CN112273444A - Milk base material and dairy product containing same and capable of improving immunity - Google Patents

Milk base material and dairy product containing same and capable of improving immunity Download PDF

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Publication number
CN112273444A
CN112273444A CN202010985410.1A CN202010985410A CN112273444A CN 112273444 A CN112273444 A CN 112273444A CN 202010985410 A CN202010985410 A CN 202010985410A CN 112273444 A CN112273444 A CN 112273444A
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Prior art keywords
milk
dairy product
parts
milk base
base material
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CN202010985410.1A
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Inventor
刘正冬
史苏华
房洪涛
李冬艳
王怀勇
李泽乐
裴晨红
江雨婷
庄建鹏
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Inner Mongolia Oushi Mengniu Dairy Product Co ltd
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Inner Mongolia Oushi Mengniu Dairy Product 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
    • 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
    • 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
    • A23V2400/11—Lactobacillus
    • A23V2400/113—Acidophilus
    • 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
    • A23V2400/531—Lactis

Abstract

The invention relates to a milk base material and a milk product containing the same and capable of improving immunity. The milk base material comprises the following components in parts by mass: 4500-6500 parts of milk material, 0.5-10 parts of phospholipid, 5-15 parts of composite mineral substance, 80-200 parts of sugar and 15-40 parts of dextrin. The invention mixes phospholipid and dextrin with a proper amount of milk material, and then forms a milk base material with a specific formula together with the compound mineral substance and sugar. The product prepared by adding prebiotics into the milk base material of the formula has the advantages of difficult deliquescence and good storage performance. Furthermore, the inventors have surprisingly found that probiotics are able to maintain good activity in the product. Meanwhile, the product prepared by adding prebiotics or/and probiotics into the milk base material of the formula has the effect of enhancing the immunity of the organism.

Description

Milk base material and dairy product containing same and capable of improving immunity
Technical Field
The invention relates to the technical field of dairy products, in particular to a milk base material and a dairy product containing the milk base material and capable of improving immunity.
Background
The balance of intestinal microecology is vital to human health, not only affects metabolic activity, but also participates in defense against invading pathogens. However, the global environment is continuously worsened, the pressure of people on life and work is getting stronger, living habits are irregular, food intake is unbalanced, exercise and sleep are lack, and antibiotics are abused, so that the intestinal micro-ecology of a human body is unbalanced, the intestinal flora is unbalanced, the body immunity is reduced, and various diseases are further caused.
Prebiotics can improve host health by stimulating the growth or activity of one or a few bacteria in the colon, improving the gut, promoting metabolism, and beneficially affecting the host. Particularly, after the probiotic product containing the compound components of the probiotics and the prebiotics is taken into the body, the probiotics and the prebiotics can act synergistically to maintain the micro-ecological balance of the intestinal tract of the body and resist invasive pathogens together. Overall, the synergistic effect of probiotics and prebiotics can promote health from two aspects, namely: improving health status and reducing disease occurrence. In particular, such probiotic products may promote the regulation of biological metabolism (including central and peripheral regulation), enhance appetite, promote nutrient absorption, strengthen biological defenses (including suppressing allergic reactions), induce immune stimulation and prevent hypertension, diabetes, cancer, high cholesterol, anemia, platelet aggregation and geriatric diseases, among others.
Based on the important regulatory effects of prebiotics on the body, a variety of food or health care formulations containing prebiotics have been developed. After the traditional probiotic product formula is ingested into the body, the body function can be obviously improved, and the health is promoted. However, the addition of prebiotics often results in increased water activity and moisture absorption of the resulting product.
Disclosure of Invention
Based on the above, the main object of the present invention is to provide a milk base and a milk product containing the milk base and capable of enhancing immunity. The dairy product prepared by adding prebiotics into the milk base material of the invention is not easy to deliquesce.
The milk base material comprises the following preparation raw materials in parts by mass:
Figure BDA0002689000300000021
in some of these embodiments, the milk base is prepared from:
Figure BDA0002689000300000022
in some of these embodiments, the phospholipid is selected from at least one of soy phospholipid and sunflower phospholipid.
In some of these embodiments, the dextrin is selected from at least one of maltodextrin and solid corn syrup.
In some of these embodiments, the dairy material comprises raw milk, whey powder, and whey protein powder; the milk base material comprises 4400-6400 parts of raw milk, 25-80 parts of whey powder and 5-45 parts of whey protein powder by mass.
