CN112239509A - Method for optimizing sweet potato polysaccharide extraction conditions based on response surface method - Google Patents

Method for optimizing sweet potato polysaccharide extraction conditions based on response surface method Download PDF

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CN112239509A
CN112239509A CN202011039184.4A CN202011039184A CN112239509A CN 112239509 A CN112239509 A CN 112239509A CN 202011039184 A CN202011039184 A CN 202011039184A CN 112239509 A CN112239509 A CN 112239509A
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王春玲
杜船
张倩
周韬
刘爽
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Tianjin University of Science and Technology
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Abstract

The invention relates to a method for optimizing sweet potato polysaccharide extraction conditions based on a response surface method, which comprises the following steps: peeling sweet potato, cutting into strips, freeze-drying, oven-drying, pulverizing, and sieving; extracting sweet potato polysaccharide by a water extraction and alcohol precipitation method, and determining variable parameters of sweet potato polysaccharide extraction by a response surface method; adding petroleum ether for degreasing, and adding ethanol into the precipitate; adding water, and extracting; centrifuging, concentrating, adding anhydrous ethanol, standing overnight, centrifuging, and making into crude polysaccharide solution; removing protein, decolorizing, concentrating, and dialyzing to obtain sweet potato crude polysaccharide. The invention adopts a response surface method to determine the variable parameters of sweet potato polysaccharide extraction, designs a four-factor three-level response surface analysis test, further optimizes the optimal extraction process conditions of sweet potato polysaccharide, and provides reference basis for comprehensive development and utilization of sweet potatoes. Where the actual value is very close to the theoretical predicted value. Therefore, the Design Expert software is reliable in response surface optimization of sweet potato polysaccharide extraction process conditions, and has practical value.

Description

Method for optimizing sweet potato polysaccharide extraction conditions based on response surface method
Technical Field
The invention belongs to the technical field of food, and particularly relates to a method for optimizing sweet potato polysaccharide extraction conditions based on a response surface method.
Background
Sweet potato (academic name: Dioscorea esculenta (Lour.) Burkill) herbaceous plant, which belongs to sweet potato species of Ipomoea of Convolvulaceae in classification, also known as sweet potato, sweet potato and sweet potato. The sweet potato is rich in nutrition and also contains various bioactive substances such as protein, polysaccharide, polyphenol, dietary fiber, vitamins and minerals. The sweet potato active polysaccharide also has the functions of resisting oxidation, resisting tumor, resisting mutation, enhancing immunity, reducing blood fat and the like.
The principle of water extraction and alcohol precipitation is to precipitate polysaccharide with ethanol by utilizing the property that the polysaccharide is insoluble in ethanol; the crude polysaccharide is obtained by taking water as an extraction solvent under the heating condition, and concentrating, precipitating with ethanol, centrifuging, drying and the like.
The Response Surface Method (RSM) can obtain a fitting equation, calculate optimal process parameters, solve the problems of interaction between multiple variables, and the like, and has a 3D surface map and a contour map that can visually reflect data conditions. The method adopts a more scientific and reasonable experimental design, so that researchers can not only effectively reduce the experimental times and time, but also obtain more comprehensive and scientific research results, and can quickly and effectively obtain the optimal experimental conditions.
In the prior art, the extraction rate of sweet potato polysaccharide is low, and the waste of raw materials is easily caused.
Through searching, the following patent publications relevant to the present invention application are found:
the extraction method of purple sweet potato polysaccharide (CN104592406A) comprises extracting polysaccharide from purple sweet potato by biological enzymolysis, centrifuging, precipitating with ethanol, removing protein, and decolorizing to obtain high purity purple sweet potato polysaccharide. The method has simple process, strong operability and simple required equipment, greatly improves the industrial value of the purple sweet potato, and the obtained purple sweet potato polysaccharide has obvious antioxidant activity in-vitro tests, can be applied to preparing antioxidant functional foods and has wide market prospect; moreover, the obtained purple sweet potato polysaccharide can change the structure of RANKL protein, thereby having potential anti-tumor effect. If anion exchange resin column separation and purification is adopted, the obtained purple sweet potato polysaccharide has higher purity and more obvious biological function.
