CN107874257B - Preparation method of euphausia superba oil nanoparticles with high embedding rate and small particle size - Google Patents
Preparation method of euphausia superba oil nanoparticles with high embedding rate and small particle size Download PDFInfo
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- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 claims abstract description 10
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Images
Classifications
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- 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
- A23L33/115—Fatty acids or derivatives thereof; Fats or oils
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23P—SHAPING OR WORKING OF FOODSTUFFS, NOT FULLY COVERED BY A SINGLE OTHER SUBCLASS
- A23P10/00—Shaping or working of foodstuffs characterised by the products
- A23P10/30—Encapsulation of particles, e.g. foodstuff additives
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
Abstract
The invention provides a preparation method of euphausia superba oil nanoparticles with high embedding rate and small particle size, which adopts the following specific technical scheme: mixing sodium caseinate, arabic gum and tween-20 as stabilizer, and dissolving in phosphate buffer solution to obtain water phase; dissolving antarctic krill oil in a mixed solvent of dichloromethane and acetone to obtain an oil phase; pouring the oil phase into the water phase to obtain a coarse nano emulsion; homogenizing the coarse nano emulsion, homogenizing under high pressure, and removing the organic solvent in the emulsion under vacuum state to obtain nano emulsion; and (3) carrying out vacuum drying on the obtained nanoemulsion to obtain the small-particle-size euphausia superba oil nanoparticles with high embedding rate. According to the invention, by regulating and controlling the circulating pressure in the high-pressure homogenization process of the emulsion, the form uniformity and the embedding rate of the euphausia superba oil nanoparticles are improved, and the particle size of the nanoparticles is reduced.
Description
Technical Field
The invention relates to the field of preparation of euphausia superba oil nanoparticles, in particular to a preparation method for preparing euphausia superba oil nanoparticles with high uniformity, high embedding rate and low nanoparticle size.
Background
Antarctic krill is the largest single biological resource on earth and has huge reserves. The antarctic krill oil mainly comprises glyceride, phospholipid, free fatty acid, sterol and astaxanthin. The Antarctic krill oil is rich in phospholipid type Omega-3 polyunsaturated fatty acids and astaxanthin, and various biological activities are endowed by the characteristics of the astaxanthin, but the oxidation stability of the phospholipid type polyunsaturated fatty acids is obviously lower than that of neutral oil such as glyceride, sterol and the like; the free fatty acid has an oxidation promoting effect, can reduce the oxidation stability of other coexisting lipid components, and accelerates the degradation of the grease; moreover, phospholipid swells when meeting water, so that the solubility is poor, and the application of the phospholipid in food is influenced.
In recent years, nanoparticle technology has attracted attention. Compared with the common microcapsule, the nano-particles can improve the physical and chemical properties of the embedded object such as stability, solubility, dispersity and the like, and also show unique advantages in the aspects of increasing the absorption of the embedded object, controlling the release of the embedded object and the like. At present, there are various methods for preparing nanoparticles, such as: high pressure homogenization, microemulsion, nano-precipitation, emulsion solvent evaporation, and the like. The high-pressure homogenization method is the most commonly used method for preparing nanoparticles at present, and is divided into a hot emulsion homogenization method and a cold emulsion homogenization method, wherein a core material and a surfactant are inevitably required to be heated at a high temperature to form a molten state in the preparation process, so that heat-sensitive substances such as astaxanthin, unsaturated fatty acid and the like contained in the core material are influenced in the preparation process, and the content of the heat-sensitive substances is reduced. Also, the microemulsion method requires heating of the substance during the preparation process, thereby reducing the content of the heat-sensitive substance. Although the nano precipitation method does not generate high temperature in the preparation process as a method for preparing nanoparticles, the precipitation is generated in the preparation process, and the generated precipitation cannot be homogenized under high pressure, so that the particle size and uniformity of the nanoparticles cannot be improved, and the method is less used in the actual production. In the emulsion solvent evaporation method, a solvent which is mutually soluble with water is used as a water phase, an organic solvent which is not mutually soluble with water is used as an oil phase, nano-scale emulsion droplets are formed when the water phase is contacted with the oil phase, and then the nano-scale emulsion droplets are solidified and separated to obtain nano-particles; because the high temperature can not be generated in the preparation process, the thermosensitive substances can be better retained, so the preparation method can be applied to the preparation of the antarctic krill oil nanoparticles, and the thermosensitive substances such as astaxanthin, polyunsaturated fatty acid and the like in the krill oil can be better preserved.
