Method for extracting boschniakia rossica polysaccharide

Abstract

The invention belongs to the technical field of polysaccharide extraction, and particularly relates to a boschniakia rossica polysaccharide extraction method. The boschniakia rossica polysaccharide extract is obtained by dispersing boschniakia rossica powder in an ethanol solution, then adding a compound enzyme, adjusting the pH value to 5-7, and carrying out ultrasonic treatment, enzyme deactivation and centrifugation. Compared with a traditional single extraction method, the extraction method has the advantages that the extraction process of the boschniakia rossica polysaccharide is optimized through the response surface method, and compared with the traditional single extraction method, the extraction method is practical and reliable, can be used for extracting the boschniakia rossica polysaccharide, and can remarkably improve the polysaccharide yield which can reach 13.67%.

Description

Technical Field

[0001] The invention belongs to the technical field of polysaccharide extraction, and more specifically relates to a method for extracting polysaccharide from Cistanche rossica. Background Art

[0002] Cistanche deserticola is a perennial herb of the genus Cistanche in the family Orobanchaceae. The whole plant is used as medicine. It is sweet, sour and moist in nature. It is mainly used to treat kidney deficiency, impotence, cold pain in the waist and knees, urine retention, metrorrhagia, constipation due to dry intestines, cystitis, etc. It contains multiple active ingredients such as polysaccharides, Cistanche deserticola glycosides, Cistanche deserticola phenylpropanoid glycosides, Cistanche deserticola iridoid glycosides, oleanolic acid, ursolic acid and chlorogenic acid.

[0003] Polysaccharides are a type of macromolecular carbohydrates composed of monosaccharide units. They are mainly composed of monosaccharides. At present, it has been found that many polysaccharides have the effects of anti-tumor, anti-inflammatory, immune regulation, wound healing, etc., and have little toxicity and side effects, which has attracted widespread attention from researchers. Polysaccharides are also one of the main active ingredients of Cistanche deserticola. Cistanche deserticola polysaccharides also have various activities, such as enhancing immunity, anti-aging, anti-cancer, anti-inflammatory, etc.

[0004] The existing methods for extracting polysaccharides mainly include acid-base extraction, ultrasound-assisted extraction, microwave-assisted extraction, enzyme-assisted extraction, and low eutectic solvent extraction. Although these methods have been widely used, the yield of a single extraction method is low and unstable. How to improve the yield and stability of polysaccharides has become a difficult problem that technicians in this field need to solve urgently. Summary of the invention

[0005] The purpose of the present invention is to provide a method for extracting polysaccharides from Cistanche rossica to solve the problems existing in the prior art.

[0006] To achieve the above object, the present invention provides the following solutions:

[0007] One of the technical solutions of the present invention is to provide a method for extracting polysaccharides from Cistanche deserticola, comprising the following steps:

[0008] dispersing the Cistanche rossica powder in an ethanol solution to obtain a Cistanche rossica solution;

[0009] Adding complex enzyme into the Cistanche rossica solution, adjusting the pH value to 5-7, ultrasonic treatment, enzyme inactivation and centrifugation to obtain the Cistanche rossica polysaccharide extract.

[0010] Furthermore, the Cistanche deserticola powder is a sieve that passes through a 60-100 mesh sieve, preferably 80 mesh.

[0011] Furthermore, the solid-liquid ratio of the Cistanche rossica powder to the ethanol solution is 1:10-30 (g / mL), preferably 1:25 (g / mL).

[0012] Furthermore, the pH value is adjusted to 5.

[0013] Furthermore, the volume fraction of the ethanol solution is 60-100%.

[0014] Furthermore, the complex enzyme includes cellulase, pectinase and papain.

[0015] Preferably, the amount of cellulase used is 3-5% of the mass of Cistanche rossica powder; the amount of pectinase used is 3-5% of the mass of Cistanche rossica powder; the amount of papain used is 3-5% of the mass of Cistanche rossica powder.

[0016] More preferably, the amount of the cellulase is 5% of the mass of the Cistanche rossica powder; the amount of the pectinase is 4% of the mass of the Cistanche rossica powder; and the amount of the papain is 4% of the mass of the Cistanche rossica powder.

