CN108967699A - Laver amylose Zn complex and its preparation method and application - Google Patents
Laver amylose Zn complex and its preparation method and application Download PDFInfo
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- CN108967699A CN108967699A CN201811051775.6A CN201811051775A CN108967699A CN 108967699 A CN108967699 A CN 108967699A CN 201811051775 A CN201811051775 A CN 201811051775A CN 108967699 A CN108967699 A CN 108967699A
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/163—Sugars; Polysaccharides
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/20—Inorganic substances, e.g. oligoelements
- A23K20/30—Oligoelements
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/30—Feeding-stuffs specially adapted for particular animals for swines
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/60—Feeding-stuffs specially adapted for particular animals for weanlings
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Abstract
The present invention provides laver amylose Zn complexes and its preparation method and application.Laver amylose Zn complex of the present invention, which is reacted by laver amylose with zinc salt, to be prepared.Laver amylose Zn complex of the present invention can not only improve the immunity and anti-oxidation stress of weanling pig, even more have effects that improve Intestinal Microflora of Weanling Piglets richness, be that a kind of good nutritional supplementation and function improve additive.Simultaneously, preparation method of the present invention is easy to operate, and raw material seaweed abundance, laver amylose is prepared using seaweed as raw material and laver amylose Zn complex is further obtained by reaction, it is able to solve absorption problem present in traditional zinc additive, in the production of current strict control pig in the case where feed Zinc supplementation, laver amylose Zn complex can be used as a kind of novel, environmentally friendly, healthy zinc additive substitute.
Description
Technical field
The present invention relates to feed additive field, in particular to laver amylose Zn complex and preparation method thereof and
Using.
Background technique
Zinc is of great significance to the healthy growth of animal body and a kind of essential microelement,
All contain zinc in almost all of animal tissue's organ.Meanwhile also containing zinc at least 300 kinds of enzymes in animal body, these zinc
Different biological functions is carry, plays important work in various aspects such as the growth and development of animal, immune, metabolism and breedings
With.
Seaweed is the common aquatic products of coastal area of china, abundance.Numerous studies find laver amylose (Porphyra
Polysaccharide, PP) there is the various biologicals such as anticoagulation, reducing blood lipid, anti-mutation, anti-oxidant and anti-radiation activity.
In order to improve the growth performance of animal, mostly fed with the feed added with zinc preparation.However, animal is raised at present
What is added in material is mostly the inorganic zincs such as zinc oxide, zinc sulfate, and the zinc of high concentration can generate toxic side effect for body, while inorganic
The absorption rate of zinc is poor, and 90% zinc excretes in excrement, pollutes environment, and country strictly limits in pannage at present
The additive amount of zinc.
Therefore, efficient, nontoxic, the free of contamination zinc additive product of one kind is found out for large scale of pig farm with particularly significant
Meaning.
In view of this, the present invention is specifically proposed.
Summary of the invention
The first object of the present invention is to provide a kind of laver amylose Zn complex, and laver amylose Zn complex of the present invention exists
Animal body internal absorption factor is high, discharge is few, nonhazardous effect, and will not generate heavy metal pollution.
The second object of the present invention is to provide the preparation method of laver amylose Zn complex described in one kind.
The third object of the present invention is to provide the application of the cooperation of laver amylose described in one kind.
In order to realize above-mentioned purpose of the invention, the following technical scheme is adopted:
A kind of laver amylose Zn complex, the laver amylose Zn complex, which is reacted by laver amylose with zinc salt, to be prepared into
It arrives.
Meanwhile the present invention also provides the preparation methods of the laver amylose Zn complex, comprising: to more dissolved with seaweed
Lye is added in the solution of sugar and zinc salt with regulation system pH, by reaction solution alcohol precipitation and collects sediment after heating reaction；Precipitating
Object is concentrated and dried after purification to obtain the final product.
Preferably, in preparation method of the present invention, the zinc salt is zinc sulfate, zinc nitrate, or corresponding hydrate
At least one of.
Preferably, in preparation method of the present invention, the mass ratio of laver amylose and zinc salt is 1:3~3:1；More preferably
, the mass ratio of laver amylose and zinc salt is 1:2~2:1.
Preferably, in preparation method of the present invention, the lye includes: sodium hydroxide/potassium hydroxide solution, and
Ammonium hydroxide；
And/or the regulation system pH is the pH to 6.5~7.5 of regulation system；It is furthermore preferred that the regulation system pH
For the pH to 7 of regulation system.
