Preparation method and application of sodium alginate-chitosan modified pomelo oil liposome

Abstract

The invention relates to the technical field of pomelo oil delivery, in particular to a preparation method and application of sodium alginate-chitosan modified pomelo oil liposome. The method comprises the following steps: dissolving grapefruit oil, egg yolk phospholipid, tween-80 and beta-sitosterol in absolute ethyl alcohol according to a certain proportion, uniformly mixing, adding deionized water, heating and stirring to obtain a mixture; carrying out rotary evaporation on the mixture to remove the organic solvent, and carrying out homogenization and ultrasonic treatment to obtain a pomelo oil liposome; and adding the chitosan solution into the pomelo oil liposome, uniformly stirring, then adding the sodium alginate solution, and uniformly stirring to obtain the sodium alginate-chitosan modified pomelo oil liposome. According to the invention, the encapsulation efficiency and the bioavailability can be obviously improved.

Description

Technical Field

[0001] The invention relates to the technical field of grapefruit oil delivery, and in particular to a preparation method and application of sodium alginate-chitosan modified grapefruit oil liposomes. Background Art

[0002] Fat-soluble active substances such as polyunsaturated fatty acids, fat-soluble vitamins, polyphenols, etc. have a variety of health benefits, but due to their poor water solubility and low bioavailability, they are not easily absorbed by the human body and cannot fully exert their functional characteristics, limiting their application in the fields of food and medicine. In order to overcome these obstacles, emulsions and liposomes are usually used to deliver fat-soluble active substances. Liposomes are amphiphilic phospholipids dispersed in the aqueous phase to self-assemble into closed vesicles with a bilayer structure. Compared with other delivery systems, the closed vesicle structure of liposomes can simultaneously encapsulate fat-soluble and water-soluble substances, thereby improving the bioavailability and stability of active substances.

[0003] Camellia oil is made from tea seeds through physical or chemical processing. Tea oil processed from tea oil grown locally in Hainan is called "grapefruit oil". Grapefruit oil contains polyphenols, squalene, phytosterols, vitamin E, tea saponins and other active substances, which have lipid-lowering, anti-inflammatory, antibacterial, anti-tumor, antioxidant activity and cardiovascular disease prevention effects. However, grapefruit oil has poor water solubility and is prone to oxidative decomposition, which greatly limits its application in food, medicine and other fields.

[0004] Traditional liposomes have not been modified in any way, and their ability to resist changes in the external environment is still relatively weak. They are prone to aggregation, causing leakage of embedded materials, resulting in low bioavailability. Modifying liposomes by biomacromolecules can effectively improve the embedding rate, stability, and bioavailability of liposomes. However, there are few reports on grapefruit oil liposomes and their modification in the prior art. How to prepare layer-by-layer modified liposomes to improve the encapsulation rate and bioavailability of grapefruit oil has become an urgent problem to be solved. Summary of the invention

[0005] The purpose of the present invention is to overcome the deficiencies of the prior art and provide a method for preparing and applying a sodium alginate-chitosan modified grapefruit oil liposome, so as to significantly improve the encapsulation rate and bioavailability of grapefruit oil.

[0006] In order to achieve the above object, the present invention provides the following technical solutions:

[0007] The present invention provides a method for preparing sodium alginate-chitosan modified grapefruit oil liposomes, comprising the following steps:

[0008] Step 1, grapefruit oil, egg yolk lecithin, Tween-80 and β-sitosterol are mixed and dissolved in anhydrous ethanol in a mass ratio of 1:5:1:1, and after mixing evenly, 4 times the volume of deionized water is added to the mixture, and magnetic stirring is performed to obtain a grapefruit oil liposome suspension;

[0009] Step 2, the grapefruit oil liposome suspension obtained in step 1 is subjected to rotary evaporation treatment to remove ethanol, and then homogenized and ultrasonically treated to obtain grapefruit oil liposomes;

[0010] Step 3, adding an equal volume of 1.5 mg / mL chitosan solution to the grapefruit oil liposomes obtained in step 2, stirring evenly to obtain chitosan-modified grapefruit oil liposomes;

[0011] Step 4: Add an equal volume of 4 mg / mL sodium alginate solution to the chitosan-modified grapefruit oil liposomes obtained in step 3, and stir evenly to obtain sodium alginate-chitosan-modified grapefruit oil liposomes.

