CN106616139A - Children's functional drink - Google Patents
Children's functional drink Download PDFInfo
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- CN106616139A CN106616139A CN201610788381.3A CN201610788381A CN106616139A CN 106616139 A CN106616139 A CN 106616139A CN 201610788381 A CN201610788381 A CN 201610788381A CN 106616139 A CN106616139 A CN 106616139A
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- Prior art keywords
- enzymolysis
- children
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- product
- enzyme
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 47
- 235000015510 Cucumis melo subsp melo Nutrition 0.000 claims abstract description 10
- FJJCIZWZNKZHII-UHFFFAOYSA-N [4,6-bis(cyanoamino)-1,3,5-triazin-2-yl]cyanamide Chemical compound N#CNC1=NC(NC#N)=NC(NC#N)=N1 FJJCIZWZNKZHII-UHFFFAOYSA-N 0.000 claims abstract description 10
- 235000021552 granulated sugar Nutrition 0.000 claims abstract description 10
- 239000000047 product Substances 0.000 claims description 53
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- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 24
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- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 20
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- 239000004480 active ingredient Substances 0.000 abstract 1
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- 206010018910 Haemolysis Diseases 0.000 description 2
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 2
- AGPKZVBTJJNPAG-WHFBIAKZSA-N L-isoleucine Chemical compound CC[C@H](C)[C@H](N)C(O)=O AGPKZVBTJJNPAG-WHFBIAKZSA-N 0.000 description 2
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- CDBYLPFSWZWCQE-UHFFFAOYSA-L sodium carbonate Substances [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 2
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Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/38—Other non-alcoholic beverages
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/70—Clarifying or fining of non-alcoholic beverages; Removing unwanted matter
- A23L2/84—Clarifying or fining of non-alcoholic beverages; Removing unwanted matter using microorganisms or biological material, e.g. enzymes
-
- 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
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
Abstract
The invention provides children's functional drink. According to a technical scheme of the invention, the children's functional drink is prepared from, by weight, 10-30 parts of white granulated sugar, 11-15 parts of active component B, 9-13 parts of active ingredient, 2-6 parts of sodium hydrogen phosphate, 0.3-0.5 part of Hami melon essence, 3-5 parts of glucose, and 105-135 parts of water. The children's functional drink has rich nutrition and good taste, contains immune factors required by human bodies, can promote the growth and development of bodies, adjust gastrointestinal function, improve bodily immunity, refresh milk and enhance fatigue resistance, has significant health effect for the diseases such as heart, blood vessels and cervical vertebra; the children's functional drink is suitable for general people, especially children, and can significantly improve the growth and development of children and supplement energy.
Description
Technical field
The invention belongs to medicines and health protection field, and in particular to a kind of children's drinks.
Background technology
At present, children's beverage great majority on the market do not possess health care, and some sodas drink for a long time meeting
Many negative impacts are caused to the healthy of children.And such as calcium milk drink, also mainly supplementing calcium activated and vitamin
A, C, D etc. are main purpose, and its action effect is single, and assimilation effect is slow, and real meaning is not also belonged to a certain extent
Heath-function beverage.
The content of the invention
In view of this, the present invention provides a kind of children's drinks, and the drinks of the present invention are nutritious, in good taste,
Immune-regulating factor rich in needed by human body, can promote body growth to develop, and adjust gastrointestinal function, improve immunity of organisms, carry
Refreshing restoring consciouness, strengthens anti-fatigue ability, has obvious health-care effect to diseases such as the heart, the cerebrovascular and cervical vertebras.The present invention is adapted to vast
Crowd is particularly children and drinks, and to children's growth, supplementary physical efficiency significant facilitation is played.
The technical scheme is that：A kind of children's drinks, it is in parts by weight, composed of the following components：Activearm
Divide A 40-50, potassium chloride 1-3, xylitol 5-8, sodium citrate 2-4, white granulated sugar 10-30, active component B 11-15, activearm
Divide C 9-13, dibastic sodium phosphate 2-6, Hami melon essence 0.3-0.5, glucan 3-5, water 105-135.
Further, described children, it is in parts by weight, composed of the following components：Active component A 43, potassium chloride 2, wood
Sugar alcohol 6, sodium citrate 3, white granulated sugar 14, active component B 13, active component C 11, dibastic sodium phosphate 4, Hami melon essence 0.4,
Bextran 45, water 118.