The early-stage investigation finds that in the process of preparing the nanoparticles, predecessors directly perform high-pressure homogenization under a certain pressure to obtain the nanoemulsion after preliminarily mixing an oil phase and a water phase to obtain a coarse emulsion, so that the prepared nanoparticles have the problems of nonuniform particle size, low embedding rate, large particle size and the like.
Disclosure of Invention
In order to solve the problems, the invention provides a preparation method of euphausia superba oil nanoparticles with high embedding rate and small particle size, which comprises four steps of preparation of water phase, preparation of oil phase, high-pressure homogenization of emulsion, drying of nanoparticles and the like, and the specific technical scheme is as follows:
s1, preparing a water phase: mixing the following raw materials in percentage by weight to prepare a water phase:
the method comprises the following steps of taking a mixture of sodium caseinate, Arabic gum and Tween-20 as a stabilizer, wherein the stabilizer accounts for 1-3% of the total mass of a water phase, the sodium caseinate accounts for 60-70% of the total mass of the stabilizer, the Arabic gum accounts for 5-10% of the total mass of the stabilizer, and the Tween-20 accounts for 20-25% of the total mass of the stabilizer;
dissolving the stabilizer in a phosphate buffer solution with the pH value of 7, and stirring at 20-70 ℃ for 0.5-2 h to obtain a water phase; preferably, the stirring temperature is 40-60 ℃, and the stirring time is 0.5-1 h;
in a preferable mode, the stabilizer accounts for 2-3% of the total mass of the water phase, sodium caseinate and arabic gum are used as wall materials of the antarctic krill oil nanoparticles, and tween-20 is used as an emulsifier; during preparation of the water phase, the solubility of the sodium caseinate and the arabic gum is increased along with the increase of the temperature, but the viscosity of the arabic gum is reduced along with the increase of the temperature;
preparing an oil phase: dissolving antarctic krill oil in a mixed solvent of dichloromethane and acetone, wherein the volume ratio of the dichloromethane to the acetone is 1 (1-5), and the antarctic krill oil accounts for 5-10 (g/100ml) of the organic solvent;
wherein, the volume ratio of the dichloromethane to the acetone is preferably 1: (1-3);
s2, slowly pouring the oil phase prepared in the step S1 into the water phase, and stirring for 0.5-2 hours at 25-45 ℃ to obtain a coarse nano emulsion; homogenizing the coarse nano emulsion at the rotating speed of 6000-10000 rpm/min for 5-10 min; then homogenizing the emulsion at high pressure, sequentially homogenizing at pressures of 0-50 bar, 200-400 bar and 550-650 bar for 1 time each, circularly homogenizing at 700-900 bar for 3 times, and finally removing the organic solvent in the emulsion in a vacuum state to obtain the nano-emulsion;
and (3) carrying out vacuum drying on the obtained nanoemulsion to obtain the small-particle-size euphausia superba oil nanoparticles with high embedding rate.
Wherein the stirring time is preferably 1-1.5 h.
The invention adopts low-temperature stirring when mixing the oil phase, thereby avoiding the oxidative degradation of heat-sensitive substances in the shrimp sauce, such as astaxanthin, caused by high temperature.
In a preferable mode, the crude nano emulsion is homogenized for 6-8 min at the rotating speed of 7000-8000 rpm.
Preferably, the vacuum drying conditions in step S2 are as follows:
pre-freezing for 1-3 h at-60 to-50 ℃; keeping the temperature of minus 45 ℃ to minus 40 ℃ for 1 to 3 hours, and vacuumizing to keep the temperature of 1 Par; keeping the temperature at minus 35 ℃ to minus 25 ℃ for 1 to 3 hours; keeping the temperature of the mixture at minus 15 ℃ to minus 5 ℃ for 4 to 8 hours; keeping the temperature at 0 ℃ to-5 ℃ for 4-8 h, and keeping the temperature at 10 ℃ to 15 ℃ for 4-8 h. The invention adopts a gradient vacuum drying process, so that the shrimp sauce nanoparticles can better keep the original nanoparticle morphology.
The technical innovation of the invention is as follows:
1. the preparation method of the euphausia superba oil nanoparticles is based on an emulsion solvent evaporation method, and adopts multi-stage gradient circulation high-pressure homogenization, so that the uniformity and the embedding rate of the euphausia superba oil nanoparticles are improved, and the particle size of the euphausia superba oil nanoparticles is reduced. The product prepared by the invention has the dispersion index reduced from 0.659 to 0.302, and the reduction amplitude is 118 percent; the embedding rate is increased from 61.83% to 81.82%, and the amplification reaches 33.33%; the particle size is reduced from 313nm to 132nm, and the reduction amplitude reaches 137%.
