CN115769841B - Formula milk powder and preparation method thereof - Google Patents

Formula milk powder and preparation method thereof Download PDF

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Publication number
CN115769841B
CN115769841B CN202211529126.9A CN202211529126A CN115769841B CN 115769841 B CN115769841 B CN 115769841B CN 202211529126 A CN202211529126 A CN 202211529126A CN 115769841 B CN115769841 B CN 115769841B
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parts
formula
powder
milk
lnnt
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CN115769841A (en
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郝威
刘彪
孔小宇
周名桥
肖竞舟
李放
王逸伦
闫雅璐
段素芳
司徒文佑
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Inner Mongolia Yili Industrial Group Co Ltd
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Inner Mongolia Yili Industrial Group Co Ltd
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    • 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
    • A23C21/00—Whey; Whey preparations
    • A23C21/06—Mixtures of whey with milk products or milk components
    • 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
    • A23C21/00—Whey; Whey preparations
    • A23C21/02—Whey; Whey preparations containing, or treated with, microorganisms or enzymes
    • A23C21/026—Whey; Whey preparations containing, or treated with, microorganisms or enzymes containing, or treated only with, lactic acid producing bacteria, bifidobacteria or propionic acid bacteria
    • 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
    • A23C21/00—Whey; Whey preparations
    • A23C21/04—Whey; Whey preparations containing non-milk components as source of fats or proteins
    • 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
    • A23C21/00—Whey; Whey preparations
    • A23C21/08—Whey; Whey preparations containing other organic additives, e.g. vegetable or animal products
    • 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
    • A23C21/00—Whey; Whey preparations
    • A23C21/10—Whey; Whey preparations containing inorganic additives

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  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Food Science & Technology (AREA)
  • Polymers & Plastics (AREA)
  • Inorganic Chemistry (AREA)
  • Microbiology (AREA)
  • Zoology (AREA)
  • Dairy Products (AREA)

Abstract

The invention provides formula milk powder and a preparation method thereof. The preparation raw materials of the formula milk powder comprise: 0-4.5 parts of breast milk oligosaccharide, 890-3800 parts of raw milk, 0-480 parts of lactose, 0-540 parts of desalted whey powder, 0-350 parts of skim milk powder, 80-80 parts of whey protein powder WPC, 340-120 parts of whey protein powder WPC, 0-225 parts of vegetable oil, 0-45 parts of alpha-whey protein, 0-23 parts of beta-casein, 1-2.5 parts of phospholipid, 0-30 parts of fructo-oligosaccharide, 0-90 parts of galacto-oligosaccharide syrup, 0-0.18 parts of probiotics, 0-5 parts of anhydrous cream, 0-0.8 parts of nucleotide, 1-5 parts of vitamin nutrition and the like. The formula milk powder provided by the invention provides a solution for improving intestinal microenvironment health, such as reducing hydrogen sulfide.

Description

Formula milk powder and preparation method thereof
Technical Field
The invention relates to the field of dairy products, in particular to formula milk powder and a preparation method thereof.
Background
Breast milk oligosaccharides are approximately 12-14g/L in mature milk and 20-24g/L in colostrum. Breast milk oligosaccharides are mainly composed of three major classes: (1) Fucoidan-based oligosaccharides, represented by 2' -fucoidan and 3-fucoidan; (2) Sialyl-based oligosaccharides, represented by 3 '-sialyllactose and 6' -sialyllactose; (3) Oligosaccharides having a core sugar chain structure containing no fucosyl group or sialic acid group are represented by lactose-N-tetraose (LNT) and lactose-N-neotetraose (LNnT).
As the three major functions of breast milk oligosaccharides are gradually reported and discovered: (1) inhibiting the attachment and infection of specific pathogens; (2) As a prebiotic, promoting the growth of bacteria in the intestinal symbiotic system; (3) Directly reducing the inflammatory response of mucous membranes under toxic stimuli the presence and amount of breast milk oligosaccharides varies from individual to individual and is related to the lewis secretory composition of the lactating mother.
lactose-N-neotetraose (LNnT) is one of the highest neutral sugar chain core human milk oligosaccharides in human milk, and is a tetraose composed of two D-galactose, one N-acetyl-D glucosamine and one D-glucose. Currently, commercially available LNnT is prepared via microbial fermentation, and the structure is determined by mass spectrometry and nuclear magnetic resonance to have the same structure as that of oligosaccharides found in human milk.
Breast milk oligosaccharides can be regarded as a microecological management tool for improving the health of the body, and can alter, regulate and recombine the already existing intestinal flora. Anaerobic bacteria, bifidobacteria, eubacteria, streptococcus, lactobacillus and the like in the intestinal flora can release metabolite short chain fatty acids mainly comprising acetic acid, propionic acid, butyric acid, valeric acid and the like through fermenting carbohydrates, proteins, lipids and the like.
The microbial flora in the intestinal tract of infants can also ferment to generate gases such as carbon dioxide, methane and the like, so that the infants have symptoms such as abdominal distension, intestinal flatulence and the like. In addition, during degradation of the intestinal mucosa, harmful flora can invade the inside of the intestinal mucosa, and mucopolysaccharide can be rapidly degraded into thiosulfate and free sulfate radical through intermediate reaction, and finally toxic gas hydrogen sulfide is generated. In inflammatory reactions, intestinal homeostasis is disrupted, thiosulfate can be oxidized to tetrathionate, and further attack by harmful bacteria such as salmonella is promoted. Under these conditions, the intestinal wall integrity is compromised and the symptoms of intestinal leakage can lead to reduced production of short chain fatty acids. At present, in the fields of infant formula powder, complementary food, nutritional supplements and the like, the breast milk oligosaccharide and the composition formed by the breast milk oligosaccharide and probiotics provide a scheme capable of improving intestinal microenvironment health such as reducing toxic gas hydrogen sulfide.