Further, the active component A be long tail anchovy enzymolysis product, the preparation method of the long tail anchovy enzymolysis product
For：S1. long tail anchovy minced fillet is taken, makes material-water ratio be 1:2-1:8 (w/v), 2.0-3.5 is adjusted to 2 mol/L hydrochloric acid by solution ph
So that stomach acid or alkali environment is presented, pepsin is added by 1200-1500U/g of enzyme-to-substrate concentration ratio, in water bath with thermostatic control vibration
37 DEG C of holding, 100-130 r/min vibration enzymolysis 1.5-2.5 h, obtain with this understanding the first of Jing pepsins enzymolysis in device
Product；S2. the enzymolysis liquid pH value Jing after pepsin enzymolysis is adjusted into 8.0-9.0 with the NaOH of 2 mol/L, keeps 37
DEG C hydrolysis temperature, complex enzyme addition is 4500-6000 U/g, trypsase and chymotrypsin in the complex enzyme
Mass ratio is 4:1-6:1, the material-water ratio 1:3-1:6, enzymolysis time is 3-4.5 h, after enzymolysis terminates, in 100 DEG C of boiling water baths
In go out enzyme 10min；By the enzymolysis product in S2 by vacuum freeze drying, settings drying condition is -30 DEG C of precooling temperature, precooling
Time 2h, heating-up temperature starts slowly to be heated up after 18h from -10 DEG C, reaches 25 DEG C, and remains to and be dried terminal, obtains long tail anchovy
Enzymolysis product.
Further, the active component A be long tail anchovy enzymolysis product, the preparation method of the long tail anchovy enzymolysis product
For：S1. long tail anchovy minced fillet is taken, makes material-water ratio be 1:3 (w/v), solution ph with 2 mol/L hydrochloric acid be adjusted to 2.5 stomach is presented
Portion's acid or alkali environment, by 1300 U/g of enzyme-to-substrate concentration ratio pepsin is added, and 37 are kept in thermostatic control oscillator vibration
DEG C, 110 r/min vibration 2 h of enzymolysis, the head product of Jing pepsins enzymolysis is obtained with this understanding；S2. with the hydrogen of 2 mol/L
Enzymolysis liquid pH value Jing after pepsin enzymolysis is adjusted to 8.5 by sodium oxide molybdena, keeps the 37 DEG C of hydrolysis temperatures, complex enzyme addition to be
5200 U/g, trypsase and the mass ratio of chymotrypsin are 5 in the complex enzyme:1, the material-water ratio 1:4.5, enzyme
The solution time is 3.5 h, and after enzymolysis terminates, go out enzyme 10min in 100 DEG C of boiling water baths；Enzymolysis product in S2 is cold by vacuum
Lyophilized dry, setting drying condition is -30 DEG C of precooling temperature, and pre-coo time 2h, heating-up temperature starts slowly to be risen after 18h from -10 DEG C
Temperature, reaches 25 DEG C, and remains to and be dried terminal, obtains long tail anchovy enzymolysis product.
Further, active component B is mud blood clam enzymolysis product, and the preparation method of the mud blood clam enzymolysis product is：S1.
Take mud blood clam whole viscera and blend into meat gruel, make material-water ratio be 1:1-1:2 (w/v), 2.0 are adjusted to 2 mol/L hydrochloric acid by solution ph
So that stomach acid or alkali environment is presented, pepsin is added by 800-1000U/g of enzyme-to-substrate concentration ratio, in thermostatic control oscillator vibration
It is middle to keep 37 DEG C, 75-105 r/min vibration enzymolysis 1-1.5 h, the head product of Jing pepsins enzymolysis is obtained with this understanding；
S2. the enzymolysis liquid pH value Jing after pepsin enzymolysis is adjusted into 8.0-9.0 with the NaOH of 2 mol/L, keeps 37 DEG C of enzymolysis
Temperature, complex enzyme addition is 2500-3500 U/g, the mass ratio of trypsase and chymotrypsin in the complex enzyme
For 6:1-8:1, the material-water ratio 1:2-1:4, enzymolysis time is 2.5-3.5 h, after enzymolysis terminates, is gone out in 100 DEG C of boiling water baths
Enzyme 10min；By the enzymolysis product in S2 by way of being spray-dried, setting drying condition is 160 DEG C of EAT, and sample introduction is fast
Spend for 50ml/min, centrifugal atomizer rotating speed is 25000r/min, obtains mud blood clam enzymolysis product.
Further, active component B is mud blood clam enzymolysis product, and the preparation method of the mud blood clam enzymolysis product is：
S1. take mud blood clam whole viscera and blend into meat gruel, make material-water ratio be 1:1.5 (w/v), 2.0 are adjusted to 2 mol/L hydrochloric acid by solution ph
So that stomach acid or alkali environment is presented, pepsin is added by 880U/g of enzyme-to-substrate concentration ratio, protected in thermostatic control oscillator vibration
37 DEG C, 85 r/min vibration 1.3 h of enzymolysis are held, the head product of Jing pepsins enzymolysis is obtained with this understanding；S2. with 2
Enzymolysis liquid pH value Jing after pepsin enzymolysis is adjusted to 8.2 by the NaOH of mol/L, keeps 37 DEG C of hydrolysis temperatures, complex enzyme
Addition is 2900 U/g, and trypsase and the mass ratio of chymotrypsin are 7 in the complex enzyme:1, the material-water ratio
1:2.5, enzymolysis time is 2.8 h, and after enzymolysis terminates, go out enzyme 10min in 100 DEG C of boiling water baths；Enzymolysis product in S2 is led to
The mode being spray-dried is crossed, setting drying condition is 160 DEG C of EAT, and sample introduction speed is 50ml/min, and centrifugal atomizer turns
Speed is 25000r/min, obtains mud blood clam enzymolysis product.