Preferably, in preparation method of the present invention, the purifying includes: after washing gained sediment with solvent,
Carry out dialysis treatment.
Meanwhile the present invention also provides the laver amylose Zn complexes to increase weanling pig intestinal flora abundant
Application in degree；And/or laver amylose Zn complex is increasing the application in weanling pig enteron aisle in the quantity of beneficial bacterium；With/
Or, laver amylose Zn complex alleviate because of immunological stress caused by Intestinal Microflora of Weanling Piglets it is unbalance in application.
Likewise, present invention provides the laver amylose Zn complexes to enhance weanling pig immunity of organisms,
Enhance the application in resisting stress capability.
Further, the application the present invention also provides the laver amylose Zn complex in feed addictive；It is excellent
Choosing, the feed addictive is feed additive for weaning piglet.
Meanwhile present invention provides the feeds comprising laver amylose Zn complex of the present invention.
Compared with prior art, the invention has the benefit that
Preparation method of the present invention is easy to operate, and raw material seaweed abundance, prepares laver amylose, simultaneously by raw material of seaweed
Laver amylose Zn complex is further obtained by reaction, absorption problem present in traditional zinc additive is able to solve, in mesh
In preceding strict control pig production in the case where feed Zinc supplementation, laver amylose Zn complex can be used as a kind of novel, environmentally friendly, strong
The zinc additive substitute of health.
Meanwhile laver amylose Zn complex of the present invention can not only improve the immunity and anti-oxidation stress of weanling pig
Property, even more have effects that improve Intestinal Microflora of Weanling Piglets richness, is that a kind of good nutritional supplementation and function improvement add
Add agent.
Detailed description of the invention
In order to more clearly explain the embodiment of the invention or the technical proposal in the existing technology, to embodiment or will show below
There is attached drawing needed in technical description to be briefly described.
Fig. 1 is that laver amylose Zn complex influences testing result figure to microorganism richness in intestine of young pigs.
Specific embodiment
Embodiment of the present invention is described in detail below in conjunction with embodiment, but those skilled in the art will
Understand, the following example is merely to illustrate the present invention, and is not construed as limiting the scope of the invention.It is not specified in embodiment specific
Condition person carries out according to conventional conditions or manufacturer's recommended conditions.Reagents or instruments used without specified manufacturer is
The conventional products that can be obtained by commercially available purchase.
Laver amylose Zn complex provided by the present invention is a kind of novel fodder zinc additive, is able to solve inorganic
Zinc additive is the problems of in terms of absorbing and animal.
Meanwhile laver amylose complex of the present invention can also play the blood antioxidant ability for improving weanling pig body,
Alleviate weanling pig because of the effect of lipopolysaccharides (LPS) immune produced immunological stress；But also there is the improvement micro- life of intestine of young pigs
Object richness increases the quantity of beneficial bacterium in enteron aisle, intestine of young pigs flora caused by alleviating because of immunological stress (LPS immunological stress)
Unbalance effect.
Specifically, laver amylose complex of the present invention can be prepared by the following method:
(a) respectively by laver amylose and zinc salt (zinc sulfate, hydrate of zinc sulfata, zinc nitrate, one in zinc nitrate hydrate
Kind is a variety of) stirring and dissolving in distilled water is added；
Wherein, mass ratio 1:3~3:1 of laver amylose and zinc salt, preferably 1:2~2:1, more preferably 2:1.
(b) lye is added, into acquired solution with the pH to 6.5~7.5 (preferably 7) of regulation system；
Wherein, lye used is NaOH/KOH solution and ammonium hydroxide；Preferably NaOH solution and ammonium hydroxide；
Specifically, can first with concentration for the NaOH solution regulation system of 0.5mol/L pH to close to neutrality, then with
Concentration is the pH to 7 of the ammonia water conditioning system of 1% (m/m).
(c) after adjusting pH, reaction system reacts 60min under 70 DEG C of heating conditions, after reaction, naturally cools to room
Temperature；
Then, it is overnight that ethyl alcohol (concentration is greater than 95%, preferably dehydrated alcohol) alcohol precipitation is added, filters/be collected by centrifugation precipitating
Object.
(d) gained sediment is washed with organic solvent, preferably respectively with dehydrated alcohol, 95% ethyl alcohol, acetone,
Ether is washed, and can specifically be washed using elution or by the way of impregnating；
After washing, by the way of filtering or centrifugation, precipitating is collected.