Disclosure of Invention
It is an object of the present invention to provide a formula.
The invention also aims at providing a preparation method of the formula milk powder.
In order to achieve the above purpose, in one aspect, the present invention provides a formula, wherein the formula comprises the following raw materials in parts by weight based on 1000 parts by weight of the total formula: 0-4.5 parts of breast milk oligosaccharide (HMOs), 890-3800 parts of raw milk, 0-480 parts of lactose, 0-540 parts of desalted whey powder, 0-350 parts of skim milk powder, 80-80 parts of whey protein powder WPC, 34-120 parts of whey protein powder WPC, 0-225 parts of vegetable oil, 0-45 parts of alpha-whey protein, 0-23 parts of beta-casein, 1-2.5 parts of phospholipid, 0-30 parts of fructo-oligosaccharide (fructo-oligosaccharide powder), 0-90 parts of galacto-oligosaccharide syrup, 0-0.18 part of probiotics, 0-5 parts of anhydrous cream, 0-0.8 part of nucleotide, 1-5 parts of vitamin nutrient, 0-3 parts of choline chloride, 0-2 parts of mineral A and 1-12 parts of mineral B; wherein, the mineral A is selected from the combination of mineral salts of one or more of copper, iron, magnesium, zinc, selenium, manganese and iodine, and the mineral B is selected from the combination of mineral salts of one or more of calcium, phosphorus, sodium and potassium.
According to some embodiments of the invention, wherein the human milk oligosaccharide comprises human milk oligosaccharide LNnT, wherein the human milk oligosaccharide LNnT is present in an amount of 0.01 to 4.5 parts by weight.
According to some embodiments of the invention, wherein the vegetable oil is selected from one or more of the group consisting of structural oil OPO, high oleic sunflower oil, corn oil, canola oil, and soybean oil; when the vegetable oil components are independent, the total weight of the formula milk powder is 1000 parts, and the dosages of the vegetable oil components are independent respectively: 0 to 170 parts of structural grease OPO, 0 to 150 parts of high oleic sunflower seed oil, 0 to 50 parts of corn oil, 0 to 50 parts of low erucic acid rapeseed oil and 0 to 80 parts of soybean oil.
According to some embodiments of the invention, wherein the vitamin nutrient is selected from the group consisting of vitamin a, vitamin D, vitamin E, vitamin K1, vitamin B2, vitamin B6, vitamin B12, nicotinamide, folic acid, pantothenic acid, and biotin.
The vitamin nutrient and various mineral substances in the formula can adopt a composite nutrient composition of nutrient components meeting the national standard, different addition amounts can be used according to different formulas, and the specific addition mode can be carried out by referring to the conventional operation in the field.
Preferably, the formula milk powder can selectively adopt any one or any combination of the following compound nutrient components (per gram content; each index is calculated according to the middle value of the adding range; basically, the condition that the content of all indexes is simultaneously maximum) if the compound nutrient is added according to the requirement:
vitamin a: 1300-4000. Mu.gRE
Vitamin D: 24-60 mug
Vitamin E: 13-41 mg alpha-TE
Vitamin K1: 200-550 mug
Vitamin B1: 2600-6500 mug
Vitamin B2: 600-3000 mug
Vitamin B6: 1200-4000 mug
Vitamin B12: 2-14.0 mug
Nicotinamide: 12000-30000 mug
Folic acid: 300-653 mu g
Pantothenic acid: 8750-16230 mug
Biotin: 56-150 mug.
According to some embodiments of the invention, wherein the mineral content (per gram content) is:
sodium: 30-400 mg
Potassium: 63-400 mg
Copper: 2200-5540 mug
Magnesium: 133-400 mg
Iron: 23-95 mg
Zinc: 25-52 mg
Calcium: 110-300 mg
Phosphorus: 75-200 mg
Iodine: 200-1592 mug
Selenium: 16-48 mu g
Manganese: 16-547 mug.
According to some embodiments of the invention, wherein the probiotic is selected from: one or more of Bifidobacterium animalis subspecies BB-12, bifidobacterium infantis YLGB-1496, bifidobacterium animalis subspecies HN019 and Bifidobacterium lactis BL-99.
According to some embodiments of the invention, the bifidobacterium infantis YLGB-1496 is bifidobacterium infantis with a accession number of CGMCC No. 21109.
According to some embodiments of the invention, the bifidobacterium lactis BL-99 is bifidobacterium lactis with a preservation number of CGMCC No. 15650.
According to some embodiments of the invention, wherein the probiotic is bifidobacterium lactis BL-99.
According to some embodiments of the invention, wherein the formula contains breast milk oligosaccharide LNnT in an amount of 142.9-428.6mg/100g of powder per 100g of formula; preferably 178.6-392.9mg/100g of powder.
