Method for extracting polysaccharides from bryophyllum pinnatum

Abstract

The application belongs to the technical field of polysaccharide extraction, and more particularly relates to a method for extracting polysaccharide from desertliving cistanche. The application obtains the polysaccharide extract of desertliving cistanche by dispersing the powder of desertliving cistanche in an ethanol solution, adding a composite enzyme, adjusting the pH to 5-7, and performing ultrasonic treatment, enzyme inactivation and centrifugation. Compared with the traditional single extraction method, the extraction method of the application is practical and reliable, can be used for extracting polysaccharide from desertliving cistanche, can significantly improve the polysaccharide yield, and the polysaccharide yield can reach 13.67%.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of polysaccharide extraction, and more particularly to a method for extracting polysaccharides from Cistanche. BACKGROUND

[0002] Cistanche is a perennial herbaceous plant of the family of Orobanchaceae, and the whole plant is used as medicine. Cistanche has the effects of treating kidney deficiency, impotence, cold pain in the waist and knees, incontinence, metrorrhagia, constipation, and cystitis. Cistanche contains polysaccharides, cistanche glycosides, cistanche phenylpropanoid glycosides, cistanche iridoid glycosides, oleanolic acid, ursolic acid, and chlorogenic acid, and other active ingredients.

[0003] Polysaccharides are a class of macromolecular carbohydrates composed of monosaccharide units, which are mainly composed of monosaccharides. At present, a variety of polysaccharides have been found to have the effects of anti-tumor, anti-inflammatory, immune regulation, and wound healing promotion, and have little toxic and side effects, which has attracted widespread attention of researchers. Polysaccharides are one of the main active ingredients of Cistanche, and Cistanche polysaccharides also have various activities, such as enhancing immunity, anti-aging, anti-cancer, and anti-inflammatory.

[0004] The existing polysaccharide extraction methods mainly include acid-base extraction, ultrasonic-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, and how to improve the yield and stability of polysaccharides has become a difficult problem that needs to be solved by the technical personnel in the field. SUMMARY

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

[0006] To achieve the above-mentioned purpose, the present application provides the following solutions.

[0007] One of the technical solutions of the present application provides a method for extracting polysaccharides from Cistanche, which comprises the following steps:

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

[0009] adding a composite enzyme to the Cistanche solution, adjusting the pH to 5-7, and then performing ultrasonic treatment, enzyme inactivation, and centrifugation to obtain a Cistanche polysaccharide extract.

[0010] Further, the Cistanche powder is undersize material passing through a 60-100 mesh sieve, preferably 80 mesh.

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

[0012] Further, the adjusted pH value is 5.

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

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

[0015] Preferably, the cellulase is used in an amount of 3-5% of the mass of the O. acanthium powder; the pectinase is used in an amount of 3-5% of the mass of the O. acanthium powder; and the papain is used in an amount of 3-5% of the mass of the O. acanthium powder.

[0016] More preferably, the cellulase is used in an amount of 5% of the mass of the O. acanthium powder; the pectinase is used in an amount of 4% of the mass of the O. acanthium powder; and the papain is used in an amount of 4% of the mass of the O. acanthium powder.

[0017] Further, the ultrasonic treatment is performed at a power of 300-500 W, a temperature of 40-80°C and for a time of 0.5-3 h.

[0018] Further, the enzyme inactivation is performed by boiling water bath treatment for 10 min.

