CN113116820B - Astaxanthin pickering emulsion and preparation method thereof - Google Patents
Astaxanthin pickering emulsion and preparation method thereof Download PDFInfo
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- CN113116820B CN113116820B CN202110485240.5A CN202110485240A CN113116820B CN 113116820 B CN113116820 B CN 113116820B CN 202110485240 A CN202110485240 A CN 202110485240A CN 113116820 B CN113116820 B CN 113116820B
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- astaxanthin
- chitosan
- guar gum
- emulsion
- pickering emulsion
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- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
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- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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Abstract
The invention belongs to the technical field of preparation of active substance preparations, and particularly relates to an astaxanthin pickering emulsion and a preparation method thereof. Comprises the steps of preparing a chitosan-guar gum nanoparticle emulsifier and preparing astaxanthin pickering emulsion by taking nanoparticles as the emulsifier; the preparation method comprises the following steps of preparing a chitosan-guar gum nanoparticle emulsifier, namely slowly dripping a chitosan acetic acid solution into a guar gum solution under the condition of high-speed stirring, continuously stirring, and adjusting the pH value to 3-6 to obtain the chitosan-guar gum nanoparticle emulsifier. The preparation process is mild, the raw materials are green and safe, and no surfactant is used; the prepared pickering emulsion has high physical and chemical stability, has no obvious phase separation phenomenon after being stored for 2 months, has obviously lower content of lipid oxidation products after being subjected to accelerated oxidation for one month at 37 ℃, and has higher astaxanthin retention rate than the traditional emulsion.
Description
Technical Field
The invention belongs to the technical field of preparation of active substance preparations, and particularly relates to an astaxanthin pickering emulsion and a preparation method thereof.
Background
Astaxanthin is a carotenoid widely found in fish, shrimp, crab, phaffia rhodozyma and haematococcus pluvialis. Due to its antioxidant properties, it is widely used in cosmetics, health products and aquaculture industries. Astaxanthin is a strong singlet oxygen quencher and a high-efficiency scavenger, and plays an important role in the antioxidant of superoxide anions and hydroxyl radicals. Regular administration of astaxanthin has many health benefits, such as a unique role in boosting and regulating the immune system, reducing cardiovascular disease, delaying aging, enhancing resistance and preventing and treating cancer. Therefore, the astaxanthin has great market prospect as a novel functional active factor. However, the hydrophobic structure of astaxanthin causes extremely poor water solubility, and the application of astaxanthin in aqueous medium is limited. In addition, the highly unsaturated conjugated double chains in the molecular structure of astaxanthin are easily oxidized and degraded under the conditions of illumination, oxygen, high temperature and ultraviolet rays, so that the activity function of the astaxanthin is lost, the stability of a sample is influenced, and the application and consumption of astaxanthin are greatly limited. Therefore, how to improve the water solubility and stability of astaxanthin is a key technical problem in the market of large-scale natural astaxanthin products.
Emulsions are generally stable systems consisting of oil, water, surfactants, co-surfactants, electrolytes, and the like. The system in which the dispersed phase is oil and the dispersion medium is water is called O/W type emulsion, and vice versa is called W/O type microemulsion. Can be used as a carrier for encapsulating and delivering fat-soluble active substances and is widely applied to the fields of food, cosmetics, medicines and the like. At present, various emulsion types exist, and materials for preparing the emulsion are various, and how to select proper emulsion types and materials, the prepared astaxanthin emulsion system which is green, safe, good in stability and simple in process is a problem to be overcome in the field.
Emulsion systems stabilized by solid particles instead of surfactants are known as "Pickering emulsions" (Pickering emulsions). The stabilizing mechanism is mainly that solid particles are irreversibly adsorbed on an oil-water interface and form a monolayer or multilayer film of the solid particles so as to stabilize the emulsion. By selecting the size of the stabilizing particles, the thickness of the surface layer can be easily controlled and the effective density of the emulsion droplets can be adjusted to improve stability. However, the surfactants used in the traditional emulsions often have certain side effects, and the use of the traditional emulsions in the food field is limited to a certain extent. Therefore, the development of "green tag" Pickering emulsion stabilizers of natural origin is becoming increasingly important. Like surfactants, inorganic particles often cause irritation and are not suitable for use in food.