According to some embodiments of the invention, wherein the formula contains probiotics in an amount of 10 per gram of formula 6 -2×10 12 CFU; preferably 10 8 -2×10 10 CFU。
According to some embodiments of the invention, the formula further comprises the following components: 0.20 to 2.04 parts by weight of inositol, 0 to 0.65 part by weight of taurine, 0.06 to 0.5 part by weight of L-carnitine, 3 to 18 parts by weight of DHA and 3 to 22 parts by weight of ARA.
According to some embodiments of the invention, the formula is an infant formula for reducing hydrogen sulfide in the infant's intestinal tract.
According to a specific embodiment of the invention, the infant formula of the invention may be an infant formula or a baby formula.
According to a specific embodiment of the present invention, the infant formula of the present invention may further comprise conventional components of infant formula in addition to the lactobacillus and the breast milk oligosaccharide.
In the formula milk powder of the invention, the proteins and fats of raw milk can be replaced by whole milk powder and/or skim milk powder; whey protein powder WPC80 and whey protein powder WPC34 may be partially replaced by alpha-lactalbumin.
In another aspect, the invention also provides a preparation method of the formula milk powder, which comprises the following steps: wet compounding, homogenizing, concentrating, sterilizing, spray drying, dry mixing and packaging.
According to some specific embodiments of the present invention, the wet-process batching includes homogenizing and sterilizing raw milk, and sequentially adding powder raw materials, grease raw materials, vitamin nutrients, mineral raw materials and breast milk oligosaccharides to obtain a mixed feed liquid; the powder raw material is other solid raw materials except breast milk oligosaccharide, vitamin nutrient and mineral raw materials.
According to some embodiments of the invention, wherein the wet-process formulation comprises the steps of:
1) Rough filtration of cow milk: filtering raw milk, degassing, preheating, and separating impurities by a separator;
2) Homogenizing and sterilizing cow milk: homogenizing one part of raw milk after removing impurities in a homogenizer and the other part of raw milk after removing impurities in the homogenizer, mixing the two parts in a mixing tank after homogenizing, and sterilizing;
3) Powder adding: adding various powder raw materials into a mixing tank according to a formula;
4) Melting and oil preparing: after the grease raw materials are dissolved, adding the grease raw materials into a mixing tank;
5) Nutrient dissolution and addition: respectively dissolving vitamin nutrients and mineral raw materials with purified water, and sequentially adding the dissolved vitamin nutrients and mineral raw materials into a mixing tank to obtain mixed feed liquid;
6) LNnT dissolution addition: and 5) dissolving the LNnT raw material with part of the mixed liquor in the step 5), and adding the dissolved LNnT raw material into a mixing tank to obtain the mixed liquor containing LNnT.
According to some embodiments of the invention, wherein the wet-process formulation comprises the steps of:
1) Rough filtration of cow milk: the cow milk is subjected to coarse filtration and degassing by a balance cylinder, is preheated by a plate heat exchanger, and is separated from impurities by a separator;
2) Homogenizing and sterilizing cow milk: part of raw milk after removing impurities enters a homogenizer for homogenization and the other part of raw milk is not homogenized, and the raw milk and the homogenized raw milk are mixed and enter a sterilization system for sterilization;
3) Powder adding: the powder raw materials are metered according to the formula, then are added into a powder mixing tank in a unified way through an air conveying system, and are sucked into the mixing tank through a vacuum system;
4) Melting and oil preparing: placing the grease raw materials specified in the formula into an oil melting room according to the formula requirement, keeping the temperature of the oil melting room at 50-90 ℃, after the oil is melted, pumping the mixed oil into a mixed oil storage tank, and pumping the mixed oil into the mixed oil tank through an oil pump according to the formula requirement;
5) Nutrient dissolution and addition: respectively dissolving vitamin nutrients and mineral raw materials with purified water, and sequentially adding the dissolved vitamin nutrients and mineral raw materials into a mixing tank to obtain mixed feed liquid;
6) LNnT dissolution addition: and 5) dissolving the LNnT raw material with part of the mixed liquor in the step 5), and adding the dissolved LNnT raw material into a mixing tank to obtain the mixed liquor containing LNnT.
According to some embodiments of the invention, the method further comprises the step of filtering the resulting LNnT-containing mixed liquor.
According to some embodiments of the invention, the homogenizing comprises homogenizing the LNnT-containing mixed liquor with a homogenizer.
According to some embodiments of the invention, the homogenizing further comprises cooling the homogenized mixed liquor to below 20 ℃.
According to some embodiments of the invention, the concentrating and sterilizing includes sterilizing the mixed liquor (preferably the homogenized mixed liquor or the cooled mixed liquor) containing LNnT at 83 ℃ for 25 seconds or more to obtain concentrated and sterilized milk.
According to some embodiments of the invention, the concentrating and sterilizing includes double-effect concentrating the mixed liquor (preferably homogenized mixed liquor or cooled mixed liquor) containing LNnT, and sterilizing at not less than 83 ℃ for 25 seconds to obtain concentrated sterilized milk.
According to some embodiments of the invention, wherein the parameters of the spray drying include: the air inlet temperature is 170-190 ℃, the air exhaust temperature is 80-90 ℃, the pump pressure is 150-250bar, and the negative pressure is-4.5-3.5 mbar.
According to some embodiments of the invention, wherein the parameters of the spray drying include: the air inlet temperature is 180 ℃, the air outlet temperature is 86 ℃, the pump pressure is 200bar, and the negative pressure is-4.0 mbar.
