Method for increasing aloe polysaccharide content in aloe product by utilizing electric field

Abstract

The invention provides a method for increasing the content of aloe polysaccharide in an aloe product by utilizing an electric field, and relates to the technical field of biological pharmacy. According to the method for increasing the content of the aloe polysaccharide in the aloe product, the content of biological macromolecular polysaccharide generated by polymerization reaction can be controlled by adjusting proper current and electric field treatment time, and the yield of the aloe polysaccharide in the aloe product is increased. The method for increasing the content of the aloe polysaccharide in the aloe product has the advantages that the aloe polysaccharide is a main active ingredient in the aloe, and the content of the aloe polysaccharide is crucial, so that the method for increasing the content of the aloe polysaccharide in the aloe product has important significance on development of the medicinal value of the aloe polysaccharide product, and the method is worthy of popularization and application. The method has important value for improving the polysaccharide yield and reducing the unit cost of the polysaccharide.

Description

Technical Field

[0001] The present invention relates to the field of biopharmaceutical technology, and in particular to a method for increasing the content of aloe polysaccharides in aloe products by using an electric field. Background Art

[0002] Aloe polysaccharide is the main bioactive component of aloe vera gel, and is mostly found in fresh aloe leaf juice. Aloe polysaccharide is a polymer derived from aloe vera. Aloe polysaccharide has a special two-way function: it inhibits the activity of diseased cells while promoting the activity of normal cells. At the same time, a large number of studies have shown that aloe polysaccharide has biological activities such as regulating blood sugar, anti-microbial, anti-tumor, regulating immunity, anti-virus, and anti-oxidation.

[0003] Therefore, how to increase the content of aloe polysaccharides in aloe gel is a technical problem that needs to be solved urgently in this field. Summary of the invention

[0004] The purpose of this application is to provide a method for increasing the content of aloe polysaccharides in aloe products by using an electric field. To achieve the above application purpose, the technical solution adopted by this application includes the following steps: Take the treated aloe leaves and cultivate them with pure water; The cultivated aloe leaves are peeled and pulped to obtain aloe gel; The obtained aloe vera gel is placed in a reaction tank, the temperature of the aloe vera gel in the reaction tank is maintained at 4-12° C., and the aloe vera gel is stirred; The positive and negative electrode plates are placed in the reaction tank, and the aloe gel is treated with direct current, and the current is maintained between 0.5A and 1A. After the treatment, an aloe product with increased aloe polysaccharide content is obtained.

[0005] In one or more embodiments of the present application, the step of taking the processed aloe leaves includes: removing the blank parts of the tip and tail of the aloe and retaining the aloe leaves.

[0006] In one or more embodiments of the present application, the step of taking the processed aloe leaves further includes: removing 15 to 20 cm of the tip of the aloe.

[0007] In one or more embodiments of the present application, the selected aloe vera is Aloe vera, and the growth period of the selected aloe vera is more than 10 months.

[0008] In one or more embodiments of the present application, in the step of cultivating with pure water, the cultivation time of aloe vera with pure water is 24 hours.

[0009] In one or more embodiments of the present application, the step of peeling and pulping the cultivated aloe leaves to obtain aloe gel also includes: the amount of gelling each time does not exceed 40L, and the pulping time is 20 to 30 minutes.

[0010] In one or more embodiments of the present application, the volume of the reaction tank is 300L, the diameter of the reaction tank is 85cm, the depth of the reaction tank is 75cm, a circulating water jacket is provided inside the reaction tank, and the circulating water jacket is connected to an external cold water circulation unit so that the circulating water temperature is set to 4~12°C.

[0011] In one or more embodiments of the present application, in the step of stirring the aloe vera gel, the stirring rate is configured to make the ripples caused by the stirring disappear just at the edge of the reaction tank.

[0012] In one or more embodiments of the present application, when the aloe vera gel is treated with direct current, the direct current treatment time is 3 to 12 hours.

[0013] In one or more embodiments of the present application, the electrodes of the positive and negative electrode plates are ruthenium iridium titanium electrodes.

[0014] Based on the above technical solution, the method for increasing aloe polysaccharides in aloe products provided in the present application has at least the following beneficial technical effects: The method for increasing aloe polysaccharides in aloe products of the present application can control the content of biomacromolecular polysaccharides produced by polymerization reaction by adjusting appropriate current and electric field treatment time, thereby successfully increasing the content of aloe polysaccharides in aloe vera gel by using an external electric field. Since aloe polysaccharides are the main active ingredients in aloe vera, their content is crucial. Therefore, the method for increasing aloe polysaccharides in aloe products of the present invention is of great significance to the development of the medicinal value of aloe polysaccharide products, and is of great value in increasing polysaccharide yield and reducing the unit cost of polysaccharides. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] In order to more clearly illustrate the technical solutions in the embodiments of the present application, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present application. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0016] Figure 1 It is the chromatogram of the sample provided in Comparative Example 1 of the present application.