Chitosan is a water-insoluble polysaccharide at neutral pH, consisting of repeating units of β - (1 → 4) D-glucosamine and N-acetylglucosamine. At pH <6.5, the amino group will be protonated, allowing chitosan to interact with negatively charged compounds. The chitosan with common size can not really show the excellent performance and biological activity of the chitosan, and the introduction of the nanotechnology along with the development of the nanotechnology is an effective way. Chitosan (CS) has been used in many countries as a dietary food additive and food processing aid with antibacterial, mucoadhesive and gelling properties. And has cholesterol lowering activity and obesity controlling effect. Guar gum is a nonionic galactomannan extracted from endosperm of guar of Leguminosae, and guar gum and its derivatives have good water solubility. And Guar Gum (GG) is a hydrophilic polysaccharide for oral controlled delivery, having sensitivity to large intestine microbial degradation and sustained drug release properties.
Many researchers report the preparation method of astaxanthin emulsion at present, but some defects still exist. For example, chinese patent 201410665629.8 discloses a high stability astaxanthin ester self-microemulsion and a preparation method thereof. However, in the preparation method, the emulsifier such as tween and the emulsifying assistant such as ethanol, 1, 2-propylene glycol, 1, 3-propylene glycol and glycerol are required to be added, which can not completely meet the current green and safe consumption requirements of the market for food and health care products. The chinese patent 201610004295.9 provides a method for preparing astaxanthin derivatives which can be dissolved in water by chemical transformation, but the method has complex operation process and high cost, and the chemical transformation method is not favorable for the development of green food. The Chinese patent 201911110978.2 provides an astaxanthin emulsion and a preparation method thereof, but the astaxanthin content of the astaxanthin emulsion prepared by the method is low and only accounts for about 3% of the total mass, and the storage stability of the emulsion is not considered. Therefore, how to obtain an astaxanthin emulsion with high astaxanthin retention rate and strong stability is a problem which needs to be solved urgently at present.
Disclosure of the invention
The technical problem to be solved by the invention is that a plurality of researchers report preparation methods of astaxanthin emulsion at present, but the defects of more addition of additives, incapability of meeting green food, low astaxanthin content and the like still exist, and research needs to be carried out on obtaining the astaxanthin emulsion with high astaxanthin retention rate and strong stability.
In order to solve the problems, the invention provides an astaxanthin pickering emulsion and a preparation method thereof. The preparation process is mild, the raw materials are green and safe, and no surfactant is used. The pickering emulsion prepared by the method has high physical and chemical stability, and has no obvious phase separation phenomenon after being stored for 2 months. The content of lipid oxidation products is obviously lower than that of Tween 80 emulsion after accelerated oxidation for one month at 37 ℃. The astaxanthin retention rate is significantly higher than that of the traditional emulsion.
In order to achieve the purpose, the invention is specifically realized by the following technical scheme: a preparation method of astaxanthin pickering emulsion comprises the steps of preparing chitosan-guar gum nanoparticle emulsifier, and preparing the astaxanthin pickering emulsion by taking nanoparticles as the emulsifier; the preparation method comprises the following steps of preparing a chitosan-guar gum nanoparticle emulsifier, namely slowly dripping a chitosan acetic acid solution into a guar gum solution under the condition of high-speed stirring, continuously stirring, and adjusting the pH value to 3-6 to obtain the chitosan-guar gum nanoparticle emulsifier. At acidic pH, the amino groups of chitosan are protonated, thus making it water-soluble and positively charged. This property enables it to interact with negatively charged GG through intermolecular electrostatic interactions, thereby forming a polyelectrolyte complex. When the pH value is 3-6, the chitosan solution has positive charges, the guar gum solution has negative charges, the electrostatic interaction of the chitosan solution and the guar gum solution is strongest, and the emulsification effect is best.