According to some embodiments of the invention, wherein the spray drying is spray drying with a drying tower.
According to some embodiments of the invention, wherein the high pressure pump has a pressure of 150-250bar (preferably 200 bar), the column negative pressure is-4.5 to-3.5 mbar (preferably-4.0 mbar).
According to some embodiments of the invention, wherein the spray drying comprises preheating the concentrated sterilized milk to 50-70deg.C (preferably 60-65deg.C), filtering (preferably with a filter having a pore size of 1-2 mm), and spray drying.
According to some embodiments of the invention, wherein the spray drying further comprises a secondary drying (preferably with a fluidized bed) after the spray drying, followed by cooling to 25-35 ℃ (preferably 30 ℃).
According to some embodiments of the invention, the dry mix comprises a homogeneous mixing of DHA and ARA with milk powder obtained after spray drying.
In summary, the invention provides a formula milk powder and a preparation method thereof. The formula milk powder has the following advantages:
the formula milk powder provided by the invention provides a solution for improving intestinal microenvironment health, such as reducing hydrogen sulfide. Meanwhile, the LNnT and probiotics combination provided by the invention is applied to preparing food for reducing hydrogen sulfide in intestinal tracts. The breast milk infant formula milk powder containing LNnT and probiotics (bifidobacterium lactis BL-99) has the effect of reducing hydrogen sulfide in intestinal tracts.
Drawings
Figure 1 is a graph of results of combined small batch fermentation of LNnT and probiotics to produce total short chain fatty acids in a simulated infant gut environment.
Figure 2 is a graph of results simulating the combined small batch fermentation of LNnT and probiotics to produce hydrogen sulfide as a percentage of total gas production in an infant intestinal environment.
Detailed Description
The following detailed description of the invention and the advantages achieved by the embodiments are intended to help the reader to better understand the nature and features of the invention, and are not intended to limit the scope of the invention.
Experimental method
The method for measuring the air pressure, the gas components and the short chain fatty acid content of the product after sample fermentation specifically comprises the following steps:
first, sample collection is carried out: a faecal sample of a 3-6 month old breast milk or formula fed infant was selected. Collecting one part of oral swab of each mother in the breast-feeding group, one part of fresh breast milk and one part of corresponding infant feces during the month-old period; one stool was collected for each infant in the artificial feeding group. Fresh feces were obtained from donors, transported to the laboratory in 4 hours with ice bags, fermented, and the short chain fatty acids of the fermented products were measured.
The measuring process is specifically as follows:
1. preparation of culture medium
(1) Preparing YCFA anaerobic basic culture medium, and packaging each 30ml of the culture medium into an anaerobic penicillin bottle with the total volume of 50ml for standby.
The YCFA anaerobic basal medium has the following formula (g/L): tryptone 10, yeast extract 2.5, L-cysteine hydrochloride 1, naCl 0.9, caCl 2 ·6H 2 O 0.009，KH 2 PO 4 0.45，K 2 HPO 4 0.45，MgSO 4 ·7H 2 O0.09；
The composition also comprises the following components: 1mL of resazurin (1 mg/mL), 2mL of heme (5 mg/mL), 200 μl of vitamin I solution;
wherein the vitamin I solution comprises (mg/mL): biotin (VH) 0.05, cobalamin (VB 12) 0.05, p-aminobenzoic acid 0.15, folic acid 0.25, pyridoxamine (VB 6) 0.75.
(2) The culture medium required in the examples of the present invention was formulated.
Before fermentation experiments, probiotics or prebiotics are added into the YCFA culture medium as required to form the culture medium required by the project. The final concentration of the added prebiotics in the experiment is 4 per mill; the final concentration of each added probiotic is 10 9 CFU/mL。
The probiotics and prebiotics added as referred to in the experiment are shown in table 1. Wherein each medium was divided into two cases of adding and not adding ETEC, and the total of 42 fermentation conditions was obtained. The final concentration of ETEC added was 10 10 CFU/mL。
TABLE 1 fermentation condition list
2. Strain activation and identification
Respectively taking bacterial strains BB12, YLGB-1496, HN019 and BL-99 bacterial powder, preparing into 10 in an anaerobic workstation 7 CFU/mL, concentration was measured using plate counting. Before preparing the culture medium, taking out the glycerol tube strain kept in a refrigerator at-80 ℃ and inoculating the glycerol tube strain into the MRS culture medium for activation, and then inoculating the activated strain liquid into the corresponding culture medium by using a syringe.
Taking bacterial strain ETEC (ATCC 35401) bacterial powder, preparing to 10 in an anaerobic workstation 10 CFU/mL, concentration was measured using plate counting.
3. In vitro fermentation
(1) Preparation of samples before fermentation:
accurately weighing 0.800+/-0.010 g of fresh feces, placing the fresh feces into one side of a stirring spoon of a feces pretreatment box, calculating according to the mass-volume ratio of 10%, and supplementing PBS buffer solution with corresponding volume. Vortex for about 5-10 minutes to thoroughly break up fecal debris and mix well with PBS buffer to prepare a uniform 10% fecal suspension (w/v). And standing the fecal pretreatment box on a table top, and filtering the suspension by two layers of filter screens for later use.