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

[0020] Compared with the traditional single extraction method, the ultrasonic-assisted enzyme extraction method of the present application is practical and reliable, can be used for O. acanthium polysaccharide extraction, and can significantly improve the polysaccharide yield, which can reach 13.67%. BRIEF DESCRIPTION OF DRAWINGS

[0021] The accompanying drawings, which form a part of the present application, are included to provide further understanding of the present application and are incorporated herein in conjunction with the present application. The schematic embodiments of the present application and the descriptions thereof 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 a glucose standard curve;

[0023] Figure 2 is the O. acanthium polysaccharide yield using the method of Example 6 and Comparative Examples 1-2;

[0024] Figure 3 is the influence of the ethanol mass fraction on the O. acanthium polysaccharide yield;

[0025] Figure 4 is the influence of the ultrasonic treatment time on the O. acanthium polysaccharide yield;

[0026] Figure 5 is the influence of the ultrasonic temperature on the O. acanthium polysaccharide yield;

[0027] Figure 6Effect of ultrasonic power on the yield of crude polysaccharides from Cistanche deserticola Y.C.Ma;

[0028] Figure 7 Contour and three-dimensional surface plots of the effect of ethanol concentration and extraction time on the yield of polysaccharides, wherein the left plot is a contour plot and the right plot is a three-dimensional surface plot;

[0029] Figure 8 Contour and three-dimensional surface plots of the effect of ethanol concentration and ultrasonic power on the yield of polysaccharides, wherein the left plot is a contour plot and the right plot is a three-dimensional surface plot;

[0030] Figure 9 Contour and three-dimensional surface plots of the effect of ethanol concentration and ultrasonic power on the yield of polysaccharides, wherein the left plot is a contour plot and the right plot is a three-dimensional surface plot;

[0031] Figure 10 Contour and three-dimensional surface plots of the effect of extraction time and extraction temperature on the yield of polysaccharides, wherein the left plot is a contour plot and the right plot is a three-dimensional surface plot;

[0032] Figure 11 Contour and three-dimensional surface plots of the effect of extraction time and ultrasonic power on the yield of polysaccharides, wherein the left plot is a contour plot and the right plot is a three-dimensional surface plot;

[0033] Figure 12 Contour and three-dimensional surface plots of the effect of extraction temperature and ultrasonic power on the yield of polysaccharides, wherein the left plot is a contour plot and the right plot is a three-dimensional surface plot;

[0034] Figure 13 SEM images of Cistanche deserticola powder after treatment by the method of Example 6 and Comparative Examples 1-2, wherein (a) is Example 6, (b) is Comparative Example 1, and (c) is Comparative Example 2. DETAILED DESCRIPTION

[0035] The various illustrative embodiments of the present application will now be described in detail below. This description is not intended to be a limitation on the present application but rather a description of certain aspects, features, and embodiments of the present application. Variations and modifications can be made to the described embodiments within the scope of the present application.

[0036] It should be understood that the terms used herein are for the purpose of describing particular embodiments and are not intended to limit the present application. Additionally, for numerical ranges that are expressed in a lower limit and an upper limit, every intermediate value of the range is specifically included herein. In addition, any smaller ranges falling within the inclusive ranges are also specifically included. The upper and lower limits of these smaller ranges can independently be included or excluded in the range.

[0037] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. While only preferred methods and materials have been described herein, any methods and materials similar or equivalent to those described herein may be used in the implementation or testing of this invention. All references to this specification are incorporated by way of citation to disclose and describe methods and / or materials associated with those references. In the event of any conflict with any incorporated reference, the content of this specification shall prevail.

[0038] Various modifications and variations can be made to the specific embodiments described in this specification without departing from the scope or spirit of the invention, as will be apparent to those skilled in the art. Other embodiments derived from this specification will also be readily apparent to those skilled in the art. This specification and embodiments are merely exemplary.

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

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

[0041] Example 1

[0042] A method for extracting polysaccharides from Cistanche deserticola, comprising the following steps:

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

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

[0045] Example 2

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

[0047] Example 3

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

[0049] Example 4

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

[0051] Example 5

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

[0053] Example 6

[0054] A method for extracting polysaccharides from Cistanche deserticola, comprising the following steps:

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

[0056] S2. Weigh 0.05g of cellulase, 0.04g of pectinase and 0.04g of papain and add them to the Cistanche deserticola solution in step S1. Adjust the pH to 5 and then perform ultrasonic treatment (power 450W, temperature 73℃, time 100min). After ultrasonic treatment, inactivate the enzymes in a boiling water bath for 10min. The supernatant obtained by centrifugation is the Cistanche deserticola polysaccharide extract.