[0017] Furthermore, the ultrasonic treatment has a power of 300-500W, a temperature of 40-80°C, and a time of 0.5-3h.

[0018] Furthermore, the enzyme inactivation is carried out by treating the mixture in a boiling water bath for 10 minutes.

[0019] The present invention discloses the following technical effects:

[0020] The ultrasound-assisted enzyme extraction of crude Cistanche rossica polysaccharide of the present invention is practical and reliable compared with a traditional single extraction method, can be used for Cistanche rossica polysaccharide extraction, and can significantly improve the polysaccharide yield, which can reach 13.67%. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] The drawings constituting a part of the present application are used to provide a further understanding of the present application. The illustrative embodiments and descriptions of the present application are used to explain the present application and do not constitute an improper limitation on the present application. In the drawings:

[0022] Figure 1 is the glucose standard curve;

[0023] Figure 2 The yield of crude polysaccharide from Cistanche rossica using the methods of Example 6 and Comparative Examples 1-2;

[0024] Figure 3 is the effect of ethanol mass fraction on the yield of crude polysaccharides from Cistanche rossica;

[0025] Figure 4 The effect of ultrasonic treatment time on the yield of crude polysaccharides from Botrytis cistanche;

[0026] Figure 5 The effect of ultrasonic temperature on the yield of crude polysaccharides from Cistanche rossica;

[0027] Figure 6The effect of ultrasonic power on the yield of crude polysaccharides from Cistanche rossica;

[0028] Figure 7 The contour map and three-dimensional surface map of the effect of ethanol concentration and extraction time on polysaccharide yield, where the left map is a contour map and the right map is a three-dimensional surface map;

[0029] Figure 8 The figure below shows the contour map and three-dimensional surface map of the effect of ethanol concentration and ultrasonic power on polysaccharide yield, where the left figure is the contour map and the right figure is the three-dimensional surface map;

[0030] Fig. 9 The figure below shows the contour map and three-dimensional surface map of the effect of ethanol concentration and ultrasonic power on polysaccharide yield, where the left figure is the contour map and the right figure is the three-dimensional surface map;

[0031] Fig.10 The contour map and three-dimensional surface map of the effects of extraction time and extraction temperature on polysaccharide yield, where the left picture is a contour map and the right picture is a three-dimensional surface map;

[0032] Fig.11 The contour map and three-dimensional surface map of the effects of extraction time and ultrasonic power on polysaccharide yield, where the left picture is a contour map and the right picture is a three-dimensional surface map;

[0033] Fig.12 The contour map and three-dimensional surface map of the effect of extraction temperature and ultrasonic power on polysaccharide yield, where the left picture is a contour map and the right picture is a three-dimensional surface map;

[0034] Fig.13 The SEM images of the Cistanche deserticola powder treated by the methods of Example 6 and Comparative Examples 1-2 are shown in FIG. 6 , wherein (a) is Example 6, (b) is Comparative Example 1, and (c) is Comparative Example 2. DETAILED DESCRIPTION

[0035] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0036] It should be understood that the terms described in the present invention are only for describing a particular embodiment and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. The intermediate value in any stated value or stated range, and each smaller range between any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0037] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0038] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to those skilled in the art. The present invention description and examples are exemplary only.

[0039] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0040] The raw materials and reagents used in the specific embodiment of the present invention are all commercially available products, among which, cellulase is 400 U / g, pectinase is 500 U / g, and papain is 800 U / g.

[0041] Example 1

[0042] A method for extracting polysaccharides from Cistanche rossica comprises the following steps:

[0043] S1. Weigh 1 g of Cistanche rossica powder (80 mesh) into a 50 mL centrifuge tube, add 25 mL of 70% ethanol solution at a solid-liquid ratio of 1:25 (g / mL), and disperse evenly to obtain a Cistanche rossica solution;

[0044] S2. Weigh 0.05 g of cellulase, 0.04 g of pectinase and 0.04 g of papain and add them to the Cistanche rossica solution of step S1, adjust the pH to 5, and then perform ultrasonic treatment (power 500 W, temperature 50 ° C, time 1 h). After ultrasonic treatment, inactivate the enzyme in a boiling water bath for 10 min, and the supernatant obtained by centrifugation is the Cistanche rossica polysaccharide extract.