[0017] Figure 2 This is the chromatogram of the sample provided in Example 3 of the present application.

[0018] Figure 3This is the chromatogram of the sample provided in Example 1 of the present application. DETAILED DESCRIPTION

[0019] In order to make the technical problems, technical solutions and beneficial effects to be solved by the present application more clearly understood, the present application is further described in detail below in conjunction with the embodiments. It should be understood that the specific embodiments described herein are only used to explain the present application and are not used to limit the present application.

[0020] In this application, the term "and / or" describes the association relationship of associated objects, indicating that there may be three relationships. For example, A and / or B can mean: A exists alone, A and B exist at the same time, and B exists alone. A and B can be singular or plural. The character " / " generally indicates that the associated objects are in an "or" relationship.

[0021] In this application, "at least one" means one or more, and "plurality" means two or more. "At least one of the following" or similar expressions refers to any combination of these items, including any combination of single items or plural items. For example, "at least one of a, b, or c", or "at least one of a, b, and c" can all mean: a, b, c, ab (i.e., a and b), ac, bc, or abc, where a, b, and c can be single or multiple.

[0022] It should be understood that in the various embodiments of the present application, the size of the serial numbers of the above-mentioned processes does not mean the order of execution, some or all of the steps can be executed in parallel or sequentially, and the execution order of each process should be determined by its function and internal logic, and should not constitute any limitation on the implementation process of the embodiments of the present application.

[0023] The terms used in the embodiments of the present application are only for the purpose of describing specific embodiments, and are not intended to limit the present application. The singular forms "a", "said" and "the" used in the embodiments of the present application and the appended claims are also intended to include plural forms, unless the context clearly indicates other meanings.

[0024] The weight of the relevant components mentioned in the embodiment description of the present application can not only refer to the specific content of each component, but also indicate the proportional relationship between the weights of the components. Therefore, as long as the content of the relevant components is proportionally enlarged or reduced according to the embodiment description of the present application, it is within the scope disclosed in the embodiment description of the present application. Specifically, the mass described in the embodiment description of the present application can be a mass unit known in the chemical industry such as µg, mg, g, kg, etc.

[0025] At present, people are increasingly interested in developing advanced polymerization technologies using external fields. Such as: temperature, light, electricity, magnetic field, ultrasound, etc. Since chemically regulated polymerization reactions may contain foreign substances, they may lead to the accumulation of impurities and even adverse side reactions. The polymerization reaction technology regulated by external fields is a non-invasive technology. It has excellent and unique capabilities in process intensification and spatial and temporal regulation. The parameters of the external field can be adjusted to adjust the start and rate of the polymerization reaction. Therefore, polymerization reactions through external field regulation are a new polymerization technology. So far, there has been no report on the technology of using external fields such as electric fields to regulate the polymerization reaction of aloe polysaccharides to increase the content of aloe polysaccharides.

[0026] Based on the above considerations, in order to solve the technical problem of increasing the polysaccharide content in aloe products, the present application embodiment provides a method for increasing the aloe polysaccharide content in aloe products by using an electric field, comprising the following steps: (1) Remove the tip and the blank part of the aloe vera, and keep the aloe vera leaves. Preferably, the length of the aloe vera tip removed is 15 to 20 cm. The selected aloe vera is Aloe vera, and the growth period of the selected aloe vera is more than 10 months.

[0027] (2) Take the treated aloe leaves and cultivate them with pure water. Preferably, the cultivation time of the aloe with pure water is 24 hours.

[0028] (3) Peeling and pulping the cultivated aloe leaves to obtain aloe gel. Preferably, the amount of glue applied each time does not exceed 40 L, and the glue application time is 20 to 30 minutes. Experimental verification shows that if the glue application time exceeds 30 minutes in this step, the viscosity of the aloe gel will decrease, and the polysaccharide content in the final test aloe gel will decrease. Therefore, the glue application time in the present invention needs to be within 30 minutes.

