CN111887297A - Preparation method of infant formula milk powder with natural bioactivity - Google Patents
Preparation method of infant formula milk powder with natural bioactivity Download PDFInfo
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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
- A23C9/16—Agglomerating or granulating milk powder; Making instant milk powder; Products obtained thereby
-
- 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
- A23C9/14—Milk preparations; Milk powder or milk powder preparations in which the chemical composition of the milk is modified by non-chemical treatment
- A23C9/142—Milk preparations; Milk powder or milk powder preparations in which the chemical composition of the milk is modified by non-chemical treatment by dialysis, reverse osmosis or ultrafiltration
- A23C9/1422—Milk preparations; Milk powder or milk powder preparations in which the chemical composition of the milk is modified by non-chemical treatment by dialysis, reverse osmosis or ultrafiltration by ultrafiltration, microfiltration or diafiltration of milk, e.g. for separating protein and lactose; Treatment of the UF permeate
-
- 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
- A23C2210/00—Physical treatment of dairy products
- A23C2210/25—Separating and blending
- A23C2210/256—Removal or separation of bacteria, or a fraction containing bacteria, by centrifugation; Bactofugation
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- Engineering & Computer Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Water Supply & Treatment (AREA)
- Dairy Products (AREA)
Abstract
The invention belongs to the field of dairy processing, and discloses a preparation method of infant formula milk powder with natural bioactivity. The whole process from milking to powdering is controlled within 2 hours, the heat treatment strength in the production process is reduced, the activities of lactoferrin, IgG, peroxidase and lysozyme in fresh milk are kept above 80%, the biological activity of immune components in the milk is effectively kept, and the immune index of the infant formula milk powder is improved. The invention is used for producing infant formula milk powder.
Description
Technical Field
The invention belongs to the field of dairy processing, relates to milk processing, and particularly relates to a preparation method of infant formula milk powder with natural bioactivity.
Background
About 1600-1800 ten thousand babies are born in China every year, and 200 ten thousand newborns are newly added every year along with the gradual release of the two-fetus policy. Wherein the pure breast-milk feeding rate of infants in 0-6 months is 27.8%, the pure breast-milk feeding rate in rural areas is 30.3%, the pure breast-milk feeding rate in cities is only 15.8%, and the infant formula milk powder becomes the best choice for tens of millions of parents except breast milk. According to the AC Nielson monitoring data, about 600 million yuan of infant formula milk powder is sold in the Chinese market in 2016, the infant formula milk powder is expanded to 1200 million yuan in 2016-2019, the annual growth rate is about 20%, and China replaces the United states to become the first world large infant formula milk powder consuming country. How to provide safe, nutritional and healthy infant formula milk powder for Chinese babies still remains a major problem to be urgently solved at present.
Commercially available infant formula milk powder is divided into two types of milk-based infant formula milk powder and soy-based infant formula milk powder, wherein the milk-based infant formula milk powder accounts for more than 90 percent. The cow milk not only contains rich nutritional ingredients such as protein, fat, lactose, minerals and vitamins, but also can provide immune active ingredients such as immunoglobulin, lactoferrin, lysozyme, peroxidase and the like for infants. The immune system of the newborn is not developed perfectly, the immunoglobulin synthesized by the newborn is not enough to resist the invasion of pathogen and is easy to be infected by bacteria and virus, and the immune active ingredients in the cow milk can provide passive immunity for the newborn to a certain extent and improve the disease resistance of the newborn. Therefore, the biological activity of immune components in the cow milk is maintained in the processing process of the infant formula milk powder, the infant formula milk powder is beneficial to improving the infant immunity and reducing the risks of infection and diseases.
The biological activity of the immunological components in cow's milk is susceptible to the intensity of heat treatment and storage time. Immunoglobulin, lactoferrin, peroxidase and lysozyme in cow milk belong to heat-sensitive protein substances, and are easy to denature to different degrees in the heat treatment process at the temperature of more than 70 ℃. In the process of storing cow milk for a long time (0-6 ℃), extracellular protease secreted by partial bacteria can decompose and destroy the structure of an immunocompetent substance, so that the bioactivity of the immunocompetent substance is lost. Therefore, reduction of heat treatment intensity, reduction of storage time, and effective control of growth and reproduction of microorganisms during processing are important measures for maintaining biological activity of immune components.
