Extraction process optimization method, extraction method and application of duchesnea indica polysaccharide

Abstract

The invention discloses a duchesnea indica polysaccharide extraction process optimization method, an extraction method and application. The extraction process optimization method comprises the following steps: extracting and preparing duchesnea indica polysaccharide dry powder; drawing a glucose standard curve; weighing a proper amount of duchesnea indica polysaccharide dry powder to prepare a duchesnea indica polysaccharide solution, and calculating the duchesnea indica polysaccharide content in combination with the glucose standard curve Taking the solid-liquid ratio, the extraction temperature, the extraction time and the extraction times as dependent variables, taking the duchesnea indica polysaccharide extraction rate as a measurement index, measuring the content of the extracted duchesnea indica polysaccharide for multiple times, and determining the process condition under the maximum extraction amount as the optimal extraction condition; the optimal extraction process is subjected to reproducible operation. Through duchesnea indica polysaccharide extraction process optimization, a glucose standard curve is formulated, the content of a duchesnea indica polysaccharide solution is measured, optimal process parameters are determined through an orthogonal experiment, finally reproducible operation is performed under the condition of the optimal process parameters, and overall optimization is accurate, convenient and fast.

Description

Technical Field

[0001] The invention relates to the technical field of extracting polysaccharides from snakeberry, and in particular to an extraction process optimization method, an extraction method and an application of polysaccharides from snakeberry. Background Art

[0002] Duchesnea indica (Andr.) Focke, a plant of the genus Duchesnea in the Rosaceae family, is widely distributed in China. At present, the research directions of Duchesnea indica mainly focus on chemical component analysis, gardening application and plant cloning, while the pharmacological effects and mechanisms of Duchesnea indica are less studied. Snakeberry was first recorded in the "Famous Doctors' Records" and was recorded in the "Compendium of Materia Medica" that it can be used for "typhoid fever, creek poison, and spit poison, which is very good. It can regulate menstruation and apply to snake bites. As a traditional Chinese medicine, the decoction of snakeberry can be used to treat diseases such as cough, vomiting blood, sore throat, and dysentery. Inflammation is the body's first line of defense against injury or infection. Inflammatory reactions are involved in many diseases. It can remove the source of infection to maintain tissue homeostasis, but excessive inflammatory reactions can cause autoimmune damage. Modern medicine mainly uses antibiotics and steroid drugs to treat inflammation. These drugs will produce many toxic side effects during the treatment process, and will also produce drug resistance during long-term use. In recent years, traditional Chinese medicine (TCM) has many advantages in the treatment of inflammation, such as low toxicity and side effects and a wide range of effects. It has also become a new hotspot in the world's research. Its recognition worldwide is also increasing, and its clinical and experimental research has made gratifying progress. However, at present, the extraction of active ingredients of many natural drugs has been studied in depth, but there is no optimal process for the extraction of snakeberry polysaccharides. Summary of the invention

[0003] The purpose of the present invention is to provide a method for optimizing the extraction process of snakeberry polysaccharide, an extraction method and application thereof in order to solve the above problems.

[0004] The present invention achieves the above-mentioned purpose through the following technical solutions:

[0005] A method for optimizing the extraction process of snakeberry polysaccharide comprises the following steps:

[0006] S1, treating the raw material of Duchesnea indica hay to extract and prepare Duchesnea indica polysaccharide dry powder;

[0007] S2, weighing an appropriate amount of glucose to prepare a glucose solution, taking different volumes of glucose solution to prepare glucose standard solutions of different gradients, treating the glucose standard solutions with phenol-concentrated sulfuric acid, measuring the absorbance of each treated glucose standard solution, and drawing a glucose standard curve;

[0008] S3, weighing an appropriate amount of Duchesnea edulis polysaccharide powder to prepare a Duchesnea edulis polysaccharide solution, taking an appropriate amount of the Duchesnea edulis polysaccharide solution and treating it with phenol-concentrated sulfuric acid, measuring its absorbance, and calculating the content of Duchesnea edulis polysaccharide in combination with a glucose standard curve;

[0009] S4, taking the solid-liquid ratio, extraction temperature, extraction time and extraction times as dependent variables, taking the extraction rate of snakeberry polysaccharide as a measurement index, repeatedly measuring the extracted snakeberry polysaccharide content, and determining the process conditions under the maximum extraction amount as the optimal extraction conditions;

[0010] S5. Perform reproducible operation on the optimal extraction process.