Use of a milk base as described above in the preparation of a food or health product containing prebiotics.
In some of these embodiments, the prebiotic is an oligosaccharide or a polysaccharide.
A dairy product comprising a milk base as described above and a prebiotic.
In some embodiments, 3-12g of the prebiotic is present per 100g of the dairy product.
In some of these embodiments, the dairy product further contains a probiotic.
In some embodiments, the dairy product contains not less than 4 × 10 per 100g9The probiotic of CFU.
In some of these embodiments, a pH adjuster is added to the milk product to adjust the milk product to a suitable pH.
The invention has the following beneficial effects:
the invention mixes phospholipid and dextrin with a proper amount of milk material, and then forms a milk base material with a specific formula together with the compound mineral substance and sugar. The product prepared by adding prebiotics into the milk base material of the formula has the advantages of difficult deliquescence and good storage performance. Furthermore, the inventors have surprisingly found that probiotics are able to maintain good activity in the product. Meanwhile, the product prepared by adding prebiotics or/and probiotics into the milk base material of the formula has the effect of enhancing the immunity of the organism.
Drawings
FIG. 1 is a graph of the caking of the dairy products of example 1, comparative example 1 and comparative example 2 when left for 0 weeks;
FIG. 2 is a graph of the caking of the dairy products of example 1, comparative example 1 and comparative example 2 after being left for 1 week;
FIG. 3 is a graph of the caking of the dairy products of example 1, comparative example 1 and comparative example 2 when left for 2 weeks.
Detailed Description
In order that the invention may be more fully understood, reference will now be made to the following description. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. As used herein, the term "and/or" includes any and all combinations of one or more of the associated listed items.
The milk base material comprises the following preparation raw materials in parts by mass:
Figure BDA0002689000300000041
preferably, the milk base is prepared from the following raw materials:
Figure BDA0002689000300000042
preferably, the phospholipid is selected from at least one of soybean phospholipid and sunflower seed phospholipid.
Preferably, the dextrin is selected from at least one of maltodextrin and solid corn syrup.
Preferably, the dairy material comprises raw milk, whey powder, and whey protein powder; the milk base material comprises 4400-6400 parts of raw milk, 25-80 parts of whey powder and 5-45 parts of whey protein powder by mass.
Preferably, the complex mineral comprises calcium, iron, and zinc salts; the milk base material comprises, by mass, 6-13 parts of calcium salt, 0.3-1 part of iron salt and 0.05-0.6 part of zinc salt.
Preferably, the sugar is selected from at least one of fructose, lactose, sucrose, glucose, and maltose.
The embodiment of the invention also provides the application of the milk base material in preparing foods or health care products containing prebiotics.
The "prebiotics" in the present invention may be commonly used species such as oligosaccharides including fructo-oligosaccharide, galacto-oligosaccharide, xylo-oligosaccharide, isomalto-oligosaccharide, soy oligosaccharide, inulin, etc., some microalgae may also be used as prebiotics such as spirulina, arthrospira, etc., and furthermore polysaccharides (such as polysaccharose, nitrogen-containing polysaccharide of carrot), protein hydrolysates (such as hydrolysate of casein, alpha-lactalbumin, lactoferrin, etc.), vegetables in natural plants, Chinese herbal medicines, wild plants, etc. may also be used as prebiotics.
Preferably, the prebiotic is an oligosaccharide or a polysaccharide.
A dairy product comprising a milk base as described above and a prebiotic.
Preferably, 3-12g of the prebiotics are present per 100g of the dairy product.
Preferably, each 100g of the dairy product contains 3-10g of fructo-oligosaccharide and 0.0.45-2g of xylo-oligosaccharide.
Preferably, the dairy product further contains a probiotic.
The probiotics is active microorganisms which are beneficial to a host and change the composition of a flora at a certain part of the host by colonizing in a human body, and mainly comprises saccharomycetes, probiotic bacillus, clostridium butyricum, lactobacillus, bifidobacterium, actinomycetes and the like.
Preferably, the probiotics are lactobacillus acidophilus and bifidobacterium lactis.
Preferably, the dairy product contains not less than 4 x 10 per 100g9The probiotic of CFU.
Preferably, the dairy product contains not less than 2 x 10 per 100g9CFU of Bifidobacterium lactis of not less than 2X 109CFU lactobacillus acidophilus. The product prepared by adding prebiotics or/and probiotics into the milk base material has the effect of enhancing the immunity of the organism.