[0045] Example 2

[0046] Compared with Example 1, the difference is that the mass fraction of ethanol is 60%, 80%, 90% or 100%.

[0047] Example 3

[0048] Compared with Example 1, the difference is that the ultrasonic treatment time is 0.5h, 1.5h, 2h or 3h.

[0049] Example 4

[0050] Compared with Example 1, the difference is that the temperature of ultrasonic treatment is 40°C, 60°C, 70°C or 80°C.

[0051] Example 5

[0052] Compared with Example 1, the difference is that the power of ultrasonic treatment is 300W, 350W, 400W or 450W.

[0053] Example 6

[0054] A method for extracting polysaccharides from Cistanche rossica comprises the following steps:

[0055] S1. Weigh 1 g of Cistanche rossica powder (80 mesh) into a 50 mL centrifuge tube, add 25 mL of 80% ethanol solution at a solid-liquid ratio of 1:25 (g / mL), and disperse evenly to obtain a Cistanche rossica solution;

[0056] S2, weigh 0.05g of cellulase, 0.04g of pectinase and 0.04g of papain and add them to the Cistanche rossica solution of step S1, adjust the pH to 5, and then perform ultrasonic treatment (power 450W, temperature 73°C, time 100min), inactivate the enzyme in a boiling water bath for 10min after ultrasonic treatment, and the supernatant obtained by centrifugation is the Cistanche rossica polysaccharide extract.

[0057] Comparative Example 1

[0058] The method for enzyme-assisted extraction (EAE) of Cistanche rossica polysaccharide comprises the following steps:

[0059] S1. Weigh 1 g of Cistanche rossica powder (80 mesh) into a 50 mL centrifuge tube, add 25 mL of 80% ethanol solution at a solid-liquid ratio of 1:25 (g / mL), and disperse evenly to obtain a Cistanche rossica solution;

[0060] S2. Weigh 0.05 g of cellulase, 0.04 g of pectinase and 0.04 g of papain and add them to the Cistanche rossica solution of step S1, adjust the pH to 5, and after enzymatic hydrolysis for 1 hour, inactivate the enzyme in a boiling water bath for 10 minutes. The supernatant obtained by centrifugation is the Cistanche rossica polysaccharide extract.

[0061] Comparative Example 2

[0062] The method for ultrasonic-assisted extraction (UAE) of Cistanche deserticola polysaccharide comprises the following steps:

[0063] S1. Weigh 1 g of Cistanche rossica powder (80 mesh) into a 50 mL centrifuge tube, add 25 mL of 80% ethanol solution at a solid-liquid ratio of 1:25 (g / mL), and disperse evenly to obtain a Cistanche rossica solution;

[0064] S2. The Cistanche rossica solution was subjected to ultrasonic treatment (power 450 W, temperature 73° C., time 100 min). The supernatant obtained by centrifugation after ultrasonic treatment was the Cistanche rossica polysaccharide extract.

[0065] Test example

[0066] Drawing of standard curve:

[0067] Accurately measure 1 mg of glucose standard, dilute to 10 mL with water, and prepare 100ug / mL glucose mother solution. Then, take 0 mL, 1 mL, 2 mL, 4 mL, 8 mL, and 10 mL of the mother solution and place them in test tubes, dilute to 10 mL with distilled water, and take 0.2 mL of glucose standard solution from each tube. Add 0.1 mL of 6% phenol solution, let stand for 5 minutes, add 0.4 mL of concentrated sulfuric acid and mix for 30 minutes. Use the solution with a glucose concentration of 0 as the zero solution, and use a 1 mL cuvette to measure the absorbance at a wavelength of 487 nm. Use glucose concentration as the horizontal coordinate and absorbance A as the vertical coordinate to draw a standard curve. The linear regression equation of the standard curve is Y=0.0109X+0.043, and the correlation coefficient R 2 =0.995. The yield of crude polysaccharide in the sample was determined as above. The absorbance of the sample solution was measured at a wavelength of 487nm, and the polysaccharide yield was calculated by substituting it into the standard curve equation. Figure 1 .

