Ginseng polysaccharide as well as preparation method and application thereof

Abstract

The invention discloses ginseng polysaccharide and a preparation method and application thereof.The ginseng polysaccharide is ginseng polysaccharide GP60 and is composed of glucose, galactose and arabinose, the molar ratio of glucose to galactose to arabinose is 77.1: 16.8: 6.1, a new preparation method is adopted, soaking and two times of decoction are conducted, 95% ethyl alcohol of different proportions is added into filter liquor in batches, and the ginseng polysaccharide GP60 is obtained after filtration. Adding water into the finally obtained precipitate to prepare a solution with the relative density of 1.1 g / ml; the preparation method comprises the following steps: taking a certain amount of solution, adding trichloromethane and n-butyl alcohol in a certain proportion to prepare a polysaccharide aqueous solution, adding 95% ethanol into the polysaccharide aqueous solution until the alcohol content reaches 60%, standing for 12 hours, carrying out centrifugal separation, collecting precipitates, washing the collected precipitates with 95% ethanol, absolute ethanol, isopropanol and diethyl ether in sequence, and volatilizing the solvent to obtain the ginseng polysaccharide GP60. The ginseng polysaccharide GP60 disclosed by the invention is non-toxic, has an anti-photoaging effect, and can be widely applied to the fields of foods or health foods, anti-photoaging skin care products, wash supplies and anti-photoaging medicines.

Description

Technical Field

[0001] The invention belongs to the technical field of ginseng polysaccharide extraction, and particularly relates to ginseng polysaccharide, a preparation method and application thereof. Background Art

[0002] With the global warming caused by greenhouse effects such as carbon dioxide, the atmosphere has been damaged to a certain extent, leading to an increase in ultraviolet intensity. In addition to aggravating skin aging, the increase in ultraviolet intensity has also gradually increased the number of patients with light-related skin diseases such as solar dermatitis and sunburn, especially those working outdoors. This phenomenon has received increasing attention from all walks of life. Acute skin light damage, commonly known as sunburn, is one of the most common skin injuries. The clinical manifestations are burning, redness, swelling, pain, and even blisters, burning pain, and skin desquamation. Although sunburn patients will almost always recover after a period of time, the discomfort caused by sunburn will still cause certain pain and distress to patients, and there is evidence that 70% of skin cancers are directly or indirectly caused by sunburn. In addition, ultraviolet radiation in sunlight can also cause skin aging, causing the skin to become dry and rough, wrinkles and various pigment deposits, which have a negative impact on people's appearance and even cause a variety of skin diseases or skin tumors.

[0003] Public information shows that ginseng polysaccharide is one of the main components of the precious Chinese medicine ginseng. It is a kind of natural high molecular polymer composed of multiple monosaccharides. It has small toxic side effects and rich biological activity. It has anti-tumor, immunomodulatory, antioxidant, hypoglycemic, anti-radiation and other biological activities. It is widely used in food, health care products and clinical aspects. At present, the separation and extraction technology of ginseng polysaccharides is relatively mature, and there have been many literature reports. However, there are differences in the structure and activity of ginseng polysaccharides obtained by different extraction methods. In recent years, researchers have found that many Chinese herbal extracts with antioxidant effects can effectively alleviate skin photodamage. Ginseng polysaccharides have been reported to have obvious antioxidant damage effects, but there are few studies on the protective effect of ginseng polysaccharides on photoaging.

[0004] Xu Yangsen et al.'s invention patent for a ginseng polysaccharide extraction production process (publication number CN110862464A) discloses a process for industrial production of ginseng polysaccharide solid beverages, including slicing ginseng through a slicing device, the thickness of the slices is 0.1-1.0 cm, soaking for 1-10 hours, soaking temperature 60-100 ° C, material-liquid ratio 1:10-1:80 g / ml, controlling the temperature, membrane filtration, reverse osmosis membrane concentration, spray drying, and automatic packaging. The invention patent with publication number CN117487035A discloses a ginseng polysaccharide and its preparation method and application: after ginseng is crushed and extracted by hot water reflux, it is concentrated and centrifuged, the supernatant is precipitated with ethanol solution, the obtained precipitate is dialyzed with distilled water, and the non-dialyzed fluid is freeze-dried to obtain ginseng polysaccharide; the dialysis molecular weight cutoff is 3.5 kDa. The ginseng polysaccharide prepared by the above method can be used to regulate intestinal flora, antidepressants, and epilepsy drugs. Publication No. 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: the ginseng extracellular vesicles are prepared by gradient centrifugation, and ginseng juice is obtained by squeezing ginseng juice. Ginseng or ginseng residues are mixed with water for extraction, and the obtained extract is mixed with ethanol / ethanol aqueous solution for alcohol precipitation, centrifugation, collection of precipitates, and drying. The composition is used to improve skin anti-aging and inflammation after mechanical damage to the skin. Summary of the invention