[0057] Comparative Example 1

[0058] The method for enzyme-assisted extraction (EAE) of polysaccharides from Cistanche deserticola includes the following steps:

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

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

[0061] Comparative Example 2

[0062] The method for ultrasound-assisted extraction (UAE) of polysaccharides from Cistanche deserticola includes the following steps:

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

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

[0065] Test case

[0066] Plotting the standard curve:

[0067] Accurately measure 1 mg of glucose standard and dilute to 10 mL with water to prepare a 100 μg / mL glucose stock solution. Then, sequentially pipette 0 mL, 1 mL, 2 mL, 4 mL, 8 mL, and 10 mL of the stock solution into separate test tubes, and dilute to 10 mL with distilled water for each. Pipette 0.2 mL of the glucose standard solution from each test tube, add 0.1 mL of 6% phenol solution, let stand for 5 minutes, then add 0.4 mL of concentrated sulfuric acid and mix well. React for 30 minutes. Use the solution with a glucose concentration of 0 as the zeroing solution, and measure the absorbance at 487 nm using a 1 mL cuvette. Plot a standard curve with glucose concentration on the x-axis and absorbance A on the y-axis. The linear regression equation for the standard curve is Y = 0.0109X + 0.043, and the correlation coefficient R0 is [missing value]. 2 =0.995. The determination of crude polysaccharide yield in the sample was the same as above. The absorbance of the sample solution was measured at a wavelength of 487 nm, and the result was substituted into the standard curve equation to calculate the polysaccharide yield. The glucose standard curve is shown below. Figure 1 .

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

[0069] Figure 2 The figure shows the yield of crude polysaccharides from Cistanche deserticola using the methods of Example 6 and Comparative Examples 1-2 (UAEE represents Example 6, UAE represents Comparative Example 1, and EAE represents Comparative Example 2). As can be seen from the figure, the yield of crude polysaccharides from Cistanche deserticola 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 extracts of Cistanche deserticola obtained from the single-factor experiments (Examples 1-5) was determined using the phenol-sulfuric acid method. The results are as follows: Figures 3 to 6 As shown.

[0071] Figure 3The effect of ethanol mass fraction on the yield of crude polysaccharides from Cistanche deserticola was shown in the figure. As the ethanol concentration increased between 60% and 70%, the polysaccharide yield increased, but decreased when the concentration exceeded 70%. BRPS (polysaccharide yield) reached its highest value (11.45% ± 0.04) at 70%. The effect of ethanol concentration on polysaccharide yield is mainly reflected in changes in solubility and hydrogen bonding interactions. When the ethanol concentration is moderate, it can effectively reduce the interactions between polysaccharide chains, increase the solubility of polysaccharides, and promote extraction. However, excessively high concentrations lead to a relative decrease in water molecules, thereby weakening the hydrogen bonding interactions with the hydroxyl groups of polysaccharides, resulting in polysaccharide precipitation or sedimentation and reducing the yield. Furthermore, the competition for hydrogen bonds between ethanol molecules and polysaccharide functional groups is also an important factor. At appropriate concentrations, ethanol can form hydrogen bonds with hydroxyl groups, increasing solubility, while excessively high concentrations inhibit hydrogen bond formation, further affecting the extraction effect.

