CN1164186C - Milk with probiotics and its production process - Google Patents
Milk with probiotics and its production process Download PDFInfo
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- CN1164186C CN1164186C CNB011127996A CN01112799A CN1164186C CN 1164186 C CN1164186 C CN 1164186C CN B011127996 A CNB011127996 A CN B011127996A CN 01112799 A CN01112799 A CN 01112799A CN 1164186 C CN1164186 C CN 1164186C
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Abstract
The present invention relates to milk with beneficial bacteria, which contains fat, protein and nonfat milk solid. The present invention is characterized in that the milk with beneficial bacteria also contains 1 wt% to 10 wt% of oligosaccharide and beneficial bacteria with the concentration of 1.0*10<6>-3.6*10<8> colony number / milliliter; the acidness of the milk with beneficial bacteria is 14 to 25 titration gradient. The production method of the milk with beneficial bacteria comprises: nonresistant milk is added to the oligosaccharide, stirred for 5 to 10 minutes, heated until the temperature reaches 60 to 80DEG C, homogenized, sterilized at the temperature of 65 to 140DEG C for 3 seconds to 30 minutes and cooled until the temperature reaches 4 to 25DEG C; beneficial bacteria are added to the nonresistant milk and the oligosaccharide, and finally stirred for 5 to 10 minutes. The milk with beneficial bacteria of the present invention has the functions of regulating intestinal bacteria, degrading cholesteryl, regulating immunity, etc., and tastes favorable.
Description
The present invention relates to a kind of milk product and manufacture method thereof, particularly a kind of milk product and manufacture method thereof of making carrier, adding probio and compound sugar with milk.
Probiotic composition profitable probliotics sour milk, probio oral liquid and the probio pulvis in the existing market, sold.Probiotic yogurt needs to add batchings such as certain amount of stabilizer, white granulated sugar on prescription, and being the consumer, these auxiliary materials really do not want to take in, and because the pH value lower (generally below pH4.5) of sour milk, so probio is not easy existence, the viable count instability of product does not reach the Expected Results of product.The probio oral liquid generally all needs some base culture medium (as moyashi), and composition is more single, and normal temperature preserves oral liquid and have viable bacteria hardly, so also do not reach the Expected Results of product.The probio pulvis contains a small amount of dextrin, starch, skimmed milk power etc., and nutrient is very limited, and the probio in the pulvis all is in resting state basically, also is difficult to reach the Expected Results of product.Aspect the mouthfeel of product, general milk has milk fragrance, but mouthfeel is more flat; Probiotic yogurt flavor acrid (flavor is like acetic acid) omits the tool bitter taste, and therefore, it is in order to overcome the mouthfeel defective, and the probio content in the general probiotic yogurt is all very low, so the product effect is bad; Probio oral liquid flavor as " soya bean soup ", mouthfeel is very poor.
The objective of the invention is at the defective of probiotic composition in the market, a kind of suitable probio existence is provided, improve mouthfeel, the milk with probiotics of enriched health care function and manufacture method thereof.
The invention provides a kind of milk with probiotics, contain fat, protein, non-fat solid, its characteristics are also to contain the compound sugar that percentage by weight is 1%-10% (one or more mixing) and 1.0 * 10
6-3.6 * 10
8The probio of clump count/milliliter (one or more mixing), and the acidity of this milk with probiotics is 14-25 titration gradient;
The percentage by weight of contained compound sugar is preferably 1.2%-7.6%;
Contained probio is preferably 6.0 * 10
7-3.2 * 10
8Clump count/milliliter;
The acidity of this milk with probiotics is preferably 17-21 titration gradient.
The present invention also provides a kind of manufacture method of milk with probiotics, and its raw material is the antibiotic-free milk of 90%-98.5%, the probio of 0.02%-0.5%, and the compound sugar of 1.4%-9.7%, above percentage is weight percentage; This manufacture method is as follows: antibiotic-free milk added in the compound sugar stirred 5-10 minute, be heated to 60-80 ℃, and homogeneous again, then 65-140 ℃ of sterilization 3 seconds-30 minutes, cooling causes 4-25 ℃ again, adds probio, stirs at last 5-10 minute;
Homogeneous has two kinds of methods: one-level homogeneous and double-stage homogenization, one-level homogenization pressure are 15295500-30591000Pa, and double-stage homogenization is 3059100-5098500Pa and 18354600-25492500Pa.
The definition of antibiotic-free milk: commonly do not contain antibiotic milk or do not contain antibiotic recombined milk or milk-contained drink with what milk powder, whey powder, rare cream or other milk ingredients were mixed with.