Further, mixing the chitosan-guar gum nanoparticle emulsifier with the astaxanthin oil solution (oil phase), and then shearing at high speed to obtain the astaxanthin pickering emulsion. Wherein the astaxanthin is a mixture of natural free astaxanthin, astaxanthin monoester and astaxanthin diester derived from one or more of Haematococcus pluvialis, Phaffia rhodozyma, Euphausia superba, Penaeus vannamei and Salmon. At this time, CS/GG nano-particles are wrapped around oil droplets to form a rigid and stable emulsifying system.
Furthermore, the addition amount of the chitosan in the chitosan acetic acid solution is 0.1-2% of the acetic acid aqueous solution.
Further, the addition amount of the guar gum in the guar gum solution is 0.05-1% of the ultrapure water.
Further, the rotation speed of mixing and stirring the chitosan acetic acid solution and the guar gum solution is 400-1200rpm, and the time is 0.5-24 h. If the stirring time is too short, the polyelectrolyte complex is not completely formed; excessive stirring may cause the complex to break up.
Further, in the mixed stirring solution of chitosan and guar gum, the mass ratio of chitosan to guar gum is (5:1) - (1: 2). The proportion is determined by optimizing and changing the concentration of the chitosan under the condition of keeping the concentration of the guar gum unchanged. Too high a chitosan solution viscosity is too high to facilitate the formation of chitosan and guar gum complexes, and too low a chitosan concentration results in too low a concentration of CS/GG nanoparticles formed to form a stable Pickering emulsion.
Further, the astaxanthin oil solution is a mixture of astaxanthin and animal and vegetable oil. The animal and vegetable oil is corn oil, soybean oil, sunflower seed oil, linseed oil, rapeseed oil, olive oil, rice oil, peanut oil, fish oil and algae oil.
Further, the content of the astaxanthin accounts for 1-99% of the total oil mass. The animal and vegetable oil is used as a primary carrier of astaxanthin, so that the bioavailability and the effective absorption rate of the astaxanthin are improved. In addition, the high concentration of astaxanthin sample leads to a reduction of the emulsifying effect of the Pickering emulsion.
Further, the oil phase accounts for 20-70% of the total mass of the Pickering emulsion. The Pickering emulsion formed by too low oil fraction is easy to have phase separation phenomenon, the viscosity of the emulsion is low, the gel-like characteristic is difficult to form, and the stability of the emulsion is poor; too high an oil fraction, however, may result in a portion of the grease being difficult to encapsulate by the CS/GG nanoparticles. The result was a mixture of fat and oil with an emulsion.
Further, the rotating speed of the high-speed shearing is 6000-25000rpm, and the time is 1-3 min. Too low a shear rate is insufficient to rapidly form a Pickering emulsion; and the rotation speed is too high, the time is too long, the emulsion is overheated, and the emulsion breaking phenomenon occurs.
An astaxanthin Pickering emulsion prepared by the method. After 2 months of storage, there was no significant phase separation. The content of lipid oxidation products is obviously lower than that of Tween 80 emulsion after accelerated oxidation for one month at 37 ℃. The astaxanthin retention rate is significantly higher than that of the traditional emulsion.
The invention has the following beneficial effects:
(1) the preparation method of the astaxanthin Pickering emulsion provided by the invention is simple and efficient in preparation process, green and safe in formula, and does not add a surfactant, so that an astaxanthin emulsion system is expanded.
(2) The invention provides an astaxanthin emulsion product with high stability, which has high physical and chemical stability, no obvious phase separation phenomenon after being stored for 2 months, and lower content of lipid oxidation products. The astaxanthin retention rate is obviously superior to that of the traditional emulsion.
(3) The astaxanthin Pickering emulsion can be applied to the fields of food, cosmetics and medicines.
