CN1883505A - Method for extracting active substances from cow colostrums and equipment therefor - Google Patents
Method for extracting active substances from cow colostrums and equipment therefor Download PDFInfo
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- CN1883505A CN1883505A CNA2005100501665A CN200510050166A CN1883505A CN 1883505 A CN1883505 A CN 1883505A CN A2005100501665 A CNA2005100501665 A CN A2005100501665A CN 200510050166 A CN200510050166 A CN 200510050166A CN 1883505 A CN1883505 A CN 1883505A
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Abstract
The present invention relates to a method for extracting active substances from cow colostrums and its equipment. The invention uses continuous ion exchange equipment to extract active substances from cow colostrums. The process simplifies cow colostrums separation steps, increases its yield and purity, reduces separation cost for industrialization and improves cow colostrums resource utility ratio. The present invention also provides a kind of equipment, which realizes continuous ion exchange, increases yield and purity, reduces separation cost for industrialization and reaches the target of comprehensive utilization of cow colostrums.
Description
Technical field
The present invention relates to a kind of extracting method and extract the used equipment of active substance, relate in particular to a kind of method and equipment thereof that extracts the cow colostrum active material.
Background technology
Scientific research confirms, cattle colostrums is nature immune factor one of the living resources of enrichment the most, comprising important immune factor and somatomedin, as immunoglobulin (IgG), lactoferrin (Lf) and insulin like growth factor (IGF) etc., content than normal newborn high 10-100 doubly.These physiologically active ingredients have immunomodulating, slow down aging, promotion growth promoter, suppress a series of biological function such as tumor, not only can manufacturing function food, but also have the great potential of exploitation natural activity bio-pharmaceutical.But, the exploitation of colostrum resource at present just begins, mainly be some health food, the valid density of active substance is not high enough in the product, its immunocompetence is lost bigger in the course of processing, and also be short of aspect the colostrum comprehensive utilization of resources, the potential value that colostrum itself is possessed can not get sufficient reflection.Therefore, high-efficiency comprehensive utilization cattle colostrums resource is developed rational isolation technics route to extract the natural active protein of high-purity high-activity, and particularly therefrom developing natural food additive raw material will be of great practical significance.
China's milch cow total quantity just reaches more than 420 ten thousand at present, the annual 200000 tons of colostrums of still can having more than needed except that feeding milk cattle.But these resources are not well utilized, and wherein the rich functions active substance does not obtain due utilization.The development and use of cattle colostrums at present still are in the starting stage, and problem demanding prompt solution is also a lot, mainly is:
(1) product variety is comparatively single, mainly is exploitation immunoglobulin (IgG).
(2) be still waiting in the comprehensive utilization of resource to improve.Functional component is a lot of in the cattle colostrums, and the technology that adopts generally all is at a certain target protein that will extract at present, as immunoglobulin, and other functional components that be rich in first Ruzhong is considered less, has caused the wasting of resources.
(3) product specification is also lower, and as the domestic IgG that extracts from cattle colostrums, because the problem of isolation technics, its purity and activity are also lower, can only be used for bread and cheese, and this greatly reduces the value of product itself.
The extracting method that activated protein is general all is to adopt ion exchange chromatography, and ion exchange and absorption are one of important chemical unit operationss, uses the component in solid absorbent separation and the purification mixture to obtain application in many industrial circles.But owing to lack a kind of real successive Technology feasible in industry, the potentiality of ion exchange and adsorbing separation are fully played as yet.Traditional ion exchange and adsorption process are carried out in fixed bed.Along with carrying out from the process of friendship, bed then is divided into three zones---saturation region (or exhausting the district), mass-transfer zone and fresh resin district, in fixed bed, have only the resin of mass-transfer zone in running order, a large amount of resins in saturation region and fresh district then are in idle and waiting state.After whole resin bed penetrates, just enter the operation of next step, as washing, regeneration.In actual production,, need 3 to 4 large-scale fixed beds and complicated valve, pipeline switched system usually in order to implement this operation.
Summary of the invention
The object of the present invention is to provide a kind of method of from cattle colostrums, extracting active substance, this method simplified cattle colostrums separating step, improved productive rate and purity, reduction separation costs to realize industrialization, improved the resource utilization of cattle colostrums.
Another object of the present invention is to provide used equipment in a kind of said method.
In order to realize the foregoing invention purpose, the present invention adopts following technical scheme:
A kind of method of extracting active substance from cattle colostrums of the present invention comprises the steps:
(1) chooses raw material: choose the galactopoiesis of cow in 6 days puerperal;
(2) filter;
(3) defat;
(4) place continuous ionic displacement chromatography equipment to extract the cattle colostrums active substance.