Preferably, a pH regulator for regulating the milk product to a proper pH is added into the milk product.
It is understood that the pH adjusting agent may be one or more of sodium hydroxide, potassium hydroxide, calcium hydroxide, citric acid, sodium citrate, potassium citrate. The pH regulator is added to adjust the pH of the milk product to 6.6-7.
The raw materials involved in the embodiments of the present invention can be obtained commercially. Wherein: bifidobacterium lactis, having the trade name Bifidobacterium lactis Bi-07, deposited under the number SD5220 with the American Type Culture Collection (ATCC). Lactobacillus acidophilus, under the name NCFM, has been deposited with the ATCC, American type culture Collection, with the number SD 5221.
Example 1
This example provides a dairy product prepared from a nutritional composition (also referred to as "milk base") having anti-deliquescent efficacy, prebiotics, probiotics, and a pH modifier;
the nutritional composition with the anti-deliquescence effect comprises the following components in parts by mass (in g) (specifically shown in a table 1): 5585g of milk (5500g of raw milk, 55g of whey powder and 30g of whey protein), 2g of phospholipid (soybean lecithin), 11.061g of composite mineral (10.53g of calcium carbonate, 0.398g of ferrous sulfate and 0.133g of zinc sulfate), 120g of sugar (10g of fructose and 110g of white granulated sugar) and 28.4g of maltodextrin;
the prebiotics are xylo-oligosaccharide and fructo-oligosaccharide, wherein the mass of the xylo-oligosaccharide and the fructo-oligosaccharide in each 100g of the dairy product is 0.45g and 6g respectively, and the prebiotics are added according to the content, and the total content of the prebiotics in the dairy product is 6.45 percent;
the probiotic bacteria are selected from Bifidobacterium lactis and Lactobacillus acidophilus, and the content of Bifidobacterium lactis in 100g dairy product is not less than 2 × 109CFU containing Lactobacillus acidophilus of not less than 2 × 109CFU, adding probiotics according to the content, wherein the probiotics content is not less than 4 multiplied by 10 in every 100g of dairy products9CFU；
The pH regulator is citric acid, and the addition amount of the citric acid is based on the regulation of the pH of the dairy product to 6.6-7.
Example 2
This example provides a dairy product comprising a nutritional composition having anti-deliquescent efficacy (also referred to as a "dairy base"), prebiotics, probiotics, and a pH adjusting agent;
the nutritional composition with the anti-deliquescence effect comprises the following components in parts by mass (in g) (specifically shown in a table 1): 5885g milk (5800g raw milk, 60g whey powder and 25g whey protein), 5g phospholipid (soya lecithin), 8.1g composite minerals (7.55g calcium carbonate, 0.35g ferrous sulphate and 0.2g zinc sulphate), 150g sugar (30g fructose and 120g white granulated sugar), and 20g maltodextrin;
the prebiotics are xylo-oligosaccharide and fructo-oligosaccharide, wherein each 100g of dairy product contains 0.5g and 10g of xylo-oligosaccharide and fructo-oligosaccharide, and the total content of prebiotics in the dairy product is 10.5%;
the probiotic bacteria are selected from Bifidobacterium lactis and Lactobacillus acidophilus, and the content of Bifidobacterium lactis in 100g dairy product is not less than 2 × 109CFU containing Lactobacillus acidophilus of not less than 2 × 109CFU, adding probiotics according to the content, wherein the probiotics content is not less than 4 multiplied by 10 in every 100g of dairy products9CFU；
The pH regulator is potassium citrate, and is added to regulate the pH of the milk product to 6.6-7.
Example 3
This example provides a dairy product comprising a nutritional composition having anti-deliquescent efficacy (also referred to as a "dairy base"), prebiotics, probiotics, and a pH adjusting agent;
the nutritional composition with the anti-deliquescence effect comprises the following components in parts by mass (in g) (specifically shown in a table 1): 4525g of milk material (4400g of raw milk, 80g of whey powder and 45g of whey protein), 10g of phospholipid (soybean lecithin), 6.35g of composite mineral (6g of calcium carbonate, 0.3g of ferrous sulfate and 0.05g of zinc sulfate), 80g of sugar (40g of fructose and 40g of white granulated sugar), and 40g of maltodextrin;
the prebiotics are xylo-oligosaccharide and fructo-oligosaccharide, wherein each 100g of dairy product contains 2g and 3g of xylo-oligosaccharide and fructo-oligosaccharide, and the total content of prebiotics in the dairy product is 5%;
the probiotic bacteria are selected from Bifidobacterium lactis and Lactobacillus acidophilus, and the content of Bifidobacterium lactis in 100g dairy product is not less than 2 × 109CFU containing Lactobacillus acidophilus of not less than 2 × 109CFU, in accordance withThe amount of the added probiotics is not less than 4 multiplied by 10 per 100g of dairy products9CFU；
The pH regulator is sodium citrate, and is added to regulate the pH of the milk product to 6.6-7.