Probio mainly comprises: Bifidobacterium, lactobacillus acidophilus (lactobacillus.acidophilus), Lactobacillus casei (lactobacillus.casei), Lactobacillus salivarius (lactobacillus salivarius), streptococcus lactis (Strept.lactis), streptococcus faecalis (Enterococcus faecium), LGG (lactobacillus GG), Saccharomyces boulardii and Saccharomyces Cerecisiae etc.
Bifidobacterium comprises: bifidobacterium adolescentis (bifid.adolescentis), bifidobacterium infantis (bifid.infantis), bifidobacterium longum (bifid.longum), bifidobacterium breve (bifid.breve), animal bifidobacteria (bifid.breve), bifidobacterium bifidum (bifid.bifidum) and bifidobacterium thermophilum (bifid.themophilus) etc.
Compound sugar comprises: oligomeric (different) maltose, FOS, galactooligosaccharide, xylo-oligosaccharide, soyabean oligosaccharides, gossypose, lactulose etc.
Fig. 1 is a process chart of the present invention.
Fig. 2 is the another kind of process chart of the present invention.
Fig. 3 is another process chart of the present invention.
Fig. 4 is another process chart of the present invention.
Fig. 5 is another process chart of the present invention.
Fig. 6 is another process chart of the present invention.
Provide specific embodiment below the present invention made detailed description:
Embodiment 1
Prescription:
| Raw material | Material purity | Addition (gram) |
| Antibiotic-free milk | F≥2.0％，P≥1.8％，SNF≥7.0％ | 900 |
| Oligomeric maltose | 95％ | 80 |
| Xylo-oligosaccharide | 90％ | 17 |
| Lactobacillus casei | 1×10 11cfu/g | 1.0 |
| Bifidobacterium | 5×10 10cfu/g | 2.0 |
Production technology as shown in Figure 1.
Assay
| The finished product component | Content (percentage by weight) |
| Fat | 1.76％ |
| Protein | 1.62％ |
| Non-fat solid | 6.00％ |
| Compound sugar (comprising FOS and galactooligosaccharide) | 9.13％ |
| Probio (comprising lactobacillus acidophilus and Bifidobacterium) | 1.5×10 8cfu/ml |
| Acidity | 21 |
Embodiment 2
Prescription:
| Raw material | Material purity | Addition (gram) |
| Antibiotic-free milk | F≥4.2％，P≥4.0％，SNF≥11.5％ | 960 |
| FOS | 75％ | 20 |
| Galactooligosaccharide | 80％ | 15 |
| Lactobacillus acidophilus | 1×10 9cfu/g | 2.5 |
| Bifidobacterium | 1×10 9cfu/g | 2.5 |
Production technology as shown in Figure 2.
Assay
| The finished product component | Content (percentage by weight) |
| Fat | 3.95％ |
| Protein | 3.90％ |
| Non-fat solid | 11.00％ |
| Compound sugar (comprising FOS and galactooligosaccharide) | 2.20％ |
| Probio (comprising lactobacillus acidophilus and Bifidobacterium) | 1.0×10 6cfu/ml |
| Acidity | 14 |
Embodiment 3
Prescription:
| Raw material | Material purity | Addition (gram) |
| Antibiotic-free milk | F≥2.5％，P≥2.0％，SNF≥7.5％ | 977 |
| FOS | 75％ | 12 |
| Xylo-oligosaccharide | 100％ | 10 |
| Lactobacillus acidophilus | 3×10 11cfu/g | 0.5 |
| Bifidobacterium | 3×10 11cfu/g | 0.5 |
Production technology as shown in Figure 3.
Assay
| The finished product component | Content (percentage by weight) |
| Fat | 2.35％ |
| Protein | 1.90％ |
| Non-fat solid | 7.40％ |
| Compound sugar (comprising FOS and galactooligosaccharide) | 1.70％ |
| Probio (comprising lactobacillus acidophilus and Bifidobacterium) | 3.2×10 8cfu/ml |
| Acidity | 17 |
Embodiment 4
Prescription:
| Raw material | Material purity | Addition (gram) |
| Antibiotic-free milk | F≥2.0％，P≥1.7％，SNF≥7.0％ | 945 |
| Inulin | 75％ | 40 |
| Galactooligosaccharide | 80％ | 15 |
| Lactobacillus acidophilus | 5×10 11cfu/g | 0.1 |
| Bifidobacterium | 5×10 11cfu/g | 0.1 |
Production technology as shown in Figure 4.
Assay
| The finished product component | Content (percentage by weight) |
| Fat | 1.15％ |
| Protein | 1.20％ |
| Non-fat solid | 7.00％ |
| Compound sugar (comprising FOS and galactooligosaccharide) | 4.20％ |
| Probio (comprising lactobacillus acidophilus and Bifidobacterium) | 3.6×10 8cfu/ml |
| Acidity | 15 |
Embodiment 5
Prescription:
| Raw material | Material purity | Addition (gram) |
| Antibiotic-free milk | F≥3.4％，P≥3.2％，SNF≥10.5％ | 985 |
| Xylo-oligosaccharide | 90％ | 7 |
| Galactooligosaccharide | 80％ | 7 |
| Lactobacillus casei | 5×10 11cfu/g | 0.5 |
| Bifidobacterium | 5×10 11cfu/g | 0.5 |
Production technology as shown in Figure 5.
