CN115400080A - Stable Pickering emulsion prepared from luteolin, and preparation method and application thereof - Google Patents
Stable Pickering emulsion prepared from luteolin, and preparation method and application thereof Download PDFInfo
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- CN115400080A CN115400080A CN202211005668.6A CN202211005668A CN115400080A CN 115400080 A CN115400080 A CN 115400080A CN 202211005668 A CN202211005668 A CN 202211005668A CN 115400080 A CN115400080 A CN 115400080A
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- luteolin
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- pickering emulsion
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- 239000000839 emulsion Substances 0.000 title claims abstract description 93
- MWDZOUNAPSSOEL-UHFFFAOYSA-N kaempferol Natural products OC1=C(C(=O)c2cc(O)cc(O)c2O1)c3ccc(O)cc3 MWDZOUNAPSSOEL-UHFFFAOYSA-N 0.000 title claims abstract description 70
- IQPNAANSBPBGFQ-UHFFFAOYSA-N luteolin Chemical compound C=1C(O)=CC(O)=C(C(C=2)=O)C=1OC=2C1=CC=C(O)C(O)=C1 IQPNAANSBPBGFQ-UHFFFAOYSA-N 0.000 title claims abstract description 70
- LRDGATPGVJTWLJ-UHFFFAOYSA-N luteolin Natural products OC1=CC(O)=CC(C=2OC3=CC(O)=CC(O)=C3C(=O)C=2)=C1 LRDGATPGVJTWLJ-UHFFFAOYSA-N 0.000 title claims abstract description 70
- 235000009498 luteolin Nutrition 0.000 title claims abstract description 70
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Abstract
The invention relates to the technical field of emulsion preparation, and particularly relates to a stable Pickering emulsion prepared from luteolin, and a preparation method and application thereof. The method of the invention comprises the following steps: s1, preparation of an oil phase: dissolving fat-soluble active substances in antarctic krill oil; s2, preparation of a water phase: dissolving luteolin in anhydrous ethanol, slowly injecting deionized water into the solution, and performing rotary evaporation to completely volatilize the anhydrous ethanol; s3, mixing the oil phase with the water phase, and dispersing at a high speed; and S4, homogenizing to obtain the stable Pickering emulsion containing the luteolin. The invention adopts luteolin as an emulsifier, selects specific edible oil as an oil phase, adopts fat-soluble active substances as an embedded material, can encapsulate the oil phase through Pickering emulsion, and finally prepares the Pickering emulsion stabilized by luteolin by integrating various indexes. The invention has simple operation, and the prepared emulsion has strong light stability, small average particle size, large viscosity, high retention rate of fat-soluble active substances and better stability.
Description
Technical Field
The invention relates to the technical field of emulsion preparation, and particularly relates to a stable Pickering emulsion prepared from luteolin, and a preparation method and application thereof.
Background
Fat-soluble active substances such as phospholipid, fat-soluble vitamins, polyunsaturated fatty acids and the like have various health-beneficial effects, but are not easy to be absorbed by human bodies due to the characteristics of poor water solubility, easy oxidative degradation, low bioavailability and the like, cannot fully exert the functional characteristics of the active substances, and seriously limit the application of the active substances in the fields of food, medicine and the like. To overcome these obstacles, emulsions are often chosen to deliver lipid soluble actives and perform well in active delivery system applications.
Emulsions stabilized with solid particles instead of surfactants are collectively referred to as Pickering emulsions. Compared with the traditional emulsion, the Pickering emulsion has the performances of lipid oxidation resistance, strong stability, low toxicity, high safety, low cost and the like, is more favorable for loading fat-soluble active substances, and is widely applied to an embedding delivery system. In recent years, food-grade solid particles commonly used for stabilizing Pickering emulsion are protein, polysaccharide, polyphenol and the like. With the continuous expansion of the application of the emulsion in the food industry, the research and development of the food-grade Pickering emulsion become a hot point of research.
