CN105981829B - Sterile liquid bovine colostrum product and preparation method and application thereof - Google Patents
Sterile liquid bovine colostrum product and preparation method and application thereof Download PDFInfo
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Abstract
The invention provides a sterile liquid bovine colostrum product, a preparation method and application thereof. The method comprises the following steps: degreasing raw material liquid bovine colostrum; removing part of casein from the degreased emulsion and controlling the content of the casein in the emulsion to be 10-20% of the total content of protein in the emulsion; and (3) adjusting the pH value of the emulsion after removing part of casein to 4.5-5.5 by using weak alkaline salt, and sequentially carrying out microfiltration by using polypropylene filter membranes of 0.8 mu m, 0.45 mu m and 0.2 mu m to obtain filtrate, namely the sterile liquid bovine colostrum product. The method of the invention can avoid the denaturation and inactivation of the immunoglobulin caused by high-temperature sterilization, and well ensure the content of the immunoglobulin.
Description
Technical Field
The invention relates to a sterile liquid bovine colostrum product, a preparation method and application thereof, in particular to a method for preparing the sterile liquid bovine colostrum product by utilizing a polypropylene filter membrane, the obtained sterile liquid bovine colostrum product and related application thereof, and belongs to the field of dairy product preparation.
Background
The bovine colostrum is milk secreted by healthy cows within 72 hours after farrowing, contains precious active immunoglobulin and abundant nutritional ingredients such as milk calcium, protein, various trace elements and the like, and is known as the king of immunity in the 21 st century. Compared with normal milk, the colostrum milk has the advantages that the content is obviously different, the protein content of colostrum is higher, the fat and sugar content is lower, the iron content is 10-17 times of that of the normal milk, and the vitamin D and the vitamin A are 3 times and 10 times of that of the normal milk respectively.
The colostrum of cow has attracted much attention because it naturally contains various immune factors, such as active immunoglobulin, lactoferrin, lactoperoxidase, lysozyme, etc., of which the most important substance is active immunoglobulin, and the content of which is more than 10 times that of colostrum. The bovine colostrum contains five types of immunoglobulin, namely IgA, IgD, IgE, IgM and IgG, wherein the content of the IgG is more than 80%, and the characteristics of the immunoglobulin are more stable than those of other four types of immunoglobulin, and the immunoglobulin is the most main functional substance. A large number of researches and clinical trials prove that IgG in the bovine colostrum has the immunoregulation function, can effectively prevent and improve diseases, enhances the disease resistance, and can inhibit various pathogenic microorganisms. In addition, the bovine colostrum has other functions, such as promoting growth and development, improving intelligence, improving body condition, improving athletic performance, relieving fatigue, delaying aging, regulating intestinal flora, etc. Bovine colostrum was listed as the most promising non-herbal natural health food in the 21 st century by the American food technology Association (IFT) in 2000.
However, bovine colostrum contains some sources of sensitization, especially if applied to infant food, with a large potential risk. A great deal of research at present shows that the sensitization source is mainly in casein, so that the sensitization risk can be greatly reduced if the casein in the bovine colostrum is removed. On the other hand, since IgG in bovine colostrum is a strong heat sensitive substance, sterilization cannot be performed using a conventional high temperature sterilization technique. These factors all affect the use of bovine colostrum to some extent.
For the solution containing heat-sensitive substances, the membrane filtration is a good sterilization process, particularly static filtration (parallel flow filtration), the temperature rise cannot be caused in the membrane filtration process, the heat-sensitive substances cannot be damaged, meanwhile, the particle size of microorganisms in the solution is larger than 0.2 μm, and the complete sterilization effect can be achieved when the microorganisms pass through a 0.2 μm microfiltration membrane. However, the bovine colostrum solution is very viscous and contains more substances with larger particle size, and if the bovine colostrum solution is directly passed through a membrane, the membrane flux is very low.