Further, active component C be Asiatic plantain thalline crude extract, the preparation side of the Asiatic plantain thalline crude extract
Method is：Root, stem, the leaf for taking fresh plant is cleaned respectively to without obvious impurity, after surface sterilization is processed, is seeded to PDA
（Separate endogenetic fungus）With Gause I culture medium（Separate endogeny rayungus）Upper culture 5-7 d, are then peeled off obtaining bacterium colony；
Picking inoculated by hypha block puts 30 DEG C, 150 r/ in the conical flask equipped with 100 mL fermentation mediums from activated inclined plane
The d of min shaking tables shaken cultivation 3；Fermentation culture medium removes thalline, and filtrate is extracted 3 times with equal-volume ethyl acetate, is merged organic
Phase, 45 DEG C of reduced pressure concentrations are evaporated, as mixed bacteria liquid crude extract；Centrifugation and the thalline being filtrated to get, with appropriate 95% ethanol
Condensing reflux 3 times, merges ethanol phase, and reduced pressure concentration is evaporated, as thalline crude extract.
In above-mentioned technical proposal, the preparation method of described motor function beverage is：By proportioning, each component is mixed, stirred
Finished product is obtained by mixing after being uniformly dispersed.
Raw material of the present invention are easy to buying, make simple；And rich in rich in protein, vitamin and mineral matter；Just
In carrying, conveniently drink, quickly eliminate hunger, mouthfeel is soft, taste is unique；Especially suitable drinking of children.Experience of the present invention
Card, drinks for a long time the effect that can reach strengthen immunity and whitening, based on main active component in the present invention using simulation human body
The mode of gastro-intestinal digestion is obtained, and coordinates the active drive member extracted from plant body, active drive member to enter in enteron aisle,
On the one hand can colonize in enteron aisle, maintain the balance of intestinal microflora；On the other hand it is active drive member and phoenix tail
Fish enzymolysis product and mud blood clam enzymolysis product collective effect induce intestine immunity in the immune system of host, and stimulate thymus gland, spleen
It is dirty to wait immune organ, promote macrophage activity, by strengthening reactivity of B, T lymphocyte to antigenic stimulus, play special
Property immunocompetence, so as to strengthen the immunologic function of body, it is easy to be absorbed by the body, the low age crowd long-term taking of condition 98% is invariably
Suitable symptom.
Specific embodiment
Below in conjunction with specific embodiment, the invention will be further described.
Embodiment 1
A kind of children's drinks, it is in parts by weight, composed of the following components：Active component A 43, potassium chloride 2, xylitol 6,
Sodium citrate 3, white granulated sugar 14, active component B 13, active component C 11, dibastic sodium phosphate 4, Hami melon essence 0.4, glucan
4, water 118.
Further, the active component A be long tail anchovy enzymolysis product, the preparation method of the long tail anchovy enzymolysis product
For：S1. long tail anchovy minced fillet is taken, makes material-water ratio be 1:3 (w/v), solution ph with 2 mol/L hydrochloric acid be adjusted to 2.5 stomach is presented
Portion's acid or alkali environment, by 1300 U/g of enzyme-to-substrate concentration ratio pepsin is added, and 37 are kept in thermostatic control oscillator vibration
DEG C, 110 r/min vibration 2 h of enzymolysis, the head product of Jing pepsins enzymolysis is obtained with this understanding；S2. with the hydrogen of 2 mol/L
Enzymolysis liquid pH value Jing after pepsin enzymolysis is adjusted to 8.5 by sodium oxide molybdena, keeps the 37 DEG C of hydrolysis temperatures, complex enzyme addition to be
5200 U/g, trypsase and the mass ratio of chymotrypsin are 5 in the complex enzyme:1, the material-water ratio 1:4.5, enzyme
The solution time is 3.5 h, and after enzymolysis terminates, go out enzyme 10min in 100 DEG C of boiling water baths；Enzymolysis product in S2 is cold by vacuum
Lyophilized dry, setting drying condition is -30 DEG C of precooling temperature, and pre-coo time 2h, heating-up temperature starts slowly to be risen after 18h from -10 DEG C
Temperature, reaches 25 DEG C, and remains to and be dried terminal, obtains long tail anchovy enzymolysis product.
Further, active component B is mud blood clam enzymolysis product, and the preparation method of the mud blood clam enzymolysis product is：S1.
Take mud blood clam whole viscera and blend into meat gruel, make material-water ratio be 1:1.5 (w/v), with 2 mol/L hydrochloric acid by solution ph be adjusted to 2.0 with
Stomach acid or alkali environment is presented, by 880U/g of enzyme-to-substrate concentration ratio pepsin is added, kept in thermostatic control oscillator vibration
37 DEG C, 85 r/min vibration 1.3 h of enzymolysis, obtain with this understanding the head product of Jing pepsins enzymolysis；S2. 2 mol/L are used
NaOH the enzymolysis liquid pH value Jing after pepsin enzymolysis is adjusted into 8.2, keep 37 DEG C of hydrolysis temperatures, complex enzyme addition
Measure as 2900 U/g, trypsase and the mass ratio of chymotrypsin are 7 in the complex enzyme:1, the material-water ratio 1:
2.5, enzymolysis time is 2.8 h, and after enzymolysis terminates, go out enzyme 10min in 100 DEG C of boiling water baths；Enzymolysis product in S2 is passed through
The mode of spray drying, setting drying condition be 160 DEG C of EAT, sample introduction speed be 50ml/min, centrifugal atomizer rotating speed
For 25000r/min, mud blood clam enzymolysis product is obtained.