[0012] Furthermore, in step 1, the heating temperature of the magnetic stirring is 45° C., and the stirring rate is 600 r / min.

[0013] Furthermore, in step 2, the mass concentration of egg yolk lecithin in the grapefruit oil liposome is 10 mg / mL.

[0014] Furthermore, in step 2, the conditions for the rotary evaporation are: heating temperature 45° C., rotation speed 200 r / min, and time 40 min.

[0015] Furthermore, in step 2, the homogenization is carried out under ice bath conditions, the homogenization speed is 10000 r / min, and the homogenization time is 2 min; the power of the ultrasonic treatment is 180 W, and the ultrasonic treatment time is 6 min.

[0016] Furthermore, in step three, the preparation method of the 1.5 mg / mL chitosan solution is: dissolving chitosan in a 1% v / v acetic acid aqueous solution, stirring at 600 r / min for 45 min to prepare a 1.5 mg / mL chitosan solution; the chitosan has a deacetylation degree of >95% and a viscosity of 100-200 mp.s.

[0017] Furthermore, in step 4, the preparation method of the 4 mg / mL sodium alginate solution is: dissolving sodium alginate completely in deionized water, adjusting the pH to 6, and magnetically stirring at 600 r / min for 45 minutes to obtain a 4 mg / mL sodium alginate solution.

[0018] The invention also provides a sodium alginate-chitosan modified grapefruit oil liposome, which is prepared by the method for preparing the sodium alginate-chitosan modified grapefruit oil liposome.

[0019] The present invention also provides an application of sodium alginate-chitosan modified camellia oil liposomes in the preparation of camellia oil delivery preparations.

[0020] Advantages of the present invention:

[0021] The present invention uses egg yolk lecithin as the wall material and camellia oil as the core material to prepare camellia oil liposomes; sodium alginate and chitosan are used to modify the camellia oil liposomes. The modified sodium alginate-chitosan camellia oil liposomes have good stability, high encapsulation efficiency, and high bioavailability.

[0022] (1) Improve the encapsulation efficiency of camellia oil: The camellia oil liposomes are modified by sodium alginate and chitosan. Experimental data show that the encapsulation efficiency can be significantly improved. After modification in Examples 1-5, the encapsulation efficiency is significantly higher than that of Examples 6-8 which are not fully modified, up to 92.25±2.72%, effectively encapsulating camellia oil in the liposomes and reducing its exposure in the external environment.

[0023] (2) Enhance bioavailability: The closed vesicle structure of liposomes can originally improve the bioavailability and stability of active substances, and is further optimized after modification with sodium alginate-chitosan. This modification improves the performance of liposomes, enabling camellia oil to be better absorbed and utilized after entering the human body, and exerting its various effects such as reducing blood lipids, anti-inflammatory, and antibacterial.

[0024] (3) Solve the application limitations of camellia oil: Camellia oil is limited in applications in the fields of food, medicine, etc. due to its poor water solubility and easy oxidation and decomposition. The modified liposome technology of the present invention improves the stability of camellia oil, reduces the risk of oxidation and decomposition, solves the problem of poor water solubility, and expands the application scope of camellia oil in multiple fields.

[0025] (4) Optimize the performance of liposomes: Traditional liposomes have weak resistance to changes in the external environment and are prone to aggregation, resulting in leakage of the embedded substances. The present invention uses biological macromolecules for modification, enhancing the liposomes' resistance to environmental changes, reducing aggregation and leakage of the embedded substances, and improving the overall performance of the liposomes.

[0026] (5) Identify key factors in the preparation: Through multiple examples and experimental comparisons, the importance of various factors in the preparation process is clarified. For example, the structure of chitosan, the integrity of the modification steps, and the preparation conditions of camellia oil liposomes (such as ultrasonic treatment, homogenization, etc.) have a significant impact on the formation and performance of the final liposomes, providing key references for subsequent large-scale production. Specific embodiments

[0027] The specific implementation modes of the present invention are described below so that those skilled in the art can understand the present invention. However, it should be clear that the present invention is not limited to the scope of the specific implementation modes. For those of ordinary skill in the art, as long as various changes are within the spirit and scope of the present invention as defined and determined by the attached claims, these changes are obvious, and all inventions and creations utilizing the concept of the present invention are protected.