(e) precipitating after washing is redissolved with distilled water, is then purified again by way of dialysis, by dialysing
After obtaining product concentrate drying, product laver amylose Zn complex is obtained.
The obtained laver amylose Zn complex of method as above can be further used as feed addictive use, especially make
It is used for the zinc additive of feed for weaned piglet, so that playing improves piglet immunological function, improves piglet anti-oxidation stress ability,
And the effect of improving intestinal flora richness.
Embodiment 1
Weigh 2g laver amylose and 1g ZnSO4·7H2O is dissolved in 10ml distilled water, is filled in 70 DEG C of stirred in water bath
Point, 3000r/min is centrifuged 10min, and slow Aspirate supernatant removes the impurity in solution.Prepare the NaOH solution of 0.5mol/L
And mass concentration is 1% ammonium hydroxide, is sequentially added in reaction system, under the conditions of pH meter accurate measurements, by pH value of reaction system
It is adjusted to 7.
Reaction system reacts 60min under 70 DEG C of water bath conditions.After reaction, cooled to room temperature, with anhydrous second
Alcohol alcohol precipitation is stayed overnight, and precipitating is collected by centrifugation, and is impregnated precipitating with dehydrated alcohol, 95% ethyl alcohol, acetone, ether respectively, and is used glass bar
Stirring, which is swung, washes sediment, and precipitating is collected by centrifugation.
Bag filter processing: bag filter is put into 2%NaHCO3It is boiled in EDTA (pH=8) mixed liquor of 1mmol/L
Then 10min is cleaned with distilled water, is placed into and boil 10min in distilled water, cooling to be placed in spare (the bag filter processing of 4 DEG C of refrigerators
It needs to wear gloves, guarantees that bag filter is immersed in after processing and save in liquid).
Resulting laver amylose Zn complex crude product will be centrifuged sufficiently to be redissolved with distilled water, be fitted into bag filter and clamp, magnetic
For 24 hours, sample is concentrated through rotary evaporation after dialysis, is freeze-dried 48h for power stirring dialysis, obtains brown white laver amylose zinc cooperation
Object, yield 55.45%
Embodiment 2
Weigh 3g laver amylose and 1g ZnSO4·7H2O is dissolved in 10ml distilled water, is filled in 70 DEG C of stirred in water bath
Point, 3000r/min is centrifuged 10min.The NaOH solution and mass concentration for preparing 0.5mol/L are 1% ammonium hydroxide, are sequentially added anti-
It answers in system, under the conditions of pH meter accurate measurements, pH value of reaction system is adjusted to 7.
Then, referring to the method for embodiment 1, the laver amylose Zn complex of embodiment 2, yield are as follows: 52.46 are obtained.
Embodiment 3
Weigh 2g laver amylose and 1g ZnSO4·7H2O is dissolved in 10ml distilled water, is filled in 70 DEG C of stirred in water bath
Point, 3000r/min is centrifuged 10min.The NaOH solution and mass concentration for preparing 0.5mol/L are 1% ammonium hydroxide, are sequentially added anti-
It answers in system, under the conditions of pH meter accurate measurements, pH value of reaction system is adjusted to 6.5.
Then, referring to the method for embodiment 1, the laver amylose Zn complex of embodiment 3, yield are as follows: 51.73% are obtained.
Embodiment 4
Weigh 2g laver amylose and 1g Zn (NO3)2·6H2O is dissolved in 10ml distilled water, is stirred in 70 DEG C of water-baths
It mixes sufficiently, 3000r/min is centrifuged 10min.Prepare 0.5mol/L NaOH solution and mass concentration be 1% ammonium hydroxide, successively plus
Enter in reaction system, under the conditions of pH meter accurate measurements, pH value of reaction system is adjusted to 7.5.
Then, referring to the method for embodiment 1, the laver amylose Zn complex of embodiment 4, yield are as follows: 48.65% are obtained.
Experimental example 1
Raw material laver amylose and embodiment 1-4 each 50mg of laver amylose Zn complex are accurately weighed, according to GBT
13885-2003 handles sample and configuration standard curve, and sample sample detection is said according to the operation of acetylene torch atomic absorption instrument
It is bright, the zinc content in sample is obtained, the Zn content percentage in sample is calculated.
