient formulation.
The content of each component of the compound nutrient is to strengthen the adding amount of the nutrient substance, and does not include the content of nutrient components in other raw materials of milk powder,
according to a particular embodiment of the invention, in the infant formula of the invention, the probiotic is bifidobacteria. Preferably, the bifidobacterium is added in an amount of 0.1 to 0.2 parts by weight based on 1000 parts by weight of the infant formula; and more preferably 0.18 to 0.2 parts by weight. More preferably, the bifidobacterium powder contains 3 x 10 bifidobacteria per weight part10Above CFU.
According to some preferred embodiments of the present invention, an infant formula of the present invention comprises the following raw materials:
Figure BDA0003385201190000081
it will be appreciated that the specific amounts of the ingredients in the infant formula of the present invention are adjusted to meet the specifications required for the formula product. In the infant formula of the present invention, product performance criteria not specified or listed should be performed in accordance with the national standards of infant formula (infant formula powder) and the regulations of the relevant standards and regulations.
In the infant formula food, all raw materials can be purchased commercially, and the selection of all raw materials meets the requirements of relevant standards. In addition, the expressions "compounded", "combined" and "combined" used in the present invention are merely for convenience of description, and do not mean that the components must be mixed together and then applied. All raw materials are added and used on the premise of meeting relevant regulations.
In another aspect, the invention also provides a method of preparing the infant formula, the method comprising:
the infant formula is prepared by mixing breast milk oligosaccharides with other raw materials in the infant formula using a wet or dry process manufacturing process. The preparation process mainly comprises the following steps: preparing materials, homogenizing, concentrating, sterilizing, spray drying, and dry mixing to obtain the final product. In the invention, the breast milk oligosaccharide can be mixed together during the compounding process, and can also be added in the post-mixing process or the dry mixing process. The specific method conditions can be carried out according to the conventional method in the field of formula milk powder production.
According to a particular embodiment of the invention, the method of preparing the infant formula of the invention comprises:
1) adding powder: various powder raw materials are metered according to the formula and then uniformly added into a powder preparation tank through an air conveying system for storage.
2) Vacuum powder absorption: various powder raw materials in the powder mixing tank are sucked into the vacuum mixing tank through a vacuum system.
3) Dissolving and oil blending: and (3) putting the grease specified in the formula into an oil dissolving chamber according to the formula requirement, keeping the temperature of the oil dissolving chamber at 50-90 ℃, and pumping the oil into a mixed oil storage tank according to the formula proportion requirement through an oil pump and a flowmeter after the oil is dissolved.
4) And (3) mixed oil storage: and (3) storing the mixed oil in an oil storage tank in a heat-preservation way at the temperature of 40-50 ℃ for less than 12 hours to prevent fat oxidation.
5) Weighing: and pumping the mixed oil into a mixing tank through an oil pump according to the formula requirement.
6) Dissolving and adding nutrients: respectively adding calcium powder, mineral substances, vitamins and the like, respectively dissolving 100-200 kg of purified water, and then pumping into a wet mixing cylinder, wherein after each time of stirring, the adding tank and the pipeline are washed by 100kg of purified water.
7) Dissolving and adding breast milk oligosaccharide: and (4) dissolving the breast milk oligosaccharide raw material by using part of the mixed material liquid in the step (6), and adding the dissolved breast milk oligosaccharide raw material into a mixing tank to obtain the mixed material liquid containing the breast milk oligosaccharide.
8) And (3) filtering: filtering the mixed feed liquid by a filter screen to remove physical impurities possibly brought in the raw materials.
9) Homogenizing: homogenizing the mixed material liquid with a homogenizer at a first-stage pressure of 105 + -5 bar and a first-stage pressure of 32 + -3 bar, and mechanically processing the fat globules to disperse them into uniform fat globules.
10) Cooling and storing: and (3) feeding the homogenized material liquid into a plate heat exchanger for cooling: cooling to below 20 ℃, temporarily storing in a pre-storage cylinder, entering the next procedure within 6 hours, and starting the stirrer according to the set requirement.
11) Concentration and sterilization: double-effect concentration is adopted during production, the sterilization temperature is more than or equal to 83 ℃, and the sterilization time is 25 seconds. The discharged material concentration is 48-52% dry matter.
12) Storing concentrated milk, preheating, filtering and spray drying: the concentrated milk is temporarily stored in a concentrated milk balancing tank. Preheating to 60-70 ℃ by a scraper preheater, filtering the preheated material by a filter with the aperture of 1mm, pumping the filtered material into a drying tower by a high-pressure pump for spray drying, and agglomerating fine powder at the tower top or a fluidized bed as required. Air inlet temperature: 165-180 ℃, the exhaust temperature is 75-90 ℃, the high-pressure pump pressure is 160-210 bar, and the tower negative pressure is-4 to-2 mbar.