Assay
| The finished product component | Content (percentage by weight) |
| Fat | 3.35％ |
| Protein | 3.15％ |
| Non-fat solid | 9.60％ |
| Compound sugar (comprising FOS and galactooligosaccharide) | 1.20％ |
| Probio (comprising lactobacillus acidophilus and Bifidobacterium) | 7.1×10 7cfu/ml |
| Acidity | 20 |
Embodiment 6
Prescription:
| Raw material | Material purity | Addition (gram) |
| Antibiotic-free milk | F≥3.2％，P≥3.0％，SNF≥12.5％ | 920 |
| Xylo-oligosaccharide | 100％ | 68 |
| Galactooligosaccharide | 80％ | 10 |
| Lactobacillus casei | 8×1010cfu/g | 1.0 |
| Bifidobacterium | 8×1010cfu/g | 1.0 |
Production technology as shown in Figure 6.
Assay
| The finished product component | Content (percentage by weight) |
| Fat | 3.05％ |
| Protein | 2.90％ |
| Non-fat solid | 8.40％ |
| Compound sugar (comprising FOS and galactooligosaccharide) | 7.60％ |
| Probio (comprising lactobacillus acidophilus and Bifidobacterium) | 6.0×10 7cfu/ml |
| Acidity | 20 |
The source of raw material in the foregoing description:
The source of probio: French Crhodia, Japanese Wisby, Denmark Hansen etc.
Compound sugar: Guangdong Jiangmen city rivers and mountains bioengineering Co., Ltd, Tianyuan Health Food Co., Ltd., Yunnan Prov, Danisco-Ke Te (Shanghai) company
Antibiotic-free milk: Shanghai Bright Dairy ﹠ Food Co., Ltd the 7th, nine pastures
The antibiotic-free milk acceptance criteria: check general index according to GB6914-86, antibiotic index test method is as follows:
Get raw material milk 1000ml, insert 20-30ml working stock culture (Bacillus acidi lactici, lactobacillus acidophilus, streptococcus thermophilus fermentation agent),, test the titratable acidity of this sample,, then declare defective, otherwise qualified if acidity is lower than 65 ° of T in 40-42 ℃ of fermentation 3 hours.
Good effect of the present invention is with the milk of the nearly neutrality carrier as probio, and probiotic (compound sugar) arranged again, realized the best store method of probio, thereby the activity of probio keeps better, so the present invention has functions such as the gut flora of adjusting, degraded cholesterol, adjusting immunity.In addition, mouthfeel of the present invention is little sweet, and strong milk fragrance is arranged again, and the mouthfeel of more general probiotic composition is much better.
Be laboratory sample with the foregoing description 6 below, provide effect embodiment.
Effect embodiment 1 functions of loosening bowel relieving constipation
Sample proterties and processing: sample is a white liquid, is mixed with each concentration with distilled water, for examination.
Dosage design: this product human body RD is 20ml/60kg every day, and basic, normal, high three dosage groups are established in this experiment, is respectively 17,33,100ml/kg, and other establishes the blank group and gives blank milk liquid and the diphenoxylate constipation model control group that manufacturer provides.
Give sample loading mode: irritate stomach (irritating stomach in the high dose group 24h several times).
Animal used as test: Kunming kind small white mouse, the 18-22 gram, the cleaning level animal (raising of our station SPF Animal House) that provides by animal used as test portion of Shanghai Medical Univ, the quality certification number: 02-22-1, temperature 20 ± the 2C of laboratory animal breeding room, relative humidity 50-70%, the Animal House quality certification number: 02-28, the animal feeding material, technology ﹠ development Co. provides by the Su Hang animal used as test.
Cause emplastic: R-1132 (state-run Wujin pharmaceutical factory produces, specification 2.5mg/ sheet)
Experimental technique and result:
Sample is to the influence of mouse small intestine ahead running:
Get 50 of Kunming kind small white mouses, weigh, by body weight at random layering be divided into 5 groups, 10 every group, according to dosage method for designing was fed 10 days continuously.End day fasting was weighed after 24 hours, and oral sample or solvent are once, after 30 minutes, oral diphenoxylate 50mg/kg is except the blank group, press 0.2ml/10g with charcoal end suspension after 60 minutes and irritate stomach, take off cervical vertebra after 20 minutes and put to death animal, open abdomen and get small intestine, cut off the upper end to pylorus, the lower end is to the intestinal tube of ileocecus, pull into straight line gently, measure the charcoal end and advance length and small intestine total length, calculate the charcoal end and advance mobility.