2. The method used by the invention is simple and reliable, is convenient to operate, and can achieve ideal effect only by adopting multi-stage gradient circulation high-pressure homogenization.
In conclusion, the method improves the uniformity and the embedding rate of the euphausia superba oil nanoparticles and reduces the particle size of the nanoparticles.
Drawings
FIG. 1 is a graph of particle size of the nano emulsion obtained by high pressure homogenization at 0-50 bar;
FIG. 2 is a graph of particle size of the nano emulsion obtained by high pressure homogenization at 200-400 bar;
FIG. 3 is a graph of particle size of the nanoemulsion obtained by high-pressure homogenization at 550-650 bar;
FIG. 4 is a graph of particle size of the nanoemulsion obtained by homogenizing under high pressure for 1 time under pressures of 0-50 bar, 200-400 bar and 550-650 bar, and then circularly homogenizing for 3 times under pressures of 700-900 bar;
fig. 5 is an EPA standard curve.
Detailed Description
The present invention will be described in further detail with reference to examples, but the embodiments of the present invention are not limited thereto.
Example 1:
preparation of crude emulsifier
0.3g of antarctic krill oil was dissolved in 20ml of a dichloromethane and acetone solution in a ratio of 1: 2, as an oil phase. And (2) dissolving a stabilizer (sodium caseinate, Arabic gum and Tween-20) into a phosphate buffer solution with the pH value of 7, wherein the concentration of the stabilizer in the water phase is 2-3%, the adding temperature is 30-60 ℃, and the stirring time is 0.5-1 h. The mixing ratio of the oil phase and the water phase is 1: and 9, pouring the oil phase into the water phase, and uniformly mixing the oil phase and the water phase by using a magnetic stirring and homogenizing machine.
High pressure homogenization
And (3) homogenizing the uniformly mixed emulsion under high pressure, wherein the homogenizing force is 0-50 bar, and circulating once. And then the nano emulsion is subjected to rotary evaporation in a vacuum state to remove the organic solvent. And carrying out vacuum freeze drying on the obtained nano emulsion to obtain the nano particles.
Detection of particle size
Dissolving the nano emulsion obtained by high-pressure homogenization into deionized water, wherein the proportion of the nano emulsion is 1; 15-1: 25. the detection is carried out by a Malvern laser particle sizer, and the result is shown in figure 1.
Surface oil extraction
Weighing 2-5 g of freeze-dried nanoparticles and a conical flask, adding 30-50 ml of n-hexane, extracting at 37 ℃ for 0.5h, collecting supernatant, and removing an organic solvent to obtain surface oil.
Detection of embedding Rate
The method takes EPA (eicosapentaenoic acid) as a standard substance and detects the embedding rate of the euphausia superba oil nanoparticles by an external standard method. Firstly, an EPA (eicosapentaenoic acid) standard curve (shown in figure 5) is obtained by GC-MS (6890N GC-5973 MSD Agilent), and then the content of EPA in the antarctic krill oil and the antarctic krill oil nanoparticle surface oil is calculated according to the standard curve. The embedding rate of the antarctic krill oil nanoparticles is measured by the following formula:
the nanoparticles obtained by the above method have particle diameter of 313.35 + -19.12 nm, dispersion index of 0.659, and embedding rate of 61.83 + -1.96%
Example 2:
preparation of crude emulsifier
0.3g of antarctic krill oil was dissolved in 20ml of a dichloromethane and acetone solution in a ratio of 1: 2, as an oil phase. And (2) dissolving a stabilizer (sodium caseinate, Arabic gum and Tween-20) into a phosphate buffer solution with the pH value of 7, wherein the concentration of the stabilizer in the water phase is 2-3%, the adding temperature is 30-60 ℃, and the stirring time is 0.5-1 h. The mixing ratio of the oil phase and the water phase is 1: and 9, pouring the oil phase into the water phase, and uniformly mixing the oil phase and the water phase by using a magnetic stirring and homogenizing machine.
High pressure homogenization
And (3) homogenizing the uniformly mixed emulsion at high pressure, wherein the homogenizing pressure is 200-400 bar, and circulating once. And then the nano emulsion is subjected to rotary evaporation in a vacuum state to remove the organic solvent. And carrying out vacuum freeze drying on the obtained nano emulsion to obtain the nano particles.
