Nanocrystallization preparation method and application of small-molecular panax notoginseng polysaccharide

Abstract

The invention relates to a nanocrystallization preparation method and application of micromolecular panax notoginseng polysaccharide, and belongs to the field of biological medicine. The method comprises the following steps: (1) weighing a certain amount of micromolecular panax notoginseng polysaccharide, lecithin, lanolin and triton X-100, dissolving in absolute ethyl alcohol, carrying out rotary evaporation to remove ethyl alcohol, and forming a uniform and transparent film on a bottle wall; (2) adding a PBS (Phosphate Buffer Solution) into the round-bottom flask to wash the film on the bottle wall so as to obtain a film-washed solution; (3) carrying out ultrasonic treatment on the membrane-washed liquid, and standing for a certain period of time to obtain a small-molecular panax notoginseng polysaccharide nano raw material; (4) placing in an ultrafiltration centrifuge tube, centrifuging, and collecting filtrate; and (5) concentrating the filtrate, and freeze-drying to obtain the micromolecular nano pseudo-ginseng polysaccharide extract. When the micromolecular nano pseudo-ginseng polysaccharide extract prepared by the preparation method disclosed by the invention is used for preparing cosmetics with a moisturizing function, the moisturizing performance is remarkable.

Description

Technical Field

[0001] This application relates to the field of biomedicine, and particularly to a method for preparing nanosized small molecule notoginseng polysaccharide and its application. Background Art

[0002] Polysaccharide is one of the main active substances in notoginseng, and the content of polysaccharide in notoginseng is about 10%. Research shows that notoginseng polysaccharide plays an important role in promoting skin health, delaying skin aging, and maintaining the balance of skin immunogenic tolerance. The polysaccharide extract mainly composed of notoginseng disclosed in CN101461763A "Preparation Method and Use of Cosmetics Containing Notoginseng Polysaccharide" can be made into cosmetics such as facial milk, ionic water, essence, facial mask, and cream by adding auxiliaries such as surfactants and emulsifiers, which can play a role in moisturizing and hydrating.

[0003] However, the notoginseng polysaccharide prepared by traditional methods is often limited by factors such as particle size, irregular shape, and dispersibility to a certain extent, which restricts its moisturizing effect. Summary of the Invention

[0004] To solve or partially solve the problems existing in the related art, this application provides a method for preparing nanosized small molecule notoginseng polysaccharide. Through this preparation method, the particle size of the prepared small molecule nanosized notoginseng polysaccharide extract is 20.00 - 450.00 nm, the shape is more regular, generally spherical granular, and the dispersion is more uniform. Compared with the non-nanosized small molecule notoginseng polysaccharide, when used in cosmetics with moisturizing functions, the moisturizing performance is significantly improved.

[0005] A method for preparing nanosized small molecule notoginseng polysaccharide in this application includes the following steps:

[0006] (1) Weigh a certain amount of small molecule notoginseng polysaccharide, lecithin, lanolin, and Triton X-100, dissolve them in absolute ethanol, pour the mixed solution into a round-bottom flask and perform rotary evaporation to remove ethanol, and form a uniform and transparent film on the inner wall of the flask;

[0007] (2) Add PBS buffer solution to the round-bottom flask to wash the film on the inner wall of the flask to obtain the solution after film washing;

[0008] (3) After ultrasonic treatment of the solution after film washing and standing for a certain period of time, obtain the small molecule notoginseng polysaccharide nano raw material;

[0009] (4) Place the small molecule notoginseng polysaccharide nano raw material in the inner tube of an ultrafiltration centrifuge tube with a molecular weight cut-off of 10 kDa, centrifuge, and collect the filtrate in the outer tube of the ultrafiltration centrifuge tube; repeat the above centrifugation operation several times and add an appropriate amount of distilled water to the inner tube each time;

[0010] (5) The collected filtrate is concentrated and then freeze-dried to obtain the small molecule nano notoginseng polysaccharide extract.

[0011] Furthermore, the mass ratio of the small molecule notoginseng polysaccharide, lecithin, lanolin, Triton X-100 and absolute ethanol is 1:(5-15):(5-15):(1-5):(18-22).

[0012] Furthermore, step (4) is repeated 3 times;

[0013] Furthermore, when performing rotary evaporation in step (4), the temperature is lower than 60 °C.

[0014] Furthermore, the proportion of the small molecule notoginseng polysaccharide with a molecular weight of 164-1034 Da is 90%.