[0072] Extraction time (ultrasonic treatment time) directly affects the degree of polysaccharide solubility. During the extraction process, there is a dynamic equilibrium between the dissolution and precipitation of polysaccharides. Figure 4 The effect of ultrasonic treatment time on the yield of crude polysaccharides from Cistanche deserticola was investigated. As shown in the figure, in the initial stage of extraction, the polysaccharide yield increased with increasing extraction time, reaching its maximum at 90 min. Shorter extraction times may result in insufficient release of polysaccharides, leading to a low yield. Appropriately extending the extraction time can improve the solubility of polysaccharides, and continuous cavitation and microbubble implosion exacerbate cell wall rupture in the plant material, reducing the restriction of cell structure on mass transfer processes and thus increasing the yield. However, with further extension of ultrasonic treatment time, the BRPS at 120 min and 150 min decreased, possibly due to the prolonged ultrasonic treatment time leading to brief periods of 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 deserticola is mainly reflected in the promoting effect of heat energy on polysaccharide solubility and extraction efficiency. Figure 5 The effect of ultrasonic temperature on the yield of crude polysaccharides from Cistanche deserticola was investigated. As shown in the figure, the BRPS value increased significantly with increasing temperature, reaching its maximum at 70℃. However, the extraction rate decreased after exceeding 70℃ with further increases in temperature. This may be because at higher temperatures, the surface tension decreases, resulting in a lower cavitation intensity threshold required for ultrasonication, which facilitates the extraction process. However, further increasing the temperature above 80℃ leads to the degradation of the polysaccharide structure due to the thermal effect, offsetting the increased mass transfer rate due to the increased temperature, thus resulting in a decrease in the yield of crude polysaccharides from Cistanche deserticola.

[0074] The effect of ultrasonic power on the yield of crude polysaccharide from Cistanche deserticola is mainly reflected in the cavitation effect generated by ultrasound. This effect enhances the interaction between the solvent and the solid by forming microbubbles and then rapidly collapsing, thereby affecting the solubility and precipitation efficiency of polysaccharides. Figure 6 The effect of ultrasonic power on the yield of crude polysaccharides from Cistanche deserticola was investigated. As shown in the figure, the yield of BRPS increased significantly with increasing ultrasonic power from 300 W to 450 W, and then gradually decreased. With increasing ultrasonic power, cavitation, thermal, and mechanical effects contributed to the improvement of BRPS yield. However, due to the influence of ultrasound, excessive ultrasonic power may lead to the degradation of polysaccharides in the solvent.

[0075] Based on the results of the single-factor experiments, the factors with the greatest influence were selected, with the yield of crude polysaccharide from Cistanche deserticola as the response value. Following the Box-Behnken design principle, response surface methodology was used to optimize the extraction conditions. The response surface experimental factors and their coding levels are shown in Table 1, the results of the response surface experiments are shown in Table 2, and the analysis of variance for the response surface model is shown in Table 3.

[0076] Table 1

[0077]

[0078] Table 2

[0079]

[0080] Table 3

[0081]

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

[0083] The experimental data were fitted using multiple regression analysis with Design Expert DX-8 software, yielding the response surface equation: 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. Table 3 shows that the F-value of this model is 83.20, and the root mean square error (p < 0.0001), indicating that the regression model of the response surface fitting equation is highly significant. The lack of fit (p = 0.0728 > 0.05) indicates that the lack of fit is not statistically significant, meaning the model fit is good. Based on the above analysis, this model can be used to analyze and predict the total polysaccharide yield of *Cistanche deserticola*. Furthermore, Table 7 shows that the linear terms C and D, and the quadratic term A^... 2 B^ 2 C^ 2 D^ 2 The p-values ​​were all less than 0.0001, indicating a highly significant level, thus demonstrating the rationality of the single-factor experiment. The p-value of the interaction term AB was less than 0.05, reaching a significant level, while the remaining terms were not significant, indicating that the interaction between ethanol concentration and extraction time had a significant impact on the polysaccharides of Cistanche deserticola.

[0084] Response surface methodology was used to obtain 3D surface plots and contour plots. The interaction between various factor variables was analyzed by observing the slope of the surface and the shape of the contour lines. Figures 7 to 12 As shown.