In the prior art, research reports on sterilization of bovine colostrum by adopting a microfiltration membrane filtration technology have been reported, wherein most of the microfiltration membranes are ceramic membranes. For example, CN101263839A discloses that milk containing bovine colostrum powder is filtered and sterilized by a ceramic microfilm with a pore diameter of 0.8-1.4 microns. The literature, "analysis of pollution resistance during the filtration of bovine colostrum by ceramic membrane" (Wangzhigao, Chen Wei hong, Xunan Ping, membrane science and technology, 2.2004, vol. 24, No. 1) examines the influence of pore size on the process of membrane filtration of bovine colostrum, and analyzes the formation of membrane filtration resistance with pore size of 0.2 μm, 0.5 μm and 0.8 μm and the influence thereof on permeation flux and protein retention rate. The ceramic filter membrane is prepared from materials such as alumina, titanium oxide, zirconia and the like through a special process, has high mechanical strength and strong pressure resistance, however, when the ceramic filter membrane is used for microfiltration, the operation mode is usually cross flow operation (namely dynamic filtration), has high tangential flow velocity, and can enable bovine colostrum to better pass under the conditions of high pressure and high rotating speed, thereby achieving the membrane-passing effect. But higher temperatures are generated during the filtration process, which causes denaturation and inactivation of IgG in the bovine colostrum; meanwhile, the ceramic membrane has high cost and needs other supporting facilities, so that the overall cost is too high during production, and the ceramic membrane is not beneficial to popularization.
CN 103039630A discloses a method for preparing sterile bovine colostrum whey with natural immunocompetence by a membrane filtration technology, wherein fat is removed from bovine colostrum by centrifugation, casein is precipitated by adopting an isoelectric point method or a chymosin method, obtained whey liquid is pre-filtered by adopting a hollow fiber membrane or a ceramic membrane with the aperture of 0.4-1.2 mu m, and residual fat, casein and most of thalli in the whey are removed; and filtering and sterilizing by adopting a polytetrafluoroethylene membrane filter with the pore diameter of 0.1-0.4 mu m. By using the method, the treatment capacity of the feed liquid is improved, and the loss rate of IgG can be reduced to a certain degree. However, the prefiltration in this method also employs dynamic filtration, as mentioned above, where the tangential flow rate allows the bovine colostrum to pass through the filter membrane well, but during the filtration process the temperature rises causing denaturation inactivation of IgG in the bovine colostrum. In addition, the method needs to adopt two membranes made of different materials to filter the bovine colostrum, the number of matched filter equipment is large, and the prices of the hollow fiber membrane, the ceramic membrane and the polytetrafluoroethylene membrane are relatively high, so that the practical application of the technology is limited.
Furthermore, the literature "influence of pH on the isolation and purification of insulin-like growth factor-1 (IGF-1) from bovine colostrum" (Xiaoyan, journal of food and Biotechnology, 7.2005, Vol. 24, No. 4) showed that IGF-1 permeability increased under alkaline conditions during ultrafiltration. It is believed that at pH >7, whey proteins acquire a net negative charge, increasing the electrostatic repulsion between molecules, eliminating the aggregation of protein molecules and increasing the permeability. However, the reference does not specify the filter material used.
On the other hand, the polypropylene micro-filtration membrane is a filtration membrane commonly used in the food industry, is prepared by hot-melting and bonding polypropylene superfine fibers together, belongs to a membrane material for deep filtration, and has the following characteristics: food-grade isotactic polypropylene is used as a raw material, and no additive is added in the whole production process; the physical and chemical properties are stable, and the compatibility is good; has serial pore sizes, high porosity, large pollutant carrying capacity, capacity of being back flushed and high temperature disinfected, relatively high pressure resistance and low cost. However, there is no report in the prior art that polypropylene filters are used for microfiltration of bovine colostrum for sterilization.
Disclosure of Invention
The invention mainly aims to develop a method for preparing a sterile liquid bovine colostrum product by microfiltration of bovine colostrum with a polypropylene filter membrane, so that the loss of IgG in the processing process is reduced, and the application range of the bovine colostrum is widened.
It is another object of the present invention to provide a liquid, sterile bovine colostrum product.