Example 4
This example provides a dairy product comprising a nutritional composition having anti-deliquescent efficacy (also referred to as a "dairy base"), prebiotics, probiotics, and a pH adjusting agent;
the nutritional composition with the anti-deliquescence effect comprises the following components in parts by mass (in g) (specifically shown in a table 1): 6430g of milk (6400g raw milk, 25g whey powder and 5g whey protein), 0.5g of phospholipid (soybean lecithin), 16.6g of composite minerals (15g calcium carbonate, 1g ferrous sulfate and 0.6g zinc sulfate), 200g of sugar (50g fructose and 150g white granulated sugar), and 15g of maltodextrin;
the prebiotics are xylo-oligosaccharide and fructo-oligosaccharide, wherein each 100g of dairy product contains 1g and 5g of xylo-oligosaccharide and fructo-oligosaccharide, and the total content of prebiotics in the dairy product is 6%;
the probiotic bacteria are selected from Bifidobacterium lactis and Lactobacillus acidophilus, and the content of Bifidobacterium lactis in 100g dairy product is not less than 2 × 109CFU containing Lactobacillus acidophilus of not less than 2 × 109CFU, adding probiotics according to the content, wherein the probiotics content is not less than 4 multiplied by 10 in every 100g of dairy products9CFU；
The pH regulator is potassium citrate, and is added to regulate the pH of the milk product to 6.6-7.
Table 1, Dairy product formulas of examples 1 to 4 (parts by mass)
Figure BDA0002689000300000081
Figure BDA0002689000300000091
Comparative example 1
The comparative example is a comparative example of example 1, the main difference with respect to example 1 being that the dairy product comprises a milk base of a different composition, the milk base of the comparative example not comprising lecithin and maltodextrin, in particular the milk auxiliary of the comparative example comprises: 5615.4g of milk material (5530.4g of raw milk, 55g of whey powder and 30g of whey protein), 0g of soybean lecithin, 11.061g of compound mineral (10.53g of calcium carbonate, 0.398g of ferrous sulfate and 0.133g of zinc sulfate), 120g of sugar (10g of fructose and 110g of white granulated sugar) and 0g of maltodextrin; the rest of the components and their contents in the milk product were the same as in example 1.
Comparative example 2
The comparative example is the comparative example of example 1, and the main difference with respect to example 1 is that the dairy product contains different compositions of milk base, the milk base of the comparative example does not contain lecithin and maltodextrin, and the dosage of the milk base is low, and specifically, the milk auxiliary material of the comparative example contains: 4360g milk material (4100g raw milk, 250g whey powder and 10g whey protein), 0g soybean lecithin, 11.061g composite mineral (10.53g calcium carbonate, 0.398g ferrous sulfate and 0.133g zinc sulfate), 120g sugar (10g fructose and 110g white granulated sugar), and 0 maltodextrin; the rest components and the content thereof in the dairy product are the same as in example 1.
Performance test one, storage test
Taking the dairy product samples of the example 1, the comparative example 1 and the comparative example 2, opening the tank and placing the samples under the conditions of the temperature of 37.5 +/-2 ℃ and the humidity RH 75% +/-5%, observing the state of the samples every day, recording the caking condition of the samples and sampling the samples to test the moisture content.
The results are shown in the following table:
TABLE 2
Figure BDA0002689000300000101
As can be seen from the above table and fig. 1, 2, and 3: the dairy product of example 1 was not deliquescent and had better storage properties, whereas comparative example 1 and comparative example 2 were relatively poor, which demonstrates that the use of phospholipids and dextrin was effective in improving the deliquescent problem of dairy products when prebiotics were added. With respect to comparative example 1 and comparative example 2, comparative example 2 is relatively more deliquescent, which illustrates that the regulation of the milk content helps to ameliorate the deliquescence problem of the dairy formula of the invention. The dairy products of examples 2 to 4 were tested with reference to the above method, and the results show that the results of examples 1 and 2 are relatively good, which indicates that there is a preferred scheme for the dairy product formulation of the present invention.