Sample is to the influence of small intestine propulsion trial the weight of animals
As seen from the above table, sample does not have obvious influence to the weight of animals
| Group | Number of animals (only) | Body weight (gram) | Weightening finish | ||
| At the beginning of the experiment | In the experiment | Experiment eventually | |||
| Dosage high dose in the blank diphenoxylate contrast low dosage | 10 10 10 10 10 | 19±0.7 19±0.7 19±0.7 19±0.7 19±0.7 | 25±0.5 25±0.9 25±0.6 25±0.8 25±0.6 | 27±0.7 27±0.7 27±0.5 27±0.8 27±0.7 | 8±0.9 8±0.9 8±0.8 8±0.7 8±0.7 |
Sample antagonism diphenoxylate suppresses the effect that intestines advance
| Group | Number of animals (only) | The charcoal end advances mobility (%) |
| Blank | 10 | 65.7±13.4 * |
| The diphenoxylate contrast | 10 | 24.5±7.6 |
| Low dosage | 10 | 26.8±8.6 |
| Middle dosage | 10 | 34.0±11.6 |
| High dose | 10 | 44.4±15.2 * |
*(through variance analysis) compared with the diphenoxylate control group in P＜0.05
As seen from the above table, the blank group has been compared significant difference with the diphenoxylate control group, and the establishment of constipation model is described.Per os gives the sample 10 days of various dose, can resist the intestines inhibitory action that diphenoxylate causes, compares with the diphenoxylate control group, and high dose group has significant difference.
The mensuration of defecation grain number, defecation time and defecation gross weight:
Get 50 of the heavy mouse in Kunming, be divided into 5 groups, 10 every group by the body weight stratified random.According to dosage method for designing was fed 10 days continuously, end day fasting 24 hours, oral sample or solvent once, after 30 minutes, oral administered compound diphenoxylate 10mg/kg (0.2ml/10g) is except the blank group.Give compound diphenoxylate after 30 minutes, each group is pressed 0.2ml/10g with charcoal end suspension and is irritated stomach 1 time, after the administration mouse is put into little mouse cage respectively, be lined with filter paper in the cage, the all single cage of 5 treated animals is raised, normal drinking-water feed, first grain row's melena time of record mouse, in 6 hours, observe and write down the defecation grain number of every group of mouse, defecation gross weight.
Sample is to the influence of constipation experimental animal body weight
| Group | Number of animals (only) | Body weight (gram) | Weightening finish | ||
| At the beginning of the experiment | In the experiment | Experiment eventually | |||
| Dosage high dose in the blank diphenoxylate contrast low dosage | 10 10 10 10 10 | 19±0.8 19±0.7 19±0.7 19±0.7 19±0.7 | 25±0.7 25±0.9 25±0.9 25±0.7 25±0.7 | 27±1.0 27±0.8 27±0.8 27±0.7 27±0.7 | 8±1.2 8±0.8 8±0.5 8±0.5 8±0.6 |
As seen from the above table, sample does not have obvious influence to the weight of animals
Sample is to the influence of constipation animal pattern fecal grains
| Group | Number of animals (only) | Fecal grains (grain) |
| Blank | 10 | 38.4±4.9 * |
| The diphenoxylate contrast | 10 | 15.3±7.9 |
| Low dosage | 10 | 20.2±9.5 |
| Middle dosage | 10 | 20.6±6.8 |
| High dose | 10 | 26.4±9.0 * |
*(through variance analysis) compared with the diphenoxylate control group in P＜0.05
As seen from the above table, the blank group has been compared significant difference with the diphenoxylate control group, and the establishment of constipation model is described.Per os gives the sample 10 days of various dose, and sample high dose group fecal grains is compared with the diphenoxylate control group, and significant difference is arranged.
Sample is to the influence of constipation animal pattern defecation time
| Group | Number of animals (only) | Defecation time (branch) |
| Blank | 10 | 69.0±18.4 * |
| The diphenoxylate contrast | 10 | 167.0±60.9 |
| Low dosage | 10 | 154.1±43.7 |
| Middle dosage | 10 | 127.0±25.4 |
| High dose | 10 | 85.0±24.2 * |
*(through variance analysis) compared with the diphenoxylate control group in P＜0.05
As seen from the above table, the blank group has been compared significant difference with the diphenoxylate control group, and the establishment of constipation model is described.Per os gives the sample 10 days of various dose, and sample high dose group defecation time is compared with the diphenoxylate control group, and significant difference is arranged.
