Ginseng polysaccharide, preparation method and application thereof

Abstract

The application discloses a ginseng polysaccharide, a preparation method and application, and the ginseng polysaccharide is ginseng polysaccharide GP60, which is composed of glucose, galactose and arabinose, and the molar ratio of each monosaccharide is glucose: galactose: arabinose = 77.1: 16.8: 6.1; a new preparation method is adopted; after soaking and twice decocting, different proportions of 95% ethanol are added into the filtrate in batches; the obtained precipitate is added with water to prepare a solution with a relative density of 1.1 g / ml; a certain amount of the solution is taken, a certain proportion of chloroform and n-butanol is added, and a polysaccharide aqueous solution is prepared; the polysaccharide aqueous solution is added with 95% ethanol until the alcohol content reaches 60%; the solution is statically placed for 12 hours; centrifugal separation is carried out; the precipitate is collected; the collected precipitate is sequentially washed with 95% ethanol, anhydrous ethanol, isopropyl alcohol and ether; the solvent is volatilized; and the ginseng polysaccharide GP60 is obtained. The ginseng polysaccharide GP60 is non-toxic, has the effect of preventing light aging, and can be widely applied in the fields of food or health food, anti-light aging skin care products, washing and caring products and anti-light aging medicines.

Description

Technical Field

[0001] This invention belongs to the field of ginseng polysaccharide extraction technology, and specifically relates to a ginseng polysaccharide, its preparation method, and its application. Background Technology

[0002] Global warming caused by the greenhouse effect, including carbon dioxide emissions, has damaged the atmosphere to some extent, leading to an increase in ultraviolet (UV) radiation. Besides accelerating skin aging, this increased UV intensity has also led to a rise in sun-related skin conditions such as photodermatitis and sunburn, especially among outdoor workers. This phenomenon is receiving increasing attention. Acute photodamage, commonly known as sunburn, is one of the most common skin injuries, clinically manifesting as burning, redness, swelling, pain, and even blisters, burning pain, and skin peeling. Although sunburn almost always heals on its own after a period of time, the discomfort caused by sunburn can still cause considerable pain and distress. Furthermore, evidence suggests that 70% of skin cancers are directly or indirectly caused by sunburn. In addition, UV radiation from sunlight can also cause skin aging, leading to dryness, roughness, wrinkles, and various pigment deposits, negatively impacting appearance and even triggering various skin diseases or tumors.

[0003] Public information shows that ginseng polysaccharides are one of the main components of ginseng, a precious traditional Chinese medicine. They are natural high-molecular polymers composed of various monosaccharides, with low toxicity and rich bioactivity, including anti-tumor, immunomodulatory, antioxidant, hypoglycemic, and anti-radiation activities, and are widely used in clinical applications. Currently, the separation and extraction technology of ginseng polysaccharides is relatively mature, and there are many published reports. However, the structure and activity of ginseng polysaccharides obtained by different extraction methods vary. In recent years, researchers have found that many traditional Chinese medicine extracts with antioxidant effects can effectively alleviate photodamage to the skin. Ginseng polysaccharides have been reported to have significant antioxidant effects; however, research on the protective effects of ginseng polysaccharides against photoaging is scarce.

[0004] A patent for the extraction and production process of ginseng polysaccharides by Xu Yangsen et al. (publication number CN110862464A) discloses an industrial-scale process for producing ginseng polysaccharide solid beverages. The process includes slicing ginseng using a slicing device (slice thickness 0.1-1.0 cm), soaking for 1-10 hours at a temperature of 60-100℃, a material-to-liquid ratio of 1:10-1:80 g / ml, temperature control, membrane filtration, reverse osmosis membrane concentration, spray drying, and automatic packaging. Patent publication number CN117487035A discloses a ginseng polysaccharide, its preparation method, and its applications: ginseng is pulverized, extracted by hot water reflux, concentrated, centrifuged, and the supernatant is precipitated with ethanol solution. The precipitate is dialyzed with distilled water, and the non-dialyzed solution is freeze-dried to obtain ginseng polysaccharides; the molecular weight cutoff for dialysis is 3.5 kDa. The ginseng polysaccharides prepared by the above methods can be used in regulating intestinal flora, as well as in antidepressant drugs and anti-epileptic drugs. Patent CN116036142A discloses an invention patent for an anti-aging composition containing ginseng extracellular vesicles and its application. The mass ratio of ginseng extracellular vesicles to ginseng polysaccharide extract is [missing information]. The ginseng extracellular vesicles are obtained by gradient centrifugation. Ginseng juice is obtained by ginseng extraction. Ginseng or ginseng residue is mixed with water and extracted. The extract is mixed with ethanol / ethanol-water solution, precipitated, centrifuged, the precipitate is collected, and dried. This composition is used to improve skin anti-aging and inflammation after mechanical damage to the skin. Summary of the Invention