[0005] The present invention aims to propose a ginseng polysaccharide GP60, a preparation method and an application thereof, clarify the molecular weight and composition of the ginseng polysaccharide GP60, the proportions of glucose, galactose and arabinose in the ginseng polysaccharide GP60, confirm that the ginseng polysaccharide GP60 obtained by the preparation method has a good protective effect on UVA-induced NIH3T3 cell photoaging, and provide 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, and the molecular weight distribution is a peak molecular weight of 10945 g / mol, a number average molecular weight of 9688 g / mol, and a weight average molecular weight of 79108 g / mol.

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

[0008] A method for preparing ginseng polysaccharide, wherein the ginseng polysaccharide is obtained by the method, and the method steps are as follows: Step 1, decoction twice: the weight ratio of ginseng slices to water is 1:15, soaking time is 1 hour, then heating to boiling, decoction for 1.5 hours, filtering, and collecting the filtrate; adding 10 times the amount of water to the filter residue for secondary decoction, decoction for 1 hour, and collecting the filtrate; Step 2, adding alcohol once: Combine the filtrates collected by filtering twice 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, secondary alcohol addition: 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; Step 4, adding alcohol three times: add 95% pure ethanol to the supernatant collected in step 3 until the alcohol content reaches 60%, let it stand for 12 hours, then centrifuge and collect the precipitate; Step 5, preparing a precipitate water solution: adding 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, preparing 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, aspirate the upper layer, add chloroform and n-butanol in the same proportion as in step 6.1, shake at 140 r / min for 10 min, let stand for 12 h, aspirate the upper layer, repeat this process for at least 5 times, and finally aspirate the upper layer to obtain a polysaccharide aqueous solution; Step 7, preparing a precipitate: adding 95% ethanol to the polysaccharide aqueous solution until the alcohol content reaches 60%, standing for 12 hours, centrifuging, and collecting the precipitate; Step 8, washing the precipitate: washing the precipitate collected in step 7 with 95% ethanol, anhydrous ethanol, isopropanol, and ether in sequence, evaporating the solvent to obtain the ginseng polysaccharide.

[0009] Furthermore, in step 6.1, the volume ratio of the solution, chloroform and n-butanol is 5:0.8:0.2.

[0010] Furthermore, the process in step 6.2 is repeated 5 times.

[0011] Furthermore, the centrifugal separation described in steps 2-4 and 7 is carried out at a speed of 3000 r / min and a time of 10 min.

[0012] Furthermore, the ginseng polysaccharide can be used for anti-photoaging food or health food.

[0013] Furthermore, the ginseng polysaccharide can be used in anti-photoaging skin care products.

[0014] Furthermore, the ginseng polysaccharide can be used in anti-photoaging drugs.

[0015] The beneficial effects of the present invention are as follows: a ginseng polysaccharide GP60 having a protective effect on UVA-induced NIH3T3 cell photoaging is obtained, and the molecular weight distribution of the ginseng polysaccharide GP60 is 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 each monosaccharide of glucose: galactose: arabinose = 77.1: 16.8: 6.1, which provides theoretical support for the application of ginseng polysaccharides in photoaging.

[0016] The ginseng polysaccharide GP60 obtained by the preparation method of the present invention is non-toxic and has anti-photoaging properties, has broad application prospects, and can be widely used in the fields of food or health food, anti-photoaging skin care products, toiletries and anti-photoaging medicines.

[0017] The present invention is further described below in conjunction with the accompanying drawings and specific implementation methods. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 Schematic diagram of the process flow of the preparation method of ginseng polysaccharide GP60; Figure 2 Overlay of HPLC analysis results of monosaccharide reference substance PMP derivatization products; Figure 3 The results of monosaccharide composition analysis of ginseng polysaccharide GP60; Figure 4 Cytotoxicity test results of ginseng polysaccharide GP60; Figure 5 Effect of ginseng polysaccharide GP60 on the survival rate of NIH3T3 cells damaged by UVA irradiation. DETAILED DESCRIPTION

[0019] Embodiment 1: This embodiment is a method for preparing ginseng polysaccharide. Figure 1 As shown, the preparation method steps are as follows: Step 1, decoction twice: the weight ratio of ginseng slices to water is 1:15, the soaking time is 1 hour, then heated to boiling, decocted for 1.5 hours, filtered, and the filtrate is collected; the filter residue is added with water 10 times the weight of the ginseng slices for secondary decoction, decocted for 1 hour, and the filtrate is collected.