Another object of the present invention is to provide the use of said liquid bovine colostrum product.
As mentioned above, polypropylene microfiltration membranes have many advantages such as safety, stability, and low cost, and have been widely used for microfiltration sterilization in the food industry, however, no report of using polypropylene microfiltration membranes for sterilization of bovine colostrum has been found in the prior art. The inventor finds in research that the mechanical strength of polypropylene microfiltration membranes widely sold in the market is much lower than that of ceramic membranes, particularly the polypropylene microfiltration membranes have high low-temperature brittleness and a pressure-resistant range generally not higher than 3Bar, and if the polypropylene microfiltration membranes are used for static filtration of bovine colostrum, even if the bovine colostrum is degreased and decarburized in advance, the emulsion after degreasing and decarburized cannot smoothly pass through the polypropylene microfiltration membranes under low pressure, and the membrane flux is low. The inventor of the present invention finds, through a large number of experimental studies in research, that the membrane flux can be significantly increased by controlling the casein content in the defatted bovine colostrum emulsion and adjusting the emulsion to a specific pH value, so that the bovine colostrum emulsion can smoothly pass through a 0.2 mu m polypropylene filter membrane in a static filtration manner under a lower pressure. The above operation is carried out in a sterile environment to obtain a sterile bovine colostrum solution, which can avoid denaturation and inactivation of immunoglobulin (mainly IgG) due to high temperature.
Thus, in one aspect, the present invention provides a process for the preparation of a sterile liquid bovine colostrum product, which comprises:
degreasing raw material liquid bovine colostrum;
removing part of casein from the degreased emulsion and controlling the content of the casein in the emulsion to be 10-20% of the total content of protein in the emulsion;
and (3) adjusting the pH value of the emulsion after removing part of casein to 4.5-5.5 by using weak alkaline salt, and sequentially carrying out microfiltration by using polypropylene filter membranes of 0.8 mu m, 0.45 mu m and 0.2 mu m to obtain filtrate, namely the sterile liquid bovine colostrum product.
According to a specific embodiment of the invention, in the preparation method of the sterile liquid bovine colostrum product, the microfiltration operation mode is static filtration, namely parallel flow filtration, and the temperature rise cannot be caused in the membrane filtration process, and the heat-sensitive substances cannot be damaged.
According to a specific embodiment of the present invention, in the method for preparing the sterilized liquid bovine colostrum product, the raw material liquid bovine colostrum is raw fresh bovine colostrum or reconstituted milk reconstituted from bovine colostrum powder. Preferably, the raw material liquid bovine colostrum has the total solid content of 13.0-18.0%, the fat content of 3.0-5.0%, the total protein content of 4.0-6.0% and the IgG content of 10-50 mg/mL.
According to a specific embodiment of the present invention, in the method for preparing the sterilized liquid bovine colostrum product, the fat in the bovine colostrum can be completely removed by a conventional defatting process in the field.
According to a specific embodiment of the present invention, in the preparation method of the sterile liquid bovine colostrum product, the step of removing part of the casein comprises adjusting the pH of the defatted milk to 4.3-4.6 by using an acid precipitation method, precipitating the casein, and then filtering the casein by filter bag filtration (preferably using a filter bag with 140-180 meshes) and/or centrifugation (for example, centrifugation at 3000 rpm-4000 rpm and at a low temperature, for example, 4-10 ℃ for 20 min-30 min). Because the isoelectric point of the immunoglobulin IgG in the bovine colostrum is slightly basic, the content of the immunoglobulin IgG is not influenced when the immunoglobulin IgG is treated by an acid precipitation method, and compared with other methods for precipitating casein, the method has the advantages of lower cost, simpler process and convenience for actual production operation. The acid used in the acid precipitation method can be one or more of lactic acid, acetic acid and citric acid. In the invention, complete removal of casein is not pursued, but the content of casein in the filtrate is controlled to be 10-20 wt% of the total content of protein in the emulsion. Unless otherwise indicated, all percentages and ratios recited herein are by weight.