By contrast, the present patent application is intrinsically different from the above-mentioned patent publications.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provides a method for optimizing sweet potato polysaccharide extraction conditions based on a response surface method.
The technical scheme adopted by the invention for solving the technical problems is as follows:
a method for optimizing sweet potato polysaccharide extraction conditions based on a response surface method comprises the following steps:
peeling sweet potatoes, cutting the sweet potatoes into strips, freeze-drying the sweet potatoes in a freeze dryer at-70 ℃, taking the sweet potatoes out at night, putting the sweet potatoes in a drying oven at 60 ℃, continuously drying the sweet potatoes for more than 8 hours, finally crushing the sweet potatoes into sweet potato powder by a crusher, sieving the sweet potato powder by a 40-mesh sieve, and bagging the sweet potato powder in a dryer for preservation;
the sweet potato polysaccharide is extracted by a water extraction and alcohol precipitation method, variable parameters of sweet potato polysaccharide extraction are determined by a response surface method, a four-factor three-level test mode is set to perform repeated tests to obtain an experiment result, the extraction rate of crude sweet potato polysaccharide is calculated, multiple regression analysis is performed by Design Expert 8.0.6 software, and the regression equation of the influence of experiment factors on response values is obtained:
R(％)＝12.65+0.59A+0.95B-0.26C+0.065D-0.61AB-0.30AC+0.46AD+0.45BC+0.91BD+0.24CD-3.27A2-2.72B2-2.41C2-2.29D2
in the formula, the response value R is the extraction rate of sweet potato polysaccharide, the variable parameter A is the extraction temperature, the variable parameter B is the feed-liquid ratio, the variable parameter C is the extraction time, the variable parameter D is the extraction frequency, the final determination that A is 60.80 ℃, B is 1:20.86, C is 1.98h, D is 3.05 and the extraction rate of polysaccharide is 12.75 percent;
weighing sweet potato powder in the step, adding petroleum ether for degreasing, discarding supernatant on the next day, and adding 95% ethanol into the precipitate for 3 times; wherein the sweet potato powder: degreasing with petroleum ether: proportion g of 95% ethanol: mL: mL is 1: 5: 6;
and (4) pouring out the supernatant after the reaction is finished, and mixing the materials according to a material-liquid ratio of 1:20, adding water, and extracting the polysaccharide in the sweet potatoes at 61 ℃ for 2 hours; centrifuging, concentrating, adding anhydrous ethanol 4 times the volume of the concentrated solution, standing at 4 deg.C overnight, centrifuging after precipitating with ethanol, collecting precipitate, and adding distilled water into the precipitate to obtain crude polysaccharide solution; the crude polysaccharide solution was mixed with Sevag reagent at 4: 1, violently shaking, centrifuging, removing protein in the solution, pouring the solution into a separating funnel filled with AB-8 macroporous adsorption resin for decolorization for 12 hours, then washing the resin with 3 times of volume of distilled water, concentrating to 1/10 of the original volume, and dialyzing by using a 3500 dialysis bag to obtain crude sweet potato polysaccharide;
wherein the Sevag reagent is a mixture of chloroform and n-butanol, and the weight ratio of chloroform: the volume ratio of n-butanol is 4: 1.
and in the step II, the volume ratio of chloroform to n-butanol is 4: 1.
and in the step two, during centrifugal separation, the rotating speed of the centrifugal machine is 6000 r/min.
Furthermore, the method comprises the steps of:
preparing the crude polysaccharide in the step II into 0.1mg/mL polysaccharide solution by using distilled water, and then diluting the prepared sample solution by different times; and (5) measuring the absorbance according to a method for drawing a standard curve, and calculating the extraction rate of the sweet potato polysaccharide.
And, the standard curve is formulated as follows:
weighing an excessive anhydrous glucose standard substance, drying in a drying oven at 105 ℃ until the weight is constant, accurately weighing 100mg and fixing the volume to 100mL to obtain a 1.0mg/mL glucose solution which is called stock solution; sucking 10mL from the stock solution by using a pipette gun, and fixing the volume by using a 100mL volumetric flask to obtain a glucose standard solution of 0.1 mg/mL;
accurately measuring 0, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8 and 0.9ml of glucose standard solution in test tubes by using a pipette, supplementing each test tube to 1ml by using distilled water, adding 1.0ml of 6 percent phenol solution into each test tube, slowly adding 5ml of concentrated sulfuric acid, uniformly shaking, carrying out boiling water bath for 15min, and measuring the absorbance value at 490nm after cooling to the room temperature;
the absorbance value A is determined by taking the glucose concentration C (ug/mL) as the abscissa490The ordinate is a standard curve.