13) Drying and cooling the fluidized bed: and (3) drying the powder from the drying tower for the second time by using a fluidized bed (first stage), and cooling to 25-30 ℃ by using a fluidized bed (second stage) to obtain the main milk powder material.
14) Subpackaging: when the formula contains DHA, ARA, lactoferrin and bifidobacterium, the DHA, ARA, lactoferrin and bifidobacterium are weighed, sealed and packaged according to the formula requirements.
15) Dry mixing: and uniformly mixing the weighed DHA, ARA, lactoferrin and bifidobacteria with the milk powder main material in a dry mixer.
16) Powder sieving: the granularity of the milk powder is uniform through the vibrating screen, and the powder residue is discarded.
17) Powder discharging: and (4) receiving the powder by using a sterilized powder collecting box, and conveying the powder to a powder feeding room from a powder discharging room.
18) Powdering: pouring the milk powder into a powder storage tank on a large and small packaging machine according to the packaging requirements.
19) Packaging: and (5) filling nitrogen for packaging by an automatic packaging machine of 800 g. The oxygen content is lower than 1% when charging nitrogen. The oxygen content of the 900 g iron can in the automatic nitrogen-filled package is lower than 5 percent.
20) Boxing: and (4) filling the packaged small bags into a paper box, adding a powder spoon, and sealing by using a box sealing machine.
21) And (4) inspecting a finished product: and sampling and inspecting the packaged product according to an inspection plan.
22) And (4) warehousing and storing: and warehousing and storing the qualified product at normal temperature with the humidity less than or equal to 65 percent.
In another aspect, the invention also provides the use of the infant formula as a food product for improving the intestinal health of an infant and increasing the tolerance of the product. In particular, the improvement of gut health in infants comprises prevention of intestinal leakage and/or improvement of immune response. The prevention of intestinal leakage comprises improving intestinal permeability and/or intestinal barrier between cells, and/or reducing interstitial space between cells. The improvement of the immune response comprises improving the activity of the monocyte differentiation into macrophage and/or the improvement of the immune response comprises increasing the amount of TNF-alpha secreted by the monocyte after differentiation into macrophage in response to stimulation.
The infant formula food disclosed by the invention has the effects of reducing high risk of milk protein allergy and gastrointestinal function discomfort, promoting the intestinal microenvironment health, increasing the product tolerance, shortening the using time of an infant to a hydrolyzed formula and using the whole protein formula as soon as possible.
Drawings
Fig. 1 shows the effect of the breast milk oligosaccharides of the present invention on the molecular transport of FD 4.
FIG. 2 shows the effect of the immune response of breast milk oligosaccharides of the present invention on monocyte differentiated macrophages.
Detailed Description
For a more clear understanding of the technical features, objects and advantages of the present invention, reference is now made to the following detailed description of the technical aspects of the present invention with reference to specific examples, which are intended to illustrate the present invention and not to limit the scope of the present invention.
Unless specifically defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the relevant art. In the examples, each breast milk oligosaccharide material was from the supplier Jennewein, and the content of breast milk oligosaccharides was determined by a method conventional in the art. The operating conditions not specified in detail in the examples were carried out according to the usual procedures in the art.
Example 1
The raw material composition of the infant formula of this example was as follows (1000 kg produced):
120 kg of hydrolyzed whey protein powder (hydrolysis degree of 8), 115 kg of lactose, 435 kg of solid corn syrup, 120 kg of high-oleic acid sunflower oil, 40 kg of corn oil, 50 kg of soybean oil, 80 kg of OPO structure fat, 0.16 kg of breast milk oligosaccharide (LNT), 38 kg of compound nutrient, 9 kg of DHA, 18 kg of ARA, 0.1 kg of bifidobacterium and 0.65 kg of nucleotide.
The compound nutrient comprises about 3.0 kg of compound vitamin nutrient package, about 2.0 kg of choline chloride nutrient package, about 12 kg of calcium powder nutrient package, 16 kg of sodium potassium nutrient package, about 2 kg of mineral nutrient package and about 3.0 kg of magnesium chloride nutrient package, and the base material of each nutrient package is solid corn syrup.
The process for preparing the infant formula of this example is as follows:
1) adding powder: various powder raw materials are metered according to the formula and then uniformly added into a powder preparation tank through an air conveying system for storage.
2) Vacuum powder absorption: various powder raw materials in the powder mixing tank are sucked into the vacuum mixing tank through a vacuum system.
3) Dissolving and oil blending: and (3) putting the grease specified in the formula into an oil dissolving chamber according to the formula requirement, keeping the temperature of the oil dissolving chamber at 50-90 ℃, and pumping the oil into a mixed oil storage tank according to the formula proportion requirement through an oil pump and a flowmeter after the oil is dissolved.
4) And (3) mixed oil storage: and (3) storing the mixed oil in an oil storage tank in a heat-preservation way at the temperature of 40-50 ℃ for less than 12 hours to prevent fat oxidation.