[0068] The crude polysaccharide content of the Boehmeria rossica polysaccharide extracts obtained in Example 6 and Comparative Examples 1-2 was determined using the phenol concentrated sulfuric acid method. The results are as follows: Figure 2 shown.

[0069] Figure 2 The yield of crude polysaccharide from Cistanche rossica using the methods of Example 6 and Comparative Examples 1-2 (UAEE in the figure is Example 6, UAE is Comparative Example 1, and EAE is Comparative Example 2). It can be seen from the figure that the yield of crude polysaccharide from Cistanche rossica using the method of Example 6 of the present invention is significantly higher than that of Comparative Examples 1-2.

[0070] The polysaccharide content of the crude polysaccharide extract of Boehmeria rossica obtained by the single factor test (Examples 1 to 5) was determined by the phenol concentrated sulfuric acid method. The results are as follows: Figures 3 to 6 shown.

[0071] Figure 3The effect of ethanol mass fraction on the yield of crude polysaccharides from Cistanche deserticola. As shown in the figure, as the ethanol concentration increases between 60% and 70%, the polysaccharide yield increases, while when the concentration exceeds 70%, the polysaccharide yield decreases. BRPS (polysaccharide yield) reaches the highest value (11.45% ± 0.04) at 70%. The effect of ethanol concentration on polysaccharide yield is mainly reflected in the changes in solubility and hydrogen bond interaction. When the ethanol concentration is moderate, it can effectively reduce the interaction between polysaccharide chains, improve the solubility of polysaccharides and promote extraction. However, too high a concentration will lead to a relative reduction in water molecules, thereby weakening the hydrogen bond interaction with the polysaccharide hydroxyl group, causing the polysaccharide to precipitate or precipitate, and reducing the yield. In addition, the hydrogen bond competition between ethanol molecules and polysaccharide functional groups is also an important factor. Ethanol at an appropriate concentration can form hydrogen bonds with hydroxyl groups to increase solubility, while too high a concentration will inhibit the formation of hydrogen bonds, further affecting the extraction effect.

[0072] The extraction time (ultrasonic treatment time) directly affects the solubility of polysaccharides. During the extraction process, there is a dynamic balance between the dissolution and precipitation of polysaccharides. Figure 4 The effect of ultrasonic treatment time on the yield of crude polysaccharides from Cistanche rossica is shown in the figure. As can be seen from the figure, in the early stage of extraction, as the extraction time increases, the yield of polysaccharides from Cistanche rossica increases, reaching a maximum value at 90 minutes. Shorter extraction time may lead to the failure of the polysaccharides in Cistanche rossica to be fully released, resulting in a low yield. Proper extension of the extraction time can improve the solubility of polysaccharides, and continuous cavitation and microbubble implosion aggravate the rupture of the cell wall of the plant material, reduce the restriction of the cell structure on the mass transfer process, and thus improve the yield. However, as the ultrasonic time continues to increase, the BRPS at 120 minutes and 150 minutes decreases instead. This may be due to the extension of the ultrasonic treatment time, which leads to a short-term high temperature and released free radicals, resulting in the degradation of polysaccharides.

[0073] The effect of extraction temperature (ultrasonic temperature) on the yield of crude polysaccharides from Cistanche rossica is mainly reflected in the promotion of thermal energy on the solubility and extraction efficiency of polysaccharides. Figure 5 The effect of ultrasonic temperature on the yield of crude polysaccharides from Cistanche deserticola is shown in the figure. As can be seen from the figure, the value of BRPS increases significantly with the increase of temperature and reaches the maximum value at 70°C. However, as the temperature continues to increase, the extraction rate decreases after exceeding 70°C. This may be because at higher temperatures, the surface tension decreases, resulting in a lower threshold of cavitation intensity required for ultrasonic waves, which helps the extraction process. However, when the temperature is further increased to above 80°C, the thermal effect will cause the degradation of the polysaccharide structure, offsetting the increased mass transfer rate due to the increase in temperature, resulting in a decrease in the yield of crude polysaccharides from Cistanche deserticola.