[0011] Preferably, the acquisition of parameters in drawing the glucose standard curve in step S2 includes first accurately weighing a certain amount of standard glucose, and then diluting the glucose in a volumetric flask to obtain a glucose standard solution; then accurately aspirating different amounts of the obtained glucose standard solution and placing it in a volumetric flask, adding water to dilute the volume to obtain standard solutions with different gradients, and preparing a blank control group, quantitatively aspirating the standard solution from each standard solution and placing it in a test tube, adding phenol solution to the test tube, shaking and adding concentrated sulfuric acid, letting it stand for a period of time, and then measuring the maximum absorption wavelength at 400-700nm and specifying it as the measurement wavelength, measuring the absorbance of each standard solution at the measurement wavelength, drawing a standard curve with the absorbance as the horizontal axis and the glucose content as the vertical axis, and obtaining a linear regression equation.

[0012] Preferably, the method for determining the content of snakeberry polysaccharide in step S3 includes firstly accurately weighing a certain amount of snakeberry polysaccharide and adjusting the volume in a volumetric flask, then accurately aspirating a quantitative snakeberry polysaccharide solution and placing it in a test tube, adding a phenol solution, shaking and evenly mixing, adding concentrated sulfuric acid, standing for a period of time, and then determining the absorbance at a measurement wavelength, and calculating the content of snakeberry polysaccharide according to the linear regression equation obtained from the glucose standard curve.

[0013] Preferably, in step S4, the extraction rate of snakeberry polysaccharide is used as the measurement index, and the solid-liquid ratio, extraction temperature, extraction time and extraction times are used as dependent variables. 4 ) To optimize the orthogonal experiment of extracting polysaccharides from snakeberry by water extraction and alcohol precipitation, and to screen and determine the optimal process parameters for extracting polysaccharides from snakeberry.

[0014] Preferably, the reproducible operation method of the optimal extraction process in step S5 includes first accurately weighing the crushed snakeberry dry powder and dividing it into 3 equal parts, placing them in round-bottom flasks respectively, adding a corresponding volume of distilled water for reflux extraction according to the orthogonal test results, filtering under reduced pressure, combining the filtrate, and concentrating under reduced pressure to 1 / 2 of the original volume, taking the supernatant after centrifugation and placing it in 4 times the volume of anhydrous ethanol, standing for a period of time, centrifuging and discarding the supernatant, washing the precipitate with 2 times the volume of anhydrous ethanol, acetone, and petroleum ether for 3 times, and after the petroleum ether is evaporated, grinding it through an 80-mesh sieve to obtain snakeberry polysaccharide.

[0015] A method for extracting snakeberry polysaccharide comprises the following steps: firstly, crushing snakeberry hay and passing it through a 16-mesh sieve, weighing an appropriate amount of snakeberry whole herb powder, with a solid-liquid ratio of 1:10 (g / ml), maintaining the powder under a slightly boiling state with distilled water, and performing reflux extraction for 3 times, each time for 3 hours; subsequently, centrifuging at 5000 r / min for 7 minutes, combining the filtrate, and concentrating the mixture to 1 / 2 of the original volume under vacuum decompression; then adding 1 / 3 volume of SEVAGE reagent, stirring the mixture electrically for 30 minutes, separating the liquids, collecting the upper aqueous phase, repeating the process for 6 times to remove protein, collecting the upper aqueous phase, transferring the collected upper aqueous phase to 4 times the volume of anhydrous ethanol, standing the mixture at 4°C for 24 hours, centrifuging and discarding the supernatant, washing the precipitate with 2 times the volume of anhydrous ethanol, acetone, and petroleum ether for 3 times, and grinding the precipitate through an 80-mesh sieve to obtain the snakeberry polysaccharide.

[0016] Application of snakeberry polysaccharide extracted under the optimal process conditions of snakeberry polysaccharide extraction method in anti-inflammatory drugs.