[0085] in, Figure 7 The figures show contour plots and 3D surface plots illustrating the effects of ethanol concentration and extraction time on polysaccharide yield. The left figure is the contour plot, and the right figure is the 3D surface plot. As the figures show, the BRPS value gradually increases with increasing extraction time, indicating that a longer extraction time helps improve the extraction efficiency. This is likely because ethanol can more fully dissolve and extract the active ingredients from the sample over a longer period. Regarding ethanol concentration, the BRPS value reaches its maximum within an optimal concentration range; beyond this range, the BRPS value may decrease. This may be because excessively high or low ethanol concentrations affect the solvent polarity, thus affecting the solubility of the active ingredients and the extraction efficiency. The steep curvature of the surface plots indicates that ethanol concentration and extraction time have a significant impact on the response value. The elliptical shape of the contour lines, combined with the analysis of variance results, demonstrates a significant interaction between ethanol concentration and extraction time.

[0086] Figure 8The figures show contour plots and 3D surface plots illustrating the effects of ethanol concentration and ultrasonic power on polysaccharide yield. The left figure is the contour plot, and the right figure is the 3D surface plot. As can be seen from the figures, the BRPS value reaches its maximum at certain combinations of extraction temperature and ethanol concentration, forming an ideal extraction condition region. With increasing extraction temperature, the BRPS value gradually increases, indicating that higher temperatures contribute to improved extraction efficiency. This may be because higher temperatures accelerate the interaction between the solvent and the sample, promoting the dissolution and release of active ingredients, while the highest polysaccharide yield is still achieved within a certain optimal concentration range of ethanol. The surface plot shows a steep curve, indicating that ethanol concentration and extraction temperature have a relatively small impact on the response value. The contour lines are elliptical in shape, and combined with the analysis of variance results, this suggests that the interaction between ethanol concentration and extraction temperature is not significant.

[0087] Figure 9 The figures show contour plots and 3D surface plots illustrating the effects of ethanol concentration and ultrasonic power on polysaccharide yield. The left figure is the contour plot, and the right figure is the 3D surface plot. As can be seen from the figures, the highest polysaccharide yield is achieved only within a certain optimal concentration range for both ultrasonic power and ethanol concentration. With increasing ultrasonic power, the BRPS value gradually increases, reaches a maximum, and then decreases. This indicates that within a certain range, increasing ultrasonic power can effectively improve extraction efficiency and promote the release of active ingredients. However, when the ultrasonic power is too high, the BRPS value may decrease, possibly due to sample degradation or excessive solvent evaporation caused by excessive ultrasonic waves, thus affecting the extraction effect. The surface plot shows a relatively steep curve, indicating that ethanol concentration and extraction temperature have a significant impact on the response value. The contour lines are elliptical in shape, and combined with the analysis of variance results, this suggests that the interaction between ethanol concentration and extraction temperature is not significant.

[0088] Figure 10 The figures show contour plots and 3D surface plots illustrating the effects of extraction time and temperature on polysaccharide yield. The left figure is the contour plot, and the right figure is the 3D surface plot. As the figures show, the BRPS value gradually increases with increasing extraction time, reaches a maximum, and then decreases. This indicates that within a certain range, extending the extraction time can effectively improve extraction efficiency and promote the release of active ingredients. However, when the extraction time is too long, the BRPS value may decrease, possibly due to degradation of active ingredients caused by over-extraction or excessive evaporation of the solvent. Similarly, changes in extraction temperature also have a significant impact on BRPS. Within a moderate temperature range, the BRPS value increases significantly, but excessively high temperatures may lead to degradation of sample components, thus affecting the extraction effect. The surface plot shows a steep curve, indicating that extraction time and temperature have a relatively small impact on the response value. The contour lines are elliptical in shape, and combined with the analysis of variance results, this indicates that the interaction between extraction time and extraction temperature is not significant.