[0074] The effect of ultrasonic power on the yield of crude polysaccharides from Cistanche rossica is mainly reflected in the cavitation effect that can be produced by ultrasound. This effect enhances the interaction between the solvent and the solid by forming tiny bubbles and then collapsing rapidly, thereby affecting the solubility and precipitation efficiency of the polysaccharide. Figure 6 The effect of ultrasonic power on the yield of crude polysaccharides from Cistanche deserticola is shown in the figure. As the ultrasonic power increases from 300W to 450W, the yield of BRPS increases significantly, and then gradually decreases. As the ultrasonic power increases, cavitation effect, thermal effect and mechanical effect contribute to the improvement of BRPS. However, due to the influence of ultrasound, excessive ultrasonic power may lead to the degradation of polysaccharides in the solvent.

[0075] Then, according to the results of the single factor test, the factors with greater influence were selected, and the yield of crude polysaccharides from Cistanche deserticola was used as the response value. According to the Box-Behnken design principle, the extraction conditions were optimized by response surface analysis. The response surface test factors and their coding level settings are shown in Table 1, the response surface test results are shown in Table 2, and the variance analysis of the response surface model is shown in Table 3.

[0076] Table 1

[0077]

[0078] Table 2

[0079]

[0080] Table 3

[0081]

[0082] Note: *, significant difference at 0.05 level (p<0.05); **, significant difference at 0.01 level (p<0.01); ***, statistically significant difference at 0.001 level (p<0.001).

[0083] The experimental data were fitted by multiple regression using Design Expert DX-8 software, and the response surface fitting equation was obtained: BRPS = +13.46-0.18A+0.20B+1.20C+0.50D+0.22AB+0.13AC-0.015AD-0.042BC+0.15BD-0.16CD-0.91A^ 2 -0.84B^ 2 -2.04C^ 2 -2.65D^ 2The variance analysis of the fitted quadratic polynomial model is shown in Table 3. It can be seen from Table 3 that the F value of this model (Model) is 83.20, and the mean square error p<0.0001, indicating that the experimental response surface fitting equation regression model is extremely significant; lack of fit p=0.0728>0.05, indicating that the lack of fit is not statistically significant, that is, the model fit is good. Based on the above analysis, it can be shown that this model can be used to analyze and predict the total polysaccharide yield of Cistanche rossica. In addition, it can be seen from Table 7 that the linear terms C, D and quadratic terms A^ 2 , B^ 2 , C^ 2 , D^ 2 The p values ​​of all the factors were less than 0.0001, which was a very significant level, indicating the rationality of the single factor test. The p value of the interaction term AB was less than 0.05, reaching a significant level, and the other terms were not significant, indicating that the interaction between ethanol concentration and extraction time had a greater impact on Cistanche deserticola polysaccharides.

[0084] The response surface software was used to analyze the three-dimensional surface and contour map. The interaction between the variables of each factor was analyzed by observing the inclination of the surface and the shape of the contour line. Figures 7 to 12 shown.

[0085] in, Figure 7 The following are contour maps and three-dimensional surface maps of the effects of ethanol concentration and extraction time on polysaccharide yield. The left figure is a contour map and the right figure is a three-dimensional surface map. As can be seen from the figure, with the increase of extraction time, the BRPS value gradually increases, indicating that a longer extraction time helps to improve the extraction effect. This may be because ethanol can more fully dissolve and extract the active ingredients in the sample over a longer period of time. As for ethanol concentration, within a certain optimal concentration range, the BRPS value reaches the maximum, and beyond this range, the BRPS value may decrease. This may be because too high or too low ethanol concentration affects the polarity of the solvent, thereby affecting the solubility and extraction efficiency of the active ingredients. The surface curve is steeply curved, indicating that the ethanol concentration and extraction time have a great influence on the response value. The contour line shape is elliptical. Combined with the results of variance analysis, it shows that the interaction between ethanol concentration and extraction time is significant.