[0017] The beneficial effects of the present invention are:

[0018] 1. The present invention optimizes the extraction process of snakeberry polysaccharide by formulating a glucose standard curve, determining the content of snakeberry polysaccharide solution, and determining the optimal process parameters through orthogonal experiments. Finally, reproducible operation is performed under the optimal process parameter conditions. The overall optimization method is accurate and convenient;

[0019] 2. The present invention uses L9(3 4 ) Orthogonal experiment was used to optimize the extraction method of snakeberry polysaccharide. The anti-inflammatory effect of snakeberry polysaccharide was observed by using xylene-induced mouse ear swelling model and acetic acid-induced mouse peritoneal capillary permeability change model. Compared with the blank group, the snakeberry polysaccharide group had a good inhibitory effect on xylene-induced mouse ear swelling, and the difference was statistically significant (P<0.01). Compared with the blank group, the snakeberry polysaccharide group had an inhibitory effect on the permeability of mouse peritoneal capillaries, and the difference was statistically significant (P<0.01). Snakeberry polysaccharide has good anti-inflammatory activity for mouse ear swelling and change in peritoneal capillary permeability. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative labor.

[0021] Figure 1 It is a schematic diagram of the process of the present invention.

[0022] Figure 2 It is the glucose standard curve diagram in the present invention.

[0023] Figure 3 This is a comparative experimental diagram of the effect of snakeberry polysaccharide in the present invention on the treatment of swollen and inflammatory auricles in mice.

[0024] Figure 4 This is a comparative experimental diagram of the effect of the snakeberry polysaccharide in the present invention on the permeability of peritoneal capillaries in mice. DETAILED DESCRIPTION

[0025] The following is combined with Figure 1-4 , the technical solution of the present invention is further explained:

[0026] Embodiment 1

[0027] like Figure 2 As shown, a method for optimizing the extraction process of snakeberry polysaccharide comprises the following steps:

[0028] S1. The raw material of Duchesnea indica hay is processed to extract and prepare Duchesnea indica polysaccharide dry powder. That is, firstly, the Duchesnea indica polysaccharide dry powder is extracted by the original process as the subsequent raw material.

[0029] S2, weighing an appropriate amount of glucose to prepare a glucose solution, taking different volumes of glucose solution to prepare glucose standard solutions of different gradients, treating the glucose standard solutions with phenol-concentrated sulfuric acid, measuring the absorbance of each treated glucose standard solution, and drawing a glucose standard curve;

[0030] The acquisition of parameters in drawing the glucose standard curve in step S2 includes first accurately weighing a certain amount of standard glucose, and then adjusting the volume of glucose in a volumetric flask to obtain a glucose standard solution; then accurately aspirating different amounts of the obtained glucose standard solution and placing it in a volumetric flask, adding water to adjust the volume to obtain standard solutions with different gradients, and preparing a blank control group, quantitatively aspirating the standard solution from each standard solution and placing it in a test tube, adding phenol solution to the test tube, shaking and adding concentrated sulfuric acid, and standing for a period of time, and then measuring the maximum absorption wavelength at 400-700nm and specifying it as the measurement wavelength, measuring the absorbance of each standard solution at the measurement wavelength, drawing a standard curve with the absorbance as the horizontal axis and the glucose content as the vertical axis, and obtaining a linear regression equation.

[0031] That is to say, in this embodiment, 0.01027 g of standard glucose is accurately weighed and fixed to volume in a 100 ml volumetric flask to obtain a glucose standard solution with a concentration of 0.1027 mg / mL. 0, 4.0, 6.0, 8.0 and 10.0 mL of glucose standard solution are accurately drawn into a 10 mL volumetric flask, and water is added to fix the volume to obtain standard solutions of different gradients; 2.0 mL of each standard solution is accurately drawn into a stoppered test tube, 1.0 mL of 5% phenol solution is added, and after shaking, 5.0 mL of concentrated sulfuric acid is added, and the solution is allowed to stand for 30 minutes. Bottle "0" is used as a blank control, and the maximum absorption wavelength is measured at 400-700 nm and specified as the measurement wavelength.

[0032] like Figure 2 As shown, the absorbance of the above glucose solution is measured at the measurement wavelength, and a standard curve is drawn with the absorbance as the horizontal axis and the glucose content as the vertical axis to obtain a linear regression equation.

[0033] It should be noted that in this embodiment, the standard curve is drawn with the glucose content (mg) in the standard solution as the horizontal axis and the absorbance value (A) as the vertical axis. The result is shown in the figure, and its regression equation is: The correlation coefficient R2 = 0.9998, indicating that the standard curve has a good fit.

