A method for improving the content of aloe polysaccharide in aloe products by using an electric field

Abstract

The application provides a method for improving the content of aloe polysaccharide in aloe products by using an electric field, and relates to the technical field of biological pharmacy. The method for improving the content of aloe polysaccharide in aloe products can control the content of biological macromolecular polysaccharide generated by polymerization reaction by adjusting appropriate current and electric field treatment time, thereby successfully improving the content of aloe polysaccharide in aloe vera gel by using an external electric field. Since aloe polysaccharide is the main active ingredient in aloe, its content is very important, therefore, the method for improving the content of aloe polysaccharide in aloe products has important significance for the development of the medicinal value of aloe polysaccharide products, and has important value for improving the polysaccharide yield and reducing the unit cost of polysaccharide.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of biological pharmacy, and particularly relates to a method for improving the content of aloe polysaccharide in an aloe product by using an electric field. BACKGROUND

[0002] Aloe polysaccharide is a main bioactive component of aloe gel and is mainly present in aloe fresh leaf juice. Aloe polysaccharide is a polymer from aloe. Aloe polysaccharide exhibits special bidirectional functions: inhibiting the activity of disease cells while promoting the activity of normal cells. Meanwhile, a large number of studies show that aloe polysaccharide has biological activities such as regulating blood sugar, resisting microorganisms, resisting tumors, regulating immunity, resisting viruses, and resisting oxidation.

[0003] Therefore, how to improve the content of aloe polysaccharide in aloe gel is a technical problem that needs to be solved in the field. SUMMARY

[0004] The application aims to provide a method for improving the content of aloe polysaccharide in an aloe product by using an electric field. To achieve the above-mentioned application purpose, the technical solution adopted by the application includes the following steps:

[0005] The treated aloe leaves are planted by using pure water;

[0006] The planted aloe leaves are peeled and pulped to obtain aloe gel;

[0007] The obtained aloe gel is placed in a reaction tank, the temperature of the aloe gel in the reaction tank is maintained at 4-12°C, and the aloe gel is stirred at the same time;

[0008] The positive and negative electrode plates are placed in the reaction tank, the aloe gel is treated by passing through a direct current, and the current is maintained between 0.5A and 1A, and the aloe product with improved aloe polysaccharide content is obtained after treatment.

[0009] In one or more embodiments of the application, in the step of taking the treated aloe leaves, the following steps are included: removing the margin part of the aloe tip and tail, and retaining the aloe leaves.

[0010] In one or more embodiments of the application, in the step of taking the treated aloe leaves, the following steps are included: removing the margin part of the aloe tip and tail, and retaining the aloe leaves.

[0011] In one or more embodiments of the application, the selected aloe is Aloe barbadensis Miller, and the growth time of the selected aloe is more than 10 months.

[0012] In one or more embodiments of the application, in the step of planting by using pure water, the planting time of the aloe by using pure water is 24 hours.

[0013] In one or more embodiments of the present application, in the step of peeling and pulping the cultivated aloe leaves to obtain aloe gel, the amount of gel pulping is not more than 40L each time, and the pulping time is 20-30 minutes.

[0014] 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, the reaction tank is internally provided with a circulating water jacket, and the circulating water jacket is connected to an external cold water circulating unit to set the circulating water temperature to 4-12℃.

[0015] In one or more embodiments of the present application, in the step of stirring the aloe gel, the stirring rate is configured to make the wave brought by stirring disappear just to the edge of the reaction tank.

[0016] In one or more embodiments of the present application, when the aloe gel is treated with direct current, the direct current treatment time is 3-12 hours.

[0017] In one or more embodiments of the present application, the electrodes of the positive and negative electrode plates are ruthenium-iridium-titanium electrodes.

[0018] Based on the above technical solution, the method for increasing aloe polysaccharide in aloe products provided by the present application has at least the following beneficial technical effects:

[0019] The method for increasing aloe polysaccharide in aloe products of the present application can control the content of biological macromolecular polysaccharide generated by polymerization reaction by adjusting the appropriate current and electric field treatment time, thereby successfully increasing the content of aloe polysaccharide in aloe vera gel. Since aloe polysaccharide is the main active ingredient in aloe, its content is crucial, therefore, the method for increasing aloe polysaccharide in aloe products of the present application has important significance for the development of the medicinal value of aloe polysaccharide products, and important value for increasing polysaccharide yield and reducing the unit cost of polysaccharide. BRIEF DESCRIPTION OF DRAWINGS

[0020] In order to more clearly illustrate the technical solutions in the embodiments of the present application, the following will briefly introduce the drawings needed to be used in the embodiments or prior art description. Obviously, the drawings in the following description are only some embodiments of the present application, and other drawings can be obtained by those skilled in the art without creating any inventive labor.

[0021] Figure 1 is the chromatogram of the sample provided by Comparative Example 1 of the present application.