Sample is to the influence of constipation animal pattern defecation gross weight
| Group | Number of animals (only) | Defecation gross weight (gram) |
| Blank | 10 | 0.457±0.155 * |
| The diphenoxylate contrast | 10 | 0.251±0.075 |
| Low dosage | 10 | 0.280±0.114 |
| Middle dosage | 10 | 0.289±0.083 |
| High dose | 10 | 0.426±0.146 * |
*(through variance analysis) compared with the diphenoxylate control group in P＜0.05
As seen from the above table, the blank group has been compared significant difference with the diphenoxylate control group, and the establishment of constipation model is described.Per os gives the sample 10 days of various dose, and sample high dose group defecation gross weight is compared with the diphenoxylate control group, and significant difference is arranged.
Sum up: per os gives mouse various dose sample 10 days, has functions of loosening bowel relieving constipation.
Effect embodiment 2 immunoregulation effects
Sample proterties and processing: sample is a white liquid, is mixed with each concentration with distilled water, for examination.
Animal origin: BALB/C kind small white mouse, quality certification 02-22-1, body weight 18-22 gram, male, the cleaning level animal (raising of our station SPF Animal House) that provides by animal used as test portion of Shanghai Medical Univ.20 ± 2 ℃ of laboratory animal breeding room's temperature, relative humidity 50-70%, the Animal House quality certification number: 02-28, the animal feeding material, technology ﹠ development Co. provides by the Su Hang animal used as test.
Dosage design: this sample human body RD is 20ml/60kg every day, and basic, normal, high three dosage groups are established in this experiment, is respectively 17,33,100ml/kg, and the blank group is provided by the blank milk liquid that is provided by manufacturer.
Give sample loading mode: irritate stomach (irritating stomach in the high dose group 24h several times).
Experimental technique and result:
Body weight: animal is weighed, and by body weight layering at random, is divided into 4 groups, and 10 every group, design was according to dosage fed 28 days continuously, weighs once respectively in experiment mid-term and experiment end.
Sample (immune one group) is to the influence of the weight of animals
| The final weight gain of the initial body weight of group number of animals body weight in mid-term | |||||
| Dosage high dose in the blank low dosage | 10 10 10 10 | 19±0.7 18±0.5 20±1.3 19±1.4 | 22±0.9 23±0.9 24±1.3 23±1.3 | 24±1.5 24±0.9 26±1.6 25±1.1 | 5±1.9 6±0.7 6±1.0 6±0.8 |
As seen from the above table, each dosage group of sample is compared difference that there are no significant with control group.
Sample (immune two groups) is to the influence of the weight of animals
| The final weight gain of the initial body weight of group number of animals body weight in mid-term | |||||
| Dosage high dose in the blank low dosage | 10 10 10 10 | 19±0.6 18±0.5 20±1.3 20±1.3 | 22±1.3 23±0.4 23±1.3 24±1.3 | 25±1.4 25±0.7 26±1.3 26±1.1 | 6±1.4 7±0.9 6±0.7 6±0.9 |
As seen from the above table, each dosage group of sample is compared difference that there are no significant with control group.
Sample (immune three groups) is to the influence of the weight of animals
| The final weight gain of the initial body weight of group number of animals body weight in mid-term | |||||
| Dosage high dose in the blank low dosage | 10 10 10 10 | 19±0.7 19±0.5 20±1.3 19±1.2 | 23±1.5 23±0.8 24±1.3 23±1.3 | 25±1.1 25±1.3 26±1.3 25±0.8 | 6±1.2 6±1.3 6±1.2 6±0.6 |
As seen from the above table, each dosage group of sample is compared difference that there are no significant with control group.
Sample (immune four groups) is to the influence of the weight of animals
| The final weight gain of the initial body weight of group number of animals body weight in mid-term | |||||
| Dosage high dose in the blank low dosage | 10 10 10 10 | 20±1.3 19±0.9 19±1.4 20±1.2 | 24±0.9 23±1.2 23±1.6 24±1.3 | 26±0.8 25±1.3 25±1.6 26±1.1 | 6±1.4 6±1.1 6±1.1 6±1.1 |
As seen from the above table, each dosage group of sample is compared difference that there are no significant with control group.
The serum hemolysin test: after animal is given around the sample continuously, the eye socket blood sampling, separation of serum is put to death in the cervical vertebra dislocation.Add 25 μ l physiological saline on 96 hole Microhemagglutination plates, add 25 μ l serum first row respectively again, each row is the two-fold dilution later on, and every hole adds 1%SRBC1 drips, and vibration back room temperature was placed 3 hours, observed result after sinking appears in the blood cell contrast.
The serum hemolysin test
| Group | Number of animals (only) | Time (my god) | The antibody product |
| Blank | 10 | 28 | 42±27.6 |
| Low dosage | 10 | 28 | 55±27.1 |
| Middle dosage | 10 | 28 | 54±29.9 |
| High dose | 10 | 28 | 95±37.5 * |
*(through variance analysis) compared with the blank group in P＜0.05
Credit is analysed by statistics: the sample high dose group is compared with the blank group, and significant difference is arranged.