[0034] S3, weighing an appropriate amount of Duchesnea edulis polysaccharide powder to prepare a Duchesnea edulis polysaccharide solution, taking an appropriate amount of the Duchesnea edulis polysaccharide solution and treating it with phenol-concentrated sulfuric acid, measuring its absorbance, and calculating the content of Duchesnea edulis polysaccharide in combination with a glucose standard curve;

[0035] The method for determining the content of snakeberry polysaccharide in step S3 includes firstly accurately weighing a certain amount of snakeberry polysaccharide and adjusting the volume in a volumetric flask, then accurately aspirating a quantitative snakeberry polysaccharide solution and placing it in a test tube, adding a phenol solution, shaking and evenly mixing, adding concentrated sulfuric acid, standing for a period of time, determining the absorbance at a determination wavelength, and calculating the content of snakeberry polysaccharide according to a linear regression equation obtained from a glucose standard curve.

[0036] That is to say, accurately weigh 0.01006g of snakeberry polysaccharide, make up to volume in a 100mL volumetric flask, accurately pipette 2ml of snakeberry polysaccharide solution into a stoppered test tube, add 1.0mL of 5% phenol solution, shake well, add 5.0mL of concentrated sulfuric acid, let stand for 30min, measure the absorbance at the measurement wavelength, and calculate the content of snakeberry polysaccharide according to the standard curve.

[0037] S4, taking the solid-liquid ratio, extraction temperature, extraction time and extraction times as dependent variables, taking the extraction rate of snakeberry polysaccharide as a measurement index, repeatedly measuring the extracted snakeberry polysaccharide content, and determining the process conditions under the maximum extraction amount as the optimal extraction conditions;

[0038] In step S4, the extraction rate of snakeberry polysaccharide is used as the measurement index, and the solid-liquid ratio, extraction temperature, extraction time and extraction times are used as dependent variables. 4 ) To optimize the orthogonal experiment of extracting polysaccharides from snakeberry by water extraction and alcohol precipitation, and to screen and determine the optimal process parameters for extracting polysaccharides from snakeberry.

[0039] That is to say, taking the extraction rate of snakeberry polysaccharide as the measurement index, the main influencing factors were set as A solid-liquid ratio, B extraction temperature, C extraction time, D extraction times, and L9 (3 4 ) Optimize the orthogonal experiment of extracting polysaccharides from snakeberry by water extraction and alcohol precipitation, screen and determine the optimal process parameters for extracting polysaccharides from snakeberry. The factor levels are shown in the table below.

[0040] Table of factors and levels of orthogonal experiment for extraction process optimization

[0041]

[0042] The results of the orthogonal experiment were compared. A solid-liquid ratio, B extraction temperature, C extraction time, and D extraction times were used as variables, and the polysaccharide extraction rate was used as the evaluation index to conduct the L9 (34) orthogonal experiment. The content of the extracted snakeberry polysaccharide was determined, and the results are shown in the following table:

[0043]

[0044]

[0045] Through the orthogonal experimental results, it can be obtained that the four factors have an influence on the extraction rate of snakeberry polysaccharide from high to low, which is B>D>C>A. Among them, the extraction temperature has the greatest influence on the extraction efficiency, and the solid-liquid ratio has the least influence. According to the extraction rate of snakeberry polysaccharide, the optimal extraction process combination of polysaccharide is: A1B3C3D3, that is, the solid-liquid ratio is 1:10 (g / ml), the temperature is 95℃, and the extraction is 3 times, each time for 3 hours.

[0046] S5. Perform reproducible operation on the optimal extraction process.

[0047] The reproducible operation method of the optimal extraction process in step S5 includes first accurately weighing the crushed snakeberry dry powder and dividing it into 3 equal parts, placing them in round-bottom flasks respectively, adding corresponding volumes of distilled water for reflux extraction according to the orthogonal test results, filtering under reduced pressure, combining the filtrate, and concentrating under reduced pressure to 1 / 2 of the original volume, taking the supernatant after centrifugation and placing it in 4 times the volume of anhydrous ethanol, standing for a period of time, centrifuging and discarding the supernatant, washing the precipitate with 2 times the volume of anhydrous ethanol, acetone, and petroleum ether for 3 times, and grinding the precipitate through an 80-mesh sieve to obtain snakeberry polysaccharide after the petroleum ether is evaporated.