[0020] Step 2, adding alcohol once: Combine the filtrates collected by filtering twice in step 1, concentrate to a small volume, place at room temperature, add 95% ethanol to a concentration of 20% alcohol, let stand for 12 hours, then centrifuge at 3000 r / min for 10 minutes and collect the supernatant; Step 3, secondary alcohol addition: add 95% pure 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 minutes and collect the supernatant; Step 4, adding alcohol three times: add 95% pure ethanol to the supernatant collected in step 3 until the alcohol content reaches 60%, let it stand for 12 hours, then centrifuge at 3000 r / min for 10 min and collect the precipitate.

[0021] In the above steps 2-4, first precipitate with 20% and 40% ethanol, discard the precipitate, in order to remove the components with larger relative molecular weight in the solution, then precipitate with 60% ethanol, collect the precipitate, and remove the supernatant to remove the components with smaller relative molecular weight in the solution.

[0022] Step 5, preparing a precipitate water solution: adding 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, preparing 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, aspirate the upper layer liquid, add the same amount of chloroform and n-butanol, shake at 140 r / min for 10 min, let stand for 12 h, aspirate the upper layer liquid, repeat this process for at least 5 times, preferably 5 times in this embodiment, and finally aspirate the upper layer liquid to obtain a polysaccharide aqueous solution; Step 7, adding 95% ethanol to the polysaccharide aqueous solution until the alcohol content reaches 60%, standing for 12 hours, and then centrifuging at 3000 r / min for 10 minutes to collect the precipitate; Step 8, washing the precipitate collected in step 7 with 95% ethanol, anhydrous ethanol, isopropanol, and ether in sequence, evaporating the solvent to obtain the ginseng polysaccharide GP60.

[0023] Embodiment 2: This embodiment is an example of the preparation method of ginseng polysaccharide.

[0024] Take 2 kg of ginseng slices, add 30 L of water, soak for 1 h, boil for 1.5 h, filter, and collect the filtrate; add 20 L of water to the residue, boil for 1 h, filter, and collect the filtrate.

[0025] The two filtrates were combined, concentrated to a small volume, placed at room temperature, water was added to a total volume of 2.6 L, 95% ethanol was added to an alcohol content of 20%, and the mixture was allowed to stand for 12 h. The mixture was then centrifuged at 3000 r / min for 10 min, the supernatant was collected, 95% ethanol was added to an alcohol content of 40%, and the mixture was allowed to stand for 12 h. The mixture was then centrifuged at 3000 r / min for 10 min again, the supernatant was collected, 95% ethanol was added to an alcohol content of 60%, and the mixture was allowed to stand for 12 h. The mixture was then centrifuged at 3000 r / min for 10 min for the third time, the supernatant was collected, 95% ethanol was added to the supernatant to an alcohol content of 60%, the mixture was allowed to stand for 12 h, and the mixture was centrifuged (3000 r / min) for 10 min to collect the precipitate.

[0026] Add water to the above precipitate to dissolve it into a polysaccharide aqueous solution to 2 L, add 0.32 L of chloroform and 0.08 L of n-butanol, shake at 140 r / min for 10 min, let it stand for 12 h, absorb the upper layer, add 0.32 L of chloroform and 0.08 L of n-butanol, continue to shake at 140 r / min for 10 min, let it stand for 12 h again, absorb the upper layer, repeat this operation 5 times, and finally absorb the upper layer to obtain a polysaccharide aqueous solution.

[0027] Add 95% pure ethanol solution to the polysaccharide aqueous solution until the alcohol content reaches 60%, and let it stand for 12 hours; centrifuge for 10 minutes, collect the precipitate, and the speed of centrifugation is 3000 r / min. The collected precipitate is washed with 95% ethanol, anhydrous ethanol, isopropanol, and ether in turn, and the solvent is evaporated to obtain the ginseng polysaccharide GP60 of the present invention.