Drawings
FIG. 1 is an optical microscope image of an astaxanthin Pickering emulsion with an oil fraction of 0.6 in example 1;
FIG. 2 is a diagram showing an example of an emulsion of astaxanthin stabilized with Tween 80 in example 2 stored at 4 ℃ in the dark for 30 days;
FIG. 3 is a diagram showing an example of the astaxanthin Pickering emulsion stored at 4 ℃ for 30 days in the dark in example 2;
FIG. 4 shows the lipid oxidation stability of the CS/GG nanoparticle-stabilized astaxanthin Pickering emulsion and the Tween 80-stabilized astaxanthin emulsion in example 4 at 37 ℃ for 30 days;
FIG. 5 shows the astaxanthin stability of the CS/GG nanoparticle-stabilized astaxanthin Pickering emulsion and the Tween 80-stabilized astaxanthin emulsion in example 4 at 37 ℃ for 30 days.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention are clearly and completely described below, and it is obvious that the described embodiments are a part of the embodiments of the present invention, but not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1:
a preparation method of astaxanthin pickering emulsion comprises the following steps:
0.8g of Chitosan (CS) was weighed and dissolved in 100ml of glacial acetic acid solution to obtain a 0.8% CS solution. 0.5g of Guar Gum (GG) was weighed out and dissolved in 100ml of ultrapure water to obtain a 0.5% GG solution. The CS solution was then added dropwise to the GG solution while stirring was continued for 4h (600 rpm). And adjusting the pH value of the mixed solution to 5.0 to obtain a chitosan-guar gum nanoparticle solution based on polyelectrolyte complexation. According to oil fraction (
0.3, 0.4, 0.5, 0.6, 0.7 and 0.8), appropriate masses of CS/GG nanoparticle solution and astaxanthin oil (astaxanthin and linseed oil mixture) were weighed, put into a 20ml serum bottle, and sheared at high speed of 15000rpm for 1.5min to obtain chitosan-guar gum PiA cocking emulsion. Microscopic morphology was observed by optical microscopy and emulsion stability was investigated after 2 months of low temperature storage at 4 ℃.
Emulsification index:
the determination of the emulsification index (CI%) is used as an indicator of the stability of the emulsion to separation. It is calculated according to the following equation (Li et al, 2018; Mwangi et al, 2016):
wherein HsIs the height of the serum layer, HtIs the total height of the emulsion.
The microscopic morphology of the Pickering emulsion stabilized by chitosan-guar gum nanoparticles was observed by an optical microscope, which revealed that the size of the oil droplets of the emulsion increased with the increase of the oil volume fraction. This may be due to a reduction in the total amount of nanoparticles adsorbed at the oil/water interface, resulting in the formation of larger sized droplets. As shown in fig. 1, when the oil content is increased to 0.6, the oil droplets of the astaxanthin Pickering emulsion are large in size, uniform in distribution and strong in gel property among the oil droplets. However, as the oil content continues to increase to 0.7 and above, oil fouling and droplet flocculation can occur and the loss of sphericity begins to deform. This indicates that at too high an oil fraction, the number of nanoparticles available is not sufficient to stabilize the Pickering droplets to prevent coalescence.
Comparing the different oil fractions (
0.3, 0.4, 0.5, 0.6 and 0.7) of the emulsion of astaxanthin Pickering stored at 4 ℃ in the absence of light for 2 months. It can be seen that the separation speed is higher in the Pickering emulsion formula with lower astaxanthin oil fraction content, and the CI% value is higher, which indicates that the storage stability is poor. However, when the oil content was increased from 0.2 to 0.5, the CI% value dropped significantly from 52.05% to 10.89%, which means that with increasing oil content a more stable Pickering emulsion was formed. In addition, has high oil content
The formulations of (a) have no apparent gravitational phase separation or a very low emulsification index (CI% ═ 2.56%) over a 2 month test storage period, indicating their high stability.
Example 2:
a preparation method of astaxanthin pickering emulsion comprises the following steps:
1g of Chitosan (CS) was weighed and dissolved in 100ml of glacial acetic acid solution to obtain a 1% CS solution. 0.4g of Guar Gum (GG) was weighed out and dissolved in 100ml of ultrapure water to obtain a 0.4% GG solution. The CS solution was then added dropwise to the GG solution while stirring was continued for 2h (1100 rpm). And adjusting the pH value of the mixed solution to 4.5 to obtain a chitosan-guar gum nanoparticle solution based on polyelectrolyte complexation. 6g of CS/GG nanoparticle solution and 9g of astaxanthin oil (0.6g astaxanthin, 8.4g fish oil) were weighed into a 20ml serum bottle and sheared at 20000rpm for 1min at high speed to give chitosan-guar gum astaxanthin Pickering emulsion. And replacing the CS/GG nano-particle solution in the method with a Tween 80 solution with the same concentration to prepare the traditional emulsion of the astaxanthin serving as a control sample.