[0005] This invention aims to propose a ginseng polysaccharide GP60, its preparation method, and its application. It clarifies the molecular weight and composition of the ginseng polysaccharide GP60, the proportions of glucose, galactose, and arabinose in the ginseng polysaccharide GP60, and confirms that the ginseng polysaccharide GP60 prepared by the method has a good protective effect against UVA-induced photoaging of NIH3T3 cells, providing theoretical support for the application of ginseng polysaccharides in photoaging.

[0006] The present invention is achieved as follows: a ginseng polysaccharide, wherein the ginseng polysaccharide is ginseng polysaccharide GP60, with a molecular weight distribution of peak molecular weight of 10945 g / mol, number average molecular weight of 9688 g / mol, and weight average molecular weight of 79108 g / mol.

[0007] Furthermore, the ginseng polysaccharide is composed of glucose, galactose and arabinose, with a molar ratio of glucose:galactose:arabinose = 77.1:16.8:6.1.

[0008] A method for preparing ginseng polysaccharide, wherein the preparation method yields the ginseng polysaccharide as described above, and the method steps are as follows:

[0009] Step 1, two decoctions: The weight ratio of ginseng slices to water is 1:15. Soak for 1 hour, then heat to boiling and decoct for 1.5 hours. Filter and collect the filtrate. Add 10 times the amount of water to the residue and decoct a second time for 1 hour. Collect the filtrate.

[0010] Step 2, one-time addition of alcohol: Combine the filtrates collected from the two filtrations in Step 1, concentrate to a small volume, place at room temperature, add 95% pure ethanol until the alcohol content reaches 20%, let stand for 12 hours, then centrifuge and collect the supernatant.

[0011] Step 3, Second addition of alcohol: Add 95% pure ethanol to the supernatant collected in step 2 until the alcohol content reaches 40%, let stand for 12 hours, then centrifuge and collect the supernatant.

[0012] Step 4, three additions of alcohol: Add 95% pure ethanol to the supernatant collected in step 3 until the alcohol content reaches 60%, let stand for 12 hours, then centrifuge and collect the precipitate.

[0013] Step 5, Preparation of precipitate water dissolution: Add water to the precipitate collected in step 4 to dissolve it into a solution with a relative density of 1.1 g / ml;

[0014] Step 6, prepare polysaccharide aqueous solution:

[0015] 6.1 Take a certain amount of the solution obtained in step 5, add chloroform and n-butanol in a certain proportion, shake at 140 r / min for 10 min, and let stand for 12 h;

[0016] 6.2. Remove the supernatant, add the same proportions of chloroform and n-butanol as in step 6.1, shake at 140 r / min for 10 min, let stand for 12 h, remove the supernatant, repeat this process at least 5 times, and finally remove the supernatant to obtain a polysaccharide aqueous solution.

[0017] Step 7, Preparing the precipitate: Add 95% ethanol to the polysaccharide aqueous solution until the alcohol content reaches 60%, let stand for 12 h, centrifuge, and collect the precipitate;

[0018] Step 8, washing the precipitate: The precipitate collected in step 7 is washed sequentially with 95% ethanol, anhydrous ethanol, isopropanol and diethyl ether to evaporate the solvent and obtain the ginseng polysaccharide.

[0019] Furthermore, the volume ratio of the solution, chloroform, and n-butanol in step 6.1 is 5:0.8:0.2.

[0020] Furthermore, the process described in step 6.2 is repeated 5 times.

[0021] Furthermore, the centrifugation speed in steps 2-4 and 7 is 3000 r / min and the time is 10 min.

[0022] Furthermore, the ginseng polysaccharide can be used in anti-photoaging skincare products.

[0023] Furthermore, the ginseng polysaccharide can be used in anti-photoaging drugs.