[0028] The chitosan used in Examples 1-6 was purchased from Shanghai Yuanye Biotechnology Co., Ltd. The acetylation degree was >95% and the viscosity was 100-200 mp.s.

[0029] The chitosan used in Example 7 was purchased from Shanghai Yuanye Biotechnology Co., Ltd. with a deacetylation degree of 85% and a viscosity of ≤500 mp.s.

[0030] Example 1 Preparation method of sodium alginate-chitosan modified grapefruit oil liposomes

[0031] This embodiment provides a method for preparing sodium alginate-chitosan modified grapefruit oil liposomes, the method comprising the following steps:

[0032] (1) Weigh 250 mg of egg yolk lecithin, 50 mg of grapefruit oil, 50 mg of Tween-80 and 50 mg of β-sitosterol, add 30 mL of anhydrous ethanol and stir thoroughly to form a mixed solution. Use a syringe to inject the mixed solution into 120 mL of deionized water, stir magnetically at 45°C and 600 r / min for 45 min to obtain a grapefruit oil liposome suspension. Rotate the suspension to remove ethanol until the mass concentration of egg yolk lecithin in the grapefruit oil liposome is 10 mg / mL. The conditions for rotary evaporation are: heating temperature 45°C, rotation speed 200 r / min, time 40 min. Then, at a speed of 10000 r / min, in an ice bath, homogenization time 2 min; and at an ultrasonic power of 180 W, ultrasonic for 6 min to obtain grapefruit oil liposomes.

[0033] (2) Dissolve chitosan in 1% (v / v) acetic acid aqueous solution, stir at 600 r / min for 45 min to prepare a 1.5 mg / mL chitosan solution. Add an equal volume of 1.5 mg / mL chitosan solution dropwise to the grapefruit oil liposomes obtained in step (1), stir at 600 r / min for 45 min to obtain chitosan-modified grapefruit oil liposomes. Dissolve sodium alginate in deionized water, adjust the pH to 6, and stir magnetically at 600 r / min for 45 min to obtain a 4 mg / mL sodium alginate solution. Add an equal volume of 4 mg / mL sodium alginate solution to the chitosan-modified grapefruit oil liposomes, and continue stirring at 600 r / min for 45 min to obtain sodium alginate-chitosan grapefruit oil liposomes.

[0034] Example 2 Preparation method of sodium alginate-chitosan modified grapefruit oil liposomes

[0035] This embodiment provides a method for preparing sodium alginate-chitosan modified grapefruit oil liposomes, the method comprising the following steps:

[0036] (1) Weigh 250 mg of egg yolk lecithin, 75 mg of grapefruit oil, 50 mg of Tween-80 and 50 mg of β-sitosterol, add 30 mL of anhydrous ethanol and stir thoroughly to form a mixed solution. Use a syringe to inject the mixed solution into 120 mL of deionized water, stir magnetically at 45°C and 600 r / min for 45 min to obtain a grapefruit oil liposome suspension. Rotate the suspension to remove ethanol until the mass concentration of egg yolk lecithin in the grapefruit oil liposome is 10 mg / mL. The conditions for rotary evaporation are: heating temperature 45°C, rotation speed 200 r / min, time 40 min. Then, at a speed of 10000 r / min, in an ice bath, homogenization time 2 min; and at an ultrasonic power of 180 W, ultrasonication for 6 min to obtain grapefruit oil liposomes.

[0037] (2) Dissolve chitosan in 1% (v / v) acetic acid aqueous solution, stir at 600 r / min for 45 min to prepare a 1.5 mg / mL chitosan solution. Add an equal volume of 1.5 mg / mL chitosan solution dropwise to the grapefruit oil liposomes obtained in step (1), stir at 600 r / min for 45 min to obtain chitosan-modified grapefruit oil liposomes. Dissolve sodium alginate in deionized water, adjust the pH to 6, and stir magnetically at 600 r / min for 45 min to obtain a 4 mg / mL sodium alginate solution. Add an equal volume of 4 mg / mL sodium alginate solution to the chitosan-modified grapefruit oil liposomes, and continue stirring at 600 r / min for 45 min to obtain sodium alginate-chitosan grapefruit oil liposomes.