FIG. 2 is a physical diagram of an astaxanthin emulsion stabilized by Tween 80 stored at 4 ℃ for 30 days. FIG. 3 is a graph of chitosan-guar gum stabilized astaxanthin Pickering emulsion as a real object stored at 4 ℃ for 30 days. In figure 2 it is shown that tween 80 stabilized astaxanthin emulsion showed significant phase separation upon storage for 1 day with an emulsification index of 11.63%. After 30 days of storage, the emulsion is obviously broken, and a large amount of astaxanthin oil is floated at the last time. The astaxanthin Pickering emulsion with the stable chitosan-guar gum shown in figure 3 has no phase separation phenomenon after being stored at 4 ℃ for 30 ℃, and the emulsion is still at the bottom of a serum bottle and does not flow backwards when being inverted, which shows that the emulsion has good emulsion gel property, and the astaxanthin emulsion is more stable due to the close accumulation of liquid drops. Therefore, the stability of the astaxanthin Pickering emulsion prepared by the method is obviously superior to that of a traditional emulsion stabilized by Tween 80.
Example 3:
a preparation method of astaxanthin pickering emulsion comprises the following steps:
2g of Chitosan (CS) was weighed and dissolved in 100ml of glacial acetic acid solution to obtain a 2% CS solution. 0.5g of Guar Gum (GG) was weighed out and dissolved in 100ml of ultrapure water to obtain a 0.5% GG solution. Then, the CS solution was added dropwise to the GG solution while stirring was continued for 1h (1000 rpm). And adjusting the pH value of the mixed solution to 5.5 to obtain a chitosan-guar gum nanoparticle solution based on polyelectrolyte complexation. 5g of CS/GG nanoparticle solution and 5g of astaxanthin oil (0.1g astaxanthin, 4.9g corn oil) were weighed into a 20ml serum bottle and sheared at 25000rpm for 2min at high speed to give a chitosan-guar gum astaxanthin Pickering emulsion.
FIG. 4 shows the content changes of hydroperoxide (POV) and Malondialdehyde (MDA) in the chitosan-guar gum-stabilized astaxanthin Pickering emulsion and the tween 80-stabilized astaxanthin emulsion after 30 days of constant-temperature accelerated oxidation treatment at 37 ℃. In an accelerated oxidation experiment at 37 ℃ for 30 days, the lipid hydroperoxide and MDA in the Tween 80 stabilized emulsion increased from the initial 3.06. + -. 0.21mmol/kg emulsion and 10.55. + -. 1.03. mu. mol/kg emulsion to the final 7.04. + -. 0.65mmol/kg emulsion and 255.11. + -. 24.31. mu. mol/kg emulsion. In the CS/GG nanoparticle stabilized Pickering emulsion, the lipid hydroperoxide and MDA increased from the initial 2.90 + -0.34 mmol/kg emulsion and 10.48 + -0.92. mu. mol/kg emulsion to the final 4.76 + -0.41 mmol/kg emulsion and 160.43 + -13.85. mu. mol/kg emulsion. Fig. 5 shows the stability of astaxanthin after one month of accelerated oxidation of the two emulsions at 37 ℃, and the results show that the retention rate of astaxanthin in the CS/GG nanoparticle-stabilized astaxanthin Pickering emulsion is as high as 84.81 ± 3.96%, which is significantly better than that of tween 80-stabilized astaxanthin emulsion (55.90 ± 2.14%). In general, the contents of POV and MDA in the Pickering emulsion stabilized by CS/GG nanoparticles are obviously lower than those of a Tween 80 emulsion, and the retention rate of astaxanthin is higher. It is shown that CS/GG nanoparticles coating oil droplets can prevent lipid oxidation to reduce astaxanthin degradation more than Tween 80 coating oil droplets. This difference may be due to the emulsion droplets covered by the CS/GG nanoparticles being positively charged. Since in the CS/GG nanoparticles used in the formulation of the astaxanthin Pickering emulsion the amine groups of CS are not all attached to the carboxyl groups of GG, some of the amine groups remain free. Therefore, the method plays a role in repelling transition metal ions in the lipid oxidation process and prevents the lipid oxidation promotion effect of the metal ions. Furthermore, another reason why the oxidative stability of the astaxanthin Pickering emulsion is higher compared to the tween 80 emulsion is probably due to the fact that the Pickering emulsion has certain gel properties, forming a more dense particle-based interface layer than in the tween 80 emulsion, thus providing a physical barrier. Preventing the pro-oxidant from interacting with the internal lipids, thereby reducing the oxidative degradation of lipids.