5) Weighing: and pumping the mixed oil into a mixing tank through an oil pump according to the formula requirement.
6) Dissolving and adding nutrients: calcium powder, mineral substances, vitamins and the like are added respectively, 100-200 kg of purified water is used for dissolving respectively, then the mixture is thrown into a wet mixing cylinder, and after each product is beaten, the adding tank and the pipeline are flushed by 100kg of purified water.
7) Dissolving and adding breast milk oligosaccharide: and (4) dissolving the breast milk oligosaccharide by using part of the mixed feed liquid in the step (6), and adding the dissolved breast milk oligosaccharide into a mixing tank to obtain the mixed feed liquid containing the breast milk oligosaccharide.
8) And (3) filtering: filtering the mixed feed liquid by a filter screen to remove physical impurities possibly brought in the raw materials.
9) Homogenizing: homogenizing the mixed material liquid with a homogenizer at a first-stage pressure of 105 + -5 bar and a first-stage pressure of 32 + -3 bar, and mechanically processing the fat globules to disperse them into uniform fat globules.
10) Cooling and storing: and (3) feeding the homogenized material liquid into a plate heat exchanger for cooling: cooling to below 20 ℃, temporarily storing in a pre-storage cylinder, entering the next procedure within 6 hours, and starting the stirrer according to the set requirement.
11) Concentration and sterilization: double-effect concentration is adopted during production, the sterilization temperature is more than or equal to 83 ℃, and the sterilization time is 25 seconds. The discharged material concentration is 48-52% dry matter.
12) Storing concentrated milk, preheating, filtering and spray drying: the concentrated milk is temporarily stored in a concentrated milk balancing tank. Preheating to 60-70 ℃ by a scraper preheater, filtering the preheated material by a filter with the aperture of 1mm, pumping the filtered material into a drying tower by a high-pressure pump for spray drying, and agglomerating fine powder at the tower top or a fluidized bed as required. Air inlet temperature: 165-180 ℃, the exhaust temperature is 75-90 ℃, the high-pressure pump pressure is 160-210 bar, and the tower negative pressure is-4 to-2 mbar.
13) Drying and cooling the fluidized bed: and (3) drying the powder from the drying tower for the second time by using a fluidized bed (first stage), and cooling to 25-30 ℃ by using a fluidized bed (second stage) to obtain the main milk powder material.
14) Subpackaging: and weighing DHA, ARA and bifidobacteria, bagging and subpackaging by powder-making workshop personnel according to the formula requirement.
15) Dry mixing: and uniformly mixing the weighed DHA, ARA and bifidobacteria with the milk powder main material in a dry mixer.
16) Powder sieving: the granularity of the milk powder is uniform through the vibrating screen, and the powder residue is discarded.
17) Powder discharging: and (4) receiving the powder by using a sterilized powder collecting box, and conveying the powder to a powder feeding room from a powder discharging room.
18) Powdering: pouring the milk powder into a powder storage tank on a large and small packaging machine according to the packaging requirements.
19) Packaging: and (5) filling nitrogen for packaging by an automatic packaging machine of 800 g. The oxygen content is lower than 1% when charging nitrogen. The oxygen content of the 900 g iron can in the automatic nitrogen-filled package is lower than 5 percent.
20) Boxing: and (4) filling the packaged small bags into a paper box, adding a powder spoon, and sealing by using a box sealing machine.
21) And (4) inspecting a finished product: and sampling and inspecting the packaged product according to an inspection plan.
22) And (4) warehousing and storing: and warehousing and storing the qualified product at normal temperature with the humidity less than or equal to 65 percent.
In the product, the protein content is 9.6g/100g (the protein with the molecular weight distribution of less than 3000dal accounts for more than 85 percent of the total protein), the fat content is 29g/100g, the carbohydrate content is 55g/100g, and the lactose-N-tetrasaccharide content is 15mg/100g of powder.
Example 2
The raw material composition of the infant formula of this example was as follows (1000 kg produced):
80% of hydrolyzed whey protein powder (hydrolysis degree of 15) 200kg, 500 kg of lactose, 90 kg of high oleic acid sunflower seed oil, 10 kg of corn oil, 50 kg of soybean oil, 110 kg of OPO structural fat, 2.8 kg of breast milk oligosaccharide (LNT), 38 kg of compound nutrient, 3 kg of DHA, 6 kg of ARA, 0.1 kg of bifidobacterium and 0.65 kg of nucleotide.
The compound nutrient comprises about 3.0 kg of compound vitamin nutrient package, about 2.0 kg of choline chloride nutrient package, about 12 kg of calcium powder nutrient package, 16 kg of sodium potassium nutrient package, about 2 kg of mineral nutrient package and about 3.0 kg of magnesium chloride nutrient package, and the base material of each nutrient package is solid corn syrup.
The product preparation process was as in example 1.
In the product, the protein content is 16g/100g (the protein with molecular weight distribution below 3000dal accounts for more than 85% of the total protein), the fat content is 26g/100g, the carbohydrate content is 50g/100g, and the lactose-N-tetrasaccharide content is 270mg/100g of powder.
Example 3
The raw material composition of the infant formula of this example was as follows (1000 kg produced):
80% of hydrolyzed whey protein powder (hydrolysis degree of 15) 160 kg, 530 kg of solid corn syrup, 60 kg of high-oleic acid sunflower seed oil, 20 kg of corn oil, 70 kg of soybean oil, 140 kg of OPO structural fat, 7.8 kg of breast milk oligosaccharide (LNT), 38 kg of compound nutrient, 9 kg of DHA, 18 kg of ARA, 0.2 kg of bifidobacterium and 0.65 kg of nucleotide.
