Method for preparing oleic acid lipid vesicles by using magnetic stirrer
By using a magnetic stirrer under a heating environment, oleic lipid vesicles are prepared by mixing phosphatidylcholine and oleic acid, the problems of complex equipment and cumbersome operation of traditional methods are solved, and a simpler and more efficient preparation process is achieved.
Patent Information
- Application Number
- CN202510086940.5
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-01-20
- Publication Date
- 2025-04-18
- Estimated Expiration
- Not applicable · inactive patent
AI Technical Summary
The traditional lipid vesicle preparation methods have problems such as complex equipment, cumbersome operation, and high energy consumption, which limits their wide application.
Oleic acid lipid vesicles were prepared by dissolving phosphatidylcholine in chloroform under a heating environment using a magnetic stirrer.
It realizes simple and rapid preparation of lipid vesicles, reduces operation difficulty and equipment costs, and is more efficient than traditional methods.
Smart Images
Figure CN119838480A_ABST
Abstract
Description
Technical Field
[0001] The invention relates to the field of biotechnology, and in particular to a method for preparing oleic acid lipid vesicles by utilizing a magnetic stirrer. Background Art
[0002] Lipid vesicles are spherical structures formed by the self-assembly of lipid molecules and are widely used in drug delivery, nanomaterials, cosmetics and biological research. Among them, oleic acid lipid vesicles have become a hot topic of research due to their special physicochemical properties and biocompatibility.
[0003] However, traditional lipid vesicle preparation methods, such as thin film hydration, ultrasound, and high-pressure homogenization, all have problems such as complex equipment, cumbersome operation, and high energy consumption, which limit their widespread application. Summary of the invention
[0004] In view of the above-mentioned deficiencies in the prior art, the present invention provides a method for preparing oleic acid lipid vesicles using a magnetic stirrer, which is simple to operate and highly efficient.
[0005] In order to achieve the above-mentioned object of the invention, the technical solution adopted by the present invention is:
[0006] A method for preparing oleic acid lipid vesicles using a magnetic stirrer is provided, which comprises the following specific steps:
[0007] S1: dissolving phosphatidylcholine in chloroform to obtain a phosphatidylcholine-chloroform solution, and vacuum drying the phosphatidylcholine-chloroform solution in a glass bottle to obtain a lipid membrane;
[0008] S2: Add NaOH solution with pH 8-9 into the glass bottle where the lipid film is formed;
[0009] S3: Add oleic acid to the mixed solution, heat to maintain a constant temperature, and stir with a magnetic stirrer to obtain an oleic acid lipid vesicle solution.
[0010] Further, the concentration of phosphatidylcholine in the phosphatidylcholine-chloroform solution is 3.2 mM.
[0011] Furthermore, the volume ratio of the added oleic acid to the mixed solution is 1:1000.
[0012] Furthermore, in step S3, the stirring time of the magnetic stirrer is 2 to 4 hours.
[0013] Furthermore, the phosphatidylcholine is dipalmitoylphosphatidylcholine or distearoylphosphatidylcholine.
[0014] Furthermore, in step S3, the temperature is T+20, wherein T is the phase transition temperature of phosphatidylcholine.
[0015] The beneficial effects of the present invention are:
[0016] The present invention uses chloroform to dissolve phosphatidylcholine, adds an alkaline solution and oleic acid, and stirs under a heated environment to achieve the preparation of lipid vesicles. The whole process is simple and the stirring time is short. Compared with the existing preparation method, the operation difficulty and equipment cost are reduced. BRIEF DESCRIPTION OF THE DRAWINGS
[0017] Figure 1 This is an inverted fluorescence microscope image of the vesicle prepared in Example 1;
[0018] Figure 2 This is an inverted fluorescence microscope image of the vesicle prepared in Example 2;
[0019] Figure 3 This is an inverted fluorescence microscope image of the vesicle prepared in Example 3;
[0020] Figure 4 This is an inverted fluorescence microscope image of the vesicle prepared in Example 4;
[0021] Figure 5 This is an inverted fluorescence microscope image of the vesicles prepared in Example 5. DETAILED DESCRIPTION
[0022] 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.