Detection of particle size
Dissolving the nano emulsion obtained by high-pressure homogenization into deionized water, wherein the proportion of the nano emulsion is 1; 15-1: 25. the detection is carried out by a Malvern laser particle sizer, and the result is shown in FIG. 2.
Surface oil extraction
Weighing 2-5 g of freeze-dried nanoparticles and a conical flask, adding 30-50 ml of n-hexane, extracting at 37 ℃ for 0.5h, collecting supernatant, and removing an organic solvent to obtain surface oil.
Detection of embedding Rate
The method takes EPA (eicosapentaenoic acid) as a standard substance and detects the embedding rate of the euphausia superba oil nanoparticles by an external standard method. Firstly, an EPA (eicosapentaenoic acid) standard curve (shown in figure 5) is obtained by GC-MS (6890N GC-5973 MSD Agilent), and then the content of EPA in the antarctic krill oil and the antarctic krill oil nanoparticle surface oil is calculated according to the standard curve. The embedding rate of the antarctic krill oil nanoparticles is measured by the following formula:
the nanoparticles obtained by the above method have a particle size of 312.55 + -16.07 nm, a dispersion index of 1, and a measured embedding rate of 64.02 + -1.43%.
Example 3:
preparation of crude emulsifier
0.3g of antarctic krill oil was dissolved in 20ml of a dichloromethane and acetone solution in a ratio of 1: 2, as an oil phase. And (2) dissolving a stabilizer (sodium caseinate, Arabic gum and Tween-20) into a phosphate buffer solution with the pH value of 7, wherein the concentration of the stabilizer in the water phase is 2-3%, the adding temperature is 30-60 ℃, and the stirring time is 0.5-1 h. The mixing ratio of the oil phase and the water phase is 1: and 9, pouring the oil phase into the water phase, and uniformly mixing the oil phase and the water phase by using a magnetic stirring and homogenizing machine.
High pressure homogenization
And (3) homogenizing the uniformly mixed emulsion at high pressure, wherein the homogenizing pressure is 550-650 bar, and circulating once. And then the nano emulsion is subjected to rotary evaporation in a vacuum state to remove the organic solvent. And carrying out vacuum freeze drying on the obtained nano emulsion to obtain the nano particles.
Detection of particle size
Dissolving the nano emulsion obtained by high-pressure homogenization into deionized water, wherein the proportion of the nano emulsion is 1; 15-1: 25. the detection is carried out by a Malvern laser particle sizer, and the result is shown in FIG. 3.
Surface oil extraction
Weighing 2-5 g of freeze-dried nanoparticles and a conical flask, adding 30-50 ml of n-hexane, extracting at 37 ℃ for 0.5h, collecting supernatant, and removing an organic solvent to obtain surface oil.
Detection of embedding Rate
The method takes EPA (eicosapentaenoic acid) as a standard substance and detects the embedding rate of the euphausia superba oil nanoparticles by an external standard method. Firstly, an EPA (eicosapentaenoic acid) standard curve (shown in figure 5) is obtained by GC-MS (6890N GC-5973 MSD Agilent), and then the content of EPA in the antarctic krill oil and the antarctic krill oil nanoparticle surface oil is calculated according to the standard curve. The embedding rate of the antarctic krill oil nanoparticles is measured by the following formula:
the nanoparticle obtained by the above method has a particle size of 211.21 + -6.59 nm, a dispersion index of 0.702, and a measured embedding rate of 75.43 + -1.27%.
Example 4:
preparation of crude emulsifier
0.3g of antarctic krill oil was dissolved in 20ml of a dichloromethane and acetone solution in a ratio of 1: 2, as an oil phase.
And (2) dissolving a stabilizer (sodium caseinate, Arabic gum and Tween-20) into a phosphate buffer solution with the pH value of 7, wherein the concentration of the stabilizer in the water phase is 2-3%, the adding temperature is 30-60 ℃, and the stirring time is 0.5-1 h. The mixing ratio of the oil phase and the water phase is 1: and 9, pouring the oil phase into the water phase, and uniformly mixing the oil phase and the water phase by using a magnetic stirring and homogenizing machine.
High pressure homogenization
And (3) homogenizing the uniformly mixed emulsion at high pressure, wherein the homogenizing pressure is 0-50 bar, the homogenizing pressure is 200-400 bar and the homogenizing pressure is 550-650 bar for 1 time, and then, circularly homogenizing for 3 times at 700-900 bar. And then the nano emulsion is subjected to rotary evaporation in a vacuum state to remove the organic solvent. And carrying out vacuum freeze drying on the obtained nano emulsion to obtain the nano particles.