The compound nutrient comprises about 3.0 kg of compound vitamin nutrient package, about 2.0 kg of choline chloride nutrient package, about 12 kg of calcium powder nutrient package, 16 kg of sodium potassium nutrient package, about 2 kg of mineral nutrient package and about 3.0 kg of magnesium chloride nutrient package, and the base material of each nutrient package is solid corn syrup.
The product preparation process was as in example 1.
In the product, the protein content is 11.2g/100g (the protein with the molecular weight distribution of less than 3000dal accounts for more than 85 percent of the total protein), the fat content is 29g/100g, the carbohydrate content is 53g/100g, and the lactose-N-tetrasaccharide content is about 750mg/100g of powder.
Example 4
The raw material composition of the infant formula of this example was as follows (1000 kg produced):
80% of hydrolyzed whey protein powder (degree of hydrolysis 8) 120 kg, 115 kg of lactose, 435 kg of solid corn syrup, 120 kg of high oleic acid sunflower oil, 40 kg of corn oil, 50 kg of soybean oil, 80 kg of OPO structural fat, 0.12 kg of breast milk oligosaccharide (LNT + 3' -SL) composition, 38 kg of compound nutrient, 9 kg of DHA, 18 kg of ARA, 0.1 kg of bifidobacterium and 0.65 kg of nucleotide.
The compound nutrient comprises about 3.0 kg of compound vitamin nutrient package, about 2.0 kg of choline chloride nutrient package, about 12 kg of calcium powder nutrient package, 16 kg of sodium potassium nutrient package, about 2 kg of mineral nutrient package and about 3.0 kg of magnesium chloride nutrient package, and the base material of each nutrient package is solid corn syrup.
The product preparation process was as in example 1.
In the product, the protein content is 9.6g/100g (the protein with the molecular weight distribution of less than 3000dal accounts for more than 85 percent of the total protein), the fat content is 29g/100g, the carbohydrate content is 55g/100g, and the lactose-N-tetrasaccharide content is 85.7mg/100g of powder; the content of 3' -sialyllactose was 21.4mg/100g powder;
example 5
The raw material composition of the infant formula of this example was as follows (1000 kg produced):
80% of hydrolyzed whey protein powder (hydrolysis degree of 15) 200kg, 500 kg of lactose, 90 kg of high oleic acid sunflower seed oil, 10 kg of corn oil, 50 kg of soybean oil, 110 kg of OPO structural fat, 8.7 kg of breast milk oligosaccharide (LNT + 3' -SL) composition, 38 kg of compound nutrient, 3 kg of DHA, 6 kg of ARA, 0.1 kg of bifidobacterium and 0.65 kg of nucleotide.
The compound nutrient comprises about 3.0 kg of compound vitamin nutrient package, about 2.0 kg of choline chloride nutrient package, about 12 kg of calcium powder nutrient package, 16 kg of sodium potassium nutrient package, about 2 kg of mineral nutrient package and about 3.0 kg of magnesium chloride nutrient package, and the base material of each nutrient package is solid corn syrup.
The product preparation process was as in example 1.
In the product, the protein content is 16g/100g (the protein with the molecular weight distribution of less than 3000dal accounts for more than 85 percent of the total protein), the fat content is 26g/100g, the carbohydrate content is 50g/100g, and the content of lactose-N-tetrasaccharide is about 680mg/100g powder; the content of 3' -sialyllactose is 170mg/100g powder;
example 6
The raw material composition of the infant formula of this example was as follows (1000 kg produced):
155 kg of hydrolyzed whey protein powder (hydrolysis degree of 15), 530 kg of solid corn syrup, 60 kg of high-oleic acid sunflower seed oil, 20 kg of corn oil, 70 kg of soybean oil, 140 kg of OPO structural fat, 9.6 kg of breast milk oligosaccharide (LNT + 3' -SL) composition, 38 kg of compound nutrient, 9 kg of DHA, 18 kg of ARA, 0.2 kg of bifidobacterium and 0.65 kg of nucleotide.
The compound nutrient comprises about 3.0 kg of compound vitamin nutrient package, about 2.0 kg of choline chloride nutrient package, about 12 kg of calcium powder nutrient package, 16 kg of sodium potassium nutrient package, about 2 kg of mineral nutrient package and about 3.0 kg of magnesium chloride nutrient package, and the base material of each nutrient package is solid corn syrup.
The product preparation process was as in example 1.
In the product, the protein content is 11.2g/100g (the protein with the molecular weight distribution below 3000dal accounts for more than 85 percent of the total protein), the fat content is 29g/100g, the carbohydrate content is 53g/100g, and the lactose-N-tetrasaccharide content is about 751mg/100g of powder; the 3' sialyllactose content was approximately 188mg per 100g of powder.
Example 7
The raw material composition of the infant formula of this example was as follows (1000 kg produced):
120 kg of hydrolyzed whey protein powder (hydrolysis degree of 8), 115 kg of lactose, 435 kg of solid corn syrup, 120 kg of high-oleic acid sunflower oil, 40 kg of corn oil, 50 kg of soybean oil, 80 kg of OPO structural fat, 1.1 kg of breast milk oligosaccharide (2 '-FL + LNT + 3' -SL) composition, 38 kg of compound nutrient, 9 kg of DHA, 18 kg of ARA, 0.1 kg of bifidobacterium and 0.65 kg of nucleotide.
The compound nutrient comprises about 3.0 kg of compound vitamin nutrient package, about 2.0 kg of choline chloride nutrient package, about 12 kg of calcium powder nutrient package, 16 kg of sodium potassium nutrient package, about 2 kg of mineral nutrient package and about 3.0 kg of magnesium chloride nutrient package, and the base material of each nutrient package is solid corn syrup.
The product preparation process was as in example 1.