Luteolin is a natural polyphenol component with biological characteristics widely distributed in various plants, and belongs to weakly acidic tetrahydroxyflavonoids compounds. The structure of the antioxidant agent contains a polyhydroxy structure, so the antioxidant agent has stronger antioxidant property. Research shows that luteolin also has various pharmacological effects of resisting bacteria, inflammation and cancer, protecting liver and kidney and the like, and is a functional compound with development potential and market value. However, there are few reports in the prior art on luteolin stabilized Pickering emulsions. Moreover, since the interaction between the solid particles and the oil phase is complex, the development of a stable Pickering emulsion with better stability and stabilized by luteolin needs to overcome a great number of technical problems.
Disclosure of Invention
The technical problem is as follows: the Pickering emulsion which is stable by luteolin and has the advantages of simple operation, small average particle size of the prepared emulsion, high storage stability, strong light stability, large apparent shear viscosity and high retention rate of fat-soluble active substances and the preparation method thereof are developed.
The technical scheme is as follows:
the first purpose of the invention is to provide a preparation method of a Pickering emulsion stabilized by luteolin, which comprises the following steps:
s1, preparing an oil phase: dissolving the fat-soluble active substance in the antarctic krill oil to prepare a fat-soluble active substance-antarctic krill oil solution;
s2, preparation of a water phase: dissolving luteolin in anhydrous ethanol, slowly injecting deionized water into the solution, and performing rotary evaporation to completely volatilize the anhydrous ethanol to obtain luteolin suspension;
s3, mixing: mixing the oil phase prepared in the step S1 with the water phase prepared in the step S2, and dispersing at a high speed;
s4, homogenizing: and (4) homogenizing the product obtained in the step (S3) to obtain the Pickering emulsion stabilized by luteolin.
As an embodiment of the present invention, the fat-soluble active substance includes benzyl isothiocyanate.
In an embodiment of the present invention, the mass concentration of benzyl isothiocyanate in the oil solution of euphausia superba, which is the fat-soluble active substance in step S1, is 3 to 9mg/mL.
In one embodiment of the present invention, the mass concentration of benzyl isothiocyanate in the oil solution of euphausia superba, which is the lipid soluble active substance in step S1, is 5mg/mL.
In an embodiment of the present invention, the mass concentration of luteolin in the luteolin suspension in the step S2 is 2-6 mg/mL.
As an embodiment of the present invention, the mass concentration of luteolin in the luteolin suspension in the step S2 is 4mg/mL.
In one embodiment of the present invention, the volume of the oil phase in step S3 is 5% to 15% of the volume of the water phase.
In one embodiment of the present invention, the volume of the oil phase in step S3 is 10% of the volume of the water phase.
As an embodiment of the present invention, in the step S3, the high-speed dispersion: high-speed dispersion is carried out for 2-3 min at 9000-14000 r/min. Preferably, 10000r/min are dispersed for 2min at high speed.
As an embodiment of the present invention, in the step S4, homogenization: homogenizing under 5000-10000 psi high pressure for 3-5 times. Preferably, 7000psi high pressure homogenization is performed 3 times.
As an embodiment of the present invention, a method for preparing Pickering emulsion stabilized by luteolin comprises the following steps:
s1, preparing an oil phase: dissolving benzyl isothiocyanate in antarctic krill oil to prepare benzyl isothiocyanate-edible oil solution (oil phase) with the mass concentration of 3-9 mg/mL;
s2, preparation of a water phase: dissolving luteolin in absolute ethyl alcohol, slowly injecting deionized water with the volume of 4-8 times that of the absolute ethyl alcohol, completely volatilizing the absolute ethyl alcohol by adopting a rotary evaporator, and preparing a luteolin suspension (water phase) with the mass concentration of 2-6 mg/mL;
s3, mixing: mixing the oil phase prepared in the step S1 with the water phase prepared in the step S2 according to the addition amount of 5-15% of the volume of the water phase, and dispersing at a high speed of 9000-14000 r/min for 2-3 min;
s4, homogenizing: homogenizing under high pressure of 5000-10000 psi for 3-5 times to obtain the luteolin Pickering emulsion.