In conclusion, the preparation method of the astaxanthin Pickering emulsion provided by the invention is mild in preparation conditions, green and safe in raw materials and simple in process. The astaxanthin Pickering emulsion prepared by the method has good physical and chemical stability, and does not have obvious phase separation phenomenon after standing for 2 months at 4 ℃. The content of lipid oxidation products is obviously lower than that of Tween 80 emulsion after accelerated oxidation for one month at 37 ℃. The retention rate of the astaxanthin is obviously higher than that of the traditional emulsion, and the astaxanthin can be used in the industries of food, cosmetics, medicines and the like.
Finally, it should be noted that the above embodiments describe specific embodiments of the present invention, but do not limit the present invention; it will be understood by those skilled in the art that these are by way of example only and that the scope of the invention is defined by the appended claims. All changes, modifications and equivalents that come within the meaning and range of equivalency of the claims are to be embraced within their scope.
Claims (8)
1. A preparation method of astaxanthin pickering emulsion is characterized by comprising the following steps: comprises the steps of preparing a chitosan-guar gum nanoparticle emulsifier and preparing astaxanthin pickering emulsion by taking nanoparticles as the emulsifier; the preparation method comprises the following steps of preparing a chitosan-guar gum nanoparticle emulsifier, namely slowly dripping a chitosan acetic acid solution into a guar gum solution under the condition of high-speed stirring, continuously stirring, and adjusting the pH value to 3-6 to obtain the chitosan-guar gum nanoparticle emulsifier;
the rotation speed of mixing and stirring the chitosan acetic acid solution and the guar gum solution is 400-1200rpm, and the time is 0.5-24 h;
the mass ratio of the chitosan to the guar gum is (5:1) - (1: 2).
2. The method of preparing an astaxanthin pickering emulsion as claimed in claim 1, characterized in that: mixing the chitosan-guar gum nanoparticle emulsifier with the astaxanthin oil solution, and then shearing at a high speed to obtain the astaxanthin pickering emulsion.
3. The method of preparing an astaxanthin pickering emulsion as claimed in claim 1, characterized in that: the addition amount of chitosan in the chitosan acetic acid solution is 0.1-2% of the acetic acid aqueous solution.
4. The method of preparing an astaxanthin pickering emulsion as claimed in claim 1, characterized in that: the addition amount of the guar gum in the guar gum solution is 0.05-1% of the ultrapure water.
5. The method of preparing an astaxanthin pickering emulsion as claimed in claim 2, characterized in that: the astaxanthin oil solution is a mixture of astaxanthin and animal and vegetable oil; the content of the astaxanthin accounts for 1-99% of the total oil mass.
6. The method of preparing an astaxanthin pickering emulsion as claimed in claim 2, characterized in that: the astaxanthin oil solution accounts for 20-70% of the total mass of the pickering emulsion.
7. A method of preparing an astaxanthin pickering emulsion as claimed in claim 2, characterized in that: the high-speed shearing rotation speed is 6000-25000rpm, and the time is 1-3 min.
8. An astaxanthin pickering emulsion prepared by the method of claim 1.
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