The invention has the advantages and positive effects that:
1. the invention adopts a response surface method to determine the variable parameters of sweet potato polysaccharide extraction, selects four factors which have more obvious influence on the sweet potato polysaccharide extraction rate, namely temperature, material-liquid ratio, extraction time and extraction frequency, and designs a response surface analysis test with four factors and three levels on the basis of a single-factor test, thereby optimizing the optimal extraction process conditions of the sweet potato polysaccharide and providing reference basis for the comprehensive development and utilization of the sweet potatoes. Where the actual value is very close to the theoretical predicted value. Therefore, the conditions for response surface optimization sweet potato polysaccharide extraction by the design expert software are reliable, and the method has practical value.
2. The method has the advantages of simple extraction process, high extraction efficiency, capability of improving the extraction rate of polysaccharide, reduction of raw material loss, easiness in popularization and application, application of a response surface method for optimizing and determining optimal factors, guarantee of extraction scientificity and stability, and provision of a basis for quality standard research.
Drawings
FIG. 1 is a graph showing the effect of extraction temperature on sweet potato polysaccharide extraction rate in the present invention;
FIG. 2 is a graph showing the effect of feed liquid ratio on sweet potato polysaccharide extraction rate in the present invention;
FIG. 3 is a graph showing the effect of extraction time on sweet potato polysaccharide extraction rate in the present invention;
FIG. 4 is a graph showing the effect of the number of extractions on the extraction yield of sweetpotato polysaccharides in the present invention;
FIG. 5 is a response surface and contour plot of temperature and time versus sweet potato polysaccharide extraction rate in accordance with the present invention; wherein the upper graph is a response surface graph of temperature and time on the extraction rate of sweet potato polysaccharide, and the lower graph is a contour graph of temperature and time on the extraction rate of sweet potato polysaccharide;
FIG. 6 is a response surface and contour plot of sweet potato polysaccharide extraction rate versus temperature versus feed liquid in accordance with the present invention; wherein the upper graph is a response surface graph of the ratio of temperature to feed liquid to the extraction rate of sweet potato polysaccharide, and the lower graph is a contour graph of the ratio of temperature to feed liquid to the extraction rate of sweet potato polysaccharide;
FIG. 7 is a response surface and contour plot of temperature and extraction times on sweet potato polysaccharide extraction rate in the present invention; wherein the upper graph is a response surface graph of temperature and extraction frequency on the extraction rate of sweet potato polysaccharide, and the lower graph is a contour graph of temperature and extraction frequency on the extraction rate of sweet potato polysaccharide;
FIG. 8 is a response surface and contour plot of feed liquid ratio and time to sweet potato polysaccharide extraction rate in the present invention; wherein the upper graph is a response surface graph of the feed-liquid ratio and time to the extraction rate of sweet potato polysaccharide, and the lower graph is a contour graph of the feed-liquid ratio and time to the extraction rate of sweet potato polysaccharide;
FIG. 9 is a response surface and contour plot of feed liquid ratio and extraction frequency of the present invention to sweet potato polysaccharide extraction rate; wherein the upper graph is a response surface graph of the ratio of feed to liquid and the extraction times to the extraction rate of the sweet potato polysaccharide, and the lower graph is a contour graph of the ratio of feed to liquid and the extraction times to the extraction rate of the sweet potato polysaccharide;
FIG. 10 is a response surface and contour plot of time and extraction times on sweet potato polysaccharide extraction rate in the present invention; wherein the upper graph is a response surface graph of time and extraction frequency on the extraction rate of sweet potato polysaccharide, and the lower graph is a contour graph of time and extraction frequency on the extraction rate of sweet potato polysaccharide.
Detailed Description
The present invention is further illustrated by the following examples, which are intended to be illustrative, not limiting and are not intended to limit the scope of the invention.