Experiment for testing performance and enhancing immunity function
Taking the dairy product of the embodiment 1 as an example, the dairy product provided by the invention is tested for the immunity synergistic effect, and the specific steps are as follows:
1. dose design: the recommended human dose of the dairy product is every 50g/60 kg. Because the recommended daily dosage of the dairy product is too large, the milk base is removed to: the fructo-oligosaccharide, the xylo-oligosaccharide, the lactobacillus acidophilus and the bifidobacterium lactis are prepared according to the formula proportion of the product, and the recommended daily dosage is 3.635g/60 kg. The animal experiment is provided with three dosage groups of low, medium and high, namely 0.303g/kg, 0.606g/kg and 1.818g/kg, and is additionally provided with a distilled water blank control group.
2. Sample preparation: weighing 16130mg of fructo-oligosaccharide, 2000mg of xylo-oligosaccharide, 34mg of lactobacillus acidophilus and 16mg of bifidobacterium lactis according to the experimental dosage design requirement and the recommended dosage of a sample, adding steam feed water to 200ml, fully mixing uniformly to obtain a high-dosage test solution, diluting the high-dosage test solution with distilled water 1:2 to obtain a medium-dosage test solution, and diluting the high-dosage test solution with distilled water 1:5 to obtain a low-dosage test solution. The blank control group was given distilled water.
3. Experimental animals: clean grade ICR species mice, license number: SCXK 2007-0005 (Shanghai) with a body weight of 18-23 g, female, was supplied by Shanghai slyke laboratory animals GmbH, the central SPF animal house. The temperature of the laboratory animal feeding room is 20-25 ℃, the relative humidity is 40-70%, and the use license number of the laboratory animal is as follows: SYXK (Shanghai) 2007-0008, animal feed, provided by Suzhou Shuangshi laboratory animal feed science and technology services GmbH, registration number: su-feeding trial (2009) 05032.
4. The sample administration route comprises the following steps: the volume of the gavage is 0.4ml/20 g.BW.
5. Experimental methods
5.1 grouping: the 200 mice were randomly divided into five large groups of 40 mice each by weight (one group was immunized for NK cell activity determination and strangulation experiment, two groups were immunized for DTH, spleen index and thymus index experiment, three groups were immunized for hemolytic plaque and hemolysin experiment, four groups were immunized for clearance experiment, five groups were immunized for phagocytosis experiment). The control group and four groups of low, medium and high dose groups were set in each experimental group, and 10 groups were used in each group.
5.2 feeding and weighing: mice were continuously fed for 30 days with dose design, weighed once per week, and initial, middle (second week) and end-of-experiment body weights were calculated. The experimental results were counted using SPSS software.
5.3 organ/body weight ratio (thymus index, spleen index) determination: after the animals are continuously given samples for 30 days, cervical dislocation is killed, thymus and spleen are taken and weighed, and thymus index and spleen index are calculated respectively.
5.4 cellular immune function assay:
5.4.1 ConA-induced splenic lymphocyte transformation experiments in mice:
after the animals were continuously administered for 30 days, cervical dislocation was sacrificed, spleen was taken out and prepared into spleen cell suspension, and the cell concentration was adjusted to 3 xl 06One/ml, the cell suspension was added to a 24-well plate in two wells, 1ml per well, 75. mu.l ConA solution (equivalent to 7.5. mu.g/ml) was added to one well, and 5% CO was added to the other well as a control2And culturing at 37 ℃ for 72 h. 4h before the culture is finished, gently sucking 0.7ml of supernatant in each hole, adding 0.7ml of PRMI1640 culture solution without calf serum, simultaneously adding 50 mu 1/hole of MTT (5mg/ml), continuously culturing for 4h, adding 1ml of acidic isopropanol in each hole after the culture is finished, uniformly blowing and stirring, completely dissolving purple crystals, carrying out color comparison at the wavelength of 570nm, and counting the experimental results by SPSS15.0 software.