Experimental result shows that zinc atom content obviously increases in product after reaction, illustrates that laver amylose is chelated with zinc
Reaction, i.e. generation laver amylose Zn complex.
The test of 2 laver amylose Zn complex using effect of experimental example
Detect material: catalase (CAT) assay kit, malonaldehyde (MDA) assay kit, superoxide dismutase
Enzyme (T-SOD) assay kit, (the above kit is all from Nanjing and builds up biological work glutathione peroxidase (GSH-PX)
Journey research institute)；
Duroc × length of 28 ± 1 ages in days is white × and great Bai ternary castrates male earner pig 50.
Test method: male earner pig is randomly divided into 5 groups by weight phase approximately principle, is respectively as follows: blank control group A group, sulfuric acid
Zinc addition group group B group, laver amylose low dose group C group, middle dose group D group, high dose group E group, every group of 10 weanling pigs；
Wherein, A group feeds Basic drawing, and the Basic drawing of B group feeding addition 1000mg/kg zinc sulfate, C, D, component E are other
The Basic drawing of feeding addition 1000,3000 and 5000mg/kg laver amylose Zn complex (is raised on A, B, C, D, E group basis used
Zinc is free of in material).
Enter trial test phase 3d after on-test, with adequacy test daily ration.Then carry out experimental period 30d.Off-test is prohibited
After eating 12h, the LPS of 100 μ g/kg is injected intraperitoneally by weight for B, C, D, E group, and A group injects the physiological saline of equivalent.
Sample acquisition: the blood sampling of piglet vena cava anterior, 3500r/m are centrifuged 15min, and separation serum is used for the survey of oxidation resistance
It is fixed.
The samples of intestinal contents acquires: experimental animal jugular vein sacrificed by exsanguination opens abdominal cavity, by enteron aisle from its mesenterium
Place samples after removing on ice.Take out caecum, squeeze out 10g cecal content sample be packed into immediately it is sterilized (1 × 105Pa, 121
DEG C sterilizing 60min) processing 10mL centrifuge tube in, be put into -40 DEG C of low temperature refrigerators and save, to inspection.
CAT and GSH-Px uses xanthine oxidizing process using the measurement of phenobarbital method, T-SOD using colorimetric method for determining, MDA
Concrete operation step is measured to carry out by kit specification；Intestinal flora richness is detected by upper Shanghai's style Nuo Sen biotech firm.
" mean+SD " is used to indicate test data, SPSS17.0 software carries out single factor analysis, DuncanShi
Multiple comparative test.Superscript letters are identical or not marking-up matrix shows that difference is not significant (P > 0.05), and letter is different to indicate that difference is aobvious
It writes (P < 0.05).
The influence testing inspection result such as the following table 1 of laver amylose Zn complex to immunological stress piglet Serum antioxidant indices
It is shown:
Influence of the 1 laver amylose Zn complex of table to immunological stress piglet Serum antioxidant indices
By the experimental data of table 1 as above it is found that B, C, D, E group Content of MDA and serum T-SOD activity are compared with blank group (A
Group) there is the raising of conspicuousness, the raising of conspicuousness is also presented in CAT activity.It can be seen that piglet is after injecting LPS, in serum from
It is increased significantly by base content, body enters immunological stress state.Meanwhile serum T-the SOD of laver amylose Zn complex addition group
There are the raising of conspicuousness, the reduction of Content of MDA conspicuousness compared with zinc sulfate addition group B group with GSH-Px activity.
Comprehensive as above analysis can make enzyme radicals scavenging system it is found that adding laver amylose Zn complex in feed
System activity improves, and enhances the blood antioxidant ability of body, and the immunological stress that weanling pig is immunized by LPS is effectively relieved.
It is as shown in Figure 1 that laver amylose Zn complex influences testing result to microorganism richness in intestine of young pigs.
For testing result as shown in Figure 1 it is found that for compared to A group, B group is more single on flora composition, with
Betaproteobacteria and Clostridia is advantage Gammaproteobacteria, and abundance is respectively 40.8% and 50.9%；Simultaneously A group with
Bacilli is advantage Gammaproteobacteria, abundance 38.5%.Lactobacillaceae abundance in A group Bacilli Gammaproteobacteria is
38.5%, the Burkholderiaceae abundance much higher than 4.5%, the B group Betaproteobacteria Gammaproteobacteria of B group is
It is 46.1% that the Clostridiacea of 34.5%, Clostridia Gammaproteobacteria, which enriches,.Clostridiaceae has high temperature resistant, resists
The characteristics such as inverse property is strong；Lactobacillaceae is the beneficial bacterium in intestine of young pigs, illustrates that piglet intraperitoneal injection LPS leads to piglet
Body temperature increases, and tolerance stronger Clostridiaceae and Gram-negative bacteria Betaproteobacteria can be deposited
It is living.