(2) Inoculating: in the anaerobic station, 0.5mL of the suspension (clarified side of pretreatment cassette) was aspirated with a 1mL syringe equipped with a 5-gauge needle, the penicillin bottle butyl rubber stopper was pierced, and the medium was injected. If gas production analysis is needed, gas pressure detection and recording are carried out by using a barometer for fermentation for 0 hour.
Inoculation and dynamic sampling were all done in anaerobic workstations, with 5 biological replicates for each medium for breast feeding and artificial feeding groups, respectively.
The rest feces is as such, and the feces suspension can be split-packed as required, marked and frozen for other detection. When the stool sample is reused, after the stool sample is thawed within 30 minutes, the stool sample is gently mixed with the culture medium, is added into a batch fermentation culture medium as an initial culture substance, and the solution is continuously mixed, so that ideal mixing uniformity is maintained. Because of the consistent thawing time, the groups were similar in initial bacterial composition.
(3) Fermentation: the penicillin bottle is placed in a 37 ℃ incubator for static culture for 24 hours without disturbance. After the cultivation is finished, the small bottle is taken out, the cover is not opened, and the small bottle is directly frozen at-20 ℃ for detection.
4. Gas detection
The fermentation vials were removed, the end point of fermentation (24 hours) was checked and recorded using a barometer and the gas composition was checked using a gas analyzer (HL-QT 01, hunan halimasch biosciences).
Specifically, the apparatus is comprised of a gas sampler, valve module, vacuum generator, and gas detection chamber incorporating a plurality of gas sensors. The gas distribution module controls the gas quantity introduced by the gas detection chamber by means of a vacuum generator. The detection steps are as follows:
(1) detecting gas in the blank culture medium, and performing instrument calibration;
(2) the gas distribution module is used for adjusting the gas detection chamber to a certain vacuum level by using the vacuum generator;
(3) sucking the gas in the small bottle into a detection chamber of the instrument through a gas sampler, and adjusting the gas volume through a gas distribution module;
(4) detecting CO entering the gas detection chamber by using corresponding gas sensors respectively 2 ，H 2 ，CH 4 ，H 2 S and other 4 gases;
(5) the gas ratio is calculated by a preset software.
5. Short chain fatty acid detection
Total short chain fatty acid concentrations, including acetic acid, propionic acid, butyric acid, isobutyric acid, valeric acid, isovaleric acid, were measured using a gas chromatograph (9720, ark instruments, inc.). The method comprises the following specific steps:
(1) Preparation before sample injection: mu.L of the fermentation broth was aspirated using a sterile needle, placed in a 1.5ml centrifuge tube, 100. Mu.L of crotonic acid metaphosphoric acid solution was added, and the mixture was frozen at-30℃for 24 hours. After thawing, centrifugation was carried out at 10000rpm at 4℃for 3min, the supernatant was collected and filtered with a 0.22 μm filter (Millipore), 100. Mu.L of the sample extract was added to the cannula in the gas phase sample bottle, and after removing the bubbles by capping, the sample was loaded for analysis.
(2) The gas chromatograph conditions were as follows:
chromatographic column: agilent FFAP 30m 0.25mm 0.25 μm; column temperature: heating to 180deg.C at 75deg.C at 20deg.C/min for 1min, heating to 220deg.C at 50deg.C/min for 1min;
sample inlet: temperature: 250 ℃, sample injection amount: 1.0 μl, split ratio: (5:1); carrier gas: high purity nitrogen; flow rate: 2.5mL/min for 6.5min,2.8mL/min for 2L to 2.8mL/min for 2min;
a detector: FID; temperature: 250 ℃; tail blowing: 20mL/min; hydrogen gas: 30mL/min; air: 300mL/min.
(3) Quantitative determination is carried out by using a peak area internal standard method, and automatic calculation is carried out by using built-in software of a workstation according to a standard curve equation internal standard method.
Example 1
This example provides a simulation of the combined small batch fermentation of LNnT and probiotics in an infant intestinal environment to produce total short chain fatty acids, the results of which are shown in figure 1.
It can be seen that the combination of LNnT with probiotics is superior to the blank and four probiotics alone in all experimental groups.
The results of the group-to-group comparison of LNnT with different probiotic combinations are shown in table 2. From Table 2, it can be seen that in the absence of ETEC, the total acid production in the formula was higher for all four compositions than for the breast milk group (P < 0.05). Wherein LNnT and BL-99 still produced more short chain fatty acids in the formula than in the breast milk group in the presence of ETEC, indicating that it helped to establish a healthier intestinal environment (p=0.0229).
TABLE 2
Example 2
This example provides results simulating the combined small batch fermentation of LNnT and probiotics in an infant intestinal environment to produce hydrogen sulfide as a percentage of total gas production.
The results of simulating the combined small batch fermentation of LNnT and probiotics in an infant intestinal environment to produce hydrogen sulfide as a percentage of total gas production are shown in fig. 2. From figure 2 it can be seen that the combination of LNnT with probiotics is better than the blank and four probiotic alone groups in all experimental groups.
The results of further comparing the different groups are shown in table 3. As can be seen from table 3, the faeces of infants from the formula feeding source resulted in more hydrogen sulphide production in the gut compared to the breast feeding group, with or without ETEC. The presence or absence of ETEC in the breast milk group may produce a significant difference, whereas the absence of such difference in the formula group suggests that ETEC is not a major factor affecting hydrogen sulfide production when fed by the formula, and conversely, the presence of ETEC may lead to the production of more hydrogen sulfide gas in breast-fed infants.
TABLE 3 Table 3
Whether or not there is a significant difference P value
Breast milk without ETEC vs. Breast milk with ETEC *** 0.0008
Breast milk-ETEC-free vs. formula powder-ETEC-free **** <0.0001
Breast milk with ETEC vs. formula powder with ETEC *** 0.001
Formulation powder-ETEC-free vs. formulation powder-ETEC-free ns 0.6134
Further comparison against the breast milk ETEC-free group found that the combination of LNnT+BL-99 produced less hydrogen sulfide than LNnT monomer (P < 0.05), LNnT+BB12 and LNnT+YLGB-1496 were not significantly different from LNnT, but had a significant trend, P <0.1, indicating that the combination of LNnT and probiotics described above could produce a synergistic effect. The comparison results are shown in Table 4.
TABLE 4 Table 4
No significant difference was observed in hydrogen sulfide production under different fermentation conditions for the ETEC group of breast milk. The comparison results are shown in Table 5.
TABLE 5
Whether or not there is a significant difference P value
LNnT vs.LNnT+BB12 ns 0.7561
LNnT vs.LNnT+YLGB-1496 ns 0.4407
LNnT vs.LNnT+HN019 ns 0.6746
LNnT vs.LNnT+BL-99 ns 0.5078
LNnT+BB12vs.LNnT+YLGB-1496 ns 0.9753
LNnT+BB12vs.LNnT+HN019 ns 0.9998
LNnT+BB12vs.LNnT+BL-99 ns 0.9855
LNnT+YLGB-1496vs.LNnT+HN019 ns 0.9937
LNnT+YLGB-1496vs.LNnT+BL-99 ns >0.9999
LNnT+HN019vs.LNnT+BL-99 ns 0.9969
In the formula powder ETEC-free group, the hydrogen sulfide generated by LNnT+YLGB-1496 and LNnT+BL-99 is lower than LNnT (P < 0.05), and the LNnT+BB12, LNnT+HN019 and LNnT have no significant difference, but have significant trend, namely P <0.1, which indicates that after the LNnT is combined with probiotics, the generation of hydrogen sulfide can be synergistically increased and reduced more. The comparison results are shown in Table 6.
TABLE 6
No significant differences were observed in the formulation with ETEC group for hydrogen sulfide production under different fermentation conditions, see table 7.
TABLE 7
Whether or not there is a significant difference P value
LNnT vs.LNnT+BB12 ns 0.5117
LNnT vs.LNnT+YLGB-1496 ns 0.1455
LNnT vs.LNnT+HN019 ns 0.1728
LNnT vs.LNnT+BL-99 ns 0.1586
LNnT+BB12vs.LNnT+YLGB-1496 ns 0.5939
LNnT+BB12vs.LNnT+HN019 ns 0.6786
LNnT+BB12vs.LNnT+BL-99 ns 0.6362
LNnT+YLGB-1496vs.LNnT+HN019 ns >0.9999
LNnT+YLGB-1496vs.LNnT+BL-99 ns >0.9999
LNnT+HN019vs.LNnT+BL-99 ns >0.9999
Example 3
Infant formula 1 (preparation 1000 kg) containing LNnT and probiotic composition:
3100 kg of raw milk, 300 kg of lactose, 25 kg of whey protein powder WPC80%, 100 kg of desalted whey powder D90 kg, 72 kg of OPO structural fat, 54 kg of soybean oil, 9 kg of corn oil, 45 kg of high oleic sunflower seed oil, 27 kg of alpha-whey protein powder, 9 kg of beta-casein powder, 1 kg of anhydrous butter, 25 kg of fructo-oligosaccharide powder, 40 kg of galacto-oligosaccharide syrup, 2 kg of soybean lecithin, 2.5 kg of breast milk oligosaccharide LNnT, 17.85 kg of compound nutrient, 12 kg of DHA, 22 kg of ARA and 0.15 kg of bifidobacterium lactis BL-99.
Wherein the compound nutrients comprise about 3.5 kg of compound vitamin nutrition package, about 1.5 kg of choline chloride nutrition package, about 11 kg of calcium powder nutrition package, about 1 kg of mineral nutrition package, about 0.85 kg of magnesium chloride nutrition package, and lactose as the base material of each nutrition package.
Infant formula 2 (preparation 1000 kg) containing LNnT and probiotic composition:
1000 kg of raw milk, 250 kg of skim milk powder, 150 kg of lactose, 34% of whey protein powder WPC 50 kg, 90 kg of desalted whey powder D225 kg, 106 kg of OPO structural fat, 37 kg of soybean oil, 30 kg of corn oil, 10 kg of alpha-whey protein powder, 10 kg of beta-casein powder, 5 kg of fructo-oligosaccharide powder, 15 kg of galacto-oligosaccharide syrup, 3.5 kg of breast milk oligosaccharide LNnT, 11 kg of compound nutrient, 12 kg of DHA, 14 kg of ARA, 0.25 kg of bifidobacterium lactis BL-99 and 0.65 kg of nucleotide.
Wherein the compound nutrients comprise about 1.5 kg of compound vitamin nutrition package, about 0.75 kg of choline chloride nutrition package, about 5 kg of calcium powder nutrition package, about 1 kg of mineral nutrition package, about 0.75 kg of magnesium chloride nutrition package and about 2 kg of potassium chloride nutrition package, and the base material of each nutrition package is lactose.
Infant formula 3 (preparation 1000 kg) containing LNnT and probiotic composition:
2100 kg of raw milk, 280 kg of lactose, 230 kg of desalted whey powder D, 72 kg of OPO structural fat, 54 kg of soybean oil, 9 kg of corn oil, 45 kg of high oleic sunflower seed oil, 10 kg of alpha-lactalbumin powder, 1 kg of beta-casein powder, 6 kg of fructo-oligosaccharide powder, 22 kg of galacto-oligosaccharide slurry, 3 kg of soybean lecithin, 0.5 kg of breast milk oligosaccharide LNnT, 22.85 kg of compound nutrient, 12 kg of DHA, 14 kg of ARA, 0.45 kg of bifidobacterium lactis BL-99 and 0.65 kg of nucleotide.
Wherein the compound nutrients comprise about 1.5 kg of choline chloride nutrition package, about 16 kg of calcium powder nutrition package, about 1 kg of mineral nutrition package, about 0.85 kg of magnesium chloride nutrition package, and 3.5 kg of vitamin package, and the base material of each nutrition package is lactose.
The process is as follows:
1) Rough filtration of cow milk: the cow milk is subjected to coarse filtration and degassing by a balance cylinder, is preheated by a plate heat exchanger, and is separated from impurities by a separator.
2) Homogenizing and sterilizing cow milk: part of raw milk after removing impurities enters a homogenizer for homogenization and the other part is not homogenized, and the raw milk and the homogenized raw milk are mixed and enter a sterilizing system for sterilization.
3) Powder adding: the powder raw materials are metered according to the formula, then are added into a powder mixing tank in a unified way through an air conveying system, and are sucked into a vacuum mixing tank through a vacuum system;
4) Melting and oil preparing: placing the grease specified in the formula into an oil melting room according to the formula requirement, keeping the temperature of the oil melting room at 50-90 ℃, after the oil is melted, pumping the oil into a mixed oil storage tank, and pumping the mixed oil into the mixed oil tank through an oil pump according to the formula requirement;
5) Nutrient dissolution and addition: and respectively dissolving the nutrient packages such as calcium powder, vitamins, minerals and the like with purified water, and sequentially adding the dissolved nutrient packages into a mixing tank to obtain mixed feed liquid.
6) LNnT dissolution addition: and (3) dissolving the LNnT raw material with part of the mixed liquor in the step (5), and adding the dissolved LNnT raw material into a mixing tank to obtain the mixed liquor containing LNnT.
7) And (3) filtering: the mixed feed liquid containing LNnT is filtered by a filter screen, and physical impurities possibly brought in the raw materials are removed.
8) Homogenizing: homogenizing the mixed feed liquid by a homogenizer, mechanically treating the fat globules, and dispersing the fat globules into uniform fat globules.
9) Cooling and storing: the homogenized feed liquid enters a plate heat exchanger for cooling: cooling to below 20 ℃, temporarily storing in a pre-storing cylinder, entering the next working procedure within 6 hours, and starting the stirrer according to the set requirement.
10 Concentration sterilization: during production, double-effect concentration is used, the sterilization temperature is more than or equal to 83 ℃, and the sterilization time is 25 seconds. The discharge concentrations were 50% dry matter.
11 Concentrated milk storage, pre-heating filtration, spray drying: the concentrated milk is temporarily stored in a concentrated milk balance tank. Preheating to 60deg.C by scraper preheater, filtering with 1mm pore size filter, spray drying by high pressure pump, and agglomerating fine powder on top of tower or fluidized bed. Air inlet temperature: the temperature of exhaust is at 180 ℃ and 86 ℃, the pressure of the high-pressure pump is 200bar, and the negative pressure of the tower is about-4 mbar.
12 Fluidized bed drying and cooling: the powder from the drying tower is dried again by the fluidized bed (first stage) and then cooled to 30 ℃ by the fluidized bed (second stage).
13 Split charging: and weighing DHA, ARA or bifidobacterium longum subspecies GB1496 or bifidobacterium lactis BL-99 by powder workshop personnel according to the formula requirement, sealing bags and subpackaging.
14 Dry blending): mixing the weighed DHA, ARA or bifidobacterium longum subspecies GB1496 with milk powder in a dry mixer.
15 Screening powder: the granularity of the milk powder is uniform through the vibrating screen, and the powder slag is scrapped.
16 Powder discharge: and (3) receiving powder by using a sterilized powder collecting box, and conveying the powder from a powder outlet room to a powder feeding room.
17 Powder) is added: and pouring the milk powder into a powder storage tank on a size packaging machine according to the packaging requirement.
18 Packaging: 400 g of automatic packaging machine fills nitrogen for packaging. The oxygen content is lower than 1% when nitrogen is filled. 900 g of iron can is automatically filled with nitrogen and packaged, and the oxygen content is lower than 5%.
19 Boxing: packaging the packaged small bags into a paper box, adding a powder spoon at the same time, and sealing by a box sealing machine.
20 Inspection of the finished product: sampling and checking the packaged product according to a checking plan.
21 Warehousing and storing: the qualified products are stored in warehouse, and the storage is required at normal temperature, and the humidity is less than or equal to 65%.
Process parameters
1) Parameters of milk collecting section
Project Control parameters
Raw milk storage temperature (. Degree. C.) 4
Raw milk storage time (from start of milk collection to end of pasteurization) (h) 6
Raw milk preheating temperature (DEG C) 75
Sterilization temperature (. Degree. C.) 85
Cooling temperature (DEG C) 6
Pasteurized milk storage temperature (. Degree. C.) 4
Pasteurized milk storage time (start pasteurization until pasteurized milk empty) (h) 12
2) Parameters of pretreatment section
3) Parameters of concentration section
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4) Parameters of drying section
5) Packaging parameters
5.1 Dry mixing section
Project Control parameters
Premixing time (min) 20
Dry mixing time (min) 5
5.2 Packaging section
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Claims (9)

1. The formula milk powder comprises 1000 parts by weight of the formula milk powderThe preparation raw materials of the formula milk powder comprise: 0-4.5 parts of breast milk oligosaccharide, 890-3800 parts of raw milk, 0-480 parts of lactose, 0-540 parts of desalted whey powder, 0-350 parts of skim milk powder, 80-80 parts of whey protein powder WPC, 34-120 parts of whey protein powder WPC, 0-225 parts of vegetable oil, 0-45 parts of alpha-whey protein, 0-23 parts of beta-casein, 1-2.5 parts of phospholipid, 0-30 parts of fructo-oligosaccharide, 0-90 parts of galacto-oligosaccharide slurry, 0-0.18 parts of probiotics, 0-5 parts of anhydrous cream, 0-0.8 parts of nucleotide, 1-5 parts of vitamin nutrition, 0-3 parts of choline chloride, 0-2 parts of mineral A and 1-12 parts of mineral B; wherein, the mineral A is selected from the combination of mineral salts of one or more of copper, iron, magnesium, zinc, selenium, manganese and iodine, and the mineral B is selected from the combination of mineral salts of one or more of calcium, phosphorus, sodium and potassium; the breast milk oligosaccharide contains 0.01-3.5 parts by weight of breast milk oligosaccharide LNnT; the probiotics are selected from one or two of bifidobacterium infantis YLGB-1496 and bifidobacterium lactis BL-99; the probiotics content is 10 per gram of formula milk powder 6 -2×10 12 CFU。
2. The formula of claim 1 wherein the vegetable oil is selected from one or more of the group consisting of structural oil OPO, high oleic sunflower oil, corn oil, canola oil, and soybean oil; when the vegetable oil components are independent, the total weight of the formula milk powder is 1000 parts, and the dosages of the vegetable oil components are independent respectively: 0-170 parts of structural grease OPO, 0-150 parts of high oleic acid sunflower seed oil, 0-50 parts of corn oil, 0-50 parts of low erucic acid rapeseed oil and 0-80 parts of soybean oil.
3. The formula of claim 1 wherein the vitamin nutrient is selected from the group consisting of vitamin a, vitamin D, vitamin E, vitamin K1, vitamin B2, vitamin B6, vitamin B12, niacinamide, folic acid, pantothenic acid, and biotin.
4. The formula of claim 1 wherein the milk oligosaccharide LNnT is present in an amount of 142.9-428.6mg/100g of powder per 100g of formula.
5. The formula of claim 4 wherein the milk oligosaccharide LNnT is present in an amount of 178.6-392.9mg/100g of powder per 100g of formula.
6. Formula according to claim 1, wherein the probiotic content is 10 per gram of formula 8 -2×10 10 CFU。
7. The formula milk powder according to any one of claims 1 to 6, wherein the formula milk powder further comprises the following components: 0.20-2.04 parts of inositol, 0-0.65 part of taurine, 0.06-0.5 part of L-carnitine, 3-18 parts of DHA and 3-22 parts of ARA.
8. The formula according to any one of claims 1 to 6, wherein the formula is an infant formula for reducing hydrogen sulfide in the infant's intestinal tract.
9. The method for preparing the formula milk powder according to any one of claims 1 to 8, wherein the method comprises the following steps: wet compounding, homogenizing, concentrating, sterilizing, spray drying, dry mixing and packaging.
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CN106259953A (en) * 2015-06-01 2017-01-04 内蒙古伊利实业集团股份有限公司 Breast milk 0-12 month babies ' formula milk powder containing nucleotide and preparation method thereof
CN106259952A (en) * 2015-06-01 2017-01-04 内蒙古伊利实业集团股份有限公司 1-3 year baby formulas milk powder containing nucleotide and dietary fiber and preparation method thereof
CN110074188A (en) * 2019-05-16 2019-08-02 福建农林大学 A kind of baby formula milk powder and preparation method thereof adding human milk oligosaccharides
CN114568526A (en) * 2020-11-30 2022-06-03 内蒙古伊利实业集团股份有限公司 Mother emulsified infant formula powder for improving intestinal microenvironment health and application thereof

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CN106259938A (en) * 2015-06-01 2017-01-04 内蒙古伊利实业集团股份有限公司 Breast milk 0-12 month babies ' formula milk powder containing nucleotide and OPO structured lipid and preparation method thereof
CN106259953A (en) * 2015-06-01 2017-01-04 内蒙古伊利实业集团股份有限公司 Breast milk 0-12 month babies ' formula milk powder containing nucleotide and preparation method thereof
CN106259952A (en) * 2015-06-01 2017-01-04 内蒙古伊利实业集团股份有限公司 1-3 year baby formulas milk powder containing nucleotide and dietary fiber and preparation method thereof
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