According to a specific embodiment of the present invention, in the method for preparing the liquid bovine colostrum product under aseptic conditions, the weakly basic salt used for adjusting the pH is selected from one or more of sodium bicarbonate, potassium bicarbonate, sodium acetate, and potassium acetate, preferably sodium bicarbonate. In specific application, the pH value is adjusted by using a weak alkaline saline solution with the concentration of 5-15%, preferably 5-10%. The use of these weakly basic salts can minimize the loss of IgG content due to precipitation or denaturation inactivation of IgG caused by local overbasing, and can not bring significant adverse effects on the membrane-passing effect.
According to the specific embodiment of the invention, in the preparation method of the sterile liquid bovine colostrum product, the membrane pressure difference of each membrane is controlled to be 0.5-2 Bar during microfiltration, namely, the emulsion after pH value adjustment sequentially passes through 0.8 μm, 0.45 μm and 0.2 μm filter membranes under the pressure of 0.5-2 Bar (pressure difference on two sides of the membrane). In the filtering process, the membrane-passing pressure difference of each membrane can properly float within the range of 0.5-2 Bar, but cannot exceed the pressure-bearing range of the polypropylene filter membrane.
In the present invention, there is no special requirement for the microfiltration device, for example, a plate and frame filter commonly used in industry can be used, and other microfiltration equipment capable of realizing a pressure difference static filtration operation mode can also be used.
It will be appreciated that in the process for the preparation of the sterile liquid bovine colostrum product of the present invention, the steps should be operated at a relatively low temperature to avoid loss of active substance from the emulsion due to excessive temperatures. Preferably, the temperature of the emulsion in the whole process is controlled to be not higher than 10 ℃, and more preferably 4-8 ℃.
In another aspect, the invention provides a sterile liquid bovine colostrum product, which is prepared according to the above-described method of the invention.
According to a specific embodiment of the invention, the liquid bovine colostrum product contains 10-100 mg/mL of IgG, usually 10-50 mg/mL. Furthermore, although the sterile liquid bovine colostrum product of the present invention retains a certain amount of casein in the emulsion before membrane filtration during the preparation process, the experiment proves that no allergenic substances are present in the sterile liquid bovine colostrum product obtained after filtration. The technology of the invention can remove the possible allergic substances in the bovine colostrum and greatly reduce the potential risk while enabling the emulsion to smoothly pass through the polypropylene filter membrane in a static filtering mode, and well ensures the content of the functional components in the bovine colostrum, particularly the content of immunoglobulin IgG.
In another aspect, the invention also provides the use of the sterile liquid bovine colostrum product. The liquid bovine colostrum product prepared by the method can be used as a functional or nutritional food additive or a production raw material for preparing various foods, for example, the liquid bovine colostrum product can be applied to various neutral and acidic liquid dairy products and milk beverages, and can also be added to various yogurt products.
In conclusion, by applying the technology of the invention, the prepared liquid bovine colostrum product can achieve the aseptic effect, simultaneously, the active immune substance IgG in the bovine colostrum can be well preserved, the prepared aseptic liquid bovine colostrum product can be used as a functional raw material or a nutrient additive to be added into different beverage products, and the application range of the bovine colostrum is greatly expanded.
Detailed Description
For a more clear understanding of the technical features, objects and advantages of the present invention, reference is now made to the following detailed description of the technical aspects of the present invention with reference to specific examples, which are intended to illustrate the present invention and not to limit the scope of the present invention.
The raw material liquid bovine colostrum used in each example had a total solid content of 15.0%, a fat content of 4.0%, a total protein content of 4.5%, and an IgG content of 13.5 mg/mL; the microfiltration equipment adopted is a flat plate type microfilter, and the effective membrane area (single membrane) is 69.36cm2. Steps not described in detail in the examples can be performed according to conventional operations in the art.
Example 1
This example provides a sterile liquid bovine colostrum product, which is prepared according to the following steps:
1. degreasing:
the fat in the raw material fresh bovine colostrum is completely removed by adopting a conventional degreasing process.