[0028] The present invention adopts the above-mentioned new separation strategy to obtain a ginseng polysaccharide GP60, and analyzes its chemical characteristics as follows, and finds that the chemical characteristics are different from those of the currently disclosed ginseng polysaccharides.

[0029] 1. Molecular weight distribution analysis 1.1 Chromatographic conditions Chromatographic column: TSK® gel chromatography column guard column PWXL (40 mm × 6.0 mm) connected in series with TSK® G5000PWXL gel chromatography column (300 mm × 7.8 mm) connected in series with TSK®G3000PWXL gel chromatography column (300 ‍mm × 7.8 mm); mobile phase: 0.01% NaCl solution; column temperature: 30 ℃; detector: differential refractometer; flow rate: 0.6 ‍mL / min; injection volume: 50μL.

[0030] 1.2 Preparation of reference solution Weigh an appropriate amount of Dextran reference substance respectively, accurately weigh it, add 0.01% NaCl solution to make a solution containing 1 mg per 1 mL, filter it with a 0.45 μm microporous filter membrane, and take the filtrate to obtain the solution.

[0031] 1.3 Preparation of test solution Weigh an appropriate amount of ginseng polysaccharide GP60, accurately weigh it, add 0.01% NaCl solution to make a solution containing 1 mg per 1 mL, filter it with a 0.45 μm microporous filter membrane, and take the filtrate to obtain the product.

[0032] 1.4 Sample determination and results 50 μL of the reference solution and the test solution were accurately drawn and injected into the liquid chromatograph for determination. The GPC software was calibrated using the weight average molecular weight (y) and retention time (x) of the series of reference substances. The GPC software was used to statistically analyze the chromatogram of the test sample and 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.

[0033] 2. Monosaccharide composition analysis Monosaccharide composition and molar ratio are important parameters for the chemical characteristics of polysaccharides and are also important parameters that distinguish them from other polysaccharide phases.

[0034] 2.1 Chromatographic conditions Chromatographic column: C18 chromatographic column (250mm ×4.6mm, 5μm); column temperature: 30℃; mobile phase: acetonitrile-4.5% NaAc solution (18:82); flow rate: 1mL / min; detector: UV detector (detection wavelength is 245 nm); injection volume: 10μL.

[0035] 2.2 Preparation of reference solution Weigh about 1 mg of arabinose and place it in a stoppered test tube. Add 0.3 mL of water to dissolve it, add 0.3 mol / L NaOH solution and 0.3 mol / L PMP methanol solution, 0.4 mL each, mix well, stopper tightly, place in an oven, and keep at 70°C for 1 h; take out, cool, add 0.4 mL of 0.3 mol / L HCl solution, and mix well. Wash with chloroform 3 times, 0.6 mL each time, and discard the chloroform solution; filter the water layer with a 0.45μm microporous filter membrane, and take the filtrate to obtain. Prepare galactose, glucose, mannose, glucuronic acid, and galacturonic acid reference solutions in the same way.

[0036] 2.3 Preparation of test solution Weigh 2 mg of ginseng polysaccharide GP60 accurately and place it in a stoppered test tube. Add 2 mL of 2 mol / L TFA solution, stopper it tightly, and place it in an oven at 115°C for 6 h. Recover the solvent from the hydrolyzate under reduced pressure until it is dry, add 1 mL of water to the residue to dissolve it, and recover the solvent under reduced pressure again until the residue has no obvious sour taste. Repeat the above operation until the residue has no obvious sour taste.

[0037] Add 0.3 mL of water to the residue to dissolve it, add 0.4 mL of 0.3 mol / L NaOH solution and 0.3 mol / L PMP methanol solution, mix well, seal tightly, and place in an oven at 70°C for 1 h; take out, cool, add 0.4 mL of 0.3 mol / L HCl solution, mix well; wash with chloroform 3 times, using 0.6 mL of chloroform each time, and discard the chloroform liquid; filter the water layer with a 0.45 μm microporous filter membrane, take the filtrate, and prepare the test solution.

[0038] 2.4 Sample determination and results Accurately pipette 10 μL of the reference solution and the test solution respectively, inject into the liquid chromatograph for determination. The HPLC analysis results of the monosaccharide reference substance PMP derivatization product are superimposed as shown in the figure below. Figure 2 As shown, the chromatograms of the monosaccharide reference substances are superimposed to obtain a mixed spectrum of the reference substances. In the figure, peak 1 is D-mannose, 2 is L-rhamnose, 3 is D-glucuronic acid, 4 is D-galacturonic acid, 5 is D-glucose, 6 is D-galactose, and 7 is L-arabinose.