The raw materials used in the invention are all conventional commercial products if no special description is provided, the method used in the invention is all conventional methods in the field if no special description is provided, and the mass of all the materials used in the invention is the conventional use mass.
A method for optimizing sweet potato polysaccharide extraction conditions based on a response surface method comprises the following steps:
peeling sweet potatoes, cutting the sweet potatoes into strips, freeze-drying the sweet potatoes in a freeze dryer at-70 ℃, taking the sweet potatoes out at night, putting the sweet potatoes in a drying oven at 60 ℃, continuously drying the sweet potatoes for more than 8 hours, finally crushing the sweet potatoes into sweet potato powder by a crusher, sieving the sweet potato powder by a 40-mesh sieve, and bagging the sweet potato powder in a dryer for preservation;
the sweet potato polysaccharide is extracted by a water extraction and alcohol precipitation method, variable parameters of sweet potato polysaccharide extraction are determined by a response surface method, a four-factor three-level test mode is set to perform repeated tests to obtain an experiment result, the extraction rate of crude sweet potato polysaccharide is calculated, multiple regression analysis is performed by Design Expert 8.0.6 software, and the regression equation of the influence of experiment factors on response values is obtained:
R(％)＝12.65+0.59A+0.95B-0.26C+0.065D-0.61AB-0.30AC+0.46AD+0.45BC+0.91BD+0.24CD-3.27A2-2.72B2-2.41C2-2.29D2
in the formula, the response value R is the extraction rate of sweet potato polysaccharide, the variable parameter A is the extraction temperature, the variable parameter B is the feed-liquid ratio, the variable parameter C is the extraction time, the variable parameter D is the extraction frequency, the final determination that A is 60.80 ℃, B is 1:20.86, C is 1.98h, D is 3.05 and the extraction rate of polysaccharide is 12.75 percent;
weighing sweet potato powder in the step, adding petroleum ether for degreasing, discarding supernatant on the next day, and adding 95% ethanol into the precipitate for 3 times; wherein the sweet potato powder: degreasing with petroleum ether: proportion g of 95% ethanol: mL: mL is 1: 5: 6;
and (4) pouring out the supernatant after the reaction is finished, and mixing the materials according to a material-liquid ratio of 1:20, adding water, and extracting the polysaccharide in the sweet potatoes at 61 ℃ for 2 hours; centrifuging, concentrating, adding anhydrous ethanol 4 times the volume of the concentrated solution, standing at 4 deg.C overnight, centrifuging after precipitating with ethanol, collecting precipitate, and adding distilled water into the precipitate to obtain crude polysaccharide solution; the crude polysaccharide solution was mixed with Sevag reagent at 4: 1, violently shaking, centrifuging, removing protein in the solution, pouring the solution into a separating funnel filled with AB-8 macroporous adsorption resin for decolorization for 12 hours, then washing the resin with 3 times of volume of distilled water, concentrating to 1/10 of the original volume, and dialyzing by using a 3500 dialysis bag to obtain crude sweet potato polysaccharide;
wherein the Sevag reagent is a mixture of chloroform and n-butanol, and the weight ratio of chloroform: the volume ratio of n-butanol is 4: 1.
preferably, the volume ratio of chloroform to n-butanol in the step II is 4: 1.
preferably, the rotating speed of the centrifuge is 6000r/min during centrifugal separation in the step II.
Preferably, the method further comprises the steps of:
preparing the crude polysaccharide in the step II into 0.1mg/mL polysaccharide solution by using distilled water, and then diluting the prepared sample solution by different times; and (5) measuring the absorbance according to a method for drawing a standard curve, and calculating the extraction rate of the sweet potato polysaccharide.
Preferably, the standard curve is formulated as follows:
weighing an excessive anhydrous glucose standard substance, drying in a drying oven at 105 ℃ until the weight is constant, accurately weighing 100mg and fixing the volume to 100mL to obtain a 1.0mg/mL glucose solution which is called stock solution; sucking 10mL from the stock solution by using a pipette gun, and fixing the volume by using a 100mL volumetric flask to obtain a glucose standard solution of 0.1 mg/mL;
accurately measuring 0, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8 and 0.9ml of glucose standard solution in test tubes by using a pipette, supplementing each test tube to 1ml by using distilled water, adding 1.0ml of 6 percent phenol solution into each test tube, slowly adding 5ml of concentrated sulfuric acid, uniformly shaking, carrying out boiling water bath for 15min, and measuring the absorbance value at 490nm after cooling to the room temperature;
the absorbance value A is determined by taking the glucose concentration C (ug/mL) as the abscissa490The ordinate is a standard curve.