At present, the wet process is the mainstream of the production process of the infant formula milk powder on the global scale. The wet process is mainly the links of milking, cooling temporary storage (average 6h), transportation (average 10h), factory inspection (average 1h), milk collection (average 3h), centrifugal separation, high-temperature pasteurization (80-90 ℃ for 15s), cooling temporary storage (average 8h), wet mixed materials (40-60 ℃), cooling temporary storage (average 6h), high-temperature short-time sterilization (85-140 ℃ for 2-15 s), evaporation concentration (45-75 ℃) and spray drying (air inlet temperature 160-200 ℃ and air exhaust temperature 85-98 ℃) and fluidized bed cooling drying (25-70 ℃), wherein the average time from milking to processing state is about 20h, and the average time from milking to powder forming is about 34 h. The long-time storage and high-intensity heat treatment of the milk can seriously destroy the biological activity of immune components in the milk, which is the current situation of the global infant formula milk powder production process. Therefore, the development of a low-temperature short-time infant milk powder production process from milking to powdering is urgently needed, which is used for reducing the intermediate storage time, reducing the heat treatment intensity, effectively controlling the microbial quantity in the production process, effectively maintaining the biological activity of immune components in the milk and improving the immune index of the infant formula milk powder.
Disclosure of Invention
The invention aims to provide a preparation method of infant formula milk powder with natural bioactivity, so that the bioactivity of immune components in milk raw materials can be effectively maintained in the product.
In order to achieve the purpose, the invention adopts the technical scheme that:
a preparation method of infant formula milk powder with natural bioactivity is characterized by taking raw milk, and performing centrifugal separation, primary microfiltration sterilization, wet mixing and proportioning, secondary microfiltration sterilization, online grease addition, homogenization, evaporative concentration, spray drying and fluidized bed cooling drying on the raw milk to obtain the infant formula milk powder with natural bioactivity.
As a limitation, the temperature of the two times of microfiltration sterilization is 6-10 ℃, the aperture of the microfiltration membrane is 0.8-1.6 mu m, and the membrane area is 20-35 m2The transmembrane pressure difference is 21-41 kPa, and the circulating flow rate of the trapped fluid is 6-8 m/s; the flux of the permeate liquid membrane for the first microfiltration and sterilization is 900-1000 L.m-2·h-1The flux of the permeate liquid membrane for the second microfiltration and sterilization is 300-800 L.m-2·h-1。
As a further limitation, the two times of microfiltration sterilization are uniform transmembrane pressure difference microfiltration sterilization; the microfiltration membrane is a ceramic membrane.
As another limitation, the raw milk is fresh milk which is qualified by milking, cooling and rapid detection.
As a further limitation, the used raw material milk is temporarily stored for less than or equal to 10min after being milked, and the temperature is 6-10 ℃.
As a third limitation, the milk raw material contains 2.8-3.4% of protein, 4.5-5.2% of lactose, 3.5-4.0% of fat, 12.0-13.5% of dry matter, less than or equal to 2 ten thousand CFU/mL of colony total number, less than or equal to 20 ten thousand somatic cells/mL, and the residual contents of melamine and veterinary drugs are zero;
after the centrifugal separation, the obtained skim milk contains 2.6-3.5% of protein, 4.5-5.3% of lactose, 8.0-9.5% of dry matter, less than or equal to 0.1% of fat, less than or equal to 1100CFU/mL of total number of colonies, and zero spore content;
after the first microfiltration sterilization, the obtained penetrating fluid A contains colony total number less than or equal to 30CFU/mL, and the contents of spores and somatic cells are zero;
after the wet mixing and proportioning, the obtained mixed material liquid B contains 12-36% of solid matters by weight, less than 1000CFU/mL of total number of bacterial colonies and less than 10 spores/mL;
after the second microfiltration sterilization, the obtained penetrating fluid C contains less than 30CFU/mL of total number of bacterial colonies and zero spore content;
after evaporation and concentration, the obtained concentrated milk D contains 50-55% of solid by weight;
after the spray drying, the weight fraction of water in the obtained powder E is less than or equal to 5.0 percent;
the weight percentage of the water in the infant formula milk powder retaining natural biological activity is less than or equal to 4 percent.
As a fourth limitation, the temperature of the centrifugal separation is 6-10 ℃;
the wet mixing ingredients are prepared by adding desalted whey powder, whey protein powder, lactose and other milk-containing raw materials at 6-10 ℃ and 0.45-0.8 bar, and stirring and mixing the materials at 1400-3000rpm, wherein the total time of the wet mixing ingredients is 15-25 min;
before the grease is added on line, the penetrating fluid C needs to be preheated to 25-55 ℃;
the online addition of the grease is that the grease is metered online in equal proportion, and the online addition temperature of the grease is 25-55 ℃;
the homogenizing conditions comprise a primary homogenizing pressure of 120-160 bar and a secondary homogenizing pressure of 30-50 bar;
preheating the feed liquid F obtained after homogenization to 65-70 ℃ before evaporation and concentration;
the conditions of evaporation concentration are as follows: the temperature of the first-effect evaporation is 65-70 ℃, and the vacuum degree is 0.75-0.8 bar; the temperature of the double-effect evaporation is 60-65 ℃, and the vacuum degree is 0.8-0.85 bar; the temperature of the triple effect evaporation is 55-60 ℃, and the vacuum degree is 0.85-0.95 bar; the condensation temperature is 50-55 ℃, and the vacuum degree is 0.85-0.95 bar;
the spray drying is low-temperature spray drying, the air inlet temperature is 30-120 ℃, the air exhaust temperature is 15-70 ℃, and the air inlet temperature of a static fluidized bed is 25-60 ℃;
the fluidized bed cooling drying is four-section dynamic fluidized bed drying, the air inlet temperature of the first section and the second section is 25-45 ℃, and the air inlet temperature of the third section and the fourth section is 15-30 ℃.