[0039] Figure 3 This is the result of the monosaccharide composition analysis of the ginseng polysaccharide GP60 of the present invention. In the figure, 5 is glucose, 6 is galactose, and 7 is arabinose. The chromatogram of the test sample is compared with the mixed spectrum of the monosaccharide reference substance, and the monosaccharide composition of the polysaccharide GP60 is determined by 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 chromatographic peak area of ​​each monosaccharide in the chromatogram of the test sample, it can be known that the molar ratio of each monosaccharide is 77.1:16.8:6.1.

[0040] 2.3 Determination of GP60 specific rotation Accurately weigh a certain amount of GP60 and prepare a polysaccharide solution with a mass concentration of 1 mg / mL. ® -1 automatic polarimeter to detect the specific rotation of ginseng polysaccharide samples. The specific rotation of GP60 was measured according to the Chinese Pharmacopoeia standard. D sodium light was used for measurement, the detection wavelength was 589.4 nm, the test tube length was 20dm, and the sample temperature was 25.7℃. First, the automatic polarimeter was zeroed with ddH2O, and then the specific rotation of the sample was measured.

[0041] The results of six measurements are: The results of six measurements are:

α

α

α

α

α

α

[0042] 3. Analysis of pharmacological activity of ginseng polysaccharides The present invention uses NIH3T3 cells to study the activity of ginseng polysaccharide GP60. By establishing a NIH3T3 cell ultraviolet damage model, it is found that the ginseng polysaccharide GP60 solution of the present 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. At present, many skin-related studies at home and abroad have selected this cell line for research.

[0043] 3.1 Cytotoxicity assay of ginseng polysaccharide GP60 (1) 96-well plate: adjust the NIH3T3 cell suspension to 5×10 4 100 μL of cell suspension was added to each well to make 5000 cells per well and cultured for 24 hours.

[0044] When plating, pay attention to the grouping: 1 control group, with cells but no drug intervention; 1 blank group, with only culture medium but no cells; 3 gradient drug intervention groups, with cells and drug intervention. Set up 6 replicate wells for each group.

[0045] (2) After 24 hours, the culture medium was discarded and the gradient drug intervention groups were given 6.4×10 -4 g / L、1.6×10 - 2 g / L and 4×10 -1 g / L ginseng polysaccharide GP60 solution, 100 μL per well. The control group and blank group were added with 100 μL complete culture medium containing 10% fetal bovine serum.

[0046] (3) After the above groups were cultured for 24 h, 10 μL of CCK-8 stain solution was added to each well and incubated in a CO2 incubator for 2 h.

[0047] (4) Place the incubated 96-well plate into an ELISA reader and measure the absorbance at 450 nm.

[0048] (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%.

[0049] 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 concentration range of g / L, ginseng polysaccharide GP60 had no adverse effects on the viability of NIH3T3 cells.

[0050] 3.2 Effect of ginseng polysaccharide GP60 on the survival rate of UVA-damaged NIH3T3 cells (1) Plating in 96-well plates: Digest and centrifuge NIH3T3 cells in the logarithmic growth phase. Dilute to a density of 50,000 cells / mL with DMEM medium containing 10% fetal bovine serum, inoculate in 96-well plates, 100 μL per well, and culture for 24 hours.

[0051] (2) Cell grouping: a) control group, cells but no irradiation; b) blank group, culture medium only without cells; c) UVA group, cells, irradiation, no GP60; d) GP60+UVA group, cells, irradiation, and different concentrations of GP60. Each group had 6 replicate wells, and PBS solution was added around the 96-well plate to prevent evaporation.

[0052] (3) After 24 h of culture, aspirate the culture medium, rinse once with 100 μL PBS solution, and add 50 μL PBS solution to each well to just cover the cells.

[0053] (4) During UVA irradiation, use a shading board (aluminum foil) to cover the blank group and control group that do not need to be irradiated. The UVA group and GP60+UVA group were irradiated with UVA. After irradiation, the PBS solution in each well was discarded, and 100 μL / well DMEM complete medium was added to the blank group, control group and UVA group, and the culture was continued for 24 hours. In the GP60+UVA group, different concentrations (10 mg / L, 20 mg / L, 40 mg / L) of GP60 were added to act on the photoaged cells, and the culture was continued for 24 hours.

[0054] (5) Add 10 μL of CCK-8 reagent to each well, try to avoid light, wrap it with tin foil, and incubate it in an incubator for 2 hours.