[0038] Example 3 Preparation method of sodium alginate-chitosan modified grapefruit oil liposomes

[0039] This embodiment provides a method for preparing sodium alginate-chitosan modified grapefruit oil liposomes, the method comprising the following steps:

[0040] (1) Weigh 250 mg of egg yolk lecithin, 50 mg of grapefruit oil, 50 mg of Tween-80 and 50 mg of β-sitosterol, add 30 mL of anhydrous ethanol and stir thoroughly to form a mixed solution. Use a syringe to inject the mixed solution into 120 mL of deionized water and mix well. Rotate the suspension to remove ethanol until the mass concentration of egg yolk lecithin in the grapefruit oil liposomes is 10 mg / mL. The conditions for rotary evaporation are: heating temperature 45°C, rotation speed 200 r / min, time 40 min. Then, at a speed of 10000 r / min, in an ice bath, homogenization time 2 min; at an ultrasonic power of 180 W, ultrasonic for 6 min to obtain grapefruit oil liposomes.

[0041] (2) Dissolve chitosan in 1% (v / v) acetic acid aqueous solution, stir at 600 r / min for 45 min to prepare a 1.5 mg / mL chitosan solution. Add an equal volume of 1.5 mg / mL chitosan solution dropwise to the grapefruit oil liposomes obtained in step (1), stir at 600 r / min for 45 min to obtain chitosan-modified grapefruit oil liposomes. Dissolve sodium alginate in deionized water, adjust the pH to 6, and stir magnetically at 600 r / min for 45 min to obtain a 4 mg / mL sodium alginate solution. Add an equal volume of 4 mg / mL sodium alginate solution to the chitosan-modified grapefruit oil liposomes, and continue stirring at 600 r / min for 45 min to obtain sodium alginate-chitosan grapefruit oil liposomes.

[0042] Example 4 Preparation method of sodium alginate-chitosan modified grapefruit oil liposomes

[0043] This embodiment provides a method for preparing sodium alginate-chitosan modified grapefruit oil liposomes, the method comprising the following steps:

[0044] (1) Weigh 250 mg of egg yolk lecithin, 50 mg of grapefruit oil, 50 mg of Tween-80 and 50 mg of β-sitosterol, add 30 mL of anhydrous ethanol and stir thoroughly to form a mixed solution. Use a syringe to inject the mixed solution into 120 mL of deionized water, stir magnetically at 45°C and 600 r / min for 45 min to obtain a grapefruit oil liposome suspension. Rotate the suspension to remove ethanol until the mass concentration of egg yolk lecithin in the grapefruit oil liposome is 10 mg / mL. The conditions for rotary evaporation are: heating temperature 45°C, rotation speed 200 r / min, time 40 min. Then, successively at a speed of 10000 r / min, in an ice bath, homogenization time 2 min to obtain grapefruit oil liposomes.

[0045] (2) Dissolve chitosan in 1% (v / v) acetic acid aqueous solution, stir at 600 r / min for 45 min to prepare a 1.5 mg / mL chitosan solution. Add an equal volume of 1.5 mg / mL chitosan solution dropwise to the grapefruit oil liposomes obtained in step (1), stir at 600 r / min for 45 min to obtain chitosan-modified grapefruit oil liposomes. Dissolve sodium alginate in deionized water, adjust the pH to 6, and stir magnetically at 600 r / min for 45 min to obtain a 4 mg / mL sodium alginate solution. Add an equal volume of 4 mg / mL sodium alginate solution to the chitosan-modified grapefruit oil liposomes, and continue stirring at 600 r / min for 45 min to obtain sodium alginate-chitosan grapefruit oil liposomes.