[0015] This application also provides a small molecule nano notoginseng polysaccharide extract prepared based on the above preparation method and the application of the small molecule nano notoginseng polysaccharide extract in the preparation of cosmetics with moisturizing functions.

[0016] The mechanism of this application:

[0017] The small molecule notoginseng polysaccharide is the core for forming nanoparticles as the main component. Since polysaccharides are hydrophilic, they can form nanoparticles through hydrophobic modification or binding with other substances.

[0018] As an amphiphilic molecule, lecithin plays an emulsifying and stabilizing role in ethanol. During rotary evaporation, as ethanol is removed, lecithin combines with other components to form vesicle or liposome structures, helping the polysaccharide to form nanoparticles.

[0019] Lanolin is a hydrophobic substance and acts as a hydrophobic core to promote the aggregation of polysaccharides. When ethanol evaporates, the hydrophobicity of lanolin increases, causing polysaccharide molecules to aggregate around it to form nanoparticles. At the same time, the viscosity of lanolin can help the particles adhere to the bottle wall.

[0020] As a non-ionic surfactant, Triton X-100 reduces the interfacial tension during the dissolution process and promotes the uniform mixing of each component. During the evaporation process, as ethanol decreases, Triton X-100 forms micelles, guiding the directional arrangement of polysaccharides and lecithin to prevent aggregation. Triton X-100 acts as an emulsifier and plays a role in stabilizing the dispersion system in the medium.

[0021] Rotary evaporation accelerates the volatilization of ethanol, resulting in the rapid concentration of the solution, triggering the mutual fusion of substances such as small molecule notoginseng polysaccharide, lecithin and lanolin, forming a stable nano-structure between different components, and under the action of Triton X-100, the dispersion is more uniform.

[0022] The beneficial effects of this application are as follows:

[0023] The particle size of the small molecule nano - notoginseng polysaccharide extract prepared by the preparation method in this application is in the range of 20.00 - 450.00 nm. Its shape is more regular, generally spherical particles, and its dispersion is more uniform. Compared with the non - nano - sized small molecule notoginseng polysaccharide, when used in cosmetics with moisturizing functions, the moisturizing performance is significantly improved. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] By describing the exemplary embodiments of this application in more detail in conjunction with the attached drawings, the above - mentioned and other purposes, features, and advantages of this application will become more obvious. Among them, in the exemplary embodiments of this application, the same reference numerals generally represent the same components.

[0025] Figure 1 The SEM diagram (×2000) of the small molecule nano - notoginseng polysaccharide extract prepared in Example 1 is shown;

[0026] Figure 2 The TEM diagram of the small molecule nano - notoginseng polysaccharide extract prepared in Example 1 is shown;

[0027] Figure 3 The SEM diagram of the small molecule notoginseng polysaccharide in Example 1 is shown;

[0028] Figure 4 The diagram of the skin moisture content of the subjects is shown;

[0029] Figure 5 The molecular weight distribution diagram of the small molecule notoginseng polysaccharide in Example 1 is shown. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0030] The embodiments of this application will be described in more detail below with reference to the examples. Although the embodiments of this application are shown, it should be understood that this application can be implemented in various forms and should not be limited by the embodiments set forth herein. On the contrary, these embodiments are provided to make this application more thorough and complete, and to fully convey the scope of this application to those skilled in the art.

[0031] Example 1

[0032] A method for nano - preparing a small molecule notoginseng polysaccharide in this application includes the following steps:

[0033] (1) Weigh a certain amount of small molecule notoginseng polysaccharide, lecithin, lanolin and Triton X-100, dissolve them in absolute ethanol, pour the mixed solution into a round-bottom flask, and perform rotary evaporation at 50 °C and -0.1 MPa to remove ethanol, forming a uniform and transparent film on the inner wall of the flask. Among them, the mass ratio of small molecule notoginseng polysaccharide, lecithin, lanolin, Triton X-100 and absolute ethanol is 1:10:8:5:20;

[0034] It should be noted that the small molecule notoginseng polysaccharide in this example is prepared through the following preparation process:

[0035] 1. Add water to the notoginseng roots and extract for 2 hours, extract twice, filter, and collect the filtrate;

[0036] 2. Add ethanol to the filtrate until the final ethanol concentration reaches 40%, perform alcohol precipitation overnight, filter with filter paper, and collect the precipitate;

[0037] 3. Add water to the precipitate, stir to dissolve and dilute, add ethanol until the final ethanol concentration reaches 60%, perform alcohol precipitation overnight, filter with filter paper, and collect the precipitate;

[0038] 4. Dissolve the precipitate by adding water and stirring, add 95% ethanol to the dilution until the final ethanol concentration reaches 80%, perform alcohol precipitation overnight, and collect the precipitate;

[0039] 5. Dissolve the precipitate in water, decolorize with activated carbon, filter, collect the filtrate, and freeze-dry to obtain small molecule notoginseng polysaccharide. The molecular weight distribution range of the prepared small molecule notoginseng polysaccharide is as Figure 5 shown.

[0040] (2) Then add 20 mL of 0.05 mol / L phosphate buffer solution (PBS) with pH 6.0 to the round-bottom flask, and then continue to rotate for 30 minutes to wash the film at 50 °C and normal pressure to obtain the post-washing solution;

[0041] (3) Ultrasonically treat the post-washing solution at a power of 250 W for 5 - 15 minutes, and then let it stand at 20 °C for 2 hours to obtain small molecule notoginseng polysaccharide nano raw materials;

[0042] (4) Transfer the obtained small molecule notoginseng polysaccharide nano raw materials to the inner tube of an ultrafiltration centrifugal tube (with a molecular weight cut-off of 10 kDa), centrifuge at a speed of 3000 r / min for 20 minutes, and collect the filtrate in the outer tube of the ultrafiltration centrifugal tube;

[0043] Add 2 mL of distilled water to the inner tube, redisperse the concentrated solution of small molecule notoginseng polysaccharide nano raw materials, centrifuge again at a speed of 3000 r / min for 20 minutes, and collect the filtrate in the outer tube of the ultrafiltration centrifugal tube again. In order to separate the substances that have not formed small molecule notoginseng polysaccharide nano from the small molecule notoginseng polysaccharide nano substances, this step is repeated 3 times.

[0044] (5) After all the collected filtrates are mixed, concentrated and freeze-dried, a small molecule nano-sized Panax notoginseng polysaccharide extract is obtained.

[0045] The results showed that the small molecule nano-sized Panax notoginseng polysaccharide extract prepared in Example 1 had a particle size of 20.00 to 440.00 nm and a more regular shape, generally in the form of spherical particles. Figure 1 Shown is the SEM schematic diagram of the prepared small molecule nano-Panax notoginseng polysaccharide extract (×2000); Figure 2 Shown is a schematic diagram of the TEM of the prepared small molecule nano-Panax notoginseng polysaccharide extract. Figure 3 Shown is a SEM schematic diagram of the small molecule Panax notoginseng polysaccharide in this application. Figure 1 and Figure 3 It can be seen that the small molecule Panax notoginseng polysaccharide before nano-crystallization is rough and irregular. After nano-crystallization, the particle edges are clearer, the shape is regular, and the particle dispersion is more uniform.

[0046] Example 2

[0047] The difference between Example 2 and Example 1 is that the mass ratio of small molecule Panax notoginseng polysaccharide, lecithin, lanolin, Triton X-100 and anhydrous ethanol is 1:5:15:1:18, and the rest are the same.

[0048] The small molecule nano-sized Panax notoginseng polysaccharide extract prepared in Example 2 has a particle size of 20.00 to 450.00 nm.

[0049] Example 3

[0050] The difference between Example 2 and Example 1 is that the mass ratio of small molecule Panax notoginseng polysaccharide, lecithin, lanolin, Triton X-100 and anhydrous ethanol is 1:7:8:1:22, and the rest are the same.

[0051] The small molecule nano-sized Panax notoginseng polysaccharide extract prepared in Example 3 has a particle size of 20.00 to 450.00 nm.

[0052] Example 4

[0053] Evaluation experiment on the moisturizing performance of the small molecule nano-sized Panax notoginseng polysaccharide extract prepared in this application

[0054] Specifically, the small molecule nano - notoginseng polysaccharide extract prepared in Example 1, the raw material small molecule notoginseng polysaccharide in Example 1, and hyaluronic acid were respectively spread on the bottom of a pre - weighed weighing bottle. 1 g of purified water was added to each, and each sample had 3 parallels. They were placed in a desiccator containing drying silica gel and then in a stability test chamber (25 °C) for a moisture retention experiment. After 48 h, they were taken out and weighed. The moisture retention rate (%) was calculated. Moisture retention rate (%) = (sample mass before moisture retention - sample mass after moisture retention) / sample mass before moisture retention * 100%

[0055] The experimental results are shown in Table 1

[0056] Table 1

[0057]

[0058]

[0059] Example 5

[0060] For the skin moisture content test, the blank moisturizing cream formula is shown in Table 2:

[0061] Table 2

[0062] Raw material name Mass W(%) Tricaprylin 3.50 Isocetane 3.50 Dioctyl carbonate 2.00 Cyclopentasiloxane 2.00 Cetearyl alcohol 1.20 Polydimethylsiloxane (100 cps) 1.00 Propylparaben 0.10 M-68 emulsifier 3.00 Butylene glycol 4.00 Carbopol 940 0.40 Methylparaben 0.20 Allantoin 0.10 Betaine 1.00 Aminomethyl propanol 0.20 Phenoxyethanol 0.50 Water Up to 100

[0063] A small molecule nano - notoginseng polysaccharide solution with a mass concentration of 2 g·L -1 was prepared and added to the blank moisturizing cream to make the mass fraction of small molecule notoginseng polysaccharide in the moisturizing cream reach 5%; Moisturizing creams made by replacing the small molecule nano - notoginseng polysaccharide solution with small molecule notoginseng polysaccharide (2 g·L -1 ) and hyaluronic acid solution (2 g·L -1 ) with a mass fraction of 5% were used as controls. Twelve volunteers aged 18 - 40 years were selected as subjects. Before the experiment, the subjects washed their left and right forearms, dried them, and were equilibrated in an environment of 22 °C and relative humidity of 50% - 65% for 30 min. A 3 cm × 3 cm test area was marked on the left and right forearms of the subjects. The skin moisture content of the test area before and after applying the cream was measured using a skin moisture test probe. Three sites were taken in each area, and the results were averaged.

[0064] Six hours after applying the small molecule nano - polysaccharide moisturizing cream, the skin moisture content of the subjects was as Figure 4 shown.

[0065] From Figure 4 it can be seen that after adding small molecule notoginseng nano - polysaccharide, the moisturizing effect is significantly improved, higher than the moisturizing performance of applying small molecule notoginseng polysaccharide, and equivalent to the performance of hyaluronic acid.

[0066] The embodiments of the present application have been described above. The above description is exemplary and not exhaustive, and is also not limited to the disclosed embodiments. Many modifications and variations are obvious to those of ordinary skill in the art without departing from the scope and spirit of the described embodiments. The choice of terms used herein is intended to best explain the principles of the embodiments, practical applications, or improvements to technologies in the market, or to enable other ordinary skilled persons in the art to understand the embodiments disclosed herein.

Claims

1. A method for preparing nano-sized small molecule notoginseng polysaccharide, characterized in that It includes the following steps: (1) Weigh a certain amount of small molecule notoginseng polysaccharide, lecithin, lanolin and Triton X-100, dissolve them in absolute ethanol, pour the mixed solution into a round-bottom flask and perform rotary evaporation to remove ethanol, forming a uniform and transparent film on the inner wall of the flask; (2) Add PBS buffer solution into the round-bottom flask to wash the film on the inner wall of the flask, and obtain the solution after film washing; (3) After subjecting the solution after film washing to ultrasonic treatment, let it stand for a certain period of time to obtain small molecule notoginseng polysaccharide nano raw material; (4) Place the small molecule notoginseng polysaccharide nano raw material in the inner tube of an ultrafiltration centrifugal tube with a molecular weight cut-off of 10 kDa, centrifuge, and collect the filtrate in the outer tube of the ultrafiltration centrifugal tube; repeat the above centrifugation operation several times and supplement an appropriate amount of distilled water in the inner tube each time; (5) Concentrate and freeze-dry the collected filtrate to obtain the small molecule nano notoginseng polysaccharide extract.

2. The nano-preparation method of a small molecule notoginseng polysaccharide according to claim 1, characterized in that, The mass ratio of the small molecule notoginseng polysaccharide, lecithin, lanolin, Triton X-100 and absolute ethanol is 1:(5-15):(5-15):(1-5):(18-22).

3. A method for preparing nano-sized small molecule notoginseng polysaccharide according to claim 1, characterized in that, In step (4), it is repeated 3 times.

4. The nano-preparation method of a small molecule notoginseng polysaccharide according to claim 1, characterized in that When performing rotary evaporation in step (4), the temperature is lower than 60°C.

5. A method for preparing nanosized small molecule notoginseng polysaccharide according to claim 2, characterized in that, The proportion of the small molecule notoginseng polysaccharide with a molecular weight of 164-1034 Da is 90%.

6. The small molecule notoginseng nano polysaccharide extract prepared by the preparation method according to any one of claims 1-5.

7. The application of the small molecule notoginseng nano polysaccharide extract according to claim 1 in the preparation of cosmetics with moisturizing function.

Images