[0048]

[0049]

[0050] That is to say, the crushed snakeberry powder was accurately weighed and divided into 3 equal parts, and placed in 2000mL round-bottom flasks respectively. According to the results of the orthogonal test, the corresponding volume of distilled water was added for reflux extraction, and the filtrate was combined. The mixture was concentrated to 1 / 2 of the original volume under reduced pressure, and the supernatant was placed in 4 times the volume of anhydrous ethanol after centrifugation, and allowed to stand at 4°C for 24 hours, and the supernatant was discarded after centrifugation. The precipitate was washed 3 times with 2 times the volume of anhydrous ethanol, acetone, and petroleum ether, and after the petroleum ether was evaporated, it was ground through an 80-mesh sieve to obtain the snakeberry polysaccharide.

[0051] Embodiment 2

[0052] like Figure 1 As shown, a method for extracting snakeberry polysaccharide comprises the following steps: firstly, crushing snakeberry hay and passing it through a 16-mesh sieve, weighing an appropriate amount of whole snakeberry powder, with a solid-liquid ratio of 1:10 (g / ml), maintaining the mixture in a slightly boiling state with distilled water, and performing reflux extraction for 3 times, each time for 3 hours; then, centrifuging at 5000 r / min for 7 minutes, combining the filtrate, and concentrating the mixture to 1 / 2 of the original volume under vacuum decompression; then, adding 1 / 3 volume of SEVAGE reagent, stirring the mixture electrically for 30 minutes, separating the liquids, collecting the upper aqueous phase, repeating the process for 6 times to remove protein, collecting the upper aqueous phase, transferring the collected aqueous phase to 4 times the volume of anhydrous ethanol, standing the mixture at 4°C for 24 hours, centrifuging and discarding the supernatant, washing the precipitate 3 times with 2 times the volume of anhydrous ethanol, acetone, and petroleum ether, respectively, and grinding the precipitate through an 80-mesh sieve to obtain snakeberry polysaccharide after the petroleum ether is evaporated.

[0053] Embodiment 3

[0054] like Figure 3 and Figure 4 As shown, the application of snakeberry polysaccharide extracted under the optimal process conditions of snakeberry polysaccharide extraction method in anti-inflammatory drugs.

[0055] That is to say, this example verifies that the polysaccharide from snakeberry has an anti-inflammatory effect. The anti-inflammatory effect of the polysaccharide from snakeberry is observed by using a xylene-induced ear swelling model in mice and an acetic acid-induced change in peritoneal capillary permeability model in mice. The specific process is as follows:

[0056] 1. First, group mice and administer drugs:

[0057] Fifty Kunming mice were randomly divided into 5 groups, including a saline blank control group (n=10), a model control group (n=10), an indomethacin positive control group (dosing concentration was 0.975 mg / ml, n=10), a medium concentration experimental group of snakeberry polysaccharide (dosing concentration was 2.12 mg / ml, n=10) and a high concentration experimental group of snakeberry polysaccharide (dosing concentration was 42.4 mg / ml, n=10). On the basis of normal feeding, the mice were gavaged once a day for 5 consecutive days, with an gavage dose of 0.1 ml / 10 g.

[0058] 3. Conduct mouse ear swelling experiment

[0059] 100% xylene reagent was selected as the inflammatory agent for modeling mouse auricle swelling. One hour after the last administration, 100% xylene reagent was evenly applied on both sides of the left ear of each mouse in the other groups except the model control group to induce inflammation, and the right ear was not treated as a control. The mice were killed 30 minutes after the inflammation, and ear pieces were punched in the same part of the two ears of the mice. The ears were accurately weighed on a 1 / 100,000 balance, and the swelling rate of the mouse ears was calculated according to the formula. Mouse ear swelling rate = (left ear weight - right ear weight) / right ear weight. The swelling rates of the control group and the drug group were statistically processed.

[0060] like Figure 3 As shown in the figure, the swelling degree of the auricles of each group of mice was significantly increased compared with the model control group (P < 0.01), which indicated that the mouse auricle swelling model was successfully established. Compared with the saline blank control group, the swelling degree of the other groups was significantly reduced (P < 0.01). The therapeutic effects of high concentration and medium concentration of snakeberry polysaccharide were also significantly different (P < 0.01). With the increase of the concentration of snakeberry polysaccharide, the swelling degree gradually decreased, indicating that the anti-inflammatory activity of snakeberry polysaccharide in the treatment of mouse auricle swelling inflammation was dose-dependent. The results are shown in the figure. Figure 3 .