[0024] The beneficial effects of this invention are: a ginseng polysaccharide GP60 with protective effect against UVA-induced photoaging of NIH3T3 cells was obtained; the molecular weight distribution of the ginseng polysaccharide GP60 was determined to be a peak molecular weight of 10945 g / mol, a number-average molecular weight of 9688 g / mol, a weight-average molecular weight of 79108 g / mol, and a molar ratio of monosaccharides of glucose:galactose:arabinose = 77.1:16.8:6.1, providing theoretical support for the application of ginseng polysaccharide in photoaging.

[0025] The ginseng polysaccharide GP60 prepared by the method of this invention is non-toxic and has anti-photoaging properties, and has broad application prospects. It can be widely used in anti-photoaging skin care products, washing and care products and anti-photoaging drugs.

[0026] The present invention will be further described below with reference to the accompanying drawings and specific embodiments. Attached Figure Description

[0027] Figure 1 Schematic diagram of the process flow for preparing ginseng polysaccharide GP60;

[0028] Figure 2 Overlay of HPLC analysis results of PMP monosaccharide reference product;

[0029] Figure 3 Results of monosaccharide composition analysis of ginseng polysaccharide GP60;

[0030] Figure 4 Cytotoxicity test results of ginseng polysaccharide GP60;

[0031] Figure 5 Effect of ginseng polysaccharide GP60 on the survival rate of NIH3T3 cells damaged by UVA irradiation. Detailed Implementation

[0032] Example 1:

[0033] This embodiment describes a method for preparing ginseng polysaccharides, such as... Figure 1 As shown, the preparation method steps are as follows:

[0034] Step 1, two decoctions: The weight ratio of ginseng slices to water is 1:15. Soak for 1 hour, then heat to boiling and decoct for 1.5 hours. Filter and collect the filtrate. Add 10 times the weight of water to the ginseng slices and decoct for 1 hour. Collect the filtrate.

[0035] Step 2, one addition of alcohol: Combine the filtrates collected from the two filtrations in Step 1, concentrate to a small volume, place at room temperature, add 95% ethanol until the alcohol content reaches 20%, let stand for 12 hours, then centrifuge at 3000 r / min for 10 minutes and collect the supernatant.

[0036] Step 3, Second addition of alcohol: Add 95% ethanol to the supernatant collected in step 2 until the alcohol content reaches 40%, let stand for 12 hours, then centrifuge at 3000 r / min for 10 min and collect the supernatant.

[0037] Step 4, three additions of alcohol: Add 95% pure ethanol to the supernatant collected in step 3 until the alcohol content reaches 60%, let stand for 12 hours, then centrifuge at 3000 r / min for 10 min and collect the precipitate.

[0038] In steps 2-4 above, the first step is to precipitate with 20% and 40% ethanol, discard the precipitate, and remove the components with relatively large molecular weights from the solution. Then, precipitate with 60% ethanol, collect the precipitate, and remove the supernatant to remove the components with relatively small molecular weights from the solution.

[0039] Step 5, Preparation of precipitate water dissolution: Add water to the precipitate collected in step 4 to dissolve it into a solution with a relative density of 1.1 g / ml;

[0040] Step 6, prepare polysaccharide aqueous solution:

[0041] 6.1 Take a certain amount of the solution obtained in step 5, add chloroform and n-butanol in a certain proportion, shake at 140 r / min for 10 min, and let stand for 12 h;

[0042] 6.2. Take off the supernatant, add the same amount of chloroform and n-butanol, shake at 140 r / min for 10 min, let stand for 12 h, take off the supernatant, repeat this process at least 5 times. In this embodiment, preferably, the process is repeated 5 times. Finally, take off the supernatant to obtain a polysaccharide aqueous solution.

[0043] Step 7: Add 95% ethanol to the polysaccharide aqueous solution until the alcohol content reaches 60%, let stand for 12 h, then centrifuge at 3000 r / min for 10 min and collect the precipitate.

[0044] Step 8: Wash the precipitate collected in Step 7 sequentially with 95% ethanol, anhydrous ethanol, isopropanol, and diethyl ether to evaporate the solvent and obtain the ginseng polysaccharide GP60.

[0045] Example 2:

[0046] This embodiment is an example of the preparation method of ginseng polysaccharide in Embodiment 1.