Due to the adoption of the technical scheme, compared with the prior art, the invention has the technical progress that:
the whole process from milking to powdering is controlled within 2 hours, the microfiltration sterilization technology is adopted to replace the existing thermal sterilization process, the damage of the heat treatment to the nutrient components and the immune active components in the milk is reduced, and bacteria, somatic cells and residual spores in the skim milk are effectively removed; the low-temperature vacuum high-shear mixing technology is used for replacing the existing hot batching technology, and the low-temperature spray drying technology is combined, so that the damage of heat treatment on nutrient components and immunocompetent components in the batched milk liquid is reduced, the growth and reproduction of microorganisms in the intermediate process are effectively controlled, the heat treatment strength is reduced, the activities of lactoferrin, IgG, peroxidase and lysozyme in fresh milk are kept above 80%, and the immune index of the infant formula milk powder with natural bioactivity is improved. The invention greatly improves the quality of the global infant formula milk powder at one time, and has wide popularization significance in the global range.
The preparation method is suitable for producing infant formula milk powder.
Drawings
FIG. 1 is a flow chart of the process for producing the infant formula with natural bioactivity of example 1;
FIG. 2 is a flow chart of a conventional infant formula production process of example 7;
FIG. 3 is a standard curve for measuring IgG content by ELISA method in example 8;
FIG. 4 is a standard curve of lactoferrin content measured by ELISA method in example 8;
FIG. 5 is a standard curve for measuring peroxidase activity by the ELISA method in example 8;
FIG. 6 is a standard curve for lysozyme activity determination by the ELISA method of example 8.
Detailed Description
The present invention is further illustrated by the following specific examples, which are to be construed as merely illustrative, and not limitative of the remainder of the disclosure.
Example 1A method for preparing an infant formula with natural bioactivity
This example comprises the following steps performed in sequence, a production process flow diagram referring to figure 1,
milking, cooling and rapid detection: the self-built large high-quality standardized pasture of the company is positioned in the downwind direction of the milk powder processing factory, 5 milking turntables are simultaneously opened in the pasture, each turntable contains 80 cows, and 400 cows continuously and simultaneously milks on line. Extruding fresh milk, cooling to 6 deg.C with ice water, and adding into a buffer tank for 9 min. In the caching process, a sample of a product controller is taken and sent to a laboratory for rapid detection, a detection result is obtained after 9min, and fresh milk contains 3.25% of protein, 4.8% of lactose, 3.7% of fat, 12.5% of dry matter, 6000CFU/mL of total bacterial colony count, 150000 somatic cells/mL of body cells, and melamine and veterinary drug residues are negative.
Centrifugal separation: pumping qualified milk raw materials into a milk powder processing plant which is 200m away, filtering the milk raw materials by an 80-mesh duplex filter, then entering a centrifugal separation link, carrying out centrifugal separation at 6 ℃ and simultaneously removing impurities and slag, cream and spores to obtain skim milk, wherein the skim milk contains 3.4% of protein, 5.1% of lactose, 9.1% of dry matter, less than or equal to 0.1% of fat, less than or equal to 1100CFU/mL of total colony count and undetected spores.
A large high-quality standardized pasture built by a company is generally a pasture which is located in the downwind direction of a milk powder processing factory and is 200-1000 m away from the pasture, and 1-4 ten thousand mu of forage grass planting bases are arranged around the standardized pasture.
And (3) performing primary microfiltration sterilization: introducing the skim milk into uniform transmembrane pressure difference microfiltration sterilization equipment, wherein the temperature is 6 ℃, the aperture of a microfiltration membrane is 0.8 mu m, and the membrane area is 30m2Transmembrane pressure difference of 25kPa, retentate circulation flow rate of 6m/s and permeate membrane flux of 900 L.m-2·h-1Obtaining a permeate A1Total 20CFU/mL colonies, spores and somatic cells were not detected.
Wet mixing ingredients: the compound vitamin, the compound mineral substance and the choline chloride are respectively mixed with reverse osmosis pure water according to the weight ratio of 1: pre-dissolving at 35 deg.C to obtain vitamin complex solution, mineral complex solution and choline chloride solution;
permeate A1Pumping into high-shear vacuum mixing tank, pumping into reverse osmosis pure water, vitamin complex solution, compound mineral solution and choline chloride solution, adding lactose, lactalbumin powder, alpha-lactalbumin powder, fructo-oligosaccharide, galacto-oligosaccharide, etc., dissolving completely at 6 deg.C and 0.6bar by high-shear vacuum mixing, filtering with filter screen, and feeding into 20m3The mixing tank has the stirring speed of 1400rpm, the mixing tank and the vacuum mixing tank keep the circulation of the material liquid to ensure the uniform mixing of the material liquid, and the total wet mixing time is 20min to obtain mixed material liquid B1The solid content weight fraction was 25.0%, the total number of colonies was 300CFU/mL, and the number of spores was 4/mL.