As preferred version of the present invention, described defat is to use the frozen centrifugation defat, and the control refrigerated centrifuger carries out centrifugal degreasing under the 5000g acceleration of gravity in-4～4 ℃ of temperature ranges, and the time is 20～40 minutes.
As preferred version of the present invention, extracting the active substance that obtains is lactoferrin (Lf) and immunoglobulin (IgG).
Extracting lactoferrin (Lf) is to adopt cation exchange resin to separate, and resin is the gel type resin of methacrylic acid skeleton.
Extracting immunoglobulin (IgG) is to adopt anion exchange resin, and resin is for being the gel type resin of methacrylic acid skeleton.
The present invention preferably adopts the positive anion exchange technology of polyphone, once finishes target product and separates.
The present invention also provides used equipment in a kind of method of extracting active substance from cattle colostrums, this equipment comprises that seat in the plane PC, Programmable Logic Controller (PLC), the electromagnetic valve by Controlled by Programmable Controller, kinetic pump, pipeline, some root chromatogram columns, cabinet form, every root chromatogram column all links to each other with some electromagnetic valves, in order to realize the different links of technology.
The technological process that realizes in this equipment is divided into upper prop, impurity eluting, target product eluting, 4 continuous processs of resin regeneration, post switching, drip washing time and flow can be by the operation controls of a plurality of electromagnetic valves, electromagnetic valve is controlled by PLC, and can regulate to optimize by PLC and regulate technological parameter.
As a preferred version of the present invention, described chromatographic column is 20 or 30.
The present invention extracts the method for active substance from cattle colostrums, simplified the separating step of active substance in the cattle colostrums, has improved productive rate and purity, reduces separation costs to realize industrialization, reaches the purpose that comprehensive cattle colostrums utilizes.
Used equipment in the disclosed method of extracting active substance from cattle colostrums has saved the resin in most saturation region and fresh district among the present invention.After a part of resin is saturated, shift out system at once and sent for regeneration, regeneration back restock is to the fresh resin district, as long as regeneration and the speed of replenishing and the translational speed of mass-transfer zone are complementary, system can continuously carry out.In this equipment, do not have any inactive resin as can be seen, the service efficiency of resin is improved.This equipment also has following characteristics:
1, be a kind of ion exchange and adsorption process of really successive, steady-state operation, various logistics are with constant flow and form and pass through system without interruption.
2, reduce resin demand.Owing to got rid of inactive resin, the resin loadings can save 30～90%.
3, save regenerative agent and washings consumption.Owing to use short and small resin bed to contact with reverse flow, can make eluant/resin volume ratio less than 1, outflow concentration improves, volume reduces.
4, the motility of system.According to different technological requirements, resin column can in parallel or series connection, also can combine by parallel/series.Can change fluidic character interregional.
5, simple to operate, be easy to control.Normally used simple control loop can meet the demands.Auxiliary facilities can significantly reduce volume and consumption as pump, storage tank etc.
6, reduce investment and operating cost.Compare with fixed bed system, operating cost can reduce by 40～50% usually.
7, the present invention has used a kind of novel ion exchange resin, compares with ion-exchange gel, and day output improves 300%, and product purity is uninfluenced, and the response rate is 90% of a gel-type separating medium, and the initial stage input cost reduces 90%.
Figure of description
Fig. 1: continuous ion exchange apparatus structural representation in the embodiment of the invention
Fig. 2:; Continuous ion exchange apparatus working state schematic representation among the present invention;
Fig. 3: continuous ionic exchange flow chart in the embodiment of the invention.
The specific embodiment
The present invention will be further described by specific embodiment below in conjunction with accompanying drawing 1, Fig. 2, Fig. 3.
Embodiment: present embodiment extracts the active substance in the cattle colostrums, adopts following steps:
1, choose raw material:
Raw material adopts cow to produce behind the calf 6 times with interior milk.
Lf content in the cattle colostrums is generally high 10～100 times than common Lac Bovis seu Bubali.But cattle colostrums also has its shortcoming: the more common Lac Bovis seu Bubali of composition is more complicated, also has the blood streak sometimes; Viscosity is big, easily lumps changeableness; Abnormal flavour is arranged.Because the fat in the Lac Bovis seu Bubali can have a strong impact on the ion exchange chromatography process, therefore, cattle colostrums all will be done strict ungrease treatment.
2, the defat of colostrum
1) filter: cattle colostrums carries out the frozen centrifugation defat after filtering through 16 layers of gauze earlier again.