Further, active component C be Asiatic plantain thalline crude extract, the preparation side of the Asiatic plantain thalline crude extract
Method is：Root, stem, the leaf for taking fresh plant is cleaned respectively to without obvious impurity, after surface sterilization is processed, is seeded to PDA
（Separate endogenetic fungus）With Gause I culture medium（Separate endogeny rayungus）Upper culture 5-7 d, are then peeled off obtaining bacterium colony；
Picking inoculated by hypha block puts 30 DEG C, 150 r/ in the conical flask equipped with 100 mL fermentation mediums from activated inclined plane
The d of min shaking tables shaken cultivation 3；Fermentation culture medium removes thalline, and filtrate is extracted 3 times with equal-volume ethyl acetate, is merged organic
Phase, 45 DEG C of reduced pressure concentrations are evaporated, as mixed bacteria liquid crude extract；Centrifugation and the thalline being filtrated to get, with appropriate 95% ethanol
Condensing reflux 3 times, merges ethanol phase, and reduced pressure concentration is evaporated, as thalline crude extract.
Embodiment 2
The present embodiment provides a kind of children's drinks, in parts by weight, composed of the following components：Active component A 40, chlorination
Potassium 2, xylitol 6, sodium citrate 3, white granulated sugar 14, active component B 11, active component C 9, dibastic sodium phosphate 4, Hami melon essence
0.4, bextran 45, water 118.In the present embodiment active component A, active component B, the composition of active component C and preparation method with
Embodiment 1 is consistent.
Embodiment 3
The present embodiment provides a kind of children's drinks, in parts by weight, composed of the following components：Active component A 50, chlorination
Potassium 2, xylitol 6, sodium citrate 3, white granulated sugar 14, active component B 15, active component C 13, dibastic sodium phosphate 4, "Hami" melon is fragrant
Essence 0.4, bextran 45, water 118.Active component A, active component B, the composition of active component C and preparation method in the present embodiment
It is consistent with embodiment 1.
Embodiment 4
The present embodiment provides a kind of children's drinks, in parts by weight, composed of the following components：Active component A 50, chlorination
Potassium 2, xylitol 6, sodium citrate 3, white granulated sugar 14, active component B 15, active component C 9, dibastic sodium phosphate 4, Hami melon essence
0.4, bextran 45, water 118.
Active component A, active component B, the composition of active component C and preparation method are consistent with embodiment 1 in the present embodiment.
Embodiment 5
The present embodiment provides a kind of children's drinks, in parts by weight, composed of the following components：Active component A 40, chlorination
Potassium 2, xylitol 6, sodium citrate 3, white granulated sugar 14, active component B 11, active component C 13, dibastic sodium phosphate 4, "Hami" melon is fragrant
Essence 0.4, bextran 45, water 118.
Active component A, active component B, the composition of active component C and preparation method are consistent with embodiment 1 in the present embodiment.
Effect example
1. strengthen immunity effect experimental
To be dried by drinks thickening obtained in embodiment 1, prepare it is powdered, as test sample.
Reference substance：Its vigorous and graceful board Soybean Peptide albumen powder；Source：Beijing Tongrentang Health Pharmaceutical Co., Ltd.；Adult
Clinic recommends consumption：0.167g.kg-1.d-1。
Animal subject strain and rank：BALB/c mouse, Hartley cavys, SPF levels；Quantity and sex：BALB/c is little
Mouse 60, male；Hartley cavys 6,.Buy body weight BALB/c mouse 17-19g, Hartley cavy 250-
350g.There is provided by Guangdong Medical Lab Animal Center, animal used as test uses credit number：SYXK（Guangdong）2013-0002.Experiment
Period, mouse ad lib drinking-water observes daily mouse state.
Dose design is carried out according to the basic condition of sample, sample is basic, normal, high 5 times by adult's quantity, 10 times,
20 times of design dosage, respectively 0.15g.kg-1.d-1、 0.30g.kg-1.d-1 、0.60g.kg-1.d-1；Positive controls dosage
It is 0.835g.kg by 5 times of designs of adult's recommended dose-1.d-1.60 male mices are respectively divided into at random 5 groups（Negative control
The basic, normal, high dosage group of group, positive controls, given the test agent）, 12 per group, wherein 2 are only used as standby animal.Each group mouse is every
It presses 0.1mL/10g body weight gavage respective sample liquid, and negative control group gives equivalent pure water, once a day, successive administration 30
My god.On-test, off-test are weighed in 1 time, are weighed in weekly during test 1 time.