And (3) performing microfiltration sterilization for the second time: mixed liquid B1Pumping into uniform transmembrane pressure difference microfiltration and sterilization equipment, wherein the temperature is 6 ℃, the aperture of a microfiltration membrane is 1.4 mu m, and the membrane area is 30m2Transmembrane pressure difference of 25kPa, retention solution circulation flow rate of 6m/s, and permeate membrane flux of 600L m-2·h-1The permeate C obtained1The total number of middle colonies was 20CFU/mL, and spores were not detected.
Adding oil on line and homogenizing: permeate C1Preheating to 45 deg.C, mixing with vegetable oil preserved at 45 deg.C under nitrogen, OPO structure oil, docosahexaenoic acid oil, arachidonic acid oil and phospholipid on line, homogenizing to obtain feed liquid F1The homogenizing conditions are that the primary homogenizing pressure is 140bar and the secondary homogenizing pressure is 40 bar.
And (3) evaporation and concentration: feed liquid F1Pumping into an evaporator balance cylinder for evaporation concentration, preheating to 65 ℃ through a condenser and a tubular preheater, sequentially entering into a first-effect evaporator, a second-effect evaporator and a third-effect evaporator, wherein the first-effect evaporation temperature is 65 ℃, the vacuum degree is 0.75bar, the second-effect evaporation temperature is 60 ℃, the vacuum degree is 0.8bar, the third-effect evaporation temperature is 55 ℃, the vacuum degree is 0.85bar, the condenser temperature is 50 ℃, the vacuum degree is 0.85bar, and concentrated milk D is obtained after evaporation concentration1The weight fraction of the solid matter was 51.1%.
Spray drying and fluidized bed cooling drying: dense milk D1Preheating to 65 deg.C with tubular preheater, filtering with 80-mesh duplex filter, pumping into spray drying tower top by high pressure pump (200 + -10 bar) for low temperature spray drying with built-in static fluidized bed at bottom of spray drying tower, wherein the low temperature spray drying conditions are inlet air temperature of 140 deg.C, exhaust air temperature of 80 deg.C, inlet air temperature of static fluidized bed of 60 deg.C, and spray drying at low temperature to obtain powder E1The weight fraction of medium moisture was 4.8%.
Powder E1Drying with four-stage dynamic fluidized bed to obtain the productThe infant formula milk powder with physical activity has the temperature of 20 ℃, the moisture content of 2.7 percent by weight, the air inlet temperature of the first section and the second section of the dynamic fluidized bed is 40 ℃, and the air inlet temperature of the third section and the fourth section is 15 ℃; wherein, the first and second sections of air intake are mainly used for further drying and cooling, the third and fourth sections of air intake are used for cooling the powder, and the temperature of the obtained infant formula milk powder with natural bioactivity is 20 ℃.
The preparation method of this example was used for infant formula that retained natural biological activity.
The specific time taken for the fresh milk to flow through the processes from milking to powdering in this example is shown in table 1:
TABLE 1 summary of the time taken from milking to powdering of the fresh cow milk in the examples
The specific materials used in this example are shown in table 2:
table 2 raw material list of infant formula milk powder with natural bioactivity
| Name of raw materials | The dosage of each 3 tons of powder is per kg |
| Skimmed milk | 5000 |
| Pure water | 4500 |
| Lactose | 1425 |
| Composite vegetable oil | 525 |
| OPO structure grease | 240 |
| Whey protein powder | 72 |
| Fructo-oligosaccharide | 12 |
| Soybean lecithin | 12 |
| Arachidonic acid oil and fat (AA) | 30 |
| Docosahexaenoic acid oil (DHA) | 18 |
| Compound vitamin | 7.5 |
| Composite mineral | 7.5 |
| Choline chloride | 3 |
| Nucleotide, its preparation and use | 0.81 |
According to the requirements of GB10765 and the examination scrutiny rules of infant formula milk production permission (2013 edition), the infant formula milk powder which is obtained in the embodiment and retains natural biological activity is subjected to physicochemical and microbial index tests, the test results show that all indexes are qualified, and the test results are shown in Table 3:
TABLE 3 summary of the results of the testing of the infant formula with preserved natural bioactivity obtained in this example
Example 2-6 preparation method of infant formula milk powder with natural bioactivity
Examples 2 to 6 are methods for preparing infant formula milk powder with retained natural biological activity, which have substantially the same steps as example 1, but differ only in process parameters, as detailed in table 4:
TABLE 4 summary of the process parameters of examples 2 to 6
The contents of the other portions of examples 2 to 6 are the same as those of example 1.