[0023] Example 1
[0024] Dissolve distearoylphosphatidylcholine DSPC in chloroform to prepare a 3.2 mM DSPC-chloroform solution. Take 400 μl of the DSPC-chloroform solution and place it in a glass vial. Put it in a vacuum drying oven and dry it in vacuum to form a lipid film. Use ultrapure water and NaOH to prepare a NaOH solution with a pH value of 8.5. Take 400 μl of the NaOH solution and add it to a glass vial, then add 0.4 μl of oleic acid. Stir with a magnetic stirrer at 75°C at a stirring speed of 400 rpm for 2 hours. In specific implementation, the stirring time can also be 2.5, 3, 3.5 or 4 hours; take the product obtained after the reaction is completed and use a fluorescence microscope to photograph the result. Figure 1 As shown, a uniform vesicle structure was obtained in the reaction.
[0025] Example 2
[0026] Dissolve dipalmitoylphosphatidylcholine DPPC in chloroform to prepare a 3.2 mM DPPC-chloroform solution. Take 400 μl of the DPPC-chloroform solution and place it in a glass vial. Place it in a vacuum drying oven and dry it in vacuum to form a lipid film. Use ultrapure water and NaOH to prepare a NaOH solution with a pH value of 8.5. Take 400 μl of the NaOH solution and add it to a glass vial, then add 0.4 μl of oleic acid. Stir with a magnetic stirrer at 60°C at a stirring speed of 100 rpm for 2 hours. Take the product obtained after the reaction is completed and use a fluorescence microscope to photograph the result. The result is as shown below. Figure 2 As shown, a uniform vesicle structure was obtained in the reaction.
[0027] Example 3
[0028] The difference between this embodiment and embodiment 2 is that the magnetic stirrer is used for stirring at a stirring speed of 200 rpm.
[0029] Example 4
[0030] The difference between this embodiment and embodiment 2 is that the magnetic stirrer is used for stirring at a stirring speed of 400 rpm.
[0031] Example 5
[0032] The difference between this embodiment and embodiment 2 is that the magnetic stirrer is used for stirring at a stirring speed of 800 rpm.
[0033] In summary, when oleic acid lipid vesicles are prepared from phosphatidylcholine, uniform micron-sized vesicles can be obtained by maintaining the temperature 20° C. higher than that of phosphatidylcholine and using a stirring speed of 100 to 800 rpm.
Claims
1. A method for preparing oleic acid lipid vesicles using a magnetic stirrer, characterized in that: The specific steps include: S1: dissolving phosphatidylcholine in chloroform to obtain a phosphatidylcholine-chloroform solution, and vacuum drying the phosphatidylcholine-chloroform solution in a glass bottle to obtain a lipid membrane; S2: Add NaOH solution with pH 8-9 into the glass bottle where the lipid film is formed; S3: Add oleic acid to the mixed solution, heat to maintain a constant temperature, and stir with a magnetic stirrer to obtain an oleic acid lipid vesicle solution.
2. The method for preparing oleic acid lipid vesicles using a magnetic stirrer according to claim 1, characterized in that: The concentration of phosphatidylcholine in the phosphatidylcholine-chloroform solution was 3.2 mM.
3. The method for preparing oleic acid lipid vesicles using a magnetic stirrer according to claim 1, characterized in that: The volume ratio of the added oleic acid to the mixed solution is 1:1000.
4. The method for preparing oleic acid lipid vesicles using a magnetic stirrer according to claim 1, characterized in that: The stirring time of the magnetic stirrer in step S3 is 2 to 4 hours.
5. The method for preparing oleic acid lipid vesicles using a magnetic stirrer according to claim 1, characterized in that: The phosphatidylcholine is dipalmitoylphosphatidylcholine or distearoylphosphatidylcholine.
6. The method for preparing oleic acid lipid vesicles using a magnetic stirrer according to claim 1, characterized in that: In step S3, the temperature is T+20, wherein T is the phase transition temperature of phosphatidylcholine.