[0022] Figure 2 is the chromatogram of the sample provided by Example 3 of the present application.

[0023] Figure 3 is a chromatogram of the sample provided in Example 1 of the present application. DETAILED DESCRIPTION

[0024] In order to make the technical problems to be solved, technical solutions and beneficial effects of the present application clearer, the present application will be further described in detail below in conjunction with embodiments. It should be understood that the specific embodiments described herein are only intended to explain the present application, and not to limit the present application.

[0025] In the present application, the term "and / or" describes the association relationship of the associated objects, which means that there can be three kinds of relationships, for example, A and / or B can mean that A exists alone, A and B exist together, and B exists alone. Wherein A and B can be singular or plural. The character " / " generally represents an "or" relationship between the associated objects before and after it.

[0026] In the present application, "at least one" means one or more, and "multiple" means two or more. "At least one" or similar expressions refer to any combination of these items, including any combination of single item or multiple items. For example, "at least one of a, b, or c", or "at least one of a, b, and c", can mean a, b, c, a-b (i.e. a and b), a-c, b-c, or a-b-c, wherein a, b, and c can be single or multiple.

[0027] It should be understood that in various embodiments of the present application, the size of the sequence number of the above processes does not mean the order of execution, and part or all of the steps can be executed in parallel or in sequence, and the execution order of each process should be determined according to its function and inherent logic, and should not constitute any limitation on the implementation process of the embodiments of the present application.

[0028] 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 the plural forms, unless the context clearly indicates otherwise.

[0029] The weight of the related components mentioned in the specification of the embodiments of the present application can not only refer to the specific content of each component, but also represent the weight ratio relationship between each component, therefore, as long as the content of the related components in the specification of the embodiments of the present application is enlarged or reduced in proportion, it is within the scope disclosed in the specification of the embodiments of the present application. Specifically, the mass mentioned in the specification of the embodiments of the present application can be µg, mg, g, kg and other mass units commonly known in the chemical field.

[0030] Currently, there is a growing interest in the use of external fields to develop advanced polymerization techniques. For example, temperature, light, electricity, magnetic field, ultrasound, etc. Since the chemical regulation of polymerization reactions will exist in foreign substances, it can lead to the accumulation of impurities, and even cause adverse side reactions. While the polymerization reaction technology regulated by external fields is a non-invasive technology. It has excellent and unique ability in process intensification and space and time control. The parameters of the external field can be adjusted to regulate the initiation and rate of polymerization reaction. Therefore, the polymerization reaction regulated by external field is a new polymerization technology. So far, there is no report on the use of electric field to regulate the polymerization reaction of aloe polysaccharide to improve the content of aloe polysaccharide.

[0031] Based on the above considerations, in order to solve the technical problem of improving the content of polysaccharide in aloe products. The embodiment of the present application provides a method for improving the content of aloe polysaccharide in aloe products by using electric field, which comprises the following steps:

[0032] (1) Remove the margin part of the tip and tail of aloe, and reserve the aloe leaves. Preferably, the length of the removed aloe tip is 15-20 cm. The selected aloe is Aloe barbadensis Miller, and the growth time of the selected aloe is more than 10 months.

[0033] (2) Take the treated aloe leaves and cultivate them with pure water. Preferably, the cultivation time of aloe with pure water is 24 hours.

[0034] (3) Peel and beat the cultivated aloe leaves to obtain aloe gel. Preferably, the beating amount of aloe gel is not more than 40 L each time, and the beating time of aloe gel is 20-30 minutes. It has been verified by experiments that if the beating time of aloe gel exceeds 30 minutes in this step, the viscosity of aloe gel will decrease, and the content of polysaccharide in aloe gel will finally decrease. Therefore, the beating time of aloe gel is set to be within 30 minutes.

[0035] (4) Put the obtained aloe gel into a reaction tank, maintain the temperature of aloe gel in the reaction tank at 4-12℃, and stir the aloe gel at the same time. Preferably, the volume of the reaction tank is 300 L, the diameter of the reaction tank is 85 cm, the depth of the reaction tank is 75 cm, the reaction tank is provided with a circulating water jacket, the circulating water jacket is connected with an external cold water circulating unit, and the circulating water temperature is set to be 4-12℃. The stirring rate is such that the corrugation caused by stirring disappears at the edge of the reaction tank. If the temperature of aloe gel is higher than 12℃, aloe gel is easy to degrade or grow microorganisms. The temperature of aloe gel is set to be 4-12℃, so as to maintain the quality of aloe gel.

[0036] (5) Put the positive and negative electrode plates in the reaction tank, and treat the aloe gel with direct current and keep the current between 0.5A and 1A. After the treatment, the aloe product with increased aloe polysaccharide content is obtained. Preferably, the direct current treatment time is 3-12 hours. Preferably, the electrode of the positive and negative electrode plates is a ruthenium-iridium-titanium electrode. When the direct current is too high, the temperature in the reaction tank will rise, which is difficult to control. When the current is lower than 0.5A, the reaction speed is slower.