Mouse lymphocyte transformation experiment
Last dose 1h, the aseptic spleen that takes is placed in the culture dish for filling RPMI-1640 nutrient solutions, and places 200 eye mesh screens, with note
Emitter piston grinds above it makes splenocyte suspension, dispenses splenocyte suspension to the centrifuge tube added with lymphocyte separation medium
In.400g, is centrifuged 20min at 20 DEG C, centrifugation is finished, and takes out centrifuge tube medium size lymphocyte layer centrifuge tube, adds RPMI-1640 trainings
Nutrient solution is washed for several times, and 250g is centrifuged 10min at 4 DEG C.After washing is finished, abandoning supernatant adds complete medium, and piping and druming is equal
It is even, expect that blue dyeing counting ensures more than 95% living cell rate with platform, cell concentration is adjusted into 3 × 106Individual/ml.
Every part of splenocyte suspension point holes is added in 48 well culture plates, per hole 0.5ml, a hole adds 25 ul ConA liquid
（Equivalent to 5ug/ml）, another hole is placed in 5%CO as control2,37℃CO2Cultivate in incubator after 48h and gently suck supernatant per hole
The ml of liquid 0.3, adds 0.3 mlRPMI-1640 nutrient solutions, while adding the ul/ holes of 8 reagents of CCK 50, continues to cultivate 2 h.Knot
After beam culture, piping and druming is uniform, in being then dispensed into 96 well culture plates, 3 parallel laboratory tests is made per hole, and with ELIASA 450 nm ripples are surveyed
OD value under long.The multiplication capacity of lymphocyte is by the OD value and the OD value for being not added with ConA holes for adding ConA holes
Difference represent.
Mouse humoral immune function（Serum hemolysin）Affect
Administration 25 days, the sheep blood 3 times of fiber has been taken off with brine (2000r/min, 10min), in hematocrit SRBC plus
Enter physiological saline and be made into 2%（v/v）Cell suspension, every mouse peritoneal injection 0.2ml, when the 30th day, administration 1h after, weigh,
All mouse are extractd eyeball and take blood 0.8ml in centrifuge tube, stand 1h, and 2000r/min, 10min is centrifuged, and collect serum.Cervical vertebra takes off
Mortar puts to death mouse.The serum for diluting 300 times with SA buffer solutions takes 1ml and puts in test tube, and 10% is added again（v/v）
SRBC0.5ml, the complement 1ml that 9 times are diluted with SA buffer solutions, arrange control tube（Replace serum with SA）, 37 DEG C of waters bath with thermostatic control
20min, then ice bath, are then centrifuged for 2000r/min, 10min.Supernatant 1ml is taken in new test tube, plus 3.75ml Dou Shi reagents,
0.25ml10%（v/v）SRBC, fully mixes, and stands 10min, with control tube as blank, determines each Guan Guangmi at 540nm respectively
Angle value, the amount of hemolysin is with half hemolytic value（HC50）Represent.
Mouse humoral immune（Antibody-producting cell）Impact
Administration 25 days, the sheep blood 3 times of fiber has been taken off with brine, 2000r/min, 10min is centrifuged every time, in hematocrit
Physiological saline is added to be made into 2% in SRBC（v/v）Cell suspension, every mouse peritoneal injection 0.2ml, when the 30th day, be administered 1h
Afterwards, weigh.Cervical dislocation is put to death after mouse, takes out spleen, is placed in and is placed with Hank ' s liquid plates, through 200 mesh sieve net filtrations
Afterwards, then with Hank ' s liquid wash twice, be added in 5mL RPMI-1640 nutrient solutions and cell suspends, count cell, and adjust
Cell concentration is 5 × 106Individual/mL.
After the top layer culture medium heating for dissolving for adding distilled water to be formed to 100mL 1g agaroses, 40-45 DEG C of water bath heat preservation.
Mix with PH7.2-7.4,2 times of Hank ' s liquid equal-volumes, shake up, often pipe 0.5mL dispenses different test tubes, continue to be added to every pipe
With SA buffers into 10%SRBC (v/v）50ul and splenocyte suspension 20ul, it is quick to mix, it is poured into and scribbles one layer
On the thin slice of very thin agarose, parallel plate is done, after agarose solidification, wafer level button is placed on horse, be put into two
After carbonoxide incubator culture 1-1.5h, the complement that 9 times are diluted with SA buffer solutions is added in the groove of glass frame, continues to train
Foster 1-1.5h, then counts hemolysis plaque number.
The measure that mouse monokaryon-macrophage phagocytic function and immune organ affect
Last dose 1h, by body weight from tail vein injection physiological saline 1:The india ink of 3 dilutions（10mL/kg）, treat prepared Chinese ink
Injection, immediately timing.2,10min after injection prepared Chinese ink, respectively from orbital venous plexus 20 μ L of blood sampling, and is added to immediately 2mL 0.1%
Na2CO3In solution, with Na2CO3Solution makees blank, and OD value is determined at 600nm wavelength with spectrophotometer（OD）,
Calculate phagocytic index a.