[0055] (6) Place the 96-well plate into an ELISA reader and measure the absorbance (OD value) at 450 nm.

[0056] (7) Cell survival rate (%) = (average OD value of irradiated group - average OD value of blank group) / (average OD value of control group - average OD value of blank group) × 100% Depend on Figure 5 It can be seen that after UVA irradiation, the cell survival rate of the UVA group was significantly lower than that of the control group, indicating that the UVA-induced NIH3T3 cell photoaging model was successfully established. After treatment with a certain concentration of ginseng polysaccharide GP60 for 24 hours, the cell viability was significantly restored, and the effect was best when the drug concentration was 20 mg / L. Therefore, the results show that ginseng polysaccharide GP60 has a protective effect on UVA-induced NIH3T3 cell photoaging.

[0057] Due to the above-mentioned non-toxic and anti-photoaging properties of ginseng polysaccharide GP60, the ginseng polysaccharide GP60 of the present invention has broad application prospects and can be widely used in food or health food; anti-photoaging skin care products, such as sunscreen, sunscreen lotion, skin care milk, etc.; toiletries: such as facial cleanser, shower gel, etc.; and can also be used in anti-photoaging drugs.

[0058] Finally, it should be noted that the above is only used to illustrate the technical solution of the present invention rather than to limit it. Although the present invention has been described in detail with reference to the preferred solution, those skilled in the art should understand that the technical solution of the present invention can be modified or replaced by equivalents without departing from the spirit and scope of the technical solution of the present invention.

Claims

1. A ginseng polysaccharide, characterized in that: The ginseng polysaccharide is ginseng polysaccharide GP60, and the molecular weight distribution is as follows: the peak molecular weight is 10945 g / mol, the number average molecular weight is 9688 g / mol, and the weight average molecular weight is 79108 g / mol.

2. The ginseng polysaccharide according to claim 1, characterized in that: The ginseng polysaccharide is composed of glucose, galactose and arabinose, and the molar ratio of each monosaccharide is glucose: galactose: arabinose = 77.1: 16.8: 6.

1.

3. A method for preparing ginseng polysaccharide, characterized in that: The preparation method obtains the ginseng polysaccharide as claimed in claim 1 or 2, and the method steps are as follows: Step 1, decoction twice: the weight ratio of ginseng slices to water is 1:15, soaking time is 1 hour, then heating to boiling, decoction for 1.5 hours, filtering, and collecting the filtrate; adding 10 times the amount of water to the filter residue for secondary decoction, decoction for 1 hour, and collecting the filtrate; Step 2, adding alcohol once: Combine the filtrates collected by filtering twice 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, secondary alcohol addition: 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; Step 4, adding alcohol three times: add 95% pure ethanol to the supernatant collected in step 3 until the alcohol content reaches 60%, let it stand for 12 hours, then centrifuge and collect the precipitate; Step 5, preparing a precipitate water solution: adding 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, preparing 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, aspirate the upper layer, add chloroform and n-butanol in the same proportion as in step 6.1, shake at 140 r / min for 10 min, let stand for 12 h, aspirate the upper layer, repeat this process for at least 5 times, and finally aspirate the upper layer to obtain a polysaccharide aqueous solution; Step 7, preparing a precipitate: adding 95% ethanol to the polysaccharide aqueous solution until the alcohol content reaches 60%, standing for 12 hours, centrifuging, and collecting the precipitate; Step 8, washing the precipitate: washing the precipitate collected in step 7 with 95% ethanol, anhydrous ethanol, isopropanol, and ether in sequence, evaporating the solvent to obtain the ginseng polysaccharide.

4. The preparation method according to claim 3, characterized in that: The volume ratio of the solution, chloroform and n-butanol in step 6.1 is 5:0.8:0.

2.

5. The preparation method according to claim 3, characterized in that: The process in step 6.2 is repeated 5 times.

6. The preparation method according to claim 3, characterized in that: The centrifugal separation speed in steps 2-4 and 7 is 3000 r / min and the time is 10 min.

7. A use of ginseng polysaccharide, characterized in that: The ginseng polysaccharide is used for anti-photoaging food and health food.

8. A use of ginseng polysaccharide, characterized in that: The ginseng polysaccharide is used for anti-photoaging skin care products and toiletries.

9. A use of ginseng polysaccharide, characterized in that: The ginseng polysaccharide is used in anti-photoaging medicines.

Images