[0086] Figure 8The following are contour maps and three-dimensional surface maps of the effects of ethanol concentration and ultrasonic power on polysaccharide yield. The left figure is a contour map and the right figure is a three-dimensional surface map. It can be seen from the figure that the BRPS value reaches the highest point under certain combinations of extraction temperature and ethanol concentration, forming an ideal extraction condition area. As the extraction temperature increases, the BRPS value gradually increases, indicating that higher temperatures help improve the extraction efficiency. This may be because high temperature can accelerate the interaction between the solvent and the sample, promote the dissolution and release of the active ingredients, and the ethanol concentration still makes the polysaccharide yield the highest within a certain optimal concentration range. The curve of the surface map is steeply curved, indicating that the ethanol concentration and extraction temperature have little effect on the response value. The contour line shape is elliptical. Combined with the results of variance analysis, it shows that the interaction between ethanol concentration and extraction temperature is not significant.

[0087] Fig. 9 The following are contour maps and three-dimensional surface maps of the effects of ethanol concentration and ultrasonic power on polysaccharide yield. The left figure is a contour map, and the right figure is a three-dimensional surface map. It can be seen from the figure that the ultrasonic power and ethanol concentration are both within a certain optimal concentration range to achieve the highest polysaccharide yield. With the increase of ultrasonic power, the BRPS value gradually increases, reaches a maximum value, and then decreases. This shows that within a certain range, increasing the ultrasonic power can effectively improve the extraction efficiency and promote the release of effective ingredients. However, when the ultrasonic power is too high, the BRPS value may decrease, which may be due to the degradation of the sample or excessive evaporation of the solvent caused by excessive ultrasonic action, thus affecting the extraction effect. The curve of the surface map is relatively steep, indicating that the ethanol concentration and extraction temperature have a greater impact on the response value. The contour line shape is elliptical. Combined with the results of variance analysis, it shows that the interaction between ethanol concentration and extraction temperature is not significant.

[0088] Fig.10 The following are contour maps and three-dimensional surface maps of the effects of extraction time and extraction temperature on polysaccharide yield. The left figure is a contour map and the right figure is a three-dimensional surface map. As can be seen from the figure, with the increase of extraction time, the value of BRPS gradually increases, reaches a maximum value and then decreases. This shows that within a certain range, extending the extraction time can effectively improve the extraction efficiency and promote the release of effective ingredients. However, when the extraction time is too long, the value of BRPS may decrease, which may be due to the degradation of effective ingredients caused by over-extraction or excessive evaporation of solvent. Similarly, changes in extraction temperature also have a significant effect on BRPS. Within a moderate temperature range, the value of BRPS increases significantly, but when the temperature is too high, it may cause the degradation of sample components, thereby affecting the extraction effect. The curve of the surface map is steeply curved, indicating that the extraction time and extraction temperature have little effect on the response value. The contour line shape is elliptical. Combined with the results of variance analysis, it shows that the interaction between extraction time and extraction temperature is not significant.

[0089] Fig.11 The following are contour maps and three-dimensional surface maps of the effects of extraction time and ultrasonic power on polysaccharide yield. The left figure is a contour map and the right figure is a three-dimensional surface map. As can be seen from the figure, with the increase of ultrasonic power and extraction time, the value of BRPS gradually increases, reaches a maximum value and then decreases. This shows that within a certain range, increasing ultrasonic power and extending extraction time can effectively improve extraction efficiency and promote the release of effective ingredients. However, when the ultrasonic power or extraction time is too high, the value of BRPS may decrease. This may be due to the degradation of sample components caused by excessive ultrasonic action or too long extraction time, or excessive evaporation of solvent, thus affecting the extraction effect. The curve of the surface map is steeply curved, indicating that the ultrasonic power and extraction time have little effect on the response value. The contour line shape is elliptical. Combined with the results of variance analysis, it shows that the interaction between ultrasonic power and extraction time is not significant.