Dislocation after mouse blood sampling is put to death, and solution takes liver, spleen and thymus gland, and filter paper is blotted after internal organs peripheral blood, accurate
Weigh, calculate internal organs/body weight ratio.
NK cytoactive detections
Last dose 1h, aseptic to take spleen, the aseptic spleen that takes is placed in the culture dish for filling RPMI-1640 nutrient solutions, and places 200 mesh
Screen cloth, is ground above it with syringe piston and makes splenocyte suspension, and packing splenocyte suspension is to added with separation of lymphocytes
In the centrifuge tube of liquid.400g, is centrifuged 20min at 20 DEG C, centrifugation is finished, and takes out centrifuge tube medium size lymphocyte layer centrifuge tube, adds
RPMI-1640 nutrient solutions are washed for several times, and 250g is centrifuged 10min at 4 DEG C.After washing is finished, abandoning supernatant, addition is trained completely
Foster base, piping and druming is uniform, and with platform blue dyeing counting viable count is expected（Should be more than 95%）, finally with the complete culture solutions of RPMI 1640
Adjustment cell concentration is 2 × 107Individual/mL.
In proportion by effector cell and target cell（50:1）Each 100ul adds U-shaped 96 orifice plate；Target cell and nutrient solution are each
100ul adds target cell Spontaneous release hole；Target cell and each 100ul of 0.25%Triton add target cell maximum release aperture.Per group
3 parallel holes are all provided with, 37 DEG C, after 5%CO2 incubator culture 4h, 1500rpm, 5min are centrifuged.Per hole Aspirate supernatant 100ul in
Flat 96 orifice plate, adds LDH matrix liquids 100ul/ hole, reacts 3-10min, adds 1mol/ml HCL 30ul/ holes, enzyme mark
490nm determines OD value.
All data carry out statistical analysis using the softwares of SPSS 16.0, and test result is as shown in the table.
The impact that the embodiment sample of table 1 is converted with control group to mouse lymphocyte
Note：Statistical analysis is carried out using variance analysis method, is compared with negative control group, " a ": p<0.05;With positive controls
Relatively, " b ": p<0.01
The embodiment sample of table 2 is with control group to mouse antibodies level（Serum hemolysin）Impact
Note：Statistical analysis is carried out using variance analysis method, is compared with negative control group, " a ": p<0.05;With positive controls
Relatively, " b ": p<The embodiment sample of 0.01 table 3 and impact of the control group to mouse hemolysis plaque number
Note：Statistical analysis is carried out using variance analysis method, is compared with negative control group, " a ": p<0.05;With positive controls
Relatively, " b ": p<0.01 .
The embodiment sample of table 4 and impact of the control group to mouse phagocytic index
Note：Statistical analysis is carried out using variance analysis method, is compared with negative control group, " a ": p<0.05;With positive controls
Relatively, " b ": p<0.01 .
Impact of the embodiment sample of table 6 with control group to NK cells in mice activity
Note：Statistical analysis is carried out using variance analysis method, is compared with negative control group, " a ": p<0.05;With positive controls
Relatively, " b ": p<0.01 .
Equally, identical experiment is carried out to remaining embodiment and embodiments of the invention remaining proportionings, as a result with embodiment 1
Effect is similar to.
2. antifatigue compliance test result
To be dried by drinks thickening obtained in embodiment 1, prepare it is powdered, as test sample.By physiology
Salt solution is blank control group, with branched-amino aqueous acid as negative control group.
Experimental subjects：From SD rats, provided by Beijing Experimental Animal Center, male and female half and half, 180,220 ± 18g,
At about 3 monthly ages, after laboratory rearing one month, through preliminary swimming instruction blank control group, negative control group are randomly divided into
And sample sets.
Test method
（1) exhausting property of power test：Sample powder is taken, with distillation water dissolves, the concentration of sample is respectively 0.15g.kg-1.d-1、 0.30g.kg-1.d-1 、0.60g.kg-1.d-1 , sample sets press daily 0.1mL/10g body weight gavages；Negative control group is
Branched-chain amino acid（Leucine: isoleucine: valine in mass ratio 2：1:Obtain after 1 mixing）The aqueous solution, be made into dense
0.60g.kg-1.d-1, press 0.1mL/10g body weight gavages daily；Each group last is fed after 30min, carries out rat heavy burden
（5% body weight）Swimming test to power exhausts（Power exhausts standard：Heavy burden rats'swimming action is substantially lacked of proper care, it is impossible to is adhered to again or is sunk to
The bottom is unable to backwater surface and exhausts for power more than 3s).
（2) serum urea nitrogen test：Sample powder is taken, with distillation water dissolves, the concentration of sample is respectively
0.15g.kg-1.d-1、 0.30g.kg-1.d-1 、0.60g.kg-1.d-1 , sample sets press daily 0.1mL/10g body weight gavages；It is negative
Control group is branched-chain amino acid（Leucine: isoleucine: valine in mass ratio 2：1:Obtain after 1 mixing）The aqueous solution, match somebody with somebody
Into dense 0.60g.kg-1.d-1, press 0.1mL/10g body weight gavages daily；Each group is continuously fed after 30d, carries out rats'swimming examination
Test（Without heavy burden）, after 90min, eyeball blood sampling is pulled out, use determination of urea nitrogen kit（Diacetyl hydrazine monoxime method, Beijing Chemical Plant's life
Produce）Detection serum urea nitrogen.