Example 7A method for preparing a conventional infant formula
The conventional infant formula was produced according to the formula of example 1 using existing technology, the process flow diagram of which is shown in figure 2,
milking, cooling and temporary storage, transportation, factory inspection and milk collection: in a large high-quality standardized pasture built by a company, 6 rotary tables are started to express milk at the same time, and 80 cows are born on each rotary table. After being extruded, fresh milk is rapidly cooled to 6 ℃ by ice water, enters a 100-ton raw milk temporary storage bin for storage, and is transported to a processing plant outside 800km after being stored for 6 hours, wherein the transportation temperature is 6 ℃ and the transportation time is 10 hours. After the cow milk enters the factory, the cow milk is kept at 6 ℃, sampling is carried out for 1h, the detection result is that the cow milk is negative in protein 3.21%, fat 3.56%, lactose 4.80%, dry matter 12.6%, total number of bacterial colonies 15 ten thousand CFU/mL, somatic cells 15 ten thousand/mL and melamine and veterinary drug residues, and milk collection is started.
Centrifugal separation, pasteurization and refrigeration: filtering fresh milk by a duplex filter, centrifugally separating at 50 ℃, pasteurizing at 85 ℃ for 15s, cooling to 6 ℃ to obtain skim milk, and refrigerating in a milk warehouse, wherein the total number of bacterial colonies after pasteurization is 2 ten thousand CFU/mL. The total number of colonies increased to 5 ten thousand CFU/mL after 8h storage.
The wet mixing method comprises the following steps of mixing the composite vitamins, the composite mineral substances and choline chloride with reverse osmosis pure water respectively in a weight ratio of 1: pre-dissolving at 35 deg.C to obtain vitamin complex solution, mineral complex solution and choline chloride solution;
preheating skim milk to 45 deg.C, pumping into high-shear vacuum mixing tank, pumping into 45 deg.C reverse osmosis pure water, adding lactose, alpha-lactalbumin powder, fructo-oligosaccharide, soybean phospholipid, arachidonic acid powder and fish oil powder, mixing at 45 deg.C under high-shear vacuum, dissolving, filtering with filter screen, and making into 20m thick liquid3The mixing tank has the stirring speed of 1200rpm, the mixing tank and the vacuum mixing tank keep the circulation of the feed liquid to ensure the uniform mixing of the feed liquid, and the total wet mixing time is 1h to obtain mixed feed liquid G7The solid content was 25.0% by weight.
Adding oil on line, homogenizing, cooling and temporarily storing: feed liquid G7Mixing with vegetable oil preserved at 45 deg.C under nitrogen and OPO structure oil, and homogenizing with homogenizer under homogenizing conditionThe first-stage homogenizing pressure is 140bar, the second-stage homogenizing pressure is 40bar, and the obtained feed liquid H7And the mixture is cooled to 6 ℃ from the outlet of the homogenizer through a plate, enters the batching milk bin, has the total colony count of 3 ten thousand CFU/mL, and has the total colony count of 6 ten thousand CFU/mL after being stored for 6 hours.
Preheating, sterilizing, evaporating and concentrating, spray drying and fluidized bed drying: stored feed liquid H7Feeding to an evaporator balance cylinder, preheating feed liquid to 75 deg.C by a condenser and a tubular preheater, maintaining in a protein holding tube for 30s, transferring to a steam jet sterilizer, sterilizing at 110 deg.C for 5s, flash evaporating to 70 deg.C, cooling to 55 deg.C, feeding to a triple-effect tubular falling-film evaporator to obtain concentrated milk I7Placing into a thick milk temporary storage tank. Mixing the concentrated milk I7The straight tube type preheater is preheated to 75 ℃, filtered by a duplex filter and pumped to the top of the drying tower by a high-pressure pump (200 +/-10 bar) for spray drying, wherein the air inlet temperature of the spray drying is 170 ℃, and the air outlet temperature of the spray drying is 95 ℃. Dehydrating the feed liquid in a drying tower to form particles with the temperature of 70 ℃, dropping the particles into a built-in static fluidized bed, and allowing the particles to enter a dynamic fluidized bed from the static fluidized bed for further drying and cooling to prepare the traditional infant formula milk powder; wherein, the air inlet temperature of the first section and the second section of the dynamic fluidized bed is 40 ℃, and the air inlet temperature of the third section and the fourth section is 25 ℃. The whole process takes 34h from milking to powdering, wherein 20h from milking to entering the processing link.