Specifically, the preparation and detection are as follows:
a method for optimizing sweet potato polysaccharide extraction conditions based on a response surface method comprises the following steps:
first, sample preparation
Pulverizing dried sweet potato, weighing 100mg in a volumetric flask of 1000ml, degreasing by petroleum ether, decoloring by absolute ethyl alcohol, operating according to certain extraction temperature, material-liquid ratio, extraction time and extraction frequency, concentrating, adding 80% absolute ethyl alcohol for alcohol precipitation, standing overnight at 4 ℃, centrifuging, deproteinizing, finally dialyzing in a 3500 dialysis bag to obtain sweet potato crude polysaccharide, and calculating the extraction rate of the polysaccharide by the following formula:
Figure BDA0002706090520000051
second, experimental design and statistical analysis
1. Single factor experiment
(1) Extraction temperature: weighing 10g of sweet potato powder per part, weighing 5 parts, determining the extraction time as 2h, the material-liquid ratio is 1:20, the extraction frequency is 3 times, extracting in a constant-temperature water bath kettle at 40 ℃, 50 ℃, 60 ℃, 70 ℃, 80 ℃ and 90 ℃, centrifuging, collecting supernate, concentrating, and mixing the concentrated solution: anhydrous ethanol ═ 1: 4, adding absolute ethyl alcohol, placing in a refrigerator at 4 ℃ overnight, after alcohol precipitation, centrifugally collecting precipitate, and adding a small amount of distilled water to prepare a crude polysaccharide solution. Adding 0.014g amylase, boiling in water bath at 85 deg.C for 30min, recording volume, and measuring sugar concentration to obtain extraction rate.
(2) Extraction time: weighing 10g of sweet potato powder per part, weighing 5 parts of sweet potato powder, and mixing the raw materials according to a material-liquid ratio of 1:20. extracting for 3 times, and extracting for 3 times respectively for 1h, 1.5h, 2h, 2.5h, and 3h according to the determined optimal extraction temperature. The rest of the operation steps are the same as above.
(3) The material-liquid ratio: weighing 10g of sweet potato powder per part, weighing 5 parts, extracting for 3 times, and extracting for 3 times according to the determined optimal extraction temperature and extraction time according to the material-liquid ratio of 1:10, 1:15, 1:20, 1:25 and 1: 30. The rest of the operation steps are the same as above.
(4) The extraction times are as follows: weighing 10g sweet potato powder per part, weighing 5 parts, and extracting for 1 time, 2 times, 3 times, 4 times, and 5 times respectively according to determined optimal extraction temperature, extraction time, and material-liquid ratio. The rest operation steps are the same as the above, the content of the polysaccharide in the extracting solution is quantitatively measured by adopting a phenol-sulfuric acid method, the extraction rate of the sweet potato polysaccharide is calculated, and each experiment is carried out in parallel for three times.
2. Response surface optimization design
According to the single-factor experiment result, selecting four factors (extraction temperature, time, liquid-material ratio and extraction frequency) which have obvious influence on the extraction rate of the sweet potato polysaccharide, designing an experiment according to a four-factor three-level response surface analysis method by taking the extraction rate of the sweet potato polysaccharide as an evaluation index, calculating the extraction rate and a fitting equation, and solving the optimal condition. And verifying the optimal extraction conditions. The experimental factors and levels of the response surface analysis are shown in table 1.
TABLE 1 test factors and horizon table for response surface analysis
Figure BDA0002706090520000061
The temperature (A), the feed-liquid ratio (B), the time (C), the extraction times (D) and the polysaccharide extraction rate as the response value (R) are shown in the table 2.
TABLE 2 Experimental protocols and results table
Figure BDA0002706090520000062
Figure BDA0002706090520000071
3. Model building and statistical analysis
And performing multiple regression analysis according to the obtained data to obtain a multiple quadratic regression equation between corresponding variables (temperature, feed-liquid ratio, extraction time and extraction times) and response values (polysaccharide extraction rate).
R(％)＝12.65+0.59A+0.95B-0.26C+0.065D-0.61AB-0.30AC+0.46AD+0.45BC+0.91BD+0.24CD-3.27A2-2.72B2-2.41C2-2.29D2
The linear relation between each factor and the response value is obvious and is judged by F value test, and the smaller the P value is, the higher the significance of the explanatory variable is. Analysis of variance was performed on the data, and the results (Table 3) show P<0.0001, indicating that the regression model is significant; mismatching term P-0.0518>0.05, not significant. Regression correlation coefficient R20.9970 and coefficient of determination R2 AdjThe similarity of 0.9939 indicates that the test value and the predicted value of the test have high fitting degree and can be better used for predicting the test result.
TABLE 3 results of analysis of variance of experimental data
Figure BDA0002706090520000081
4. Analysis and optimization of experimental results
Drawing analysis is carried out according to the regression equation by using Design Expert software to obtain a response surface and contour lines of the regression equation, as shown in fig. 1 to 10.
According to model analysis, the sweet potato polysaccharide extraction optimization result is that the temperature is 60.80 ℃, the time is 1.98h, the material-liquid ratio is 1:20.86, the extraction frequency is 3.05, and the polysaccharide extraction rate is 12.75% under the condition. The relative deviation is about 0.94%, which shows that the leaching process obtained by utilizing the response surface method has higher reliability, and the model established by the experiment can be used for experimental prediction.
Although the embodiments of the present invention have been disclosed for illustrative purposes, those skilled in the art will appreciate that: various substitutions, changes and modifications are possible without departing from the spirit and scope of the invention and the appended claims, and therefore the scope of the invention is not limited to the embodiments disclosed.

Claims (5)

1. A method for optimizing sweet potato polysaccharide extraction conditions based on a response surface method is characterized by comprising the following steps: the method comprises the following steps:
peeling sweet potatoes, cutting the sweet potatoes into strips, freeze-drying the sweet potatoes in a freeze dryer at-70 ℃, taking the sweet potatoes out at night, putting the sweet potatoes in a drying oven at 60 ℃, continuously drying the sweet potatoes for more than 8 hours, finally crushing the sweet potatoes into sweet potato powder by a crusher, sieving the sweet potato powder by a 40-mesh sieve, and bagging the sweet potato powder in a dryer for preservation;
the sweet potato polysaccharide is extracted by a water extraction and alcohol precipitation method, variable parameters of sweet potato polysaccharide extraction are determined by a response surface method, a four-factor three-level test mode is set to perform repeated tests to obtain an experiment result, the extraction rate of crude sweet potato polysaccharide is calculated, multiple regression analysis is performed by Design Expert 8.0.6 software, and the regression equation of the influence of experiment factors on response values is obtained:
R(％)＝12.65+0.59A+0.95B-0.26C+0.065D-0.61AB-0.30AC+0.46AD+0.45BC+0.91BD+0.24CD-3.27A2-2.72B2-2.41C2-2.29D2
in the formula, the response value R is the extraction rate of sweet potato polysaccharide, the variable parameter A is the extraction temperature, the variable parameter B is the feed-liquid ratio, the variable parameter C is the extraction time, the variable parameter D is the extraction frequency, the final determination that A is 60.80 ℃, B is 1:20.86, C is 1.98h, D is 3.05 and the extraction rate of polysaccharide is 12.75 percent;
weighing sweet potato powder in the step, adding petroleum ether for degreasing, discarding supernatant on the next day, and adding 95% ethanol into the precipitate for 3 times; wherein the sweet potato powder: degreasing with petroleum ether: proportion g of 95% ethanol: mL: mL is 1: 5: 6;
and (4) pouring out the supernatant after the reaction is finished, and mixing the materials according to a material-liquid ratio of 1:20, adding water, and extracting the polysaccharide in the sweet potatoes at 61 ℃ for 2 hours; centrifuging, concentrating, adding anhydrous ethanol 4 times the volume of the concentrated solution, standing at 4 deg.C overnight, centrifuging after precipitating with ethanol, collecting precipitate, and adding distilled water into the precipitate to obtain crude polysaccharide solution; the crude polysaccharide solution was mixed with Sevag reagent at 4: 1, violently shaking, centrifuging, removing protein in the solution, pouring the solution into a separating funnel filled with AB-8 macroporous adsorption resin for decolorization for 12 hours, then washing the resin with 3 times of volume of distilled water, concentrating to 1/10 of the original volume, and dialyzing by using a 3500 dialysis bag to obtain crude sweet potato polysaccharide;
wherein the Sevag reagent is a mixture of chloroform and n-butanol, and the weight ratio of chloroform: the volume ratio of n-butanol is 4: 1.