[0046] Example 5 A method for preparing sodium alginate-chitosan modified grapefruit oil liposomes

[0047] This embodiment provides a method for preparing sodium alginate-chitosan modified grapefruit oil liposomes, the method comprising the following steps:

[0048] (1) Weigh 250 mg of egg yolk lecithin, 50 mg of grapefruit oil, 50 mg of Tween-80 and 50 mg of β-sitosterol, add 30 mL of anhydrous ethanol and stir thoroughly to form a mixed solution. Use a syringe to inject the mixed solution into 120 mL of deionized water, stir magnetically at 45°C and 600 r / min for 45 min to obtain a grapefruit oil liposome suspension. Rotate the suspension to remove ethanol until the mass concentration of egg yolk lecithin in the grapefruit oil liposome is 10 mg / mL. The conditions for rotary evaporation are: heating temperature 45°C, rotation speed 200 r / min, time 40 min. Then, ultrasonicate for 6 min at an ultrasonic power of 180 W to obtain grapefruit oil liposomes.

[0049] (2) Dissolve chitosan in 1% (v / v) acetic acid aqueous solution, stir at 600 r / min for 45 min to prepare a 1.5 mg / mL chitosan solution. Add an equal volume of 1.5 mg / mL chitosan solution dropwise to the grapefruit oil liposomes obtained in step (1), stir at 600 r / min for 45 min to obtain chitosan-modified grapefruit oil liposomes. Dissolve sodium alginate in deionized water, adjust the pH to 6, and stir magnetically at 600 r / min for 45 min to obtain a 4 mg / mL sodium alginate solution. Add an equal volume of 4 mg / mL sodium alginate solution to the chitosan-modified grapefruit oil liposomes, and continue stirring at 600 r / min for 45 min to obtain sodium alginate-chitosan grapefruit oil liposomes.

[0050] Example 6 Preparation method of sodium alginate-chitosan modified grapefruit oil liposomes

[0051] This embodiment provides a method for preparing sodium alginate-chitosan modified grapefruit oil liposomes, the method comprising the following steps:

[0052] (1) Weigh 250 mg of egg yolk lecithin, 50 mg of grapefruit oil, 50 mg of Tween-80 and 50 mg of β-sitosterol, add 30 mL of anhydrous ethanol and stir thoroughly to form a mixed solution. Use a syringe to inject the mixed solution into 120 mL of deionized water, stir magnetically at 45°C and 600 r / min for 45 min to obtain a grapefruit oil liposome suspension. Rotate the suspension to remove ethanol until the mass concentration of egg yolk lecithin in the grapefruit oil liposome is 10 mg / mL. The conditions for rotary evaporation are: heating temperature 45°C, rotation speed 200 r / min, time 40 min. Then, at a speed of 10000 r / min, in an ice bath, homogenization time 2 min; and at an ultrasonic power of 180 W, ultrasonic for 6 min to obtain grapefruit oil liposomes.

[0053] (2) Dissolve chitosan in 1% (v / v) acetic acid aqueous solution, stir at 600 r / min for 45 min to prepare a 1.5 mg / mL chitosan solution. Add an equal volume of the 1.5 mg / mL chitosan solution dropwise to the grapefruit oil liposomes obtained in step (1), stir at 600 r / min for 45 min to obtain chitosan-modified grapefruit oil liposomes.

[0054] Example 7 A method for preparing sodium alginate-chitosan modified grapefruit oil liposomes

[0055] This embodiment provides a method for preparing sodium alginate-chitosan modified grapefruit oil liposomes, the method comprising the following steps:

[0056] (1) Weigh 250 mg of egg yolk lecithin, 50 mg of grapefruit oil, 50 mg of Tween-80 and 50 mg of β-sitosterol, add 30 mL of anhydrous ethanol and stir thoroughly to form a mixed solution. Use a syringe to inject the mixed solution into 120 mL of deionized water, stir magnetically at 45°C and 600 r / min for 45 min to obtain a grapefruit oil liposome suspension. Rotate the suspension to remove ethanol until the mass concentration of egg yolk lecithin in the grapefruit oil liposome is 10 mg / mL. The conditions for rotary evaporation are: heating temperature 45°C, rotation speed 200 r / min, time 40 min. Then, at a speed of 10000 r / min, in an ice bath, homogenization time 2 min; and at an ultrasonic power of 180 W, ultrasonic for 6 min to obtain grapefruit oil liposomes.

[0057] (2) Chitosan was dissolved in 1% (v / v) acetic acid aqueous solution, stirred at 600 r / min for 45 min to prepare a 1.5 mg / mL chitosan solution. An equal volume of the 1.5 mg / mL chitosan solution was added dropwise to the grapefruit oil liposomes obtained in step (1), and stirred at 600 r / min for 45 min. No chitosan-modified grapefruit oil liposomes were formed.

[0058] Example 8 Preparation method of sodium alginate-chitosan modified grapefruit oil liposomes

[0059] This embodiment provides a method for preparing sodium alginate-chitosan modified grapefruit oil liposomes, the method comprising the following steps:

[0060] (1) Weigh 250 mg of egg yolk lecithin, 50 mg of grapefruit oil, 50 mg of Tween-80 and 50 mg of β-sitosterol, add 30 mL of anhydrous ethanol and stir thoroughly to form a mixed solution. Use a syringe to inject the mixed solution into 120 mL of deionized water, stir magnetically at 45°C and 600 r / min for 45 min to obtain a grapefruit oil liposome suspension. Rotate the suspension to remove ethanol until the mass concentration of egg yolk lecithin in the grapefruit oil liposome is 10 mg / mL. The conditions for rotary evaporation are: heating temperature 45°C, rotation speed 200 r / min, time 40 min. Then, at a speed of 10000 r / min, in an ice bath, homogenization time 2 min; and at an ultrasonic power of 180 W, ultrasonic for 6 min to obtain grapefruit oil liposomes.

[0061] (2) Sodium alginate was dissolved in deionized water, the pH was adjusted to 6, and magnetic stirring was performed at 600 r / min for 45 min to obtain a 4 mg / mL sodium alginate solution. An equal volume of the 4 mg / mL sodium alginate solution was added to the grapefruit oil liposomes obtained in step (1), and stirring was continued at 600 r / min for 45 min. No sodium alginate-modified grapefruit oil liposomes were formed.

[0062] Experiment 1 Determination of encapsulation efficiency of grapefruit oil

[0063] The encapsulation efficiency of grapefruit oil liposomes was determined by solvent extraction. Mix grapefruit oil liposomes (1 mL) with n-hexane (4 mL) and vortex for 30 seconds. Then, centrifuge at 5000r / min for 5 minutes to remove the unencapsulated grapefruit oil in the upper layer. Add 4 mL of ethanol to the lower layer liposomes to destroy the liposome structure, and then add 4 mL of n-hexane to extract the encapsulated grapefruit oil. Repeat the above operation twice, combine the extracts, and remove the solvent with a water bath nitrogen blowdown instrument to obtain the grapefruit oil encapsulated in the liposomes. After the grapefruit oil is methylated, it is analyzed by gas chromatography, and the amount of grapefruit oil is calculated using the amount of oleic acid. The encapsulation efficiency of grapefruit oil liposomes is calculated by the formula, and the calculation formula is as follows:

[0064] Encapsulation rate (%) = Encapsulated grapefruit oil content / Initial addition amount of grapefruit oil × 100

[0065] Fatty acid methyl esterification: add 10mg grapefruit oil to a test tube, add 4mL n-hexane to dissolve, add 200uL, 2mol / L potassium hydroxide methanol solution, vortex for 1min, add 0.5g anhydrous sodium sulfate to remove moisture. After standing for a while, use a syringe to absorb an appropriate amount of supernatant, pass through a 0.45um membrane, and wait for testing.

[0066] Gas chromatography conditions: FID detector; DB-WAX quartz capillary column (30m×0.32mm×0.25μm); programmed temperature: initial temperature 100℃, hold for 0.2min, increase to 242℃ at 10℃ / min, hold for 10min; carrier gas is nitrogen, flow rate 2.5mL / min; air flow rate 300mL / min; hydrogen flow rate 30mL / min; injection port temperature 240℃; detector temperature 260℃; no split; injection volume 1.0μL.

[0067] Table 1 Encapsulation efficiency of grapefruit oil in different groups

[0068]

[0069] Note: Different letters indicate significant differences between groups (p>0.05)

[0070] From Table 1, we can see that:

[0071] (1) Compared with Examples 6-8, chitosan and sodium alginate modification can significantly improve the encapsulation efficiency of grapefruit oil liposomes.

[0072] (2) Compared with Examples 7-8, Examples 1-6 have different chitosan structures or lack of chitosan modification steps, which are not conducive to the formation of layer-by-layer modified grapefruit oil liposomes.

[0073] (3) Comparing Experimental Example 1 with Examples 3-5, the changes in the preparation conditions of grapefruit oil liposomes are not conducive to the preparation of sodium alginate-chitosan modified grapefruit oil liposomes. Among them, the lack of the ultrasonic step is the most unfavorable for the preparation of sodium alginate-chitosan modified grapefruit oil liposomes, and the homogenization step has the second greatest impact.

[0074] (4) Compared with Example 2, the change in the ratio of grapefruit oil, egg yolk lecithin, Tween-80 and β-sitosterol is not conducive to the preparation of sodium alginate-chitosan modified grapefruit oil liposomes.

[0075] The present invention uses specific embodiments to illustrate the principles and implementation methods of the present invention. The description of the above embodiments is only used to help understand the method of the present invention and its core idea. At the same time, for those skilled in the art, according to the idea of ​​the present invention, there will be changes in the scope of the specific implementation methods. In summary, the content of this specification should not be understood as limiting the present invention.

Claims

1. A method for preparing sodium alginate-chitosan modified grapefruit oil liposomes, characterized in that: The following steps are involved: Step 1, grapefruit oil, egg yolk lecithin, Tween-80 and β-sitosterol are mixed and dissolved in anhydrous ethanol in a mass ratio of 1:5:1:1, and after mixing evenly, 4 times the volume of deionized water is added to the mixture, and magnetic stirring is performed to obtain a grapefruit oil liposome suspension; Step 2, the grapefruit oil liposome suspension obtained in step 1 is subjected to rotary evaporation treatment to remove ethanol, and then homogenized and ultrasonically treated to obtain grapefruit oil liposomes; Step 3, adding an equal volume of 1.5 mg / mL chitosan solution to the grapefruit oil liposomes obtained in step 2, stirring evenly to obtain chitosan-modified grapefruit oil liposomes; Step 4: Add an equal volume of 4 mg / mL sodium alginate solution to the chitosan-modified grapefruit oil liposomes obtained in step 3, and stir evenly to obtain sodium alginate-chitosan-modified grapefruit oil liposomes.

2. The method for preparing sodium alginate-chitosan modified grapefruit oil liposomes according to claim 1, characterized in that: In step 1, the heating temperature of the magnetic stirring is 45° C., and the stirring rate is 600 r / min.

3. The method for preparing sodium alginate-chitosan modified grapefruit oil liposomes according to claim 1, characterized in that: In step 2, the mass concentration of egg yolk lecithin in the grapefruit oil liposome is 10 mg / mL.

4. The method for preparing sodium alginate-chitosan modified grapefruit oil liposomes according to claim 1, characterized in that: In step 2, the conditions of the rotary evaporation are: heating temperature 45° C., rotation speed 200 r / min, and time 40 min.

5. The method for preparing sodium alginate-chitosan modified grapefruit oil liposomes according to claim 1, characterized in that: In step 2, the homogenization is carried out in an ice bath, the homogenization speed is 10000 r / min, and the homogenization time is 2 min; the power of the ultrasonic treatment is 180 W, and the ultrasonic treatment time is 6 min.

6. The method for preparing sodium alginate-chitosan modified grapefruit oil liposomes according to claim 1, characterized in that: In step 3, the preparation method of the 1.5 mg / mL chitosan solution is: dissolve chitosan in 1% v / v acetic acid aqueous solution, stir at 600 r / min for 45 min to prepare a 1.5 mg / mL chitosan solution; the chitosan has a deacetylation degree of >95% and a viscosity of 100-200 mp.s.

7. The method for preparing sodium alginate-chitosan modified grapefruit oil liposomes according to claim 1, characterized in that: In step 4, the preparation method of the 4 mg / mL sodium alginate solution is: dissolving sodium alginate in deionized water until it is completely dissolved, adjusting the pH to 6, and magnetically stirring at 600 r / min for 45 minutes to obtain a 4 mg / mL sodium alginate solution.

8. A sodium alginate-chitosan modified grapefruit oil liposome, characterized in that: The method is prepared according to any one of claims 1 to 8.

9. Use of the sodium alginate-chitosan modified grapefruit oil liposomes according to claim 8 in the preparation of grapefruit oil delivery preparations.

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