2. And (3) precipitating casein:
adjusting pH of bovine colostrum to 4.3 by acid precipitation, and precipitating casein.
3. And (3) filtering:
casein was roughly removed with a 140 mesh filter bag to give a filtrate. The filtrate was tested through 0.45 μm and 0.2 μm polypropylene filters under a membrane pressure differential of 2Bar, static filtration mode, and less than 20g of emulsion passed through the filter in the first 1 minute.
4. Centrifuging:
the filtrate obtained in step 3 above was centrifuged at 4000rpm for 30min to remove the precipitate and obtain a supernatant containing about 10% casein based on total protein.
5. Adjusting the pH value:
the supernatant obtained in step 4 was adjusted to pH4.5 with 8% potassium bicarbonate.
6. Preparation of sterile environment:
the microfiltration equipment and the polypropylene filter membranes with the diameters of 0.8 mu m, 0.45 mu m and 0.2 mu m are placed in a super clean bench and are irradiated for 1 hour by ultraviolet light to achieve the sterile effect.
7. Film coating:
and (3) adding the supernatant with the pH value adjusted in the step (5) into the microfiltration equipment in the step (6), enabling the supernatant to sequentially pass through polypropylene filter membranes with the diameters of 0.8 mu m, 0.45 mu m and 0.2 mu m under the pressure of about 2Bar, and then storing the solution in a sterile sealed bag to obtain a sterile liquid bovine colostrum product. Since some particulate matters still exist in the solution after centrifugal separation to affect the membrane-passing effect, the solution needs to pass through filter membranes with different pore diameters respectively to improve the membrane-passing effect.
The film-passing effect of the present example is as follows:
it took about 17.3min to produce 1000g of sterile liquid bovine colostrum product without changing the filter.
8. And (3) verifying the sterilization effect:
respectively taking 1mL of the sterile liquid bovine colostrum solution obtained in the step 7, inoculating the sterile liquid bovine colostrum solution into a total bacterial colony test piece and a mould yeast test piece, putting the total bacterial colony test piece into a 37 ℃ oven for culturing for 48 hours, putting the mould yeast test piece into a 25 ℃ oven for culturing for 120 hours, and then taking out to observe whether bacteria grow.
Example 2
This example provides a sterile liquid bovine colostrum product, which is prepared according to the following steps:
1. degreasing:
and (3) completely removing fat in the bovine colostrum by adopting a conventional degreasing process.
2. And (3) precipitating casein:
adjusting pH of bovine colostrum to 4.5 by acid precipitation, and precipitating casein.
3. And (3) filtering:
casein was roughly removed using a 180 mesh filter bag to obtain a filtrate. The filtrate was tested through a 0.45 μm polypropylene filter under a membrane pressure difference of 2Bar in a static filtration mode, and less than 20g of emulsion passed through the filter in the initial 1 minute.
4. Centrifuging:
the above solution was centrifuged at 3000rpm for 20min, and the precipitate was removed to obtain a supernatant containing 20% casein based on the total protein content.
5. Adjusting the pH value:
and (3) adjusting the pH value of the supernatant obtained in the step (4) to 5.0 by using 8% potassium acetate.
6. Preparation of sterile environment:
and (3) putting the membrane passing equipment and the polypropylene filter membranes with the diameters of 0.8 mu m, 0.45 mu m and 0.2 mu m into a super clean bench, and carrying out ultraviolet irradiation for 1h to achieve the sterile effect.
7. Film coating:
and (3) adding the supernatant with the adjusted pH value in the step (5) into the membrane passing equipment in the step (6), enabling the supernatant to sequentially pass through polypropylene filter membranes with the diameters of 0.8 mu m, 0.45 mu m and 0.2 mu m under the membrane passing pressure difference of about 2Bar, and then storing the solution in a sterile sealed bag to obtain a sterile liquid bovine colostrum product.
The film-passing effect of the present example is as follows:
it took about 20.1min to produce 1000g of sterile liquid bovine colostrum product without changing the filter.
8. And (3) verifying the sterilization effect:
respectively taking 1mL of the sterile liquid bovine colostrum solution obtained in the step 7, inoculating the sterile liquid bovine colostrum solution into a total bacterial colony test piece and a mould yeast test piece, putting the total bacterial colony test piece into a 37 ℃ oven for culturing for 48 hours, putting the mould yeast test piece into a 25 ℃ oven for culturing for 120 hours, and then taking out to observe whether bacteria grow.
Example 3
This example provides a sterile liquid bovine colostrum product, which is prepared according to the following steps:
1. degreasing:
and (3) completely removing fat in the bovine colostrum by adopting a conventional degreasing process.
2. And (3) precipitating casein:
adjusting pH of bovine colostrum to 4.3 by acid precipitation, and precipitating casein.
3. And (3) filtering:
a portion of the casein was roughly removed with a 140 mesh filter bag to obtain a filtrate.
4. Centrifuging:
the filtrate was centrifuged at 4000rpm for 25min to remove the precipitate and obtain a supernatant containing approximately 13% casein based on total protein.
5. Adjusting the pH value:
the supernatant obtained in step 4 was adjusted to pH 5.5 with 8% sodium acetate.
6. Preparation of sterile environment:
and (3) putting the membrane passing equipment and the polypropylene filter membranes with the diameters of 0.8 mu m, 0.45 mu m and 0.2 mu m into a super clean bench, and carrying out ultraviolet irradiation for 1h to achieve the sterile effect.
7. Film coating:
and (3) adding the supernatant with the adjusted pH value in the step (5) into a membrane-passing device, enabling the supernatant to sequentially pass through 0.8 mu m, 0.45 mu m and 0.2 mu m filter membranes under the membrane-passing pressure difference of about 1.5Bar, and then storing the solution in a sterile sealed bag to obtain a sterile liquid bovine colostrum product.
In this example, it took about 24.3min to produce 1000g of sterile liquid bovine colostrum product without replacing the filter membrane.
8. And (3) verifying the sterilization effect:
respectively taking 1mL of the sterile liquid bovine colostrum solution obtained in the step 7, inoculating the sterile liquid bovine colostrum solution into a total bacterial colony test piece and a mould yeast test piece, putting the total bacterial colony test piece into a 37 ℃ oven for culturing for 48 hours, putting the mould yeast test piece into a 25 ℃ oven for culturing for 120 hours, and then taking out to observe whether bacteria grow.
Example 4
This example provides a sterile liquid bovine colostrum product, which is prepared according to the following steps:
1. degreasing:
and (3) completely removing fat in the bovine colostrum by adopting a conventional degreasing process.
2. And (3) precipitating casein:
adjusting pH of bovine colostrum to 4.5 by acid precipitation, and precipitating casein.
3. And (3) filtering:
casein was roughly removed with a 140 mesh filter bag to give a filtrate.
4. Centrifuging:
the filtrate was centrifuged at 4000rpm for 20min to remove the precipitate and obtain a supernatant having a casein content of about 15%.
5. Acid adjustment:
and (3) adjusting the pH value of the supernatant obtained in the step (4) to 4.5 by using 5% potassium bicarbonate.
6. Preparation of sterile environment:
and (3) putting the membrane passing equipment and the polypropylene filter membranes with the diameters of 0.8 mu m, 0.45 mu m and 0.2 mu m into a super clean bench, and carrying out ultraviolet irradiation for 1h to achieve the sterile effect.
7. Film coating:
and (3) adding the supernatant with the pH value adjusted in the step (5) into a membrane-passing device, enabling the supernatant to sequentially pass through 0.8 mu m, 0.45 mu m and 0.2 mu m filter membranes under the membrane-passing pressure difference of about 2Bar, and then storing the solution in a sterile sealed bag to obtain a sterile liquid bovine colostrum product.
In this example, it took about 23.8min to produce 1000g of sterile liquid bovine colostrum product without replacing the filter membrane.
8. And (3) verifying the sterilization effect:
respectively taking 1mL of the sterile liquid bovine colostrum solution obtained in the step 7, inoculating the sterile liquid bovine colostrum solution into a total bacterial colony test piece and a mould yeast test piece, putting the total bacterial colony test piece into a 37 ℃ oven for culturing for 48 hours, putting the mould yeast test piece into a 25 ℃ oven for culturing for 120 hours, and then taking out to observe whether bacteria grow.
Examination of IgG content in the products of the examples
The results of examination of the IgG content in the product after the production of the sterile liquid bovine colostrum products according to the above examples are shown in table 1 (for comparison, the values in table 1 are values obtained by converting the sterile liquid bovine colostrum of each example to the same volume as the raw liquid bovine colostrum).
TABLE 1 determination of IgG content in the different examples
As can be seen from Table 1, the loss amounts of IgG in the examples of the present invention are 3.48%, 4.96%, 4.22% and 3.93%, respectively, and the loss amounts are less than 5%, and the method for preparing the sterile liquid bovine colostrum product has a good effect.
Investigation of solution effect through filter membrane under different pH conditions
Five 1000g raw materials are taken, the pH value in the step 6 is respectively adjusted to 4.5, 5.0, 5.5, 6.0 and 6.5 under the same other conditions, and the membrane passing effect of a 0.2 mu m filter membrane is respectively examined, which is shown in table 2.
TABLE 2 different pH values worth examining
| Scheme one | Scheme two | Scheme three | Scheme four | Scheme five | |
| pH value of feed liquid | 4.5 | 5.0 | 5.5 | 6.0 | 6.5 |
| By number of membranes | 1 | 1 | 1 | 4 | 6 |
| Required time of | 24.3min | 26.1min | 24.7min | 189min | 308min |
As can be seen from Table 2, under the condition of pH4.5-5.5, the time required for the same amount of bovine colostrum solution to pass through the membrane is shortest, and only 1 filter membrane is needed, while when 1000g of pH6.0 solution passes through the membrane, 4 filter membranes need to be replaced, and when 1000g of pH6.5 solution passes through the membrane, 6 filter membranes need to be replaced (in the fourth and fifth schemes, the membranes are replaced every 1 h).
Examination of the Sterilization Effect
The products prepared in the examples were verified for their sterilization effect, respectively, as shown in table 3.
TABLE 3 examination of the Sterilization Effect
As can be seen from Table 3, the products in the examples of the present invention all grew aseptically within 6 months, and the sterilization effect was good.
Examination of IgG content in the products of each example within six months
The content of IgG in the product of each example (stored at 2-6 ℃ in a dark place at a low temperature) within six months is examined, and the result is shown in Table 4.
TABLE 4 examination of IgG content (mg/mL) variation
| Time of day | Example 1 | Example 2 | Example 3 | Example 4 |
| One month | 11.95 | 11.62 | 12.01 | 12.52 |
| Two months old | 11.65 | 11.33 | 11.85 | 12.27 |
| Three months old | 11.21 | 11.14 | 11.61 | 12.05 |
| Four months old | 11.07 | 11.02 | 11.42 | 11.89 |
| Five months old | 10.99 | 10.89 | 11.29 | 11.67 |
| Six months old | 10.93 | 10.80 | 11.02 | 11.51 |
As can be seen from Table 4, the loss rate of the product in each example of the present invention is small within 6 months, and the stability is good.
Allergy study
The bovine colostrum products of examples 1 to 4, together with fully defatted and casein-free bovine colostrum and bovine colostrum only defatted and not casein-free bovine colostrum were used as test samples, and the same batch of healthy 72 mice was used for experiments, and divided into five groups of 12 mice, each group was fed with the above bovine colostrum solution, and the group was fed once every morning and evening at 100g for 30 days, and the health condition of the mice was examined every 3 days to examine the product allergy, and the content of the health condition of the mice is shown in table 5.
TABLE 5 mouse health Studies
As can be seen from Table 5, when the bovine colostrum product of the embodiments of the present invention is fed to mice, the mice have good health condition and no uncomfortable symptoms, which indicates that the bovine colostrum treated by the present process does not contain allergen and can be eaten by human body.
Claims (5)
1. A method of preparing a sterile liquid bovine colostrum product, the method comprising:
degreasing: completely removing fat in raw fresh bovine colostrum by adopting a degreasing process;
and (3) precipitating casein: adjusting pH of bovine colostrum to 4.3 by acid precipitation, and precipitating casein;
and (3) filtering: roughly removing casein by using a filter bag with 140 meshes to obtain filtrate;
centrifuging: centrifuging the obtained filtrate at 4000rpm for 30min, removing precipitate to obtain supernatant, wherein the casein content in the supernatant is 10% of the total protein content;
adjusting the pH value: regulating the obtained supernatant with 8% potassium bicarbonate to pH 4.5;
preparation of sterile environment: placing the microfiltration equipment and polypropylene filter membranes of 0.8 μm, 0.45 μm and 0.2 μm into a super clean bench, and performing ultraviolet irradiation for 1h to achieve sterile effect;
film coating: and adding the supernatant with the adjusted pH value into the sterile microfiltration equipment, sequentially passing through 0.8 mu m, 0.45 mu m and 0.2 mu m polypropylene filter membranes under the pressure of 2Bar, and storing the solution in a sterile sealed bag to obtain a sterile liquid bovine colostrum product.
2. A method of preparing a sterile liquid bovine colostrum product, the method comprising:
degreasing: completely removing fat in the bovine colostrum by adopting a degreasing process;
and (3) precipitating casein: adjusting pH of bovine colostrum to 4.5 by acid precipitation, and precipitating casein;
and (3) filtering: roughly removing casein by using a filter bag with 180 meshes to obtain filtrate;
centrifuging: centrifuging the filtrate at 3000rpm for 20min, removing precipitate to obtain supernatant containing casein 20% of total protein;
adjusting the pH value: adjusting the pH value of the obtained supernatant to 5.0 by using 8% potassium acetate;
preparation of sterile environment: putting the membrane-passing equipment and polypropylene filter membranes of 0.8 mu m, 0.45 mu m and 0.2 mu m into a super clean bench, and carrying out ultraviolet irradiation for 1h to achieve the sterile effect;
film coating: and adding the supernatant with the adjusted pH value into the sterile membrane-passing equipment, sequentially passing through 0.8 mu m, 0.45 mu m and 0.2 mu m polypropylene filter membranes under the membrane-passing pressure difference of 2Bar, and storing the solution in a sterile sealed bag to obtain a sterile liquid bovine colostrum product.
3. A method of preparing a sterile liquid bovine colostrum product, the method comprising:
degreasing: completely removing fat in the bovine colostrum by adopting a degreasing process;
and (3) precipitating casein: adjusting pH of bovine colostrum to 4.3 by acid precipitation, and precipitating casein;
and (3) filtering: roughly removing part of casein by using a filter bag with 140 meshes to obtain filtrate;
centrifuging: centrifuging the filtrate at 4000rpm for 25min, and removing precipitate to obtain supernatant containing casein 13% of total protein;
adjusting the pH value: adjusting the obtained supernatant with 8% sodium acetate to pH 5.5;
preparation of sterile environment: putting the membrane-passing equipment and polypropylene filter membranes of 0.8 mu m, 0.45 mu m and 0.2 mu m into a super clean bench, and carrying out ultraviolet irradiation for 1h to achieve the sterile effect;
film coating: and adding the supernatant with the adjusted pH value into a membrane-passing device, sequentially passing through 0.8 mu m, 0.45 mu m and 0.2 mu m filter membranes under the membrane-passing pressure difference of 1.5Bar, and storing the solution in a sterile sealed bag to obtain a sterile liquid bovine colostrum product.
4. A sterile liquid bovine colostrum product prepared according to the method of any one of claims 1 ~ 3.
5. The use of the sterile liquid bovine colostrum product of claim 4 as one of the starting materials for the preparation of IgG-containing food products.
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