Antibody generates and detects test: after animal was given around the sample continuously, after 5 days, the dislocation of animal cervical vertebra was put to death, and gets spleen, makes splenocyte suspension with the immunity of defiber sheep red blood cell (SRBC), and adjusting cell concentration is 5 * 10
6/ ml.After top layer culture medium (the 1g agarose adds distilled water 100ml) heating for dissolving, put into 45 ℃ of water bath heat preservations, mix with the Hanks liquid of equivalent PH7.2-7.2,2 times of concentration, the packing small test tube, every pipe 0.5ml, in pipe, add 50 μ l10%SRBC again, 20 μ l splenocyte suspensions, rapid mixing, be poured on the 6cm plate of brushing thin agar layer, put into CO2gas incubator incubation 1.5h, the complement (1: 100) with the dilution of SA buffer solution joins in the plate then, after continuing incubation 1.5h, counting hemolysis plaque number.The result adds up with variance analysis.
| Group | Number of animals (only) | Hemolysis plaque number (* 10 3/ full spleen) |
| Blank | 10 | 109.5±7.5 |
| Low dosage | 10 | 113.2±7.9 |
| Middle dosage | 10 | 120.8±11.9 |
| High dose | 10 | 132.9±20.0 * |
*(through variance analysis) compared with the blank group in P＜0.05
Credit is analysed by statistics: the sample high dose group is compared with the blank group, and significant difference is arranged.
The mensuration of thymus index, spleen index
After animal was given around the sample continuously, every mouse was weighed, and the cervical vertebra dislocation is put to death, and got thymus gland, spleen is weighed, and calculates thymus index and spleen index respectively.
Thymus index, spleen index
| Group | Number of animals (only) | Thymus index (%) | Spleen index (%) |
| Blank | 10 | 0.22±0.04 | 0.52±0.13 |
| Low dosage | 10 | 0.22±0.10 | 0.52±0.09 |
| Middle dosage | 10 | 0.23±0.10 | 0.52±0.10 |
| High dose | 10 | 0.22±0.08 | 0.52±0.16 |
Credit is analysed by statistics: each dosage group of sample is compared difference that there are no significant with the blank group.
The clearance test of mouse carbon
The india ink of mouse tail vein injection dilution in 1: 3 treats that prepared Chinese ink injects timing immediately.Injected behind the prepared Chinese ink 2,10 minutes, and got blood 20 μ l from the vena ophthalmica clump respectively, and it is added to 2ml Na
2CO
3In the solution, survey the OD value at 600nm wavelength place with spectrophotometer, with Na
2CO
3Solution is made blank.Weigh and spleen re-computation phagocytic index according to the weight of animals, liver, the result adds up with variance analysis.
| Group | Number of animals (only) | Phagocytic index |
| Blank | 10 | 4.44±0.56 |
| Low dosage | 10 | 7.70±2.16 |
| Middle dosage | 10 | 6.82±2.55 |
| High dose | 10 | 9.30±3.33 * |
*(through variance analysis) compared with the blank group in P＜0.05
Credit is analysed by statistics: the sample high dose group is compared with the blank group, and significant difference is arranged.
The mouse spleen lymphocyte conversion test that ConA induces
After animal is given around the sample continuously; The cervical vertebra dislocation is put to death, and gets spleen, makes splenocyte suspension, and adjusting cell concentration is 2 * 10
6/ ml divides two holes to add in 24 well culture plates cell suspension, every hole 1ml, and a hole adds 50 μ lConA liquid (being equivalent to 5 μ g/ml), and 5%CO is put in contrast in another hole
2, cultivate 72h for 37 ℃.Cultivate and finish preceding 4h, supernatant 0.7ml is inhaled in every hole gently, adding 0.7ml does not contain the PRM1640 nutrient solution of calf serum, adds MTT (5mg/ml) 50 μ l/ holes simultaneously, continues to cultivate 4h, after cultivating end, every hole adds 1ml acid isopropyl alcohol, the piping and druming mixing, purple crystal is dissolved fully after, carry out colorimetric with the 570nm wavelength, the result analyzes with variance analysis.
| Group | Number of animals | The OD value that adds ConA | The OD value that does not add ConA | Difference |
| Dosage high dose in the blank low dosage | 10 10 10 10 | 0.191±0.023 0.304±0.054 0.302±0.056 0.389±0.166 | 0.103±0.010 0.113±0.011 0.107±0.008 0.114±0.009 | 0.088±0.016 0.191±0.058 * 0.195±0.056 * 0.275±0.160 * |
*(through variance analysis) compared with the blank group in P＜0.05
Credit is analysed by statistics: each dosage group of sample is compared with the blank group, and significant difference is all arranged.