[0061] 4. Conduct mouse peritoneal capillary permeability experiment

[0062] 0.7% acetic acid solution was selected as the inflammatory agent for the modeling of changes in peritoneal capillary permeability in mice. One hour after the last administration, 0.5% Evans blue solution 0.1mL / 10g was injected into the tail vein of each group of mice. Except for the model control group, all other groups were intraperitoneally injected with 0.7% acetic acid solution 0.1mL / 10g to induce inflammation. The mice were killed 30 minutes after the inflammation, the abdomen was opened, and 5mL of normal saline was injected into the mice intraperitoneally. The mice were kneaded for one minute, and the peritoneal fluid was aspirated with a needle and transferred into a centrifuge tube. After mixing, it was centrifuged at 5000r / min for 15 minutes. The supernatant was taken for colorimetry at 590nm to measure the absorbance (A). The A values ​​of the control group and the drug-treated group were statistically processed.

[0063] Evans blue is a commonly used azo dye preparation that can bind to plasma albumin in the blood, so it is often used as a tracer in research. In this study, Evans blue was injected into the tail vein of mice to make Evans blue bind to plasma albumin in the blood and color it. Under normal circumstances, due to the permeability of the capillary wall, plasma albumin cannot pass through the capillary wall due to its large molecular weight, so the peritoneal lavage fluid of normal mice should be close to colorless. The peritoneal cavity of the studied mice was in an inflammatory state, and inflammation would increase the permeability of the capillary wall, causing plasma albumin to leak out. At this time, the peritoneal lavage fluid of the inflammatory mice should be blue. In this study, the absorbance at 540nm was used to reflect the severity of inflammation in mice.

[0064] Combined with the differences in the absorbance values ​​(A) of the peritoneal fluid of mice in each group, the following conclusions can be drawn: compared with the model control group, the A values ​​of the other groups were significantly increased (P < 0.01), proving that the model of peritoneal capillary permeability changes in mice was successfully established. Compared with the saline blank control group, the A values ​​of the other groups were significantly decreased (P < 0.01), and there was no significant difference in the A values ​​between the high-concentration group of snakeberry polysaccharide and the medium-concentration group of snakeberry polysaccharide (P = 0.139), indicating that the anti-inflammatory activity of snakeberry polysaccharide in the treatment of peritoneal capillary permeability in mice was not significantly correlated with the dose. The results are shown in Figure 4 .

[0065] It can be seen that as one of the active ingredients of snakeberry, snakeberry polysaccharide has good anti-inflammatory activity in vivo. In this study, it has been proved that the snakeberry polysaccharide experimental group has a significant anti-inflammatory effect compared with the blank group.

[0066] The above shows and describes the basic principles, main features and advantages of the present invention. Those skilled in the art should understand that the present invention is not limited to the above embodiments, and the above embodiments and descriptions are only for explaining the principles of the present invention. Without departing from the spirit and scope of the present invention, the present invention may have various changes and improvements, and these changes and improvements all fall within the scope of the present invention to be protected.

Claims

1. A method for optimizing the extraction process of polysaccharides from snakeberry, characterized in that: The following steps are involved: S1, treating the raw material of Duchesnea indica hay to extract and prepare Duchesnea indica polysaccharide dry powder; S2, weighing an appropriate amount of glucose to prepare a glucose solution, taking different volumes of glucose solution to prepare glucose standard solutions of different gradients, treating the glucose standard solutions with phenol-concentrated sulfuric acid, measuring the absorbance of each treated glucose standard solution, and drawing a glucose standard curve; S3, weighing an appropriate amount of Duchesnea edulis polysaccharide powder to prepare a Duchesnea edulis polysaccharide solution, taking an appropriate amount of the Duchesnea edulis polysaccharide solution and treating it with phenol-concentrated sulfuric acid, measuring its absorbance, and calculating the content of Duchesnea edulis polysaccharide in combination with a glucose standard curve; S4, taking the solid-liquid ratio, extraction temperature, extraction time and extraction times as dependent variables, taking the extraction rate of snakeberry polysaccharide as a measurement index, repeatedly measuring the extracted snakeberry polysaccharide content, and determining the process conditions under the maximum extraction amount as the optimal extraction conditions; S5. Perform reproducible operation on the optimal extraction process.