In the product, the protein content is 9.6g/100g (the protein with molecular weight distribution below 3000dal accounts for more than 85% of the total protein), the fat content is 29g/100g, and the carbohydrate content is 55g/100 g. The 2 '-fucosyllactose content of the product was about 57mg/100g, the amount of lacto-N-tetraose in the product was about 28.5mg/100g, and the amount of 3' -sialyllactose in the product was about 14.2mg/100 g.
Example 8
The raw material composition of the infant formula of this example was as follows (1000 kg produced):
80% of hydrolyzed whey protein powder (hydrolysis degree of 15) 200kg, 500 kg of lactose, 90 kg of high oleic acid sunflower seed oil, 10 kg of corn oil, 50 kg of soybean oil, 110 kg of OPO structural fat, 20 kg of breast milk oligosaccharide (2 '-FL + LNT + 3' -SL) composition, 38 kg of compound nutrient, 3 kg of DHA, 6 kg of ARA, 0.1 kg of bifidobacterium and 0.65 kg of nucleotide.
The compound nutrient comprises about 3.0 kg of compound vitamin nutrient package, about 2.0 kg of choline chloride nutrient package, about 12 kg of calcium powder nutrient package, 16 kg of sodium potassium nutrient package, about 2 kg of mineral nutrient package and about 3.0 kg of magnesium chloride nutrient package, and the base material of each nutrient package is solid corn syrup.
The product preparation process was as in example 1.
In the product, the protein content is 16g/100g (the protein with molecular weight distribution below 3000dal accounts for more than 85% of the total protein), the fat content is 26g/100g, and the carbohydrate content is 50g/100 g. The product contains about 1113mg/100g of 2 '-fucosyllactose, about 557mg/100g of lacto-N-tetraose and about 278mg/100g of 3' -sialyllactose.
Example 9
The raw material composition of the infant formula of this example was as follows (1000 kg produced):
80% of hydrolyzed whey protein powder (hydrolysis degree of 15) 150 kg, 530 kg of solid corn syrup, 60 kg of high-oleic acid sunflower seed oil, 20 kg of corn oil, 70 kg of soybean oil, 140 kg of OPO structural fat, 28 kg of breast milk oligosaccharide (2 '-FL + LNT + 3' -SL) composition, 38 kg of compound nutrient, 9 kg of DHA, 18 kg of ARA, 0.2 kg of bifidobacterium and 0.65 kg of nucleotide.
The compound nutrient comprises about 3.0 kg of compound vitamin nutrient package, about 2.0 kg of choline chloride nutrient package, about 12 kg of calcium powder nutrient package, 16 kg of sodium potassium nutrient package, about 2 kg of mineral nutrient package and about 3.0 kg of magnesium chloride nutrient package, and the base material of each nutrient package is solid corn syrup.
The product preparation process was as in example 1.
In the product, the protein content is 11.2g/100g (the protein with molecular weight distribution below 3000dal accounts for more than 85% of the total protein), the fat content is 29g/100g, and the carbohydrate content is 53g/100 g. The product contained about 1515mg/100g of 2 '-fucosyllactose, about 757mg/100g of lacto-N-tetraose and about 378mg/100g of 3' -sialyllactose.
Experiment for improving intestinal leakage by breast milk oligosaccharide
1. Experiment for improving cell bypass permeability
The experiment inspects the influence of different HMOs and compositions thereof on the epithelial function of the small intestine under normal conditions, and shows that the intestinal permeability and the intestinal barrier (cell bypass permeability) between cells are improved through indexes such as the intestinal barrier and FD4 (fluorescein isothiocyanate dextran; FITC labeled dextran) molecular transport condition and the like.
Normal (non-invasive) condition
Caco-2 cells were cultured in DMEM medium in a transwell system for 21 days to simulate the small intestine epithelial cell layer. The test HMO or HMO composition (final concentration 0.1mg/mL in the culture system) was added and cultured for 24 hours, and then FD4 molecule (final surface concentration 250ug/mL) was added and the culture was continued. FD4 molecular transport was measured as three biological replicates at 24 hours of continued culture. A DMEM medium control was also set without any HMO or HMO composition tested.
Data analysis
The permeability data of FD4 for molecular transport was statistically analyzed using two-tailed paired T-test. Two groups were marked with an asterisk if they were significantly different and p < 0.05. Two asterisks indicate p < 0.01. Three asterisks indicate p < 0.001.
Results of the experiment
The results are shown in FIG. 1. LNT group, mass ratio 4: the HMO composition of the LNT and the 3 '-SL of the invention 1 obviously reduces the transport of FD4 molecules (P <0.05), and shows that the composition of the LNT, the LNT and the 3' -SL of the invention can improve the intercellular intestinal permeability (reduce the intercellular intestinal permeability), improve the intestinal barrier effect, and the analysis reasons are mainly to reduce the intercellular gaps and improve the intercellular tightness, so that the tightness of the paracellular transport pathway is increased, and the intestinal leakage can be effectively reduced.
TcDA invasion assay
The experiment examines the influence of HMO compositions (the mass ratio of LNT to 3' -SL is 2: 1 and 4: 1 respectively) with different proportions on the small intestine epithelial function under the condition of invasion by microbial toxin Clostridium difficile toxin A (TcDA), and the expression of the TEER index of transmembrane resistance is determined through a TcDA invasion experiment.
Caco-2 cells were cultured in DMEM medium in a transwell system for 21 days to simulate the small intestine epithelial cell layer. The test HMO composition (final concentration in the culture system 0.1mg/mL) was added and incubated for 24 hours, followed by addition of TcDA (final surface concentration of 200ug/mL) and continued incubation. Transmembrane resistance TEER was measured at 0, 3 hours of continued culture with TcDA added, and three times were measured as biological replicates. A DMEM medium control was also set without any HMO or HMO composition tested.