2) frozen centrifugation defat: for preventing protein Yin Wendu degeneration in the cattle colostrums, refrigerated centrifuger carried out centrifugal degreasing 30 minutes under the 5000g acceleration of gravity in-4～4 ℃ of temperature ranges.Colostrum generally is light yellow before the defat, after the defat, should be milky.Colostrum is divided into three layers after centrifugal: the upper strata thickness, and density is little, should be fat, the orlop solid, shaped, density is big, is the king-sized protein of some molecular weight.The milky white liquid in intermediate layer is the defat cattle colostrums.Discard upper strata oils and fats and lower floor and just can obtain the defat cattle colostrums than the macromole precipitation, the direct upper prop of defat cattle colostrums is perhaps put into the freezing preservation of refrigerator-freezer.
3) continuous ionic displacement chromatography
Cattle colostrums is placed continuous ionic displacement chromatography equipment, carry out ion exchange, extract active substance lactoferrin and immunoglobulin.
The continuous ion exchange apparatus that present embodiment adopts comprises that seat in the plane PC 1, Programmable Logic Controller 1, the electromagnetic valve by Controlled by Programmable Controller, kinetic pump, pipeline, 20 root chromatogram columns, cabinet form.Every root chromatogram column all links to each other with electromagnetic valve, and last seat in the plane PC programmes to Programmable Logic Controller according to different process, and then Programmable Logic Controller is by the control electromagnetic valve switch, in order to realize the different links of technology.
In conjunction with Fig. 2, pretreated cattle colostrums is communicated with K
2Path, deionized water is communicated with K
1-1Path, 1.6%NaCl is communicated with K
1-2Path, 5%NaCl is communicated with K
1-3Path, target protein lactoferrin (Lf) is collected in K
3-3, K
1-1, K
1-2Path is respectively applied for collects washing foreign protein and weak adsorptivity foreign protein.Regulate the continuous ion exchange apparatus kinetic pump, linear velocity is adjusted to 500cm/hr, and the raw material processing speed is 2L/hr.
Open continuous ion exchange apparatus, time flow is as shown in table 1 in the inner predefined separation lactoferrin technology of continuous ion exchange apparatus internal programmable controller, carries out 8 cycles in one day, and the phase is handled raw material cattle colostrums 6L weekly.
Table 1: separate time flow in the lactoferrin technology
| Open the state valve | Pump | Time (min) | ||
| Last sample district | Last sample district | K 2、M、B、K 4 | P 2 | 180 |
| Elution zone | The foreign protein water wash zone | K 1-1、D、A、K 3-1 | P 1 | 60 |
| Foreign protein brine wash district | K 1-2、D、A、K 3-2 | P 1 | 40 | |
| The target protein elution zone | K 1-3、D、A、K 3-3 | P 1 | 30×2 | |
| The salt residue cleaning area | K 1-1、D、A、K 3-1 | P 1 | 20 |
Each post location sees Table 2 in one-period
Table 2: the location of each post in one-period
| Time | Last sample district | Elution zone | ||
| 1 | 2 | 3 | 4 | |
| 0 | Last sample district | The foreign protein water wash zone | The foreign protein water wash zone | |
| 60 | Foreign protein brine wash district | Foreign protein brine wash district | ||
| 100 | The target protein elution zone | D, A valves are closed | ||
| 130 | D, A valves are closed | The target protein elution zone | ||
| 160 | The salt residue cleaning area | The salt residue cleaning area | ||
| 180 | ||||
Through after one-period, 1 and 2 switch to elution zone, and 3 and 4 switch to the sample district, circulation.
Carried out continued operation in continuous 24 hours, 48 hours, 72 hours, operating result is as shown in table 3.
Table 3 different operating cycle lactoferrin separating resulting
| Consumption cattle colostrums raw material/L | Washings/L | 1.6％ NaCl/L | 5％NaCl/ L | Galactopoiesis ferritin/g | Product purity | The response rate | |
| 24 hours groups | 48 | 22 | 11 | 16 | 51.8 | 95.3％ | 34.7％ |
| 48 hours groups | 96 | 44 | 22 | 32 | 110.4 | 96.7％ | 37.3％ |
| 72 hours groups | 144 | 66 | 33 | 48 | 148.8 | 96.1％ | 33.4％ |
Present embodiment continuous ionic separation equipment separates king-post with original multistage and is designed to the replacement of connecting from beginning to end of a plurality of chromatographic columns.Inner mobile to a direction at device by pressure fluids such as current stabilization pumps, and set gradually the eluent entrance according to the on off state of fluid flow direction by by-pass valve control, the products export point, material liquid entrance and residual solution exit point, thereby all chromatographic columns are divided into four subregions, each subregion comprises a root chromatogram column at least, after having passed through a given interval, by changing the on off state of valve, make each entrance, exit point is passed along the root chromatogram column that is directed downwards of fluid flow respectively, same four subregions are also passed to next root chromatogram column thereupon, move in circles like this and just formed solid absorbent virtual reverse flow round about, thereby realize the effect that relatively moves between solid absorbent and fluid in the continuous ionic switch.