Meanwhile compared to B group for, the flora richness of laver amylose zinc addition group significantly improves, and more stable, bacterium
Group's composition is also more average, and Lactobacillaceae has apparent rising.With addition same dose C group compared with, C group with
Clostridiacea and Burkholderiaceae is dominant bacteria, but proportion is declined compared with B group, respectively 35.3%
With 18.5%, and Cordycepps is abundant compared with B group.Wherein the abundance of the Lactobacillaceae of E group is each group highest, is reached
43.1%, other floras it is abundant also more average, it is 5.2% that the abundance of Clostridiaceae, which is that each group is minimum,
The abundance that Ruminococcaceae is the Burkholderiaceae of 6.7%, Betaproteobacteria Gammaproteobacteria is 12%,
Abundance is minimum in the immunological stress each group of injection LPS.
Comprehensive as above analysis, while can it is found that the enteric microorganism richness of weanling pig can be improved in this laver amylose zinc
To increase the quantity of beneficial bacterium in enteron aisle, unbalance to intestine of young pigs flora caused by immunological stress have certain relaxation effect.
Although illustrate and describing the present invention with specific embodiment, it will be appreciated that without departing substantially from of the invention
Many other change and modification can be made in the case where spirit and scope.It is, therefore, intended that in the following claims
Including belonging to all such changes and modifications in the scope of the invention.
Claims (10)
1. a kind of laver amylose Zn complex, which is characterized in that the laver amylose Zn complex is anti-with zinc salt by laver amylose
It should be prepared.
2. the preparation method of laver amylose Zn complex described in claim 1, which is characterized in that the preparation method includes:
Lye is added into the solution dissolved with laver amylose and zinc salt with regulation system pH, after heating reaction simultaneously by reaction solution alcohol precipitation
Collect sediment；Sediment is concentrated and dried after purification, obtains laver amylose Zn complex.
3. preparation method according to claim 2, which is characterized in that the zinc salt is zinc sulfate, zinc nitrate, or corresponding
At least one of hydrate.
4. preparation method according to claim 2, which is characterized in that the mass ratio of laver amylose and zinc salt is 1:3~3:
1；
Preferably, the mass ratio of laver amylose and zinc salt is 1:2~2:1.
5. preparation method according to claim 2, which is characterized in that the lye includes: that sodium hydroxide/potassium hydroxide is molten
Liquid and ammonium hydroxide；
And/or the regulation system pH is the pH to 6.5~7.5 of regulation system；
Preferably, the regulation system pH is the pH to 7 of regulation system.
6. preparation method according to claim 2, which is characterized in that the purifying includes: by gained sediment with solvent
After washing, dialysis treatment is carried out.
7. laver amylose Zn complex described in claim 1 is increasing the application in weanling pig intestinal flora richness；
And/or laver amylose Zn complex described in claim 1 is increasing in weanling pig enteron aisle in the quantity of beneficial bacterium
Using；
And/or laver amylose Zn complex described in claim 1 weanling pig enterobacteriaceae caused by alleviating because of immunological stress
Application during group is unbalance.
8. laver amylose Zn complex described in claim 1 enhances body resisting stress in enhancing weanling pig immunity of organisms
Application in ability.
9. application of the laver amylose Zn complex described in claim 1 in feed addictive；Preferably, the feed addition
Agent is feed additive for weaning piglet.
10. the feed comprising laver amylose Zn complex described in claim 1.
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| CN201811051775.6A CN108967699A (en) | 2018-09-10 | 2018-09-10 | Laver amylose Zn complex and its preparation method and application |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116640237A (en) * | 2023-06-15 | 2023-08-25 | 广东医科大学 | Preparation method and application of rhizoma anemarrhenae polysaccharide-zinc complex |
-
2018
- 2018-09-10 CN CN201811051775.6A patent/CN108967699A/en not_active Withdrawn
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116640237A (en) * | 2023-06-15 | 2023-08-25 | 广东医科大学 | Preparation method and application of rhizoma anemarrhenae polysaccharide-zinc complex |
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Application publication date: 20181211 |