Data analysis
For the data of transmembrane resistance TEER, statistical analysis was performed using Tukey's multiple comparison test. Two groups were marked with an asterisk if they were significantly different and p < 0.05. Two asterisks indicate p < 0.01. Three asterisks indicate p < 0.001.
Results of the experiment
Table 1 shows the effect of LNT and 3' -SL combination on transmembrane resistance as measured after three hours of incubation with toxin A.
TABLE 1
Figure BDA0003385201190000171
From the results of intestinal barrier test for three hours, it can be seen that the mass ratio of 2: the LNT + 3' -SL composition of 1 significantly reduced transmembrane resistance compared to the control, whereas the mass ratio of the invention was 4: the LNT +3 '-SL composition of 1 did not significantly reduce transmembrane resistance, reflecting the tendency of the present invention to be synergistic between LNT and 3' -SL compositions in a specific ratio range.
Experiment for improving monocyte differentiation into macrophage immune response effect of breast milk oligosaccharide composition
The influence of HMO on the activity of monocyte differentiation into macrophage immune response was examined by acclimatization and activation of monocytes.
Collection and culture of monocytes
The experimental conditions for the specific blood sample collection and culture are as follows:
monocytes were isolated from blood samples from multiple healthy adult donors using the quadro macs system and CD14 microbeads magnetic bead sorting kit (Miltenyi Biotec, ledon, the netherlands) using the manufacturer's recommended protocol. Prior to blood collection, written consent was obtained from the donor.
Culturing the mononuclear cells: RPMI 1640-Glutamax medium (Gibco, Blastwick, the Netherlands) supplemented with 10% fetal bovine serum (FBS, Hyclone, Einholwen, the Netherlands), 1% MEM non-essential amino acids (Gibco Blastwick, the Netherlands), 1% sodium pyruvate (Lonza, Braudard, the Netherlands), 1% penicillin/streptomycin (Sigma, St. Louis, Mo., USA) at 1X 106Cells were cultured in 24-well plates at a concentration of 2 ml/well.
Immune response activity study experiments included pretreatment of monocytes and challenge testing of immune responses after 6 days. The specific operation is as follows:
monocytes recovered from liquid nitrogen and after 1 day of recovery, the following groups of experiments were performed, and all HMO starting materials in this experiment were from Jennewein:
group HMO: different HMO test samples (different HMO monomers or HMO compositions in different proportions, wherein the proportions of the monomers in the HMO composition are in mass ratio) were added to the monocyte culture medium at a final concentration of 0.1mg/mL, co-cultured with monocytes for 24 hours, after which the culture medium was replaced with fresh medium (to flush out the HMO under test) and continued for 6 days to differentiate the monocytes into macrophages. Then, lipopolysaccharide (lipopolysaccharide in the culture medium of 10ng/mL final concentration) stimulation for 24 hours, and from the supernatant determination of secreted TNF alpha.
Control group 1 (lipopolysaccharide group): the monocytes were pretreated with lipopolysaccharide by adding lipopolysaccharide to the monocyte culture medium at a final concentration of 0.1mg/mL, co-culturing with monocytes for 24 hours, and then replacing the fresh medium (washing away LPS) for further culturing for 6 days to differentiate monocytes into macrophages. Then, lipopolysaccharide (lipopolysaccharide in the culture medium of 10ng/mL final concentration) stimulation for 24 hours, and from the supernatant determination of secreted TNF alpha. The purpose of this control pre-lipopolysaccharide pretreatment was to reduce the ability of monocytes to respond to subsequent re-stimulation, and it was expected that lipopolysaccharide would reduce the response of monocytes to the second lipopolysaccharide stimulation.
Control group 2 (medium group): monocytes were cultured in a medium without any test substance added for 7 days to differentiate monocytes into macrophages. Then, lipopolysaccharide (lipopolysaccharide in the culture medium of 10ng/mL final concentration) stimulation for 24 hours, and from the supernatant determination of secreted TNF alpha. The experimental method refers to the method reported by Bekkering et al (2016, Clinical and Vaccine Immunology), and TNF-alpha is measured by ELISA (enzyme-linked immunosorbent assay) and a kit special for TNF-alpha, wherein the wavelength of the ELISA is 450nm, the absorption value of the ELISA is measured by a Plate Reader Spark device, the Spark sequence number is 2009003477, and the Spark sequence number is V3.1.
Data analysis
Macrophage immune response data were statistically analyzed using one way ANOVA. Two groups were marked with an asterisk if they were significantly different and p < 0.05. Two asterisks indicate p < 0.01. Three asterisks indicate p < 0.001.
Results of the experiment
See fig. 2 for experimental results. The mass ratio is 4: 2: 2' -FL of 1: LNT: the group of 3' -SL breast milk oligosaccharide compositions showed a significant increase in TNF-alpha secretion in the monocyte trained model (p < 0.05).
In addition, as expected from experiments, the test results of the lipopolysaccharide group showed that lipopolysaccharide reduced the immune response, and it was seen that the lipopolysaccharide group was significantly reduced compared to the medium group without any test substance.