[0047] Take 2 kg of ginseng slices, add 30 L of water, soak for 1 h, decoct for 1.5 h, filter, and collect the filtrate; add 20 L of water to the residue, decoct for 1 h, filter, and collect the filtrate.

[0048] Combine the two filtrates, concentrate to a small volume, place at room temperature, add water to a total volume of 2.6 L, add 95% ethanol to a concentration of 20%, let stand for 12 h, then centrifuge at 3000 r / min for 10 min, collect the supernatant, add 95% ethanol to a concentration of 40%, let stand for 12 h, then centrifuge again at 3000 r / min for 10 min, collect the supernatant, add 95% ethanol to a concentration of 60%, let stand for 12 h, then centrifuge a third time at 3000 r / min for 10 min, collect the supernatant, add 95% ethanol to the supernatant to a concentration of 60%, let stand for 12 h, then centrifuge (3000 r / min) for 10 min, collect the precipitate.

[0049] Add water to the above precipitate to dissolve it into a polysaccharide aqueous solution to a volume of 2 L. Add 0.32 L of chloroform and 0.08 L of n-butanol, shake at 140 r / min for 10 min, let stand for 12 h, and aspirate the supernatant. Add another 0.32 L of chloroform and 0.08 L of n-butanol, and continue to shake at 140 r / min for 10 min. Let stand for 12 h again, and aspirate the supernatant. Repeat this process 5 times. Finally, aspirate the supernatant to obtain the polysaccharide aqueous solution.

[0050] Add 95% ethanol solution to the above polysaccharide aqueous solution until the alcohol content reaches 60%, and let stand for 12 h; centrifuge for 10 min, collect the precipitate, and centrifuge at 3000 r / min. Wash the collected precipitate sequentially with 95% ethanol, anhydrous ethanol, isopropanol, and diethyl ether, and evaporate the solvent to obtain the ginseng polysaccharide GP60 described in this invention.

[0051] The present invention employs the above-mentioned novel separation strategy to obtain a ginseng polysaccharide GP60, and its chemical characteristics were analyzed as follows, revealing that they differ from those of existing publicly disclosed ginseng polysaccharides.

[0052] 1. Molecular weight distribution analysis

[0053] 1.1 Chromatographic conditions

[0054] Chromatographic column: TSK® PWXL guard column (40 mm × 6.0 mm) tandem with TSK® G5000PWXL gel column (300 mm × 7.8 mm) tandem with TSK® G3000PWXL gel column (300 mm × 7.8 mm); mobile phase: 0.01% NaCl solution; column temperature: 30 ℃; detector: differential refractive index detector; flow rate: 0.6 mL / min; injection volume: 50 μL.

[0055] 1.2 Preparation of reference solution

[0056] Weigh an appropriate amount of Dextran reference standard accurately, add 0.01% NaCl solution to prepare a solution containing 1 mg per 1 mL, filter through a 0.45 μm microporous membrane, and collect the filtrate.

[0057] 1.3 Preparation of test solution

[0058] Weigh an appropriate amount of ginseng polysaccharide GP60 accurately, add 0.01% NaCl solution to prepare a solution containing 1 mg per 1 mL, filter through a 0.45 μm microporous membrane, and collect the filtrate to obtain the final product.

[0059] 1.4 Sample Measurement and Results

[0060] Accurately pipette 50 μL each of the reference solution and the test solution into the liquid chromatograph and determine their molecular weights. The weight-average molecular weight (y) and retention time (x) of the series of reference standards were used to calibrate the GPC software. Statistical analysis of the chromatogram of the test sample was performed using GPC software to fit the molecular weight distribution of the polysaccharide. The peak molecular weight, number-average molecular weight, and weight-average molecular weight of ginseng polysaccharide GP60 were 10945 g / mol, 9688 g / mol, and 79108 g / mol, respectively.

[0061] 2. Monosaccharide composition analysis

[0062] Monosaccharide composition and molar ratio are important parameters for the chemical characteristics of polysaccharides, and also important parameters that distinguish them from other polysaccharide phases.

[0063] 2.1 Chromatographic conditions

[0064] Chromatographic column: C18 column (250 mm × 4.6 mm, 5 μm); column temperature: 30 ℃; mobile phase: acetonitrile - 4.5% NaAc solution (18:82); flow rate: 1 mL / min; detector: ultraviolet detector (detection wavelength 245 nm); injection volume: 10 μL.