5.4.2 determination of DTH and determination of thymus index and spleen index:
after the sensitized animals were continuously administered for 30 days, the abdomen of each mouse was unhaired in a range of about 3cm × 3cm, and 1% DNFB solution (50 μ 1) was uniformly applied to the sensitization. Generation and determination of DTH: after 5 days, the mice were challenged by spreading the mice with 10. mu.1 l% DNFB solution evenly on the right ear (both sides). After 24 hours, each mouse was weighed, the mice were sacrificed by cervical dislocation, and left and right ear pieces having a diameter of 8mm were removed with a punch, weighed and the swelling degree was calculated. Weighing thymus and spleen, and calculating thymus index and spleen index respectively.
5.4.3 humoral immune function assay:
5.4.3.1 antibody production detection assay:
continuously feeding animal for 30 days, immunizing defibered sheep red blood cell for 5 days, dislocating animal cervical vertebra, killing, collecting spleen, making splenocyte suspension, and adjusting cell concentration to 5 × 106One per ml. Heating and dissolving a surface layer culture medium (lg agarose added with double distilled water to 100ml), placing the mixture into a 45 ℃ water bath for heat preservation, mixing the mixture with an equal amount of Hanks liquid with the concentration of 2 times and the pH value of 7.2-7.4, subpackaging small test tubes, wherein 0.5ml of each tube is added, 50 mu l of 10% SRBC and 20 mu 1 spleen cell suspension are added into the tubes, quickly and uniformly mixed, poured onto a 6cm plate with a thousand agar thin layer, placed into a carbon dioxide incubator for incubation for 1.5h, then a complement (1:10) diluted by SA buffer solution is added into the plate, and after the incubation is continued for 1.5h, the number of hemolytic plaques is counted.
5.4.3.2 serum hemolysin experiment:
after animals were continuously given for 30 days, and after animals were immunized with 2% (v/v) SRBC for 5 days, blood was collected from the orbit, and serum was sacrificed by dislocation of cervical vertebrae. Adding 100 μ l of physiological saline to 96-well micro hemagglutination plate, adding 100 μ l of serum to the first row, diluting each row in multiple proportion, adding 100 μ l of 1% SRBC to each well, standing at 37 deg.C for 3 hr after shaking, and observing the result when the blood cell control is settled.
5.4.4 monocyte-macrophage function assay:
5.4.4.1 mouse carbon clearance test
After the animals were continuously dosed for 30 days, india ink diluted 1:3 was injected into the tail vein and immediately timed after the ink injection. 2, 10 minutes after the injection of the ink, 20. mu.1 of blood was collected from the venous plexus of the eyes and added to 2ml of 0.1% Na2C03In solution, the OD value is measured with a spectrophotometer at a wavelength of 600nm as Na2C03The solution was used as a blank control. Phagocytosis index was calculated from animal body weight, liver weight and spleen weight.
5.4.4.2 experiment of phagocytosis of chicken red blood cells by mouse abdominal cavity macrophages:
after animals are continuously fed for 30 days, injecting 1ml of 20% chicken erythrocyte suspension into the abdominal cavity of each mouse at intervals of 30 minutes, killing and fixing the mouse plate by dislocation of cervical vertebra, cutting the abdominal wall skin, injecting 2ml of normal saline, rotating the mouse plate for 1 minute, sucking out 1ml of abdominal washing liquid, dripping the abdominal washing liquid on 2 glass sheets, carrying out wet incubation for 30 minutes at 37 ℃, rinsing with normal saline, drying later, fixing with 1:1 acetone methanol solution, staining for 3 minutes by 4% Giemsa-phosphoric acid buffer solution, rinsing the slit dry by distilled water, carrying out oil microscopic examination, and calculating phagocytosis percentage (%) and phagocytosis index.
5.4.5NK cell Activity assay:
continuously feeding animals for 30 days, killing cervical vertebra by dislocation, collecting spleen, preparing spleen cell suspension (effector cell), adding YAC-1 cells 24h after passage into RPMI1640 complete culture solution, and adjusting cell concentration to 4 × 105Taking each 100 mu 1 of target cells and effector cells (the effective target ratio is 50:1), adding the target cells and the effector cells into a U-shaped 96-well culture plate, adding each 100 mu 1 of target cells and culture solution into natural release holes of the target cells, adding each 100 mu 1 of target cells and 1% NP 40 into maximum release holes of the target cells and the maximum release holes of the NP 40, setting three multiple wells, culturing for 4 hours in a 5% C02 incubator at 37 ℃, sucking 100 mu 1 of supernatant into each well, putting the supernatant into a flat-bottom 96-well culture plate, simultaneously adding 100 mu 1 of LDH matrix solution, reacting for 3min, adding 30 mu 1 of lmol/L HCL into each well, and measuring the optical density value (OD) at 490mm by using a microplate reader.