Claims (10)

1. An infant formula comprising human milk oligosaccharides, said human milk oligosaccharides including lacto-N-tetraose, wherein the total content of lacto-N-tetraose in the infant formula is 14.2-2273.0mg/100g powder, based on the total mass of the infant formula, or 0.02-3.0g/L, based on the converted milk;
and the total protein content in the infant formula is 9-20 g/100g based on the total mass of the infant formula, the total protein comprises hydrolyzed milk protein, and the protein with the hydrolysis degree of 8-23 and the molecular weight distribution of less than 3000dal accounts for more than 80% of the total protein.
2. The infant formula of claim 1, wherein the breast milk oligosaccharides further comprise 3 '-sialyllactose and the amount of 3' -sialyllactose used in the infant formula is from 14.2 to 1515.3mg/100g powder, based on the total mass of the infant formula, or from 0.02 to 2.0g/L, based on milk.
3. The infant formula according to claim 2, wherein the breast milk oligosaccharide has a ratio by mass of lacto-N-tetraose to 3' -sialyllactose of (3-5): 1.
4. the infant formula of claim 1 or 2, wherein the breast milk oligosaccharides further comprise 2 '-fucosyllactose, the 2' -fucosyllactose being applied in the infant formula in an amount of 14.2-3182.2mg/100g powder, or 0.02-4.2g/L on a milk basis, based on the total mass of the infant formula;
preferably, the total content of the breast milk oligosaccharides in the infant formula is below 4000mg/100g powder.
5. The infant formula according to claim 4, wherein the breast milk oligosaccharides are prepared from (1-4) by mass: the composition comprises 1 2' -fucosyllactose and lacto-N-tetraose, or comprises the following components in percentage by mass (3-5): (1.5-2.5): 1 of 2 '-fucosyllactose, lacto-N-tetraose and 3' -sialyllactose.
6. An infant formula according to claim 1, wherein the fat content is 15-29 g/100g, based on the total mass of the infant formula; the carbohydrate content is 50-58 g/100 g.
7. The infant formula of any one of claims 1-6, further comprising one or more of nutrients, DHA, ARA, nucleotides, lactoferrin, probiotics.
8. The infant formula according to claim 1, which is for:
preventing intestinal leakage, including improving intestinal permeability and/or intestinal barrier between cells, and/or reducing interstitial spaces between cells; and/or
Improving an immune response, said improving an immune response comprising improving the activity of a monocyte to differentiate into a macrophage, and/or said improving an immune response comprising increasing the amount of TNF- α secreted by a monocyte in response to a stimulus following differentiation into a macrophage.
9. A method of preparing an infant formula according to any one of claims 1 to 8, which method comprises:
mixing the breast milk oligosaccharides with other raw materials in the infant formula by adopting a wet or dry production process to prepare the infant formula;
preferably, the raw materials for providing total protein in the raw materials of the infant formula include one or more of hydrolyzed whey protein powder, hydrolyzed casein powder, hydrolyzed milk protein powder, and hydrolyzed milk fat globule membrane protein.
10. Use of an infant formula according to any one of claims 1 to 8 as a food product for improving the intestinal health and increasing the tolerance of the product in infants.
CN202111450898.9A 2021-11-30 2021-11-30 Milk protein partially hydrolyzed hypoallergenic infant formulas containing breast milk oligosaccharides Active CN114128767B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202111450898.9A CN114128767B (en) 2021-11-30 2021-11-30 Milk protein partially hydrolyzed hypoallergenic infant formulas containing breast milk oligosaccharides