Result of the test
Embodiment sample is shown in Table 7 with impact of the control group to rat walking weight load.
Impact contrast of the embodiment sample of table 7 with control group to rat walking weight load
Note：Statistical analysis is carried out using variance analysis method, is compared with negative control group, " a ": p<0.05;With positive controls
Relatively, " b ": p<0.01 .
The embodiment sample of table 8 and impact of the control group to serum urea nitrogen content after rat motor
Note：Statistical analysis is carried out using variance analysis method, is compared with negative control group, " a ": p<0.05;With positive controls
Relatively, " b ": p<0.01 .
It is obvious to a person skilled in the art that the invention is not restricted to the details of above-mentioned one exemplary embodiment, Er Qie
In the case of spirit or essential attributes without departing substantially from the present invention, the present invention can be in other specific forms realized.Therefore, no matter
From the point of view of which point, embodiment all should be regarded as exemplary, and be nonrestrictive, the scope of the present invention is by appended power
Profit is required rather than described above is limited, it is intended that all in the implication and scope of the equivalency of claim by falling
Change is included in the present invention.
Moreover, it will be appreciated that although this specification is been described by according to embodiment, not each embodiment is only wrapped
Containing an independent technical scheme, this narrating mode of specification is only that for clarity those skilled in the art should
Using specification as an entirety, the technical scheme in each embodiment can also Jing it is appropriately combined, form those skilled in the art
Understandable other embodiment.The ins and outs of all not detailed descriptions in the present invention, can be appointed by this area
One prior art is realized.
Claims (7)
1. a kind of children's drinks, it is characterised in that in parts by weight, composed of the following components：Active component A 40-50,
Potassium chloride 1-3, xylitol 5-8, sodium citrate 2-4, white granulated sugar 10-30, active component B 11-15, active component C 9-13,
Dibastic sodium phosphate 2-6, Hami melon essence 0.3-0.5, glucan 3-5, water 105-135.
2. a kind of children's drinks according to claim 1, it is characterised in that in parts by weight, by following components group
Into：Active component A 43, potassium chloride 2, xylitol 6, sodium citrate 3, white granulated sugar 14, active component B 13, active component C
11, dibastic sodium phosphate 4, Hami melon essence 0.4, bextran 45, water 118.
3. a kind of children's drinks according to claim 1, it is characterised in that the active component A is long tail anchovy
Enzymolysis product, the preparation method of the long tail anchovy enzymolysis product is：S1. long tail anchovy minced fillet is taken, makes material-water ratio be 1:2-1:8(w/
V), solution ph is adjusted into 2.0-3.5 so that stomach acid or alkali environment is presented with 2 mol/L hydrochloric acid, is with enzyme-to-substrate concentration ratio
1200-1500U/g adds pepsin, and 37 DEG C, 100-130 r/min vibration enzymolysis are kept in thermostatic control oscillator vibration
1.5-2.5 h, obtain with this understanding the head product of Jing pepsins enzymolysis；S2. with the NaOH of 2 mol/L by Jing stomach eggs
Enzymolysis liquid pH value after white enzyme enzymolysis is adjusted to 8.0-9.0, keeps 37 DEG C of hydrolysis temperatures, and complex enzyme addition is 4500-6000
U/g, trypsase and the mass ratio of chymotrypsin are 4 in the complex enzyme:1-6:1, the material-water ratio 1:3-1:6, enzyme
The solution time is 3-4.5 h, and after enzymolysis terminates, go out enzyme 10min in 100 DEG C of boiling water baths；Enzymolysis product in S2 is passed through into vacuum
Freeze-drying, setting drying condition is -30 DEG C of precooling temperature, and pre-coo time 2h, heating-up temperature starts slow after 18h from -10 DEG C
Heat up, reach 25 DEG C, and remain to and be dried terminal, obtain long tail anchovy enzymolysis product.
4. a kind of children's drinks according to claim 3, it is characterised in that the active component A is long tail anchovy enzyme
Product is solved, the preparation method of the long tail anchovy enzymolysis product is：S1. long tail anchovy minced fillet is taken, makes material-water ratio be 1:3 (w/v), use 2
Solution ph is adjusted to 2.5 so that stomach acid or alkali environment is presented by mol/L hydrochloric acid, and by 1300 U/g of enzyme-to-substrate concentration ratio stomach is added
Protease, keeps 37 DEG C, 110 r/min vibration 2 h of enzymolysis in thermostatic control oscillator vibration, and Jing stomach eggs are obtained with this understanding
The head product of white enzyme enzymolysis；S2. the enzymolysis liquid pH value Jing after pepsin enzymolysis is adjusted into 8.5 with the NaOH of 2 mol/L,
Keep 37 DEG C of hydrolysis temperatures, complex enzyme addition be 5200 U/g, trypsase and chymotrypsin in the complex enzyme
Mass ratio be 5:1, the material-water ratio 1:4.5, enzymolysis time is 3.5 h, and after enzymolysis terminates, go out enzyme in 100 DEG C of boiling water baths
10min；By the enzymolysis product in S2 by vacuum freeze drying, settings drying condition is -30 DEG C of precooling temperature, pre-coo time
2h, heating-up temperature starts slowly to be heated up after 18h from -10 DEG C, reaches 25 DEG C, and remains to and be dried terminal, obtains long tail anchovy enzymolysis
Product.