[0029] (4) The obtained aloe vera gel is placed in a reaction tank, and the temperature of the aloe vera gel in the reaction tank is maintained at 4-12°C, while the aloe vera gel is stirred. Preferably, the volume of the reaction tank is 300L, the diameter of the reaction tank is 85cm, the depth of the reaction tank is 75cm, and a circulating water jacket is provided inside the reaction tank, which is connected to an external cold water circulation unit so that the circulating water temperature is set to 4-12°C. The stirring rate is such that the ripples caused by the stirring disappear just at the edge of the reaction tank. If the temperature of the aloe vera gel is higher than 12°C, the aloe vera gel is easily degraded or microorganisms grow. Keeping the temperature of the aloe vera gel at 4-12°C can maintain the quality of the aloe vera gel.

[0030] (5) The positive and negative electrode plates are placed in the reaction tank, and the aloe gel is treated with direct current and the current is maintained between 0.5A and 1A, and an aloe product with increased aloe polysaccharide content is obtained after the treatment. Preferably, the direct current treatment time is 3 to 12 hours. Preferably, the electrodes of the positive and negative electrode plates are ruthenium iridium titanium electrodes. When the direct current is too high, the temperature in the reaction tank will increase and it will be difficult to control. When the current is lower than 0.5A, the reaction speed is slow.

[0031] The following describes the invention in conjunction with specific embodiments.

[0032] (1) Select Aloe vera grown in dry and hot river valleys, and the growth period of the Aloe vera is more than 10 months.

[0033] (2) Cut off 15-20 cm from the front end of the aloe vera obtained in step (1), remove the blank part at the tail, and incubate the treated aloe vera leaves in pure water for 24 hours.

[0034] (3) Peel and pulp the aloe vera leaves cultivated in step (2). The amount of glue in the pulping barrel is 40 L per time, and the glue-making time is 25 minutes. A comparative sample A0 without electric field treatment is obtained.

[0035] Example 1

[0036] This embodiment 1 provides a method for increasing the content of aloe polysaccharides, comprising the following steps: (1) Select Aloe vera grown in dry and hot river valleys, and the growth period of the Aloe vera is more than 10 months.

[0037] (2) Cut off 15-20 cm from the front end of the aloe vera obtained in step (1), remove the blank part at the tail, and incubate the treated aloe vera leaves in pure water for 24 hours.

[0038] (3) Peel and pulp the aloe vera leaves cultivated in step (2). The amount of glue in the pulping barrel is 40L per time, and the glueing time is 25 minutes.

[0039] (4) Place 200L of aloe vera gel obtained in step (3) in a reaction tank. The reaction tank is equipped with a cold water circulation unit, and the temperature of the circulating water is 9°C.

[0040] (5) Install a stirrer at the mouth of the reaction tank. The stirring rate is appropriate when the ripples just disappear at the edge of the pot. The stirring rate is 90 RPM.

[0041] (6) Place positive and negative electrode plates in the reaction tank and pass direct current. Adjust the distance between the positive and negative electrode plates and the direct current voltage to keep the current between 0.5A and 1.0A. Continue the process for 12 hours to obtain sample A12 of Example 1.

[0042] Example 2

[0043] (1) Select Aloe vera grown in dry and hot river valleys, and the growth period of the Aloe vera is more than 10 months.

[0044] (2) Cut off 15-20 cm from the front end of the aloe vera obtained in step (1), remove the blank part at the tail, and incubate the treated aloe vera leaves in pure water for 24 hours.

[0045] (3) Peel and pulp the aloe vera leaves cultivated in step (2). The amount of glue in the pulping barrel is 40L per time, and the glueing time is 25 minutes.

[0046] (4) Place 200L of aloe vera gel obtained in step (3) in a reaction tank. The reaction tank is equipped with a cold water circulation unit, and the temperature of the circulating water is 9°C.

[0047] (5) Set a stirrer at the mouth of the photoreaction tank. The stirring rate is 90 RPM when the ripples just disappear at the edge of the pot.

[0048] (6) Place positive and negative electrode plates in the reaction tank and pass direct current. Adjust the distance between the positive and negative electrode plates and the direct current voltage to keep the current between 0.5A and 1.0A. Continue the process for 3 hours to obtain sample A3 of Example 2.

[0049] Example 3

[0050] (1) Select Aloe vera grown in dry and hot river valleys, and the growth period of the Aloe vera is more than 10 months.

[0051] (2) Cut off 15-20 cm from the front end of the aloe vera obtained in step (1), remove the blank part at the tail, and cultivate it in pure water for 24 hours after the treatment is completed.

[0052] (3) Peel and pulp the aloe vera leaves cultivated in step (2). The amount of glue in the pulping barrel is 40L per time, and the glueing time is 25 minutes.

[0053] (4) Place 200L of aloe vera gel obtained in step (3) in a reaction tank. A cold water circulation unit is installed outside the reaction tank. The temperature of the circulating water is 9°C.

[0054] (5) Set a stirrer at the mouth of the photoreaction tank. The stirring rate is such that the ripples just disappear at the edge of the pot. This is the appropriate rate, and the stirring rate is 90 RPM.

[0055] (6) Place positive and negative electrode plates in the reaction tank and pass direct current. Adjust the distance between the positive and negative electrode plates and the direct current voltage to keep the current between 0.5A and 1.0A. Continue the process for 6 hours to obtain sample A6 of Example 3.

[0056] Example 4

[0057] (1) Select Aloe vera grown in dry and hot river valleys, and the growth period of the Aloe vera is more than 10 months.

[0058] (2) Cut off 15-20 cm from the front end of the aloe vera obtained in step (1), remove the blank part at the tail, and incubate the treated aloe vera leaves in pure water for 24 hours.

[0059] (3) Peel and pulp the aloe vera leaves cultivated in step (2). The amount of glue in the pulping barrel is 40L per time, and the glueing time is 25 minutes.

[0060] (4) Place 200L of aloe vera gel obtained in step (3) in a reaction tank. A cold water circulation unit is installed outside the reaction tank. The temperature of the circulating water is 9°C.

[0061] (5) Set a stirrer at the mouth of the photoreaction tank. The stirring rate is such that the ripples just disappear at the edge of the pot. This is the appropriate rate, and the stirring rate is 90 RPM.

[0062] (6) Place positive and negative electrode plates in the reaction tank and apply direct current. Adjust the distance between the positive and negative electrode plates and the direct current voltage to keep the current between 0.5A and 1.0A. Continue the process for 9 hours. Obtain Example Sample A9.

[0063] The samples of Comparative Example 1 and Examples 1-4 were tested: The detection method is as follows: (1) The detection method for the content of aloe polysaccharides in aloe vera gel is based on the polysaccharide detection method in the light industry standard QB / T2489-2018 of the People's Republic of China.

[0064] (2) The dry weight of alcohol-precipitated polysaccharides was obtained by taking 100 ml of aloe vera gel sample, adding 200 ml of 95% ethanol, and letting it stand for 12 hours. The precipitated part was dried in an oven at 40°C and then weighed.

[0065] (3) Sugar content is measured using a refractometer.

[0066] The test results of the aloe polysaccharide content in the aloe gels obtained by electric field treatment and without electric field treatment in Example 1 and Comparative Example 1 are shown in Table 1.

[0067] The test results of the aloe polysaccharide content in the aloe gel obtained in Example 1 and Examples 2-4 are shown in Table 2. From Table 1 and Table 2 above, we can see that: (1) The content of aloe polysaccharides in samples treated with electric field was significantly increased compared with that in samples not treated with electric field.

[0068] (2) As the electric field treatment time increases, the polysaccharide content increases significantly, with the fastest increase in the initial stage of treatment and then slowing down in the period from 9 to 12 hours.

[0069] (3) As the electric field treatment time increases, the dry weight of polysaccharides precipitated from alcohol increases significantly, with the fastest increase in the initial treatment period and a slowdown in the 9- to 12-hour period, which is consistent with the trend of the polysaccharide content results.

[0070] (4) As the electric field treatment time increases, the sugar content decreases significantly. This is because the monosaccharides in the aloe vera gel polymerize to form polysaccharides under the action of the electric field, thereby reducing the monosaccharide content and causing the sugar content to decrease.

[0071] Liquid chromatography mass spectrometry analysis: (1) Take 100 ml of aloe vera gel from samples A0, A6, and A12 respectively, precipitate with 95% alcohol, and then freeze-dry the resulting polymer.

[0072] (2) Dextran with different molecular weights was used as standard, and high performance gel permeation chromatography (HPGPC) was used for detection using a high performance gel permeation chromatography column and a differential detector. The results were analyzed using GPC software. The logarithm of the relative molecular mass of the standard was used as the ordinate, and the retention time of the corresponding chromatographic peak was used as the abscissa to develop a calibration curve equation for the determination of the molecular weight and molecular weight distribution of the polysaccharide.

[0073] (3) Experimental samples are shown in Table 3. (4) Standard substances are shown in Table 4. (5) Experimental instruments are shown in Table 5. (6) Experimental methods

[0074] a. Establishment of molecular weight calibration curve.