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202111450898.9A CN114128767B (en) 2021-11-30 2021-11-30 Milk protein partially hydrolyzed hypoallergenic infant formulas containing breast milk oligosaccharides

Publications (2)

Publication Number Publication Date
CN114128767A true CN114128767A (en) 2022-03-04
CN114128767B CN114128767B (en) 2023-12-22

Family

ID=80386770

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202111450898.9A Active CN114128767B (en) 2021-11-30 2021-11-30 Milk protein partially hydrolyzed hypoallergenic infant formulas containing breast milk oligosaccharides

Country Status (1)

Country Link
CN (1) CN114128767B (en)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN115644430A (en) * 2022-11-30 2023-01-31 内蒙古伊利实业集团股份有限公司 Milk protein deep-hydrolyzed infant formula containing breast milk oligosaccharide LNnT
CN115669732A (en) * 2022-12-29 2023-02-03 内蒙古伊利实业集团股份有限公司 Hypoallergenic infant partially hydrolyzed formula containing the breast milk oligosaccharide LNnT

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20130243797A1 (en) * 2010-11-23 2013-09-19 Norbert Sprenger Composition comprising hydrolysed proteins and oligisaccharides for treating skin diseases
US20200138836A1 (en) * 2017-05-24 2020-05-07 Societe Des Produits Nestle S.A. Hmos blends for use in infants or young children for health purposes
CN111903766A (en) * 2020-08-10 2020-11-10 沈阳农业大学 Infant formula milk powder and preparation method thereof
CN112868800A (en) * 2019-11-29 2021-06-01 内蒙古伊利实业集团股份有限公司 Infant formula milk powder containing breast milk oligosaccharide for improving immunity and preparation method thereof
CN113226062A (en) * 2018-12-21 2021-08-06 雀巢产品有限公司 Nutritional composition comprising 2 '-fucosyllactose (2' FL) for improving gastrointestinal barrier
CN113519849A (en) * 2020-04-14 2021-10-22 内蒙古伊利实业集团股份有限公司 Breast milk oligosaccharide for improving intestinal tract resistance to escherichia coli infection and application thereof

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20130243797A1 (en) * 2010-11-23 2013-09-19 Norbert Sprenger Composition comprising hydrolysed proteins and oligisaccharides for treating skin diseases
US20200138836A1 (en) * 2017-05-24 2020-05-07 Societe Des Produits Nestle S.A. Hmos blends for use in infants or young children for health purposes
CN113226062A (en) * 2018-12-21 2021-08-06 雀巢产品有限公司 Nutritional composition comprising 2 '-fucosyllactose (2' FL) for improving gastrointestinal barrier
CN112868800A (en) * 2019-11-29 2021-06-01 内蒙古伊利实业集团股份有限公司 Infant formula milk powder containing breast milk oligosaccharide for improving immunity and preparation method thereof
CN113519849A (en) * 2020-04-14 2021-10-22 内蒙古伊利实业集团股份有限公司 Breast milk oligosaccharide for improving intestinal tract resistance to escherichia coli infection and application thereof
CN111903766A (en) * 2020-08-10 2020-11-10 沈阳农业大学 Infant formula milk powder and preparation method thereof

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN115644430A (en) * 2022-11-30 2023-01-31 内蒙古伊利实业集团股份有限公司 Milk protein deep-hydrolyzed infant formula containing breast milk oligosaccharide LNnT
CN115669732A (en) * 2022-12-29 2023-02-03 内蒙古伊利实业集团股份有限公司 Hypoallergenic infant partially hydrolyzed formula containing the breast milk oligosaccharide LNnT
CN115669732B (en) * 2022-12-29 2023-04-14 内蒙古伊利实业集团股份有限公司 Hypoallergenic infant partially hydrolyzed formula containing the breast milk oligosaccharide LNnT

Also Published As

Publication number Publication date
CN114128767B (en) 2023-12-22

Similar Documents

Publication Publication Date Title
TWI391100B (en) Method for simulating the functional attributes of human milk oligosaccharides in formula-fed infants
CN114128767B (en) Milk protein partially hydrolyzed hypoallergenic infant formulas containing breast milk oligosaccharides
EP3209308A1 (en) Activated bifidobacteria and methods of use thereof
RU2600877C2 (en) Fermented infant formula
CN111838683A (en) Prebiotic composition containing human milk oligosaccharide and application thereof
CN114223729B (en) Application of bifidobacterium longum subspecies infantis and breast milk oligosaccharide composition in formula milk powder
CN114568526A (en) Mother emulsified infant formula powder for improving intestinal microenvironment health and application thereof
CN114208893A (en) Formula milk powder capable of improving intestinal microenvironment health and preparation method and application thereof
CN114145353A (en) Formula milk powder capable of improving intestinal microenvironment health and preparation method and application thereof
CN105324033A (en) Fermented nutrition high in lactose with increased iron bioavailability
CN108669563A (en) A kind of composition, its application and strengthen immunity, the product for improving allergy
RU2426438C2 (en) Synbiotic composition for correction of intestinal tract microbiocenosis abnormalities and enhancement of organism general resistivity and its production methods (versions)
CN116076728B (en) Composition for promoting growth of bifidobacteria
CN114145354B (en) Formula milk powder containing breast milk oligosaccharide and preparation method and application thereof
CN114223728B (en) Premature/low birth weight infant formula containing prebiotics and probiotics
EP2520181A1 (en) Fermented infant formula
EP4152962A1 (en) &lt;smallcaps/&gt;? ? ?l.reuteri? ? ? ? ?pre-conditioning of
CN114304651A (en) Composition for improving immunity
CN115669732B (en) Hypoallergenic infant partially hydrolyzed formula containing the breast milk oligosaccharide LNnT
CN114145356B (en) Formula milk powder capable of improving immune response and preparation method and application thereof
CN114568527A (en) Mother emulsified infant formula powder for improving intestinal microenvironment health and application thereof
WO2024114729A1 (en) Composition for resisting enteritis and/or protecting integrity of intestinal barrier, and use thereof
CN114451453B (en) Breast milk oligosaccharide composition for preventing intestinal leakage and application thereof
CN115644431B (en) Formula food containing breast milk oligosaccharide for premature infant and preparation method and application thereof
CN115769840B (en) Formula milk powder and preparation method thereof

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant