CN114259954A - Preparation method of chitosan-whey protein isolate gel for encapsulating beta-carotene - Google Patents
Preparation method of chitosan-whey protein isolate gel for encapsulating beta-carotene Download PDFInfo
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Abstract
A preparation method of chitosan-whey protein isolate gel for encapsulating beta-carotene relates to the technical field of gel preparation. Dissolving CS in an acetic acid solution to obtain a CS solution; dissolving WPI in water to obtain WPI solution; adding the CS solution into the WPI solution, mixing, and adjusting the pH of the solution to 6 to obtain a CS-WPI solution; dissolving beta-C powder in medium chain triglyceride, and carrying out ultrasonic treatment in the dark to obtain a beta-C solution; and adding the beta-C solution into the CS-WPI solution, mixing to obtain a CS-WPI solution encapsulated with the beta-C, and performing ultrahigh pressure treatment to obtain a CS-WPI gel encapsulated with the beta-C. The chitosan and the separated whey protein are utilized to form a double-layer emulsion wall material carrier, and then the emulsion is induced by ultrahigh pressure to form gel, so that the advantages of overcoming the defects of insufficient stability of a single liposome carrying wall material or an emulsion carrying system to embedded beta-C and avoiding rapid damage in a digestive environment are achieved.
Description
Technical Field
The invention relates to the technical field of gel preparation, in particular to a preparation method of chitosan-whey protein isolate gel for encapsulating beta-carotene.
Background
Vitamin A has important functions of preventing nyctalopia, maintaining normal immune function, growth, development and gastrointestinal function. However, the human body lacks the ability to synthesize vitamin a autonomously, so vitamin a can only be obtained from the daily diet. Beta-carotene (beta-C) has the highest vitamin A activity compared to other provitamin A carotenoids (alpha-carotene, cryptoxanthin), which is about twice as active as other provitamin A carotenoids, and the conversion of beta-C to vitamin A is also the highest. In addition to being the primary source of vitamin a nutrients, β -C has antioxidant capacity and, due to its unique structure, can function as a lipid radical scavenger and singlet oxygen quencher. However, the maximum uptake of beta-C from natural plant sources in humans is very low and does not reach the recommended intake (2-4 mg/day). Therefore, the separation and purification of beta-C have led to the research of a large number of scholars at home and abroad. The purified beta-C has the characteristics of poor water solubility, instability in oxygen, heat and light and rapid degradation in the gastric phase in the digestion process, so that the application value of the beta-C is greatly limited. At present, in order to improve the water phase solubility and bioavailability of beta-C, single molecular emulsifying agents are mostly adopted at home and abroad to transport and embed the beta-C. The single emulsion may cause problems of reduction of active ingredients of beta-C, insufficient stability, etc. due to movement of oil phase and diffusion of oxygen in the system during storage.
Disclosure of Invention
The invention aims to solve the problems in the prior art and provide a preparation method of chitosan-isolated whey protein gel for encapsulating beta-carotene, which can overcome the defects of insufficient stability of a single liposome carrying wall material or an emulsion carrying system on embedding beta-C and can avoid rapid damage in a digestive environment.
The invention comprises the following steps:
1) preparation of the aqueous phase: firstly, dissolving chitosan in an acetic acid solution to obtain a chitosan solution; dissolving the isolated whey protein in water to obtain an isolated whey protein solution; adding the chitosan solution into the whey protein isolate solution, mixing, and adjusting the pH value of the solution to 6 to obtain a chitosan-whey protein isolate solution;
2) preparation of oil phase: dissolving beta-carotene powder in medium-chain triglyceride, and carrying out ultrasonic treatment in the dark to obtain a beta-carotene solution;
3) mixing water and oil phases: adding the beta-carotene solution into the chitosan-whey protein isolate solution, and mixing to obtain a chitosan-whey protein isolate solution encapsulated with beta-carotene;
4) preparing gel under ultrahigh pressure: and carrying out ultrahigh pressure treatment on the chitosan-isolated whey protein solution encapsulated with the beta-carotene to obtain chitosan-isolated whey protein gel encapsulated with the beta-carotene.
In the step 1), the acetic acid solution is an acetic acid solution with the mass fraction of 1%; the mass percentage concentration of the chitosan solution can be 0.8%; the mass percentage concentration of the separated whey protein solution can be 20%; adding the chitosan solution into the whey protein isolate solution, mixing, and then placing the solution in a refrigerator at 4 ℃ overnight so as to be fully hydrated; the volume ratio of the chitosan solution to the isolated whey protein solution can be 1: 4-5.5; preferably 1: 5; after the pH value of the solution is adjusted to 6, the chitosan-whey protein isolate solution can be placed on a magnetic stirrer to be stirred for 1 hour, so that the chitosan-whey protein isolate solution and the magnetic stirrer are completely mixed.
In step 2), the beta-carotene powder is dissolved in the medium chain triglyceride so that the concentration of beta-C is 0.05%, 0.10%, 0.15% and 0.2%; the beta-C solution should be taken as it is.
In step 3), the ratio of the beta-carotene solution to the chitosan-whey protein isolate solution is 10: 1; after mixing, the mixture can be processed for 5min at the rotating speed of 600r/min by a magnetic stirrer; the concentration of the added beta-carotene is 0.05-0.2%.
In the step 4), the ultrahigh pressure treatment is to fill the solution into a soft packaging bag, carry out vacuum heat sealing, place the packaging bag filled with the solution into a pressure vessel, and carry out ultrahigh pressure treatment by using liquid as a pressure transmission medium; the soft packaging bag can adopt a No. 6 polytetrafluoroethylene sealing bag; when in vacuum heat sealing, the vacuumizing time is 20-30 s, the heat sealing time is 1-5 s, and the heat sealing temperature is 50-90 ℃. The pressure of the ultrahigh pressure treatment is 600MPa, the pressure maintaining time is 30n, and the treatment temperature is 25 ℃.
Dissolving Chitosan (CS) in an acetic acid solution, dissolving separated Whey Protein (WPI) in water, mixing the mixture in proportion, adjusting the pH of the mixed solution to 5.0-7.0 to obtain a chitosan-separated whey protein (CS-WPI) mixed solution, adding a beta-carotene (beta-C) solution dissolved in medium-chain triglyceride into the mixed solution, and stirring the mixture uniformly to obtain a chitosan-separated whey protein solution encapsulated with beta-carotene; putting the solution into an ultrahigh pressure reaction kettle for ultrahigh pressure treatment to obtain chitosan-whey protein isolate gel (CS-WPI/beta-C) encapsulated with beta-carotene.
The invention utilizes chitosan and separated whey protein to form a double-layer emulsion wall material carrier, and then induces the emulsion to form gel through ultrahigh pressure, thereby having the advantages of overcoming the defects of the single liposome carrying wall material or the emulsion carrying system on the stability of embedded beta-C and avoiding the rapid damage in the digestive environment.
Drawings
Fig. 1 is an optical picture of a chitosan-whey protein isolate gel encapsulating beta-carotene in accordance with an embodiment of the present invention.
Figure 2 is an XRD pattern according to an embodiment of the present invention.
Detailed Description
The technical solution of the present invention is illustrated by specific examples below. It is to be understood that one or more method steps mentioned in the present invention do not exclude the presence of other method steps before or after the combination step or that other method steps may be inserted between the explicitly mentioned steps; it should also be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. Moreover, unless otherwise indicated, the numbering of the various method steps is merely a convenient tool for identifying the various method steps, and is not intended to limit the order in which the method steps are arranged or the scope of the invention in which the invention may be practiced, and changes or modifications in the relative relationship may be made without substantially changing the technical content.
In order to better understand the above technical solutions, exemplary embodiments of the present invention are described in more detail below. While exemplary embodiments of the invention have been shown, it should be understood that the invention may be embodied in various forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the invention to those skilled in the art.
The test materials adopted by the invention are all common commercial products and can be purchased in the market.
The room temperature of the invention is 25 ℃.
The invention will now be described with reference to specific examples, which are intended to be illustrative only and not to be limiting in any way.
The embodiment of the invention comprises the following steps:
1) dissolving the separated whey protein powder in water, and stirring for 2 hours to obtain a separated whey protein solution;
2) dissolving chitosan in 1% acetic acid solution, and stirring for 2h to obtain chitosan solution;
3) adding the chitosan solution into the whey protein isolate solution, mixing, and adjusting the pH of the solution to 6 to obtain a chitosan-whey protein isolate solution;
4) dissolving beta-carotene powder in medium-chain triglyceride, and carrying out ultrasonic treatment for 15min in the dark to obtain a beta-carotene solution;
5) adding the beta-carotene solution into the chitosan-isolated whey protein solution and mixing to obtain a chitosan-isolated whey protein solution encapsulated with beta-carotene;
6) carrying out ultrahigh pressure treatment on the chitosan-isolated whey protein solution encapsulated with the beta-carotene to obtain chitosan-isolated whey protein gel encapsulated with the beta-carotene;
in the step 3), the adding volume ratio of the chitosan solution to the whey protein isolate solution is 1: 4-5.5; preferably 1: 5.
In step 5), the volume ratio of the added water phase to the oil phase is 10: 1.
In the step 5), the concentration of the added beta-carotene is 0.05-0.2%.
In the step 6), the solution is filled into a soft packaging bag, vacuum heat sealing is carried out, then the packaging bag filled with the solution is placed into a pressure container, and ultrahigh pressure treatment is carried out by taking liquid as a pressure transmission medium.
The soft packaging bag adopts a polytetrafluoroethylene plastic bag; the vacuum pumping time is 20 s-30 s, the heat sealing time is 1 s-5 s, and the heat sealing temperature is 50-90 ℃.
In step 6), the pressure of the ultrahigh pressure treatment is 600MPa, the pressure maintaining time is 30n, and the treatment temperature is 25 ℃.
Example 1
Dissolving the isolated whey protein powder in ultrapure water to make the isolated whey protein concentration be 20% (w/v), stirring at 600r/min for 2h, refrigerating in a refrigerator, and standing overnight in water.
Dissolving chitosan in 1% acetic acid solution to make the concentration of CS solution be 0.8%, stirring at 600r/min for 2 hr, refrigerating in refrigerator, and standing overnight for water.
Mixing the chitosan solution and the whey protein isolate solution which are recovered to room temperature in a volume ratio of 1: 5, adjusting the pH of the mixed solution to 6, and stirring for 1h on a magnetic stirrer at a rotating speed of 600 r/min.
Dissolving beta-carotene powder in medium chain triglyceride, and performing ultrasonic treatment for 15min to make the concentration of beta-carotene 0.05%.
Mixing the chitosan-whey protein isolate solution and the beta-carotene solution in a volume ratio of 10: 1, and processing for 5min at a rotating speed of 600r/min by using a magnetic stirrer to obtain a fresh mixed solution.
Removing the foam on the surface of the solution, pouring the emulsion into a polytetrafluoroethylene bag, carrying out ultra-high pressure treatment after vacuum packaging by a vacuum packaging machine, wherein the ultra-high pressure parameter is set to be 600MPa, the pressure maintaining time is 30min, and the treatment temperature is 25 ℃.
Example 2
Dissolving the isolated whey protein powder in ultrapure water to make the isolated whey protein concentration be 20% (w/v), stirring at 600r/min for 2h, refrigerating in a refrigerator, and standing overnight in water.
Dissolving chitosan in 1% acetic acid solution to make the concentration of CS solution be 0.8%, stirring at 600r/min for 2 hr, refrigerating in refrigerator, and standing overnight for water.
Mixing the chitosan solution and the whey protein isolate solution at a volume ratio of 1: 5, adjusting pH of the mixed solution to 6, and stirring for 1h at a rotation speed of 600r/min on a magnetic stirrer.
Dissolving beta-carotene powder in medium chain triglyceride, and performing ultrasonic treatment for 15min to make the concentration of beta-carotene 0.10%.
Mixing the chitosan-whey protein isolate solution and the beta-carotene solution in a volume ratio of 10: 1, and processing for 5min at a rotating speed of 600r/min by using a magnetic stirrer to obtain a fresh mixed solution.
Removing the foam on the surface of the solution, pouring the emulsion into a polytetrafluoroethylene bag, carrying out ultra-high pressure treatment after vacuum packaging by a vacuum packaging machine, wherein the ultra-high pressure parameter is set to be 600MPa, the pressure maintaining time is 30min, and the treatment temperature is 25 ℃.
Example 3
Dissolving the isolated whey protein powder in ultrapure water to make the isolated whey protein concentration be 20% (w/v), stirring at 600r/min for 2h, refrigerating in a refrigerator, and standing overnight in water.
Dissolving chitosan in 1% acetic acid solution to make the concentration of CS solution be 0.8%, stirring at 600r/min for 2 hr, refrigerating in refrigerator, and standing overnight for water.
The volume ratio of the components is 1: 5, mixing the chitosan solution which is recovered to the room temperature with the whey protein isolate solution, adjusting the pH of the mixed solution to 6, and stirring the mixed solution on a magnetic stirrer at the rotating speed of 600r/min for 1 hour.
Dissolving beta-carotene powder in medium chain triglyceride, and performing ultrasonic treatment for 15min to make the concentration of beta-carotene 0.15%.
Mixing the chitosan-whey protein isolate solution and the beta-carotene solution in a volume ratio of 10: 1, and processing for 5min at a rotating speed of 600r/min by using a magnetic stirrer to obtain a fresh mixed solution.
Removing the foam on the surface of the solution, pouring the emulsion into a polytetrafluoroethylene bag, carrying out ultra-high pressure treatment after vacuum packaging by a vacuum packaging machine, wherein the ultra-high pressure parameter is set to be 600MPa, the pressure maintaining time is 30min, and the treatment temperature is 25 ℃.
Example 4
Dissolving the isolated whey protein powder in ultrapure water to make the isolated whey protein concentration be 20% (w/v), stirring at 600r/min for 2h, refrigerating in a refrigerator, and standing overnight in water.
Dissolving chitosan in 1% acetic acid solution to make the concentration of CS solution be 0.8%, stirring at 600r/min for 2 hr, refrigerating in refrigerator, and standing overnight for water.
Mixing the chitosan solution and the whey protein isolate solution at a volume ratio of 1: 5, adjusting pH of the mixed solution to 6, and stirring for 1h at a rotation speed of 600r/min on a magnetic stirrer.
Dissolving beta-carotene powder in medium chain triglyceride, and performing ultrasonic treatment for 15min to make the concentration of beta-carotene 0.20%.
Mixing the chitosan-whey protein isolate solution and the beta-carotene solution in a volume ratio of 10: 1, and processing for 5min at a rotating speed of 600r/min by using a magnetic stirrer to obtain a fresh mixed solution.
Removing the foam on the surface of the solution, pouring the emulsion into a polytetrafluoroethylene bag, carrying out ultra-high pressure treatment after vacuum packaging by a vacuum packaging machine, wherein the ultra-high pressure parameter is set to be 600MPa, the pressure maintaining time is 30min, and the treatment temperature is 25 ℃.
Optical pictures of the beta-carotene encapsulated chitosan-whey protein isolate gels prepared in examples 1-4 are shown in FIG. 1.
Test examples
1. The effective encapsulation concentration and encapsulation efficiency of the β -carotene encapsulated chitosan-isolated whey protein gels obtained in examples 1 to 4 were measured by a microplate reader, and the results are shown in table 1.
2. XRD measurements were carried out on the chitosan-isolated whey protein gels encapsulating β -carotene obtained in examples 1 to 4, respectively: after freezing and drying the samples, taking a proper amount of fine and uniform powder, and analyzing each sample by using an X-ray diffractometer, wherein the scanning speed is 15 degrees/second, and the scanning range of the diffraction angle (2 theta) is 5-50 degrees. The results are shown in FIG. 2.
Table 1: absorbance and embedding efficiency of CS-WPI for embedding beta-carotene with different concentrations
| Initial addition concentration (g/L) | Effective envelope concentration (μ g/mL) | Encapsulation efficiency (EE%) |
| 0.05％ | 6.83 | 81.95％ |
| 0.10％ | 8.95 | 53.71％ |
| 0.15％ | 11.64 | 46.56％ |
| 0.20％ | 12.37 | 37.11％ |
In summary, according to the method of the embodiment of the present invention, the ultrahigh pressure technology is adopted, the chitosan-isolated whey protein solution loaded with β -carotene is gelatinized by a physical method at room temperature, the gelatinization process is mild, the loss of β -carotene activity is avoided, and the effective loading of β -carotene can be realized by adding different concentrations of β -carotene-chitosan-isolated whey protein gel at the beginning of the ultrahigh pressure treatment with different effective encapsulation concentrations and encapsulation efficiencies.