DTH measures
Sensitization: after animal is given around the sample continuously, every mouse belly unhairing, the about 3 * 3cm of scope, the DNFB solution 50 μ l with 1% evenly smear sensitization.
The generation of DTH and mensuration: after 5 days, evenly be applied in mouse right ear (two sides) with 1%DNFB solution 10 μ l and attack.After 24 hours, mouse is put to death in cervical vertebra dislocation, takes off the left and right sides auricle of diameter 8mm with card punch, weighs and calculates the swelling degree.
The DTH measurement result
| Group | Number of animals | Ear weight-average value (mg) | The swelling degree | |
| Right | A left side | |||
| Dosage high dose in the blank low dosage | 10 10 10 10 | 22±2.2 25±3.3 29±4.5 32±5.2 | 16±3.0 13±2.1 14±2.9 13±2.0 | 6±4.6 12±3.2 15±4.7 * 19±4.6 * |
*(through variance analysis) compared with the blank group in P＜0.05
Credit is analysed by statistics: the middle and high dosage group of sample is compared with the blank group, and significant difference is all arranged.
The NK cytoactive is measured:
After animal was given around the sample continuously, the cervical vertebra dislocation was put to death, and gets spleen, makes splenocyte suspension (effector cell), got the back 24h YAC-1 cell that goes down to posterity and added 1640 complete culture solutions, and adjusting cell concentration is 1 * 10
5/ ml (target cell), get each 100 μ l of target cell and effector cell (imitating target) than 50: 1, add U type 96 well culture plates, target cell nature release aperture adds target cell and each 100 μ l of nutrient solution, maximum release aperture adds target cell and each 100 μ l of 1%NP40, above-mentioned every three multiple holes of all establishing are in 37 ℃, 5%CO
2Cultivated 4 hours in the incubator, then with 96 well culture plates with the centrifugal 5min of 1500r/min, draw at the bottom of the supernatant 100 μ l horizontalizations in 96 well culture plates in every hole, add LDH matrix industry 100 μ l simultaneously, reaction 3min, every hole adds the HCL30 μ l of 1mol/L, measures OD value (OD) at the 490nm place with ELIASA.
The NK cytoactive is measured
| Group | Number of animals (only) | Time (my god) | The NK cytoactive |
| Blank | 10 | 28 | 38±2.2 |
| Low dosage | 10 | 28 | 40±3.0 |
| Middle dosage | 10 | 28 | 41±4.5 |
| High dose | 10 | 28 | 50±5.6 * |
*(through variance analysis) compared with the blank group in P＜0.05
Credit is analysed by statistics: the sample high dose group is compared with the blank group, and significant difference is arranged.
Turnover of Mouse Peritoneal Macrophages is engulfed the chicken red blood cell test
After animal is given around the sample continuously, every mouse lumbar injection 20% chicken erythrocyte suspension 1ml, 30 minutes at interval, the cervical vertebra dislocation is put to death, and is fixed on the mouse plate, cuts off abdominal skin, injecting normal saline 2ml, rotated the mouse plate 1 minute, sucking-off abdominal cavity washing lotion 1ml divides and drips on 2 slides, 37 ℃ of wet incubating 30 minutes of incubator, use the physiological saline rinsing, dry, fix with 1: 1 acetone methanol solution, 4%Giemsa-phosphate buffer dyeing 3 minutes, dry with the distilled water rinsing again,, calculate and engulf % and phagocytic index with oily mirror microscopy.
The macrophage phagocytic test
| Group | Number of animals (only) | Engulf % | Phagocytic index |
| Blank | 10 | 40±2.5 | 0.49±0.03 |
| Low dosage | 10 | 42±3.6 | 0.52±0.03 |
| Middle dosage | 10 | 46±5.3 | 0.55±0.06 |
| High dose | 10 | 55±7.3 * | 0.67±0.12 * |
*(through variance analysis) compared with the blank group in P＜0.05
Credit is analysed by statistics: the sample high dose group is engulfed % and is compared with the blank group with phagocytic index, all has significant difference.
Conclusion: the animal per os was fed sample after 30 days, had immunoregulation effect.
Effect embodiment 3
| Index | The probio oral liquid | The probio pulvis | Probiotic yogurt | Milk with probiotics | |
| Flavor | Milk | 5％ | 5％ | 60％ | 100％ |
| Tart flavour | 80％ | 85％ | 90％ | 25％ | |
| Bitter taste | 15％ | 10％ | 40％ | 5％ | |
| Mouthfeel | Little sweet | 30％ | 5％ | 15％ | 85％ |
| Sweet | 5％ | 0 | 55％ | 10％ | |
| Little hardship | 65％ | 35％ | 60％ | 5％ | |
| Bitter | 35％ | 15％ | 30％ | 0％ | |
| Little acid | 45％ | 5％ | 0％ | 55％ | |
| Little and acid | 25％ | 5％ | 20％ | 40％ | |
| Acetic acid | 30％ | 0 | 80％ | 5％ | |
| Overall assessment | Excellent | 0 | 5％ | 5％ | 60％ |
| Very | 15％ | 15％ | 25％ | 25％ | |
| Still can | 30％ | 50％ | 55％ | 10％ | |
| Not good | 50％ | 30％ | 10％ | 5％ | |
| Very poor | 5％ | 0％ | 5％ | 0 | |
Illustrate: 20 people participate in sense organ evaluation altogether, the evaluation content divide three parts (promptly go up table " row " and content), overall assessment is by 100%.
Claims (7)
1. a milk with probiotics contains fat, protein, non-fat solid, it is characterized in that also containing the compound sugar and 1.0 * 10 that percentage by weight is 1%-10%
6-3.6 * 10
8The probio of clump count/milliliter, and the acidity of this milk with probiotics is 14-25 titration gradient; The raw material of this milk with probiotics is 90%～98.5% antibiotic-free milk, the probio of 0.02%-0.5%, the compound sugar of 1.4%-9.7%; This milk with probiotics is to add compound sugar in antibiotic-free milk, through stirring, heating, homogeneous, sterilization, cool off, add and make after probio stirs again.
2. probio according to claim 1, the percentage by weight that it is characterized in that contained compound sugar is 1.2%-7.6%.
3. probio according to claim 1 is characterized in that contained probio is 6.0 * 10
7-3.2 * 10
8Clump count/milliliter.
4. probio according to claim 1, the acidity that it is characterized in that this milk with probiotics are 17-21 titration gradient.
5. the manufacture method of the described milk with probiotics of claim 1, its raw material is the antibiotic-free milk of 90%-98.5%, the probio of 0.02%-0.5%, the compound sugar of 1.4%-9.7%, above percentage is weight percentage; This manufacture method is as follows: antibiotic-free milk added in the compound sugar stirred 5-10 minute, be heated to 60-80 ℃, and homogeneous again, then 65-140 ℃ of sterilization 3 seconds-30 minutes, cooling causes 4-25 ℃ again, adds probio, stirs at last 5-10 minute.
6. the manufacture method of a kind of milk with probiotics according to claim 5 is characterized in that this homogeneous carries out under 15295500-30591000Pa.
7. the manufacture method of a kind of milk with probiotics according to claim 5 is characterized in that this homogeneous is a double-stage homogenization, for the first time at 3059100-5098500Pa, for the second time at 18354600-25492500Pa.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB011127996A CN1164186C (en) | 2001-04-29 | 2001-04-29 | Milk with probiotics and its production process |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB011127996A CN1164186C (en) | 2001-04-29 | 2001-04-29 | Milk with probiotics and its production process |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1383727A CN1383727A (en) | 2002-12-11 |
| CN1164186C true CN1164186C (en) | 2004-09-01 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB011127996A Expired - Lifetime CN1164186C (en) | 2001-04-29 | 2001-04-29 | Milk with probiotics and its production process |
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Families Citing this family (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| FI109602B (en) * | 2001-01-25 | 2002-09-13 | Valio Oy | Probiotkombination |
| NZ544642A (en) * | 2003-06-23 | 2009-01-31 | Nestec Sa | Infant or follow-on formula |
| ES2524344T3 (en) * | 2005-02-28 | 2014-12-05 | N.V. Nutricia | Nutritive composition with probiotics |
| WO2008054193A1 (en) * | 2006-11-02 | 2008-05-08 | N.V. Nutricia | Nutritional products that comprise saccharide oligomers |
| CN101006803B (en) * | 2007-01-10 | 2011-01-26 | 光明乳业股份有限公司 | Lipid-lowering milk containing probiotics and its preparation method |
| CN101396046B (en) * | 2007-09-24 | 2012-09-05 | 光明乳业股份有限公司 | Probiotics yoghourt and manufacture method and use thereof |
| CN102972522B (en) * | 2011-09-06 | 2014-09-10 | 光明乳业股份有限公司 | Probiotic milk and preparation method thereof |
| US20130251829A1 (en) * | 2012-03-23 | 2013-09-26 | Mead Johnson Nutrition Company | Probiotic derived non-viable material for infection prevention and treatment |
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2001
- 2001-04-29 CN CNB011127996A patent/CN1164186C/en not_active Expired - Lifetime
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Address after: 201103 Minhang District, Wuzhong Road, No. 578, Patentee after: Guangming Milks Ind Co., Ltd. Address before: 200032 Shanghai Fenglin Road No. 251 Patentee before: Shanghai Bright Dairy & Food Co., Ltd. |
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Granted publication date: 20040901 |