Counting: the results of the experiment were statistically analyzed using SPSS 13.0 software, and each dose group was compared with a control group.
And 6, judging the result:
the result is positive in any two aspects of cellular immune function, humoral immune function, monocyte-macrophage function and NK cell activity, and the tested sample can be judged to have the function of enhancing the immune function.
Wherein, the results of two experiments (namely a mouse spleen lymphocyte transformation experiment and a delayed type allergic reaction experiment) in the cellular immune function detection items are positive, or the results of two dose groups in any experiment are positive, so that the cellular immune function determination result can be judged to be positive. The results of both experiments (i.e., antibody-producing cell detection and serum hemolysin measurement) in the humoral immune function measurement item are positive, or the results of both dose groups in any one of the experiments are positive, and the positive humoral immune function measurement result can be judged. The results of two experiments (namely a mouse carbon clearance experiment and a mouse abdominal cavity phagocytic cell phagocytic chicken erythrocyte experiment) in the function determination items of the mononuclear macrophages are positive, or the results of two dose groups of any one experiment are positive, so that the function result of the mononuclear macrophages can be judged to be positive. More than one dose group of NK cell activity determination experiments have positive results, and NM cell results can be judged to be positive.
7. Results
TABLE 3 immunization panel (NK cell Activity, splenic lymphocyte transformation experiments) results
Figure BDA0002689000300000151
As can be seen from the above table, the 0.606g/kg dose combination 1.818 dose group was significantly different from the control group.
TABLE 4 immunization panel (DTH, spleen index and thymus index experiments) results
Figure BDA0002689000300000152
As can be seen from the above table, the swelling degree of the 1.818 dose group was significantly different from that of the control group in the DTH test.
In the remaining immunization groups, the dose group was not significantly different from the control group.
8. Conclusion
Under the test condition, the result of the cellular immune function is positive, the result of the NK cell activity is positive, but the results of the humoral immune function and the monocyte-macrophage function are negative. According to the result judgment principle of 'enhancing immunity' in the 2003 edition of 'health food inspection and evaluation technical Specifications', the sample is considered to have the function of enhancing immunity of mice.
The technical features of the embodiments described above may be arbitrarily combined, and for the sake of brevity, all possible combinations of the technical features in the embodiments described above are not described, but should be considered as being within the scope of the present specification as long as there is no contradiction between the combinations of the technical features.
The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.

Claims (10)

1. The milk base material is characterized by comprising the following preparation raw materials in parts by mass:
Figure FDA0002689000290000011
2. a milk base according to claim 1, characterized in that it is prepared from the following raw materials:
Figure FDA0002689000290000012
3. the milk base of claim 1, wherein the phospholipids are selected from at least one of soy phospholipids and sunflower phospholipids; and/or, the dextrin is selected from at least one of maltodextrin and solid corn syrup; and/or the milk material comprises raw milk, whey powder and whey protein powder; the milk base material comprises 4400-6400 parts of raw milk, 25-80 parts of whey powder and 5-45 parts of whey protein powder by mass.
4. The dairy base according to any one of claims 1 to 3, wherein the complex mineral comprises calcium salts, iron salts, and zinc salts; the milk base material comprises 6-15 parts of calcium salt, 0.3-1 part of iron salt and 0.05-0.6 part of zinc salt by mass; and/or, the sugar is selected from at least one of fructose, lactose, sucrose, glucose, and maltose.
5. Use of the milk base of any of claims 1 to 4 for the preparation of a prebiotic containing food or health product.
6. Use according to claim 5, wherein the prebiotic is an oligosaccharide or a polysaccharide.
7. A dairy product characterized in that the dairy product is prepared from raw materials comprising the milk base of any one of claims 1 to 4 and prebiotics.
8. The dairy product according to claim 7, characterized in that it contains 3-12g of said prebiotic per 100g of said dairy product.
9. The dairy product according to claim 7 or 8, wherein the dairy product further comprises a probiotic; and/or a pH regulator for regulating the milk product to proper pH is added into the milk product.
10. The dairy product of claim 9, comprising not less than 4 x 10 per 100g of said dairy product9The probiotic of CFU.
CN202010985410.1A 2020-09-18 2020-09-18 Milk base material and dairy product containing same and capable of improving immunity Pending CN112273444A (en)

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Application publication date: 20210129