In the description herein, references to the description of the term "one embodiment," "some embodiments," "an example," "a specific example" or "some examples" or the like are intended to mean that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the invention. In this specification, the schematic representations of the terms used above should not be understood to necessarily refer to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples. Furthermore, various embodiments or examples described in this specification can be combined and combined by those skilled in the art.
Although embodiments of the present invention have been shown and described above, it is understood that the above embodiments are exemplary and should not be construed as limiting the present invention, and that variations, modifications, substitutions and alterations can be made to the above embodiments by those of ordinary skill in the art within the scope of the present invention.
Claims (10)
1. A preparation method of chitosan-whey protein isolate gel for encapsulating beta-carotene is characterized by comprising the following steps:
1) preparation of the aqueous phase: firstly, dissolving chitosan in an acetic acid solution to obtain a chitosan solution; dissolving the isolated whey protein in water to obtain an isolated whey protein solution; adding the chitosan solution into the whey protein isolate solution, mixing, and adjusting the pH value of the solution to 6 to obtain a chitosan-whey protein isolate solution;
2) preparation of oil phase: dissolving beta-carotene powder in medium-chain triglyceride, and carrying out ultrasonic treatment in the dark to obtain a beta-carotene solution;
3) mixing water and oil phases: adding the beta-carotene solution into the chitosan-whey protein isolate solution, and mixing to obtain a chitosan-whey protein isolate solution encapsulated with beta-carotene;
4) preparing gel under ultrahigh pressure: and carrying out ultrahigh pressure treatment on the chitosan-isolated whey protein solution encapsulated with the beta-carotene to obtain chitosan-isolated whey protein gel encapsulated with the beta-carotene.
2. The method for preparing chitosan-whey protein isolate gel encapsulating β -carotene according to claim 1, wherein in step 1), said acetic acid solution is used as 1% acetic acid solution by mass fraction.
3. The method for preparing chitosan-whey protein isolate gel encapsulating β -carotene according to claim 1, wherein in step 1), the chitosan solution may have a mass percentage concentration of 0.8%; the mass percentage concentration of the whey protein isolate solution can be 20%, and the chitosan solution is added into the whey protein isolate solution and mixed, and then the solution is placed in a refrigerator at 4 ℃ overnight so as to be fully hydrated.
4. The method for preparing the chitosan-whey protein isolate gel encapsulating beta-carotene according to claim 1, wherein in the step 1), the volume ratio of the chitosan solution to the whey protein isolate solution is 1: 4-5.5; preferably 1: 5; after the pH value of the solution is adjusted to 6, the chitosan-whey protein isolate solution can be placed on a magnetic stirrer to be stirred for 1 hour, so that the chitosan-whey protein isolate solution and the magnetic stirrer are completely mixed.
5. The method of claim 1, wherein in step 2), the beta-carotene powder is dissolved in the medium-chain triglyceride to provide beta-C concentrations of 0.05%, 0.10%, 0.15%, and 0.2%.
6. The method of claim 1, wherein the ratio of the beta-carotene solution to the chitosan-whey protein isolate solution in step 3) is 10: 1.
7. The method for preparing chitosan-whey protein isolate gel encapsulating β -carotene according to claim 1, wherein in step 3), the mixture is processed for 5min at a rotation speed of 600r/min by a magnetic stirrer; the concentration of the added beta-carotene is 0.05-0.2%.
8. The method for preparing chitosan-whey protein isolate gel encapsulating β -carotene according to claim 1, wherein in step 4), the ultra-high pressure treatment is carried out by filling the solution into a soft packaging bag, vacuum heat-sealing, placing the packaging bag filled with the solution into a pressure vessel, and carrying out the ultra-high pressure treatment with the liquid as a pressure transmission medium; the soft packaging bag can adopt a No. 6 polytetrafluoroethylene sealing bag.
9. The method for preparing chitosan-whey protein isolate gel encapsulating β -carotene according to claim 1, wherein in step 4), the vacuum-heat-sealing is performed for 20s to 30s, the heat-sealing is performed for 1s to 5s, and the heat-sealing temperature is 50 ℃ to 90 ℃.
10. The method for preparing chitosan-whey protein isolate gel encapsulating β -carotene according to claim 1, wherein in the step 4), the pressure of the ultra-high pressure treatment is 600MPa, the dwell time is 30n, and the treatment temperature is 25 ℃.
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