[0065] 2.2 Preparation of reference solution

[0066] Weigh approximately 1 mg of arabinose and place it in a stoppered test tube. Add 0.3 mL of water to dissolve it, then add 0.4 mL each of 0.3 mol / L NaOH solution and 0.4 mL of 0.3 mol / L PMP methanol solution, mix well, seal tightly, and place in an oven at 70°C for 1 h. Remove, cool, and add 0.4 mL of 0.3 mol / L HCl solution, mix well. Wash three times with 0.6 mL of chloroform each time, discarding the chloroform solution; filter the aqueous layer through a 0.45 μm microporous membrane, and collect the filtrate. Prepare reference solutions of galactose, glucose, mannose, glucuronic acid, and galacturonic acid using the same method.

[0067] 2.3 Preparation of test solution

[0068] Accurately weigh 2 mg of ginseng polysaccharide GP60 and place it in a stoppered test tube. Add 2 mL of 2 mol / L TFA solution, seal tightly, and place in an oven at 115℃ for 6 h. Recover the solvent from the hydrolysate under reduced pressure until dry. Dissolve the residue in 1 mL of water, and recover the solvent again under reduced pressure until dry. Repeat the above steps until the residue has no obvious sour taste.

[0069] Dissolve the residue in 0.3 mL of water, add 0.4 mL each of 0.3 mol / L NaOH solution and 0.4 mL of 0.3 mol / L PMP methanol solution, mix well, seal tightly, and place in an oven at 70 °C for 1 h; remove, cool, add 0.4 mL of 0.3 mol / L HCl solution, mix well; wash 3 times with 0.6 mL of chloroform each time, discard the chloroform solution; filter the aqueous layer through a 0.45 μm microporous membrane, collect the filtrate, and prepare the test solution.

[0070] 2.4 Sample Measurement and Results

[0071] Accurately pipette 10 μL of both the reference solution and the test solution into the liquid chromatograph for analysis. The HPLC analysis results of the monosaccharide reference standard PMP derivatization products are superimposed as shown below. Figure 2 As shown, by overlaying the chromatograms of each monosaccharide reference standard, a mixed chromatogram of the reference standards is obtained. In the figure, peak 1 is D-mannose, peak 2 is L-rhamnose, peak 3 is D-glucuronic acid, peak 4 is D-galacturonic acid, peak 5 is D-glucose, peak 6 is D-galactose, and peak 7 is L-arabinose.

[0072] Figure 3This is the analysis result of the monosaccharide composition of ginseng polysaccharide GP60 of this invention. In the figure, 5 is glucose, 6 is galactose, and 7 is arabinose. The monosaccharide composition of polysaccharide GP60 was determined by comparing the chromatogram of the test sample with the mixed chromatogram of the monosaccharide reference standard and measuring the retention time of each chromatographic peak. The results show that ginseng polysaccharide GP60 is composed of glucose, galactose, and arabinose; by calculating the ratio of the peak areas of each monosaccharide in the chromatogram of the test sample, the molar ratio of each monosaccharide is 77.1:16.8:6.1.

[0073] 2.3 Determination of Specific Rotation of GP60

[0074] Accurately weigh a certain amount of GP60 and prepare a polysaccharide solution with a mass concentration of 1 mg / mL. Use SGW. ® -1. Specific rotation of ginseng polysaccharide samples was determined using an automatic polarimeter. The specific rotation of GP60 was determined according to the Chinese Pharmacopoeia standard. Sodium dihydrogen spectroscopy (D-D) was used for measurement at a wavelength of 589.4 nm, a test tube length of 20 dm, and a sample temperature of 25.7℃. The automatic polarimeter was first zeroed using ddH₂O before the specific rotation of the sample was measured.

[0075] The results of six measurements are as follows:

[0076] The results of six measurements are as follows:

[0077]

α

[0078]

α

[0079]

α

[0080]

α

[0081]

α

[0082]

α

[0083] The average value is [α]. 均值 = +60.354, the standard deviation is σ² = 0.605.