According to GB10765 and the requirements of the examination scrutiny (2013 edition) of infant formula milk production permission, the indexes of sense, physicochemical property, microorganisms and the like of the infant formula milk of the batch are checked, the check results show that all the indexes are qualified, and the detection results are shown in Table 5:
TABLE 5 summary of the test results obtained in example 7 for infant formulas
Example 8 detection of immunologically active substances comparison
The immune active substances in the cow milk mainly comprise four thermosensitive protein substances including immunoglobulin, lactoferrin, lactoperoxidase and lysozyme, wherein the bovine milk immunoglobulin is divided into IgG, IgM and IgA, the IgG accounts for more than 75% of the total amount of the immunoglobulin in normal milk, the relative content of IgM and IgA is low, and detection errors are easily caused.
1) Instruments and materials
11) Reagent: phosphate buffer (pH 7.4), ELISA detection kit (four of bovine immunoglobulin IgG, bovine lactoferrin, lactoperoxidase, and bovine lysozyme).
12) Collecting and storing samples: the fresh raw milk, the skim milk (after microfiltration sterilization), the blended milk (after microfiltration sterilization), the concentrated milk after evaporation concentration and the finished product powder in the embodiment 1 are respectively taken as samples to be stored, wherein the liquid is stored in a liquid nitrogen cryopreservation tube, and the liquid nitrogen cryopreservation tube is placed in a liquid nitrogen container to be frozen and stored.
Fresh cow milk, accepted raw cow milk, skim milk (after pasteurization), skim milk (after temporary storage for 8 hours), blended milk (after temporary storage for 6 hours), concentrated milk after evaporation concentration and finished products in example 7 were respectively taken as samples for storage, wherein the liquids were stored in a liquid nitrogen cryopreservation tube, and the liquid nitrogen cryopreservation tube was placed in a liquid nitrogen container for cryopreservation.
13) The instrument comprises the following steps: incubator, microplate reader (BioTek), autoclave, electronic balance (accurate to 0.1 mg).
2) Experimental procedure
21) Sample pretreatment
Powder sample: 2.5g of each sample of the infant formula powder prepared in example 1 and the infant formula powder prepared in example 7 were taken out with an autoclaved spoon, dissolved in 6mL of phosphate buffer solution, thoroughly dissolved in a vortex shaker, centrifuged at 1000rpm for 30s, and then the volume was adjusted to 10mL and stored in a refrigerator at 4 ℃.
Liquid frozen samples: wearing glasses and gloves, taking out the freezing tube from the liquid nitrogen container, and rapidly placing the freezing tube into 38 ℃ water bath, and shaking the freezing tube without time to ensure that the sample is completely melted within 1 min. Wherein, 5mL of fresh milk and skim milk samples are taken, and 5mL of phosphate buffer solution is added for dilution and uniform mixing, and the mixture is stored at 4 ℃; taking 8mL of the ingredient milk, adding 2mL of phosphate buffer solution for dilution and uniform mixing, and storing at 4 ℃; concentrating by evaporation, diluting with 2.5mL of concentrated milk, adding 7.5mL of phosphate buffer solution, mixing, and storing at 4 deg.C.
22) The ELISA kit was removed in a refrigerator at 4 ℃ and allowed to equilibrate at room temperature for 20 min.
23) And fully shaking and shaking all reagents in the kit.
24) IgG, lactoferrin, lactoperoxidase and lysozyme standards were added to corresponding wells, in triplicate for each concentration.
25) 40uL of phosphate buffer solution is added into the sample wells, and then 10uL of samples to be detected are added into the corresponding wells respectively. Three replicates were made for each sample.
26) Then 100uL of HRP-labeled detection antibody is added into each hole, the reaction holes are sealed by a sealing plate membrane, and then the reaction holes are placed in a 37 ℃ incubator for 1 h.
27) Discarding the liquid in each well, adding 200uL of washing solution into each well, standing for 1min, discarding, cleaning on toilet paper, and repeating the above plate washing operation for 5 times.
28) 50uL of tetramethylbenzidine was added to each well and incubated at 37 ℃ for 15min in the absence of light.
29) Add 2mol/L sulfuric acid 50 μ L to each well, after standing for 3min, determine each well OD value at 450nm wavelength.
210) Standard curves were made, see figures 3, 4, 5 and 6 for details, and the sample content (activity) was calculated according to the regression equation of the standard curve.
3) The result of the detection
From the above experiments, tables 6 and 7 can be obtained, which show the contents of four immunologically active substances at each stage in the production process of examples 1 and 7, respectively.
TABLE 6 summary of the test results of each sample in example 1
Note: performing significance difference analysis on each line of data by adopting span software (P is less than 0.05);
table 7 results of sample testing in example 7
Note: performing significance difference analysis (p is less than 0.05) on each line of data by adopting span data processing software;
the activity retention of four immune components from milking to powdering was calculated from the data in tables 6 and 7, and table 8 can be obtained:
TABLE 8 retention of immune component activity from milking to powdering
Note: the retention of activity of immune component a is 0.6 x 100% a/b;
wherein a is the content of an immune active component A in each 100g of finished powder;
b is the content of the immunological active component A in each 100g of fresh raw milk;
0.6 represents that the amount of skim milk required by 3 tons of infant formula milk powder per ton is 5 tons;
a is lactoferrin, IgG, peroxidase or lysozyme.
Lactoferrin is a glycoprotein and is unstable to heat, and is heated at 65, 70, 75, 80, 85, 90 and 95 ℃ for 32.68, 26.81, 23.98, 8.18, 6.07, 2.90 and 2.21min respectively, and the denaturation rate is 90%. As can be seen from Table 8, in example 1, due to the reduction of temporary storage, transportation and other links, the temperature for sterilization, concentration and spray drying is relatively low, and the retention rate of the lactoferrin activity is about 89.8% in the whole process from milking to powdering; in example 7, the lactoferrin is denatured by 35.7%, 15.5%, 11%, 13.3% and 19.1% in the processes of temporary storage transportation (0-6 ℃, 17h), pasteurization (85 ℃, 15s), temporary storage of skim milk (0-6 ℃, 8h), sterilization concentration and spray drying after extrusion of fresh raw milk, and the ferritin activity retention rate from the fresh raw milk to the powdered milk is about 32.3%.
IgG is an immune active protein which is more sensitive to heat than lactoferrin, and the IgG is respectively heated at 65, 70, 75, 80, 85 and 90 ℃ for 32.68, 26.81, 23.98, 8.18, 6.07, 2.90 and 2.21min, and the denaturation rate is 90 percent. As can be seen from Table 8, in example 1, from fresh raw milk to powder, IgG is denatured to some extent in the processes of evaporation concentration and low-temperature spray drying, but the retention rate of IgG bioactivity in the whole process is 83.3%; in example 7, the storage time and the heat treatment strength together affect the bioactivity of IgG, and the IgG bioactivity retention rate is only 18.2% in the whole process.
Peroxidase is an enzyme in milk which is relatively heat-resistant and can be used as an indicator of pasteurization, and is generally inactivated by a heat treatment at 85 ℃ for 20 seconds. As is clear from table 8, in example 1, the spray drying may affect the peroxidase activity, but since the temperature of the powder in the spray drying tower is the wet bulb temperature and is lower than the exhaust air temperature, the retention of the peroxidase activity in the whole process is about 90.1%. (ii) a In example 7, pasteurization (85 ℃, 15s) of skim milk is the main cause of peroxidase inactivation, and secondly, the storage time is too long, the decomposition effect of a part of bacteria secreted protease on peroxidase is another main cause of inactivation, and the retention rate of lactoperoxidase activity is only 0.17% from fresh milk to powder in the whole process.
Lysozyme is a simple basic globulin consisting of 129 amino acids, which is more heat stable than IgG and peroxidase. As can be seen from Table 8, in example 1, low-temperature spray drying is a main link influencing the biological activity of the lysozyme, but the activity retention rate of the lysozyme in the whole process is 91.2%; in example 7, the long storage time and high heat treatment strength resulted in an activity retention of the immunological component of only 28.4% from fresh milk to powdered milk.
In conclusion, compared with example 7, the method for shortening the transportation and temporary storage time, effectively controlling the microbial quantity in the intermediate process and reducing the heat treatment strength in example 1 can more effectively retain the immune active ingredients in the fresh milk, and the activity retention rate can reach more than 80%.
It should be noted that the embodiments 1 to 6 are only preferred embodiments of the present invention, and are not intended to limit the present invention in other forms, and any person skilled in the art may use the above technical contents as a teaching to make changes or modifications to the equivalent embodiments with equivalent changes, but all those simple changes, equivalent changes and modifications made to the above embodiments without departing from the technical spirit of the present invention, and still all those embodiments fall within the scope of the present invention.
Claims (7)
1. The preparation method of the infant formula milk powder with the natural bioactivity is characterized in that the infant formula milk powder with the natural bioactivity is obtained by taking raw milk, performing centrifugal separation, primary microfiltration sterilization, wet mixing, secondary microfiltration sterilization, online grease addition, homogenization, evaporation concentration, spray drying and fluidized bed cooling drying.
2. The method for preparing infant formula milk powder with natural bioactivity according to claim 1, wherein the temperature of the two times of microfiltration sterilization is 6-10 ℃, the aperture of the microfiltration membrane is 0.8-1.6 μm, and the membrane area is 20-35 m2The transmembrane pressure difference is 21-41 kPa, and the circulating flow rate of the trapped fluid is 6-8 m/s; the flux of the permeate liquid membrane for the first microfiltration and sterilization is 900-1000 L.m-2·h-1Said secondThe flux of the permeate liquid membrane for secondary microfiltration and sterilization is 300-800 L.m-2·h-1。
3. The method of claim 2, wherein the two microfiltration sterilization steps are uniform transmembrane pressure difference microfiltration sterilization; the microfiltration membrane is a ceramic membrane.
4. The method of claim 1, wherein the milk is milk, cooled, and rapidly tested and qualified.
5. The method for preparing the infant formula milk powder with the natural bioactivity as claimed in claim 4, wherein the raw milk is temporarily stored for less than or equal to 10min after being milked and the temperature is 6-10 ℃.
6. Process for the preparation of an infant formula retaining natural biological activity according to any one of claims 1 to 5,
the milk raw material contains 2.8-3.4% of protein, 4.5-5.2% of lactose, 3.5-4.0% of fat, 12.0-13.5% of dry matter, less than or equal to 2 ten thousand CFU/mL of colony total number, less than or equal to 20 ten thousand somatic cells/mL, and zero residual contents of melamine and veterinary drugs;
after the centrifugal separation, the obtained skim milk contains 2.6-3.5% of protein, 4.5-5.3% of lactose, 8.0-9.5% of dry matter, less than or equal to 0.1% of fat, less than or equal to 1100CFU/mL of total number of colonies, and zero spore content;
after the first microfiltration sterilization, the obtained penetrating fluid A contains colony total number less than or equal to 30CFU/mL, and the contents of spores and somatic cells are zero;
after the wet mixing and proportioning, the obtained mixed material liquid B contains 12-36% of solid matters by weight, less than 1000CFU/mL of total number of bacterial colonies and less than 10 spores/mL;
after the second microfiltration sterilization, the obtained penetrating fluid C contains less than 30CFU/mL of total number of bacterial colonies and zero spore content;
after evaporation and concentration, the obtained concentrated milk D contains 50-55% of solid by weight;
after the spray drying, the weight fraction of water in the obtained powder E is less than or equal to 5.0 percent;
the weight percentage of the water in the infant formula milk powder retaining natural biological activity is less than or equal to 4 percent.
7. Process for the preparation of an infant formula retaining natural biological activity according to any one of claims 1 to 5,
the temperature of the centrifugal separation is 6-10 ℃;
the wet mixing ingredients are prepared by adding desalted whey powder, whey protein powder, lactose and other milk-containing raw materials at 6-10 ℃ and 0.45-0.8 bar, and stirring and mixing the materials at 1400-3000rpm, wherein the total time of the wet mixing ingredients is 15-25 min;
before the grease is added on line, the penetrating fluid C needs to be preheated to 25-55 ℃;
the online addition of the grease is that the grease is metered online in equal proportion, and the online addition temperature of the grease is 25-55 ℃;
the homogenizing conditions comprise a primary homogenizing pressure of 120-160 bar and a secondary homogenizing pressure of 30-50 bar;
preheating the feed liquid F obtained after homogenization to 65-70 ℃ before evaporation and concentration;
the conditions of evaporation concentration are as follows: the temperature of the first-effect evaporation is 65-70 ℃, and the vacuum degree is 0.75-0.8 bar; the temperature of the double-effect evaporation is 60-65 ℃, and the vacuum degree is 0.8-0.85 bar; the temperature of the triple effect evaporation is 55-60 ℃, and the vacuum degree is 0.85-0.95 bar; the condensation temperature is 50-55 ℃, and the vacuum degree is 0.85-0.95 bar;
the spray drying is low-temperature spray drying, the air inlet temperature is 30-120 ℃, the air exhaust temperature is 15-70 ℃, and the air inlet temperature of a static fluidized bed is 25-60 ℃;
the fluidized bed cooling drying is four-section dynamic fluidized bed drying, the air inlet temperature of the first section and the second section is 25-45 ℃, and the air inlet temperature of the third section and the fourth section is 15-30 ℃.
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| CN115644260A (en) * | 2022-10-27 | 2023-01-31 | 内蒙古伊利实业集团股份有限公司 | Preparation method and equipment of infant formula milk powder |
| CN115777786A (en) * | 2022-12-22 | 2023-03-14 | 黑龙江飞鹤乳业有限公司 | Milk powder preparation method capable of improving milk powder dissolution state and milk powder |
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- 2020-06-24 CN CN202010584292.3A patent/CN111887297A/en active Pending
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| CN101396045A (en) * | 2007-09-26 | 2009-04-01 | 光明乳业股份有限公司 | Production method of milk and milk produced by the method |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN115644260A (en) * | 2022-10-27 | 2023-01-31 | 内蒙古伊利实业集团股份有限公司 | Preparation method and equipment of infant formula milk powder |
| CN115777786A (en) * | 2022-12-22 | 2023-03-14 | 黑龙江飞鹤乳业有限公司 | Milk powder preparation method capable of improving milk powder dissolution state and milk powder |
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Address after: 050000 No.68, Shitong Road, Luquan District, Shijiazhuang City, Hebei Province Applicant after: JUNLEBAO Dairy Group Co.,Ltd. Address before: 050000 No.68, Shitong Road, Luquan District, Shijiazhuang City, Hebei Province Applicant before: THE SHIJIAZHUANG JUNLEBAO DAIRY Co.,Ltd. |
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Application publication date: 20201106 |