A second object of the present invention is to provide a Pickering emulsion stabilized by luteolin, prepared according to the aforementioned method.
The third purpose of the invention is to provide the application of the Pickering emulsion stabilized by luteolin in preparing food, medicine or cosmetics.
The invention has the beneficial effects that:
(1) The Pickering emulsion is stabilized based on the solid particles of the luteolin, the luteolin is natural in components, wide in source and high in biological safety, has strong antioxidant property, also has various pharmacological effects of resisting bacteria, inflammation and cancer, protecting the liver and the kidney and the like, can improve the antioxidant stability and the nutritional property of the Pickering emulsion, and can be used for preparing functional foods. The invention applies the luteolin as the emulsifier to a Pickering emulsion system, can expand the application range of natural polyphenol active substances, and simultaneously provides a research basis for the effective utilization of fat-soluble active substances.
(2) In the invention, the luteolin is dispersed in the Pickering emulsion in a low-solubility particle form and is tightly adsorbed on an oil-water interface, so that an emulsion system is stable.
(3) The research of the invention finds that for a specific fat-soluble active substance, namely benzyl isothiocyanate, in the preparation process of a Pickering emulsion system embedding the benzyl isothiocyanate, the selection of solid particles and an oil phase has great influence on the stability of the system and the retention rate of the benzyl isothiocyanate: the luteolin Pickering emulsion prepared by taking the antarctic krill oil as the oil phase has smaller average particle size, higher retention rate (up to 65.25 +/-1.45 percent, as shown in figure 8) for benzyl isothiocyanate, smaller average particle size (194.27 +/-2.06 nm, as shown in figure 6) of fresh emulsion, almost no change of the average particle size of the emulsion after being stored for 14 days at 4 ℃ (compared with the change of the average particle size of the fresh emulsion which is only +1.86nm, as shown in figure 6), higher apparent shear viscosity (as shown in figure 7) and stronger light stability (figures 9-10), and comprehensively, the luteolin Pickering emulsion prepared by taking the antarctic krill oil as the oil phase is more stable.
(4) The method is simple and convenient to operate, and the prepared emulsion has the advantages of small average particle size, high storage stability, strong light stability, large apparent shear viscosity and high retention rate of benzyl isothiocyanate.
Drawings
FIG. 1 is a physical representation of product emulsions prepared in examples 1-3 of the present invention stored at 4 ℃ for 0 day;
FIG. 2 is a physical representation of the product emulsions prepared in examples 1-3 of the present invention stored at 4 ℃ for 1 day;
FIG. 3 is a physical representation of product emulsions prepared in examples 1-3 of the present invention stored at 4 ℃ for 4 days;
FIG. 4 is a physical representation of the product emulsions prepared in examples 1-3 of the present invention stored at 4 ℃ for 7 days;
FIG. 5 is a physical representation of the product emulsions prepared in examples 1-3 of the present invention stored at 4 ℃ for 14 days;
FIG. 6 is the average particle size of the product emulsions prepared in examples 1 to 3 of the present invention when stored at 4 ℃ for 0 day, 1 day, 4 days, 7 days, and 14 days, respectively;
FIG. 7 is the apparent shear viscosity of the product emulsions prepared in examples 1-3 of the present invention;
FIG. 8 is a graph showing the retention of benzyl isothiocyanate in the product emulsions prepared in examples 1 to 3 of the present invention, respectively, after storage at 4 ℃ for 14 days;
FIG. 9 is a pictorial representation of the product emulsions prepared in examples 1-3 of the present invention after being left for 2 hours in different light conditions;
FIG. 10 is a graph showing the turbidity change of the emulsion when the emulsion prepared in examples 1 to 3 according to the present invention is left for 2 hours under different light conditions.