[0090] Fig.12 The following are contour maps and three-dimensional surface maps of the effects of extraction temperature and ultrasonic power on polysaccharide yield. The left figure is a contour map and the right figure is a three-dimensional surface map. It can be seen from the figure that the BRPS value reaches the maximum at moderate ultrasonic power and extraction temperature, indicating that the optimized combination of these two factors can effectively improve the extraction efficiency. With the increase of ultrasonic power, the BRPS value gradually increases, but after reaching a certain critical point, continuing to increase the power may lead to a decrease in BRPS, which may be due to the degradation of sample components or excessive evaporation of solvents caused by excessive ultrasonic action. At the same time, the increase in extraction temperature also helps to increase BRPS, but too high a temperature may lead to the loss of effective ingredients. The curve of the surface map is steeply curved, indicating that the ultrasonic power and extraction temperature have little effect on the response value, and the contour line shape is elliptical. Combined with the results of variance analysis, it shows that the interaction between ultrasonic power and extraction temperature is not significant.

[0091] Fig.13 The SEM images of the Cistanche rossica powder treated by the methods of Example 6 and Comparative Examples 1-2 are shown, wherein (a) is Example 6, (b) is Comparative Example 1, and (c) is Comparative Example 2. As can be seen from the figure, the plant sample (Cistanche rossica powder) is better broken under the ultrasonic coenzyme condition (Example 6), which further explains why the polysaccharide yield of Cistanche rossica in the method of Example 6 is higher.

[0092] The various embodiments in this specification are described in a progressive manner, and each embodiment focuses on the differences from other embodiments. The same or similar parts between the various embodiments can be referenced to each other.

[0093] The above description of the disclosed embodiments enables one skilled in the art to implement or use the present invention. Various modifications to these embodiments will be apparent to one skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention will not be limited to the embodiments shown herein, but rather to the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A method for extracting polysaccharides from Cistanche deserticola, characterized in that the steps include: dispersing the Cistanche rossica powder in an ethanol solution to obtain a Cistanche rossica solution; After adding complex enzyme into the Cistanche rossica solution, adjusting the pH value to 5-7, ultrasonic treatment, enzyme inactivation and centrifugation are performed to obtain the Cistanche rossica polysaccharide extract.

2. The method for extracting Cistanche polysaccharide according to claim 1, characterized in that: The Cistanche deserticola powder is a material that passes through a 60-100 mesh sieve.

3. The method for extracting Cistanche polysaccharide according to claim 2, characterized in that: The Cistanche deserticola powder is a material that passes through an 80-mesh sieve.

4. The method for extracting Cistanche polysaccharide according to claim 1, characterized in that: The solid-liquid ratio of the Cistanche rossica powder to the ethanol solution is 1g:10mL-30.

5. The method for extracting Cistanche polysaccharide according to claim 1, characterized in that: The volume fraction of the ethanol solution is 60-100%.

6. The method for extracting Cistanche polysaccharide according to claim 1, characterized in that: The complex enzyme comprises cellulase, pectinase and papain.

7. The method for extracting Cistanche polysaccharide according to claim 6, characterized in that: The amount of the cellulase is 3-5% of the mass of the Cistanche deserticola powder; and / or The amount of pectinase used is 3-5% of the mass of Cistanche rossica powder; and / or The dosage of the papain is 3-5% of the mass of the Cistanche rossica powder.

8. The method for extracting Cistanche polysaccharide according to claim 7, characterized in that: The amount of the cellulase is 5% of the mass of the Cistanche deserticola powder; and / or The amount of pectinase used is 4% of the mass of Cistanche rossica powder; and / or The dosage of the papain is 4% of the mass of the Cistanche rossica powder.

9. The method for extracting Cistanche polysaccharide according to claim 1, characterized in that: The power of the ultrasonic treatment is 300-500W, the temperature is 40-80°C, and the time is 0.5-3h.

10. The method for extracting Cistanche polysaccharide according to claim 1, characterized in that: The enzyme inactivation is carried out by treating in a boiling water bath for 10 minutes.

Images