As shown in Figure 3, four subregions of this continuous ionic switch can be finished different functions respectively, and the subregion I between the outlet of extracting solution residual solution is mainly used in strong absorption object in the absorption raw material; Adsorptivity composition a little less than the subregion II between residual solution outlet and eluent 1 inlet is mainly used in eluting; Subregion III between eluent 2 inlets and eluent 3 outlets is mainly used in time strong adsorptivity composition in the adsorbed product; Subregion IV between eluent 3 inlets and feed(raw material)inlet is mainly with the strong adsorptivity product of eluting.
Continuous ion exchange apparatus among the present invention can realize simplifying technology, and the consumption of filler and solvent is little, improves separation efficiency, and the interior to greatest extent biological activity that keeps improves product purity and yield, really realizes the integrated production of seriality.Have excellent economy, the exploitation that can directly improve biological Renewable resources such as cattle colostrums is worth.
Claims (9)
1, a kind of method of from cattle colostrums, extracting active substance, it is characterized in that comprising the steps: that (1) choose raw material: (1) chooses the milk of cow in 6 days puerperal;
(2) filter;
(3) defat;
(4) place continuous ionic displacement chromatography equipment to extract the cattle colostrums active substance.
2, a kind of method of from cattle colostrums, extracting active substance according to claim 1, it is characterized in that described defat is to use the frozen centrifugation defat, the control refrigerated centrifuger carries out centrifugal degreasing under the 5000g acceleration of gravity in-4～4 ℃ of temperature ranges, degreasing time is 20～40 minutes.
3, a kind of method of extracting active substance from cattle colostrums according to claim 1, it is characterized in that extracting the active substance that obtains is lactoferrin (Lf) and immunoglobulin (IgG).
4, a kind of method of from cattle colostrums, extracting active substance according to claim 3, it is characterized in that extracting lactoferrin (Lf) is to adopt cation exchange resin to separate, resin is the gel type resin of methacrylic acid skeleton.
5, a kind of method of from cattle colostrums, extracting active substance according to claim 3, it is characterized in that extracting immunoglobulin (IgG) is to adopt anion exchange resin, resin is the gel type resin of methacrylic acid skeleton.
6, a kind of method of extracting active substance from cattle colostrums according to claim 3 is characterized in that adopting the positive anion exchange technology of polyphone, once finishes target product and separates.
7. used equipment in a kind of method of from cattle colostrums, extracting active substance according to claim 1, it is characterized in that described continuous ionic displacement chromatography equipment comprises that seat in the plane PC, Programmable Logic Controller (PLC), the electromagnetic valve by Controlled by Programmable Controller, kinetic pump, pipeline, some root chromatogram columns, cabinet form, every root chromatogram column all links to each other with some electromagnetic valves, in order to realize the different links of technology.
8, a kind of equipment according to claim 7, it is characterized in that the technological process that realizes in this equipment is divided into upper prop, impurity eluting, target product eluting, 4 continuous processs of resin regeneration, post switching, drip washing time and flow can be by the operation controls of a plurality of electromagnetic valves, electromagnetic valve is controlled by PLC, and can regulate by PLC and optimize technological parameter.
9, used equipment in a kind of method of extracting active substance from cattle colostrums according to claim 7 is characterized in that described chromatographic column is 20 or 30.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2005100501665A CN1883505A (en) | 2005-06-20 | 2005-06-20 | Method for extracting active substances from cow colostrums and equipment therefor |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2005100501665A CN1883505A (en) | 2005-06-20 | 2005-06-20 | Method for extracting active substances from cow colostrums and equipment therefor |
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| Publication Number | Publication Date |
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| CN1883505A true CN1883505A (en) | 2006-12-27 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101724013B (en) * | 2008-10-24 | 2012-05-30 | 哈尔滨康普乳品有限公司 | Method for separating and purifying immunoglobulin A, immunoglobulin G and lactoferrin from bovine colostrum in industrializing way |
-
2005
- 2005-06-20 CN CNA2005100501665A patent/CN1883505A/en active Pending
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101724013B (en) * | 2008-10-24 | 2012-05-30 | 哈尔滨康普乳品有限公司 | Method for separating and purifying immunoglobulin A, immunoglobulin G and lactoferrin from bovine colostrum in industrializing way |
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