[0084] 3. Pharmacological activity analysis of ginseng polysaccharides

[0085] This invention investigates the activity of ginseng polysaccharide GP60 in NIH3T3 cells. By establishing an NIH3T3 cell UV damage model, it was found that the ginseng polysaccharide GP60 solution described in this invention has a significant protective effect on NIH3T3 cells damaged by UVA irradiation. Mouse embryonic fibroblasts (NIH3T3) are a mouse embryonic fibroblast cell line with fibroblast morphology and characteristics established by the National Institutes of Health in the United States. Currently, many skin-related studies both domestically and internationally use this cell line.

[0086] 3.1 Cytotoxicity detection of ginseng polysaccharide GP60

[0087] (1) Laying 96-well plates: Adjusting the NIH3T3 cell suspension to 5×10⁻⁶. 4 Add 100 μL of cell suspension to each well to achieve a cell density of 5000 cells per well, and incubate for 24 h.

[0088] When plate-coating, pay attention to the following groupings: 1 control group (with cells but no drug intervention); 1 blank group (with culture medium only, no cells); 3 graded drug intervention groups (with cells and drug intervention). Each group should have 6 replicates.

[0089] (2) After 24 hours, the culture medium was aspirated, and the graded drug intervention groups were given drugs with concentrations of 6.4 × 10⁻⁶. -4 g / L, 1.6×10 - 2 g / L and 4×10 -1 100 μL of g / L ginseng polysaccharide GP60 solution was added to each well. The control group and blank group were given 100 μL of complete culture medium containing 10% fetal bovine serum.

[0090] (3) After each group was cultured for 24 h, 10 μL of CCK-8 staining solution was added to each well and incubated in a CO2 incubator for 2 hours.

[0091] (4) Place the incubated 96-well plate into an enzyme-linked immunosorbent assay (ELISA) reader and measure the absorbance at 450 nm.

[0092] (5) Cell survival rate = (OD value of drug intervention group - OD value of blank group) / (OD value of control group - OD value of blank group) × 100%.

[0093] The cytotoxicity test results of ginseng polysaccharide GP60 are as follows: Figure 4 As shown, at 6.4×10 -4 g / L~4×10 -1 Within the g / L concentration range, ginseng polysaccharide GP60 had no adverse effect on the viability of NIH3T3 cells.

[0094] 3.2 Effect of ginseng polysaccharide GP60 on the survival rate of NIH3T3 cells damaged by UVA irradiation

[0095] (1) Seeding in 96-well plates: Digest and centrifuge NIH3T3 cells in logarithmic growth phase. Dilute with DMEM medium containing 10% fetal bovine serum to a density of 50,000 cells / mL and seed in 96-well plates, 100 μL per well, and incubate for 24 h.

[0096] (2) Cell grouping: a) Control group: cells present, no irradiation; b) Blank group: culture medium added, no cells; c) UVA group: cells present, irradiated, no GP60 added; d) GP60+UVA group: cells present, irradiated, different concentrations of GP60 added. Each group was divided into 6 replicates. PBS solution was added around the perimeter of the 96-well plate to prevent evaporation.

[0097] (3) After culturing on plates for 24 hours, the culture medium was discarded, and the cells were rinsed once with 100 μL of PBS solution. 50 μL of PBS solution was added to each well to just cover the cells.

[0098] (4) During UVA irradiation, the blank and control groups that do not require irradiation are covered with a light-shielding plate (aluminum foil). The UVA group and the GP60+UVA group are irradiated with UVA. After irradiation, the PBS solution in each well is aspirated, and 100 μL / well of DMEM complete medium is added to the blank group, control group and UVA group, and cultured for another 24 h. The GP60+UVA group is treated with different concentrations (10 mg / L, 20 mg / L, 40 mg / L) of GP60 to act on photoaged cells, and cultured for another 24 h.

[0099] (5) Add 10 μL CCK-8 reagent to each well, and try to avoid light. After adding, wrap the well with aluminum foil and incubate in an incubator for 2 hours.

[0100] (6) Place the 96-well plate into the microplate reader and measure the absorbance (OD value) at 450 nm.