Detailed Description
Materials, reagents and the like used in the following examples are commercially available unless otherwise specified.
The invention provides a preparation method of a Pickering emulsion stabilized by luteolin, which comprises the following steps:
s1, preparing an oil phase: dissolving benzyl isothiocyanate in edible oil to prepare a benzyl isothiocyanate-edible oil solution (oil phase) with the mass concentration of 3-9 mg/mL;
s2, preparation of a water phase: dissolving luteolin in absolute ethyl alcohol, slowly injecting deionized water with the volume of 4-8 times that of the absolute ethyl alcohol, completely volatilizing the absolute ethyl alcohol by adopting a rotary evaporator, and preparing a luteolin suspension (water phase) with the mass concentration of 2-6 mg/mL;
s3, mixing the oil phase prepared in the step S1 with the luteolin suspension prepared in the step S2 according to the addition amount of 5-15% of the volume of the water phase, and dispersing at a high speed of 9000-14000 r/min for 2-3 min;
s4, homogenizing for 2-5 times at 5000-10000 psi under high pressure to obtain the luteolin Pickering emulsion.
Example 1 (corn oil)
A preparation method of Pickering emulsion stabilized by luteolin comprises the following steps:
s1, preparation of an oil phase: dissolving benzyl isothiocyanate in corn oil to prepare a benzyl isothiocyanate-edible oil solution (oil phase) with the mass concentration of 5 mg/mL;
s2, preparation of a water phase: dissolving luteolin in anhydrous ethanol, slowly injecting deionized water 6 times the volume of the anhydrous ethanol, and completely volatilizing the anhydrous ethanol by adopting a rotary evaporator to prepare a luteolin suspension (water phase) with the mass concentration of 4 mg/mL;
s3, mixing the oil phase prepared in the step S1 with the luteolin suspension prepared in the step S2 according to the addition amount of 10% of the volume of the water phase, and dispersing at a high speed of 10000r/min for 2min;
s4, homogenizing at 7000psi for 3 times to obtain the luteolin Pickering emulsion.
Example 2 (linseed oil)
A preparation method of a Pickering emulsion stabilized by luteolin comprises the following steps:
s1, preparing an oil phase: dissolving benzyl isothiocyanate in linseed oil to prepare a benzyl isothiocyanate-edible oil solution (oil phase) with the mass concentration of 5 mg/mL;
s2, preparation of a water phase: dissolving luteolin in anhydrous ethanol, slowly injecting deionized water 6 times the volume of the anhydrous ethanol, and completely volatilizing the anhydrous ethanol by adopting a rotary evaporator to prepare a luteolin suspension (water phase) with the mass concentration of 4 mg/mL;
s3, mixing the oil phase prepared in the step S1 with the luteolin suspension prepared in the step S2 according to the addition amount of 10% of the volume of the water phase, and dispersing at a high speed of 10000r/min for 2min;
s4, homogenizing at 7000psi for 3 times to obtain the luteolin Pickering emulsion.
Example 3 (Antarctic krill oil)
A preparation method of Pickering emulsion stabilized by luteolin comprises the following steps:
s1, preparation of an oil phase: dissolving benzyl isothiocyanate in antarctic krill oil to prepare a benzyl isothiocyanate-edible oil solution (oil phase) with the mass concentration of 5 mg/mL;
s2, preparation of a water phase: dissolving luteolin in anhydrous ethanol, slowly injecting deionized water 6 times the volume of the anhydrous ethanol, and completely volatilizing the anhydrous ethanol by adopting a rotary evaporator to prepare a luteolin suspension (water phase) with the mass concentration of 4 mg/mL;
s3, mixing the oil phase prepared in the step S1 with the luteolin suspension prepared in the step S2 according to the addition amount of 10% of the volume of the water phase, and dispersing at a high speed of 10000r/min for 2min;
s4, homogenizing at 7000psi for 3 times to obtain the luteolin Pickering emulsion.
Example 4
A preparation method of Pickering emulsion stabilized by luteolin comprises the following steps:
s1, preparing an oil phase: dissolving benzyl isothiocyanate in antarctic krill oil to prepare benzyl isothiocyanate-edible oil solution (oil phase) with the mass concentration of 3-9 mg/mL;
s2, preparation of a water phase: dissolving luteolin in absolute ethyl alcohol, slowly injecting deionized water with the volume 6 times that of the absolute ethyl alcohol, completely volatilizing the absolute ethyl alcohol by adopting a rotary evaporator, and preparing a luteolin suspension (water phase) with the mass concentration of 2-6 mg/mL;
s3, mixing the oil phase prepared in the step S1 with the luteolin suspension prepared in the step S2 according to the addition amount of 5-15% of the volume of the water phase, and dispersing at a high speed of 9000-14000 r/min for 2-3 min;
s4, homogenizing for 3-5 times at 5000-10000 psi under high pressure to obtain the luteolin Pickering emulsion.
The test method comprises the following steps:
appearance change observation and emulsion particle size measurement: taking a proper amount of the product emulsion prepared in the examples 1-3 (stored at 4 ℃ for 0 day, 1 day, 4 days, 7 days and 14 days respectively) and observing the appearance change; at the same time, an appropriate amount of the product emulsions prepared in examples 1 to 3 (stored at 4 ℃ for 0 day, 1 day, 4 days, 7 days, and 14 days, respectively) was diluted by 100 times, and the average particle size was measured using a nano-particle size analyzer. Different capital letters indicate the difference in significance of different oil phases at the same storage time, and different lower case letters indicate the difference in significance of the same oil phase at different storage times (p)<0.05). The concrete analysis and theoretical basis comprises:
(1) The appearance of the emulsion has no obvious change, and the more stable the system.
(2) The smaller the average particle size of the emulsion, the more stable the system.
Apparent shear viscosity measurement: the apparent viscosity of the emulsion was measured by taking an appropriate amount of the fresh product emulsion prepared in examples 1 to 3 using a rheometer equipped with a flat plate. And (3) sucking a proper amount of emulsion liquid to drop on the measuring platform, so that the two plates are uniformly covered with the emulsion without bubbles, and the shearing rate is 0.1-100rad/s. The change curves of the apparent viscosities of the product emulsions prepared in examples 1 to 3 were measured. The concrete analysis and theoretical basis is as follows: the greater the viscosity of the emulsion, the slower the settling rate of its oil droplets and the more stable the system.
Determination of Retention of benzyl isothiocyanate: taking a proper amount of the emulsion prepared in the embodiments 1-3, storing for 14 days at 4 ℃, extracting benzyl isothiocyanate in the emulsion by using normal hexane and methanol, analyzing the content of the benzyl isothiocyanate by using high performance liquid chromatography, and calculating the retention rate of the benzyl isothiocyanate in the emulsion according to the positive correlation between the concentration of the benzyl isothiocyanate in the solution and the peak content, wherein the retention rate (%) = the peak area of the benzyl isothiocyanate in the emulsion/the peak area of the benzyl isothiocyanate in the standard substance. Different capital letters indicate significant differences in different oil phases at the same storage time (p)<0.05). The concrete analysis and theoretical basis is as follows: the higher the embedding rate of the emulsion, the more stable the system.
Photostability analysis: the product emulsions prepared in examples 1 to 3 of the present invention were subjected to light stability analysis by different illuminations for a short time, and the light stability was reflected by the turbidity of the emulsion. Freshly prepared emulsions of examples 1 to 3 treated under different illumination conditions (natural light, ultraviolet light) were diluted 20 times with deionized water, respectively, and absorbance was measured at a wavelength of 500nm with an ultraviolet-visible spectrophotometer, followed by turbidity calculation. The turbidity calculation method comprises the following steps: r t ＝(A f -A 1 )/(A b -A 2 ) X 100%, wherein: a. The b -fresh preparation of absorbance values of the dilution medium; a. The f -absorbance of the diluted medium after operation at different lighting conditions; a. The 2 -light absorption of dilution media before experimental manipulationDegree; a. The 1 -absorbance of the diluted medium after operation at different lighting conditions; 100-conversion factor.
Different capital letters in the data indicate different oil phase significance differences under the same illumination conditions, and different lower case letters indicate the same oil phase significance differences under different illumination conditions (p < 0.05). The concrete analysis and theoretical basis comprises:
(1) The appearance shape of the emulsion has no obvious change, and the more stable the system.
(2) The greater the turbidity of the emulsion, the more stable the system.
And (4) analyzing results:
the research of constructing a luteolin Pickering emulsion conveying system by using different oil phases (corn oil, linseed oil and antarctic krill oil) and embedding benzyl isothiocyanate at the same time shows that:
compared with the luteolin Pickering emulsion prepared by taking corn oil and linseed oil as oil phases, the luteolin Pickering emulsion prepared by taking antarctic krill oil as the oil phases has smaller average particle size, higher retention rate (as high as 65.25 +/-1.45 percent) for benzyl isothiocyanate, smaller average particle size (194.27 +/-2.06 nm as shown in figure 6) of fresh emulsion and almost no change of the average particle size of the emulsion after being stored for 14 days at 4 ℃ (compared with the average particle size of the fresh emulsion which is only changed by +1.86nm as shown in figure 6), higher apparent shear viscosity (as shown in figure 7) and stronger light stability (as shown in figures 9-10), and the luteolin Pickering emulsion prepared by taking the antarctic krill oil as the oil phases is more stable in general.
The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be able to cover the technical solutions and the inventive concepts of the present invention within the technical scope of the present invention.
Claims (10)
1. A preparation method of a Pickering emulsion stabilized by luteolin is characterized by comprising the following steps:
s1, preparation of an oil phase: dissolving fat-soluble active substances in antarctic krill oil to prepare a fat-soluble active substance-antarctic krill oil solution;
s2, preparation of a water phase: dissolving luteolin in anhydrous alcohol, slowly injecting deionized water, and rotary steaming to completely volatilize anhydrous alcohol to obtain luteolin suspension;
s3, mixing: mixing the oil phase prepared in the step S1 with the water phase prepared in the step S2, and dispersing at a high speed;
s4, homogenizing: and (4) homogenizing the product obtained in the step (S3) to prepare the Pickering emulsion stabilized by luteolin.
2. The method of claim 1, wherein the lipid soluble active comprises benzyl isothiocyanate.
3. The method according to claim 1, wherein the mass concentration of the fat-soluble active substance in the oil solution of Euphausia superba in the fat-soluble active substance in step S1 is 3-9 mg/mL.
4. The method according to claim 1, wherein the mass concentration of luteolin in the luteolin suspension in the step S2 is 2-6 mg/mL.
5. The method of claim 1, wherein the volume of the oil phase in step S3 is 5-15% of the volume of the water phase.
6. The method of claim 5, wherein the volume of the oil phase in step S3 is 10% of the volume of the water phase.
7. The method according to claim 1, characterized in that in said step S3 high speed dispersion: and high-speed dispersion is carried out for 2-3 min at 9000-14000 r/min.
8. The method according to claim 1, characterized in that in step S4 homogenizing: homogenizing under 5000-10000 psi high pressure for 3-5 times.
9. A Pickering emulsion stabilized by luteolin, prepared according to the method of any one of claims 1-8.
10. Use of a luteolin stabilized Pickering emulsion according to claim 9 for the preparation of a food product, a pharmaceutical or a cosmetic product.
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