2. the method for optimizing sweet potato polysaccharide extraction process parameters based on the response surface method as claimed in claim 1, wherein: in the step II, the volume ratio of chloroform to n-butyl alcohol is 4: 1.
3. the method for optimizing sweet potato polysaccharide extraction process parameters based on the response surface method as claimed in claim 1, wherein: in the step II, during centrifugal separation, the rotating speed of the centrifugal machine is 6000 r/min.
4. The method for optimizing sweet potato polysaccharide extraction process parameters based on the response surface method as claimed in any one of claims 1 to 3, wherein: the method further comprises the steps of:
preparing the crude polysaccharide in the step II into 0.1mg/mL polysaccharide solution by using distilled water, and then diluting the prepared sample solution by different times; and (5) measuring the absorbance according to a method for drawing a standard curve, and calculating the extraction rate of the sweet potato polysaccharide.
5. The method for optimizing sweet potato polysaccharide extraction process parameters based on the response surface method as claimed in claim 4, wherein: the standard curve is formulated as follows:
weighing an excessive anhydrous glucose standard substance, drying in a drying oven at 105 ℃ until the weight is constant, accurately weighing 100mg and fixing the volume to 100mL to obtain a 1.0mg/mL glucose solution which is called stock solution; sucking 10mL from the stock solution by using a pipette gun, and fixing the volume by using a 100mL volumetric flask to obtain a glucose standard solution of 0.1 mg/mL;
accurately measuring 0, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8 and 0.9ml of glucose standard solution in test tubes by using a pipette, supplementing each test tube to 1ml by using distilled water, adding 1.0ml of 6 percent phenol solution in each test tube, slowly adding 5ml of concentrated sulfuric acid, uniformly shaking, carrying out boiling water bath for 15min, and measuring the absorbance value at 490nm after cooling to the room temperature;
the absorbance value A is determined by taking the glucose concentration C (ug/mL) as the abscissa490The ordinate is a standard curve.
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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112724276A (en) * 2021-02-01 2021-04-30 昭通学院 Ultrasonic-assisted extraction process for optimizing aralia leucotaxis stem bark polysaccharide based on response surface method
CN113461833A (en) * 2021-08-05 2021-10-01 内蒙古医科大学 Method for extracting astragalus mongholicus polysaccharide
CN116003649A (en) * 2023-02-03 2023-04-25 西南民族大学 Extraction method of polysaccharide of thalictrum hemipterum hance and antioxidation activity analysis method thereof

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110156904A (en) * 2019-05-14 2019-08-23 江苏师范大学 Purple sweetpotato polysaccharide is preparing the application in anti-lung-cancer medicament

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110156904A (en) * 2019-05-14 2019-08-23 江苏师范大学 Purple sweetpotato polysaccharide is preparing the application in anti-lung-cancer medicament

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
郭金颖: "甘薯多糖提取及性能研究", 《中国优秀博硕士学位论文全文数据库(硕士)工程科技Ｉ辑》 *

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112724276A (en) * 2021-02-01 2021-04-30 昭通学院 Ultrasonic-assisted extraction process for optimizing aralia leucotaxis stem bark polysaccharide based on response surface method
CN113461833A (en) * 2021-08-05 2021-10-01 内蒙古医科大学 Method for extracting astragalus mongholicus polysaccharide
CN116003649A (en) * 2023-02-03 2023-04-25 西南民族大学 Extraction method of polysaccharide of thalictrum hemipterum hance and antioxidation activity analysis method thereof

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Application publication date: 20210119