2. The method for optimizing the extraction process of snakeberry polysaccharide according to claim 1, characterized in that: The acquisition of parameters in drawing the glucose standard curve in step S2 includes first accurately weighing a certain amount of standard glucose, and then adjusting the volume of glucose in a volumetric flask to obtain a glucose standard solution; then accurately aspirating different amounts of the obtained glucose standard solution and placing it in a volumetric flask, adding water to adjust the volume to obtain standard solutions with different gradients, and preparing a blank control group, quantitatively aspirating the standard solution from each standard solution and placing it in a test tube, adding phenol solution to the test tube, shaking and adding concentrated sulfuric acid, and standing for a period of time, and then measuring the maximum absorption wavelength at 400-700nm and specifying it as the measurement wavelength, measuring the absorbance of each standard solution at the measurement wavelength, drawing a standard curve with the absorbance as the horizontal axis and the glucose content as the vertical axis, and obtaining a linear regression equation.

3. The method for optimizing the extraction process of snakeberry polysaccharide according to claim 1, characterized in that: The method for determining the content of snakeberry polysaccharide in step S3 includes firstly accurately weighing a certain amount of snakeberry polysaccharide and adjusting the volume in a volumetric flask, then accurately aspirating a quantitative snakeberry polysaccharide solution and placing it in a test tube, adding a phenol solution, shaking and evenly mixing, adding concentrated sulfuric acid, standing for a period of time, determining the absorbance at a determination wavelength, and calculating the content of snakeberry polysaccharide according to a linear regression equation obtained from a glucose standard curve.

4. The method for optimizing the extraction process of Duchesnea oleifera polysaccharides according to claim 1, characterized in that: In step S4, the extraction rate of snakeberry polysaccharide is used as the measurement index, and the solid-liquid ratio, extraction temperature, extraction time and extraction times are used as dependent variables. 4 ) To optimize the orthogonal experiment of extracting polysaccharides from snakeberry by water extraction and alcohol precipitation, and to screen and determine the optimal process parameters for extracting polysaccharides from snakeberry.

5. The method for optimizing the extraction process of snakeberry polysaccharide according to claim 1, characterized in that: The reproducible operation method of the optimal extraction process in step S5 includes first accurately weighing the crushed snakeberry dry powder and dividing it into 3 equal parts, placing them in round-bottom flasks respectively, adding corresponding volumes of distilled water for reflux extraction according to the orthogonal test results, filtering under reduced pressure, combining the filtrate, and concentrating under reduced pressure to 1 / 2 of the original volume, taking the supernatant after centrifugation and placing it in 4 times the volume of anhydrous ethanol, standing for a period of time, centrifuging and discarding the supernatant, washing the precipitate with 2 times the volume of anhydrous ethanol, acetone, and petroleum ether for 3 times, and grinding the precipitate through an 80-mesh sieve to obtain snakeberry polysaccharide after the petroleum ether is evaporated.

6. A method for extracting polysaccharides from Duchesnea sinensis, characterized in that: The method comprises the following steps: firstly, crushing the dried snakeberry grass and passing it through a 16-mesh sieve, weighing an appropriate amount of whole snakeberry grass powder with a solid-liquid ratio of 1:10 (g / ml), maintaining the powder under a slightly boiling state with distilled water, and performing reflux extraction for 3 times, each time for 3 hours; subsequently, centrifuging at 5000 r / min for 7 minutes, combining the filtrate, and concentrating the mixture to 1 / 2 of the original volume under vacuum decompression; then adding 1 / 3 volume of SEVAGE reagent, stirring the mixture by electric stirring for 30 minutes, separating the liquids, collecting the upper aqueous phase, repeating the process for 6 times to remove the protein, collecting the upper aqueous phase, transferring the collected phase to 4 times the volume of anhydrous ethanol, standing the phase at 4°C for 24 hours, centrifuging and discarding the supernatant, washing the precipitate 3 times with 2 times the volume of anhydrous ethanol, acetone, and petroleum ether, respectively, and grinding the precipitate through an 80-mesh sieve to obtain snakeberry polysaccharide after the petroleum ether is evaporated.

7. Use of the Duchesnea polysaccharide extracted by the extraction method as claimed in claim 6 in anti-inflammatory drugs.

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