5. a kind of children's drinks according to claim 1, it is characterised in that active component B is mud blood clam enzymolysis
Product, the preparation method of the mud blood clam enzymolysis product is：S1. take mud blood clam whole viscera and blend into meat gruel, make material-water ratio be 1:1-1:2
(w/v), solution ph is adjusted into 2.0 so that stomach acid or alkali environment is presented with 2 mol/L hydrochloric acid, with enzyme-to-substrate concentration ratio as 800-
1000U/g adds pepsin, and 37 DEG C, 75-105 r/min vibration enzymolysis 1-1.5 h are kept in thermostatic control oscillator vibration,
The head product of Jing pepsins enzymolysis is obtained with this understanding；S2. with the NaOH of 2 mol/L by Jing after pepsin enzymolysis
Enzymolysis liquid pH value be adjusted to 8.0-9.0, keep 37 DEG C of hydrolysis temperatures, complex enzyme addition is 2500-3500 U/g, described multiple
Trypsase and the mass ratio of chymotrypsin are 6 in synthase:1-8:1, the material-water ratio 1:2-1:4, enzymolysis time is
2.5-3.5 h, after enzymolysis terminates, go out enzyme 10min in 100 DEG C of boiling water baths；By the enzymolysis product in S2 by spray drying
Mode, setting drying condition is 160 DEG C of EAT, and sample introduction speed is 50ml/min, and centrifugal atomizer rotating speed is 25000r/
Min, obtains mud blood clam enzymolysis product.
6. a kind of children's drinks according to claim 5, it is characterised in that active component B is mud blood clam enzymolysis
Product, the preparation method of the mud blood clam enzymolysis product is：S1. take mud blood clam whole viscera and blend into meat gruel, make material-water ratio be 1:1.5
(w/v), solution ph is adjusted into 2.0 so that stomach acid or alkali environment is presented with 2 mol/L hydrochloric acid, is with enzyme-to-substrate concentration ratio
880U/g adds pepsin, 37 DEG C, 85 r/min vibration 1.3 h of enzymolysis is kept in thermostatic control oscillator vibration, in this condition
The head product of lower acquisition Jing pepsins enzymolysis；S2. with the NaOH of 2 mol/L by the enzymolysis liquid Jing after pepsin enzymolysis
PH value is adjusted to 8.2, keeps 37 DEG C of hydrolysis temperatures, and complex enzyme addition is 2900 U/g, in the complex enzyme trypsase with
The mass ratio of chymotrypsin is 7:1, the material-water ratio 1:2.5, enzymolysis time is 2.8 h, after enzymolysis terminates, at 100 DEG C
Go out enzyme 10min in boiling water bath；By the enzymolysis product in S2 by way of being spray-dried, setting drying condition is EAT
160 DEG C, sample introduction speed is 50ml/min, and centrifugal atomizer rotating speed is 25000r/min, obtains mud blood clam enzymolysis product.
7. a kind of children's drinks according to claim 1, it is characterised in that active component C is Asiatic plantain bacterium
Body crude extract, the preparation method of the Asiatic plantain thalline crude extract is：Root, stem, the leaf for taking fresh plant is cleaned respectively to not having
Significantly impurity, after surface sterilization is processed, is seeded to PDA（Separate endogenetic fungus）With Gause I culture medium（Life in separating is put
Line bacterium）Upper culture 5-7 d, are then peeled off obtaining bacterium colony；Picking inoculated by hypha block is in equipped with 100 mL from activated inclined plane
In the conical flask of ferment culture medium, 30 DEG C are put, the d of 150 r/min shaking tables shaken cultivation 3；Fermentation culture medium removes thalline, filtrate
Extracted 3 times with equal-volume ethyl acetate, merge organic phase, 45 DEG C of reduced pressure concentrations are evaporated, as mixed bacteria liquid crude extract；From
The heart and the thalline being filtrated to get, with appropriate 95% ethanol condensing reflux 3 times, merge ethanol phase, and reduced pressure concentration is evaporated, as bacterium
Body crude extract.
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| CN107668449A (en) * | 2017-10-26 | 2018-02-09 | 邓志程 | A kind of health drink |
| CN107691922A (en) * | 2017-10-26 | 2018-02-16 | 邓志程 | A kind of functional drink |
| CN107751693A (en) * | 2017-10-26 | 2018-03-06 | 邓志程 | A kind of health beverages |
| CN107889988A (en) * | 2017-10-26 | 2018-04-10 | 邓志程 | A kind of functional beverage |
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