Detection of particle size
Dissolving the nano emulsion obtained by high-pressure homogenization into deionized water, wherein the proportion of the nano emulsion is 1; 15-1: 25. the detection is carried out by a Malvern laser particle sizer, and the result is shown in FIG. 4.
Surface oil extraction
Weighing 2-5 g of freeze-dried nanoparticles and a conical flask, adding 30-50 ml of n-hexane, and extracting at 37 DEG C
After 0.5h, the supernatant was collected and the organic solvent was removed, thereby obtaining a surface oil.
Detection of embedding Rate
The method takes EPA (eicosapentaenoic acid) as a standard substance and detects the embedding rate of the euphausia superba oil nanoparticles by an external standard method. Firstly, an EPA (eicosapentaenoic acid) standard curve (shown in figure 5) is obtained by GC-MS (6890N GC-5973 MSD Agilent), and then the content of EPA in the antarctic krill oil and the antarctic krill oil nanoparticle surface oil is calculated according to the standard curve. The embedding rate of the antarctic krill oil nanoparticles is measured by the following formula:
the nanoparticles obtained by the above method have particle diameter of 130.93 + -2.56 nm, dispersion index of 0.302, and embedding rate of 81.82 + -1.35%
Table 1 shows the particle size and the embedding rate of nanoparticles under each high-pressure homogenization pressure, and by comparing the particle size and the embedding rate of the nanoemulsion prepared under different high-pressure homogenization pressures, we can show that the nanoparticles prepared by the present invention can effectively reduce the particle size of nanoparticles and improve the uniformity and the embedding rate of nanoparticles.
Table 1: the particle diameter and embedding rate of the nanoparticles under each high-pressure homogeneous pressure
Claims (1)
1. A preparation method of euphausia superba oil nanoparticles with high embedding rate and small particle size is characterized by comprising the following specific technical scheme:
s1, preparing a water phase: mixing the following raw materials in percentage by weight to prepare a water phase:
the method comprises the following steps of taking a mixture of sodium caseinate, Arabic gum and Tween-20 as a stabilizer, wherein the stabilizer accounts for 2-3% of the total mass of a water phase, the sodium caseinate accounts for 60-70% of the total mass of the stabilizer, the Arabic gum accounts for 5-10% of the total mass of the stabilizer, and the Tween-20 accounts for 20-25% of the total mass of the stabilizer;
dissolving the stabilizer in a phosphate buffer solution with the pH value of 7, and stirring at 40-60 ℃ for 0.5-1 h to obtain a water phase;
preparing an oil phase: the method comprises the steps of dissolving Antarctic krill oil in a mixed solvent of dichloromethane and acetone, wherein the volume ratio of the dichloromethane to the acetone is 1 (1-3), and the Antarctic krill oil accounts for 5-10 g/100ml of an organic solvent;
s2, slowly pouring the oil phase prepared in the step S1 into the water phase, and stirring for 1-1.5 hours at 25-45 ℃ to obtain a coarse nano emulsion; homogenizing the coarse nano emulsion at the rotating speed of 7000-8000 rpm/min for 6-8 min; then homogenizing the emulsion at high pressure, respectively homogenizing for 1 time under the pressure of 0-50 bar, 200-400 bar and 550-650 bar, circularly homogenizing for 3 times under the pressure of 700-900 bar, and finally removing the organic solvent in the emulsion under a vacuum state to obtain the nano-emulsion;
carrying out vacuum drying on the obtained nano-emulsion at the temperature of minus 60 ℃ to minus 50 ℃ for 1 to 3 hours; keeping the temperature of minus 45 ℃ to minus 40 ℃ for 1 to 3 hours, and vacuumizing to keep the temperature of 1 Par; keeping the temperature at minus 35 ℃ to minus 25 ℃ for 1 to 3 hours; keeping the temperature of the mixture at minus 15 ℃ to minus 5 ℃ for 4 to 8 hours; and keeping the temperature of 0-5 ℃ for 4-8 h, and keeping the temperature of 10-15 ℃ for 4-8 h to obtain the small-particle-size euphausia superba oil nanoparticles with high embedding rate.
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| CN112544731A (en) * | 2020-12-30 | 2021-03-26 | 浙江工业大学 | Preparation process of antarctic krill oil nanoemulsion |
| CN112868817B (en) * | 2021-02-04 | 2022-05-17 | 中南民族大学 | Euphausia superba oil edible product with protein as key |
| CN113287750A (en) * | 2021-04-08 | 2021-08-24 | 日照职业技术学院 | Euphausia superba oil nano-particles and preparation method thereof |
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