[0101] (7) Cell viability (%) = (mean OD value of irradiated group - mean OD value of blank group) / (mean OD value of control group - mean OD value of blank group) × 100%

[0102] Depend on Figure 5 As can be seen, after UVA irradiation, the cell viability in the UVA group was significantly lower than that in the control group, indicating that the UVA-induced photoaging model of NIH3T3 cells was successfully established. Treatment with a certain concentration of ginseng polysaccharide GP60 for 24 hours significantly restored cell viability, with the best effect observed at a drug concentration of 20 mg / L. Therefore, the results indicate that ginseng polysaccharide GP60 has a protective effect against UVA-induced photoaging of NIH3T3 cells.

[0103] Due to the aforementioned non-toxic and anti-photoaging properties of ginseng polysaccharide GP60, the ginseng polysaccharide GP60 of this invention has broad application prospects and can be widely used in anti-photoaging skin care products such as sunscreen, sun lotion, and skin care lotion; washing and care products such as facial cleanser and shower gel; and can also be used in anti-photoaging pharmaceuticals.

[0104] Finally, it should be noted that the above is only used to illustrate the technical solutions of the present invention and not to limit it. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention.

Claims

1. A method for preparing ginseng polysaccharides, characterized in that, The ginseng polysaccharide mentioned is ginseng polysaccharide GP60, composed of glucose, galactose, and arabinose, with a molar ratio of glucose:galactose:arabinose = 77.1:16.8:6.

1. Its molecular weight distribution is as follows: peak molecular weight 10945 g / mol, number-average molecular weight 9688 g / mol, and weight-average molecular weight 79108 g / mol. The preparation method is as follows: Step 1, two decoctions: The weight ratio of ginseng slices to water is 1:

15. Soak for 1 hour, then heat to boiling and decoct for 1.5 hours. Filter and collect the filtrate. Add 10 times the amount of water to the ginseng slices and decoct for 1 hour. Collect the filtrate. Step 2, one addition of alcohol: Combine the filtrates collected from the two filtrations in Step 1, concentrate to a small volume, place at room temperature, add 95% pure ethanol until the alcohol content reaches 20%, let stand for 12 hours, then centrifuge and collect the supernatant. Step 3, Second addition of alcohol: Add 95% ethanol to the supernatant collected in step 2 until the alcohol content reaches 40%, let stand for 12 hours, then centrifuge and collect the supernatant. Step 4, three additions of alcohol: Add 95% pure ethanol to the supernatant collected in step 3 until the alcohol content reaches 60%, let stand for 12 hours, then centrifuge and collect the precipitate. Step 5, Preparation of precipitate water dissolution: Add water to the precipitate collected in step 4 to dissolve it into a solution with a relative density of 1.1 g / ml; Step 6, prepare polysaccharide aqueous solution: 6.1 Take a certain amount of the solution obtained in step 5, add chloroform and n-butanol in a certain proportion, shake at 140 r / min for 10 min, and let stand for 12 h; 6.

2. Remove the supernatant, add the same proportions of chloroform and n-butanol as in step 6.1, shake at 140 r / min for 10 min, let stand for 12 h, remove the supernatant, repeat this process at least 5 times, and finally remove the supernatant to obtain a polysaccharide aqueous solution. Step 7, Preparing the precipitate: Add 95% ethanol to the polysaccharide aqueous solution until the alcohol content reaches 60%, let stand for 12 h, centrifuge, and collect the precipitate; Step 8, washing the precipitate: The precipitate collected in step 7 is washed sequentially with 95% ethanol, anhydrous ethanol, isopropanol and diethyl ether to evaporate the solvent and obtain the ginseng polysaccharide.

2. The method for preparing ginseng polysaccharides according to claim 1, characterized in that, The volume ratio of the solution, chloroform, and n-butanol in step 6.1 is 5:0.8:0.

2.

3. The method for preparing ginseng polysaccharides according to claim 1, characterized in that, The process described in step 6.2 is repeated 5 times.

4. The method for preparing ginseng polysaccharides according to claim 1, characterized in that, The centrifugation speed in steps 2-4 and 7 is 3000 r / min and the time is 10 min.

5. The use of ginseng polysaccharide GP60 based on the ginseng polysaccharide preparation method according to any one of claims 1-4, characterized in that, The ginseng polysaccharide GP60 is used in anti-photoaging skincare products and personal care products.

6. The use of ginseng polysaccharide GP60 based on the ginseng polysaccharide preparation method according to any one of claims 1-4, characterized in that, The ginseng polysaccharide GP60 is used in anti-photoaging drugs.

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