[0075] Take 5 dextran reference substances of known molecular weight, dissolve them in water and dilute them to make a solution containing 2 mg of each substance per 1 ml.

[0076] b. Preparation of test solution.

[0077] Take an appropriate amount of this product, weigh it accurately, add water to dissolve and dilute to make a solution containing about 1mg~2mg per 1ml, shake well, filter, and take the filtrate to obtain.

[0078] Note: When the sample is poorly soluble in water, dissolve it by ultrasound or heating. See Table 6 for the preparation of the test solution. c. Chromatographic conditions.

[0079] Instrument: High performance liquid chromatography.

[0080] Detector: Differential detector RID-20A.

[0081] Chromatographic column: TSK GPWXL chromatographic column (7.8mm*300mm, particle size 13μm).

[0082] Mobile phase: water.

[0083] Flow rate: 0.6 ml / min.

[0084] Column temperature: 30℃.

[0085] Detector temperature: 4°C.

[0086] Injection volume: 20μ.

[0087] (7) Experimental results.

[0088] The chromatograms of test samples A0, A6, and A12 are as follows: Figure 1 , Figure 2 , Figure 3 As shown in the chromatogram, the polysaccharide peaks between the retention time of 15-20min gradually increase significantly from A0, A6, and A12 (the thick black frame part). It can be seen that under the action of the electric field, the main increased polysaccharides are those with a molecular weight less than D4 and the retention time between 15-20min.

[0089] From the liquid phase mass spectrometry detection of the above Example 1 (sample A12), Comparative Example 1 (sample A0), and Example 3 (sample A6), it can be concluded that under the action of the electric field, the amount of monosaccharides decreases (the sugar content decreases), and the amount of polysaccharides increases. This is because monosaccharides are synthesized into polysaccharides through polymerization reactions, indicating that this method can significantly increase the polysaccharide content, and the polysaccharide content can be regulated by regulating the time of electric field treatment. The increased aloe polysaccharide is the part with a molecular weight less than the D4 standard (weight average molecular weight 13.05kDa).

[0090] The above description is only a preferred embodiment of the present application and is not intended to limit the present application. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present application should be included in the protection scope of the present application.

Claims

1. A method for increasing the content of aloe polysaccharides in aloe products by using an electric field, characterized in that: The steps include: Take the treated aloe leaves and cultivate them with pure water; The cultivated aloe leaves are peeled and pulped to obtain aloe gel; The obtained aloe vera gel is placed in a reaction tank, the temperature of the aloe vera gel in the reaction tank is maintained at 4-12° C., and the aloe vera gel is stirred; The positive and negative electrode plates are placed in the reaction tank, and the aloe gel is treated with direct current, and the current is maintained between 0.5A and 1A. After the treatment, an aloe product with increased aloe polysaccharide content is obtained.

2. The method according to claim 1, characterized in that The step of taking the processed aloe leaves includes: removing the blank parts at the tip and tail of the aloe and retaining the aloe leaves.

3. The method according to claim 2, characterized in that The step of taking the processed aloe leaves also includes: removing 15 to 20 cm of the tip of the aloe.

4. The method according to any one of claims 1 to 3, characterized in that: The selected aloe vera is Aloe vera var. Curaçao, and the growth period of the selected aloe vera is more than 10 months.

5. The method according to claim 1, characterized in that In the step of cultivating with pure water, the cultivation time of aloe vera with pure water is 24 hours.

6. The method according to claim 1, characterized in that The step of peeling and pulping the cultivated aloe leaves to obtain aloe gel also includes: the amount of gelling each time does not exceed 40L, and the pulping time is 20 to 30 minutes.

7. The method according to claim 1, characterized in that The volume of the reaction tank is 300L, the diameter of the reaction tank is 85cm, the depth of the reaction tank is 75cm, a circulating water jacket is provided inside the reaction tank, and the circulating water jacket is connected to an external cold water circulation unit so that the circulating water temperature is set to 4~12°C.

8. The method according to claim 7, characterized in that In the step of stirring the aloe vera gel, the stirring rate is configured so that the ripples generated by the stirring disappear just at the edge of the reaction tank.

9. The method according to claim 1, characterized in that: When the aloe vera gel is treated with direct current, the direct current treatment time is 3 to 12 hours.

10. The method according to claim 1, characterized in that The electrodes of the positive and negative electrode plates are ruthenium, iridium and titanium electrodes.

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