[0089] Figure 11 The figures show contour plots and 3D surface plots illustrating the effects of extraction time and ultrasonic power on polysaccharide yield. The left figure is the contour plot, and the right figure is the 3D surface plot. As the figures show, the BRPS value gradually increases with increasing ultrasonic power and extraction time, reaching a maximum value before decreasing. This indicates that within a certain range, increasing ultrasonic power and extending extraction time can effectively improve extraction efficiency and promote the release of active ingredients. However, when the ultrasonic power or extraction time is too high, the BRPS value may decrease. This could be due to excessive ultrasonic stimulation or prolonged extraction time leading to sample component degradation, or excessive solvent evaporation, thus affecting the extraction effect. The surface plot shows a steep curve, indicating that ultrasonic power and extraction time have a relatively small impact on the response value. The contour lines are elliptical in shape, and combined with the analysis of variance results, this suggests that the interaction between ultrasonic power and extraction time is not significant.

[0090] Figure 12 The figures show contour plots and 3D surface plots illustrating the effects of extraction temperature and ultrasonic power on polysaccharide yield. The left figure is the contour plot, and the right figure is the 3D surface plot. As shown, the BRPS value reaches its maximum at moderate ultrasonic power and extraction temperature, indicating that the optimized combination of these two factors can effectively improve extraction efficiency. With increasing ultrasonic power, the BRPS value gradually increases, but after reaching a certain critical point, further increases in power may lead to a decrease in BRPS. This may be due to degradation of sample components or excessive evaporation of the solvent caused by excessive ultrasonic waves. Simultaneously, increasing the extraction temperature also helps to improve BRPS, but excessively high temperatures may lead to the loss of active ingredients. The surface plot shows a steep curve, indicating that ultrasonic power and extraction temperature have a relatively small impact on the response value. The contour lines are elliptical in shape, and combined with the analysis of variance results, this indicates that the interaction between ultrasonic power and extraction temperature is not significant.

[0091] Figure 13 The images show SEM images of Cistanche deserticola powder treated according to the methods of Example 6 and Comparative Examples 1-2, where (a) is Example 6, (b) is Comparative Example 1, and (c) is Comparative Example 2. As can be seen from the images, the plant sample (Cistanche deserticola powder) was better broken down under the ultrasonic coenzyme condition (Example 6), further explaining the reason for the higher yield of Cistanche deserticola polysaccharides using the method of Example 6.

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

[0093] The above description of the disclosed embodiments enables those skilled in the art to make or use the invention. Various modifications to these embodiments will be readily apparent to those 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 invention. Therefore, the invention is not to be limited to the embodiments shown herein, but is to be accorded 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... include: Cistanche deserticola powder was dispersed in an ethanol solution to obtain a Cistanche deserticola solution; After adding a compound enzyme to the Cistanche deserticola solution, the pH was adjusted to 5, and the Cistanche deserticola polysaccharide extract was obtained by ultrasonic treatment, enzyme inactivation, and centrifugation. The ratio of the powder of Cistanche deserticola to the ethanol solution is 1g:25mL; The volume fraction of the ethanol solution is 80%. The complex enzyme includes cellulase, pectinase, and papain. The amount of cellulase used is 5% of the mass of Cistanche deserticola powder; The amount of pectinase used is 4% of the mass of Cistanche deserticola powder; The amount of papain used is 4% of the mass of Cistanche deserticola powder; The ultrasonic treatment was performed at a power of 450W, a temperature of 73℃, and a time of 100min.

2. The method for extracting Cistanche deserticola polysaccharides as described in claim 1, characterized in that, The Cistanche deserticola powder is the undersize material that has passed through a 60-100 mesh sieve.

3. The method for extracting Cistanche deserticola polysaccharides as described in claim 2, characterized in that, The Cistanche deserticola powder is the undersize material that has passed through an 80-mesh sieve.

4. The method for extracting Cistanche deserticola polysaccharides as described in claim 1, characterized in that, The enzyme inactivation was achieved by treating the enzyme in a boiling water bath for 10 minutes.

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