[0037] The following will be described in conjunction with specific examples.

[0038] (1) Select aloe vera from the dry and hot valley zone, and the growth period of the aloe vera is more than 10 months.

[0039] (2) Cut off the front end of the aloe vera in step (1) by 15-20 cm, and remove the white part at the tail. The aloe vera leaves after the treatment are cultivated with pure water for 24 hours.

[0040] (3) The aloe vera leaves cultivated in step (2) are peeled and pulped. The single pulping amount in the pulping barrel is 40L, and the pulping time is 25 minutes. The comparative sample A0 without electric field treatment is obtained.

[0041] Example 1

[0042] The present example 1 provides a method for increasing the content of aloe polysaccharide, comprising the following steps:

[0043] (1) Select aloe vera from the dry and hot valley zone, and the growth period of the aloe vera is more than 10 months.

[0044] (2) Cut off the front end of the aloe vera in step (1) by 15-20 cm, and remove the white part at the tail. The aloe vera leaves after the treatment are cultivated with pure water for 24 hours.

[0045] (3) The aloe vera leaves cultivated in step (2) are peeled and pulped. The single pulping amount in the pulping barrel is 40L, and the pulping time is 25 minutes.

[0046] (4) Put the 200L aloe gel obtained in step (3) in the reaction tank. A cold water circulating unit is provided outside the reaction tank, and the temperature of the circulating water is 9℃.

[0047] (5) Set a stirrer at the opening of the reaction tank. The stirring rate is just right when the corrugation disappears at the edge of the pot, and the stirring rate is 90RPM.

[0048] (6) Put the 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, and continuously treat for 12 hours. The sample A12 of example 1 is obtained.

[0049] Example 2

[0050] (1) Selecting Aloe vera from dry and hot valley zone, and the growth time of Aloe vera is more than 10 months.

[0051] (2) Cutting the front end of Aloe vera in step (1) by 15-20 cm, and removing the white part at the tail. After the treatment, the Aloe vera leaves are cultivated with pure water for 24 hours.

[0052] (3) Peeling and beating the Aloe vera leaves cultivated in step (2). The single beating amount in the beating barrel is 40 L, and the beating time is 25 minutes.

[0053] (4) Placing 200 L of Aloe vera gel obtained in step (3) in a reaction tank. A cold water circulating unit is arranged outside the reaction tank, and the temperature of the circulating water is 9°C.

[0054] (5) Setting a stirrer at the pot opening of the light reaction tank. The stirring rate is that the corrugation just disappears to the pot edge, which is the appropriate rate, and the stirring rate is 90 RPM.

[0055] (6) Placing positive and negative electrode plates in the reaction tank and passing direct current. By adjusting the distance between the positive and negative electrode plates and the direct current voltage, the current is maintained between 0.5 A-1.0 A, and the continuous treatment is 3 hours. Sample A3 of Example 2 is obtained.

[0056] Example 3

[0057] (1) Selecting Aloe vera from dry and hot valley zone, and the growth time of Aloe vera is more than 10 months.

[0058] (2) Cutting the front end of Aloe vera in step (1) by 15-20 cm, and removing the white part at the tail. After the treatment, the Aloe vera leaves are cultivated with pure water for 24 hours.

[0059] (3) Peeling and beating the Aloe vera leaves cultivated in step (2). The single beating amount in the beating barrel is 40 L, and the beating time is 25 minutes.

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

[0061] (5) Setting a stirrer at the pot opening of the light reaction tank. The stirring rate is that the corrugation just disappears to the pot edge, which is the appropriate rate, and the stirring rate is 90 RPM.

[0062] (6) Placing positive and negative electrode plates in the reaction tank and passing direct current. By adjusting the distance between the positive and negative electrode plates and the direct current voltage, the current is maintained between 0.5 A-1.0 A, and the continuous treatment is 6 hours. Sample A6 of Example 3 is obtained.

[0063] Example 4

[0064] (1) Selecting Aloe vera from dry-hot valley zone, and the growth time of Aloe vera is more than 10 months.

[0065] (2) Cutting the front end of Aloe vera in step (1) by 15-20 cm, and removing the tail part. The Aloe vera leaves after treatment are cultivated in pure water for 24 hours.

[0066] (3) Peeling and pulping the Aloe vera leaves cultivated in step (2). The single pulping amount in the pulping barrel is 40 L, and the pulping time is 25 minutes.

[0067] (4) Placing the obtained 200 L Aloe vera gel in step (3) in a reaction tank. A cold water circulating unit is arranged outside the reaction tank. The temperature of the circulating water is 9°C.

[0068] (5) Arranging a stirrer at the pot opening of the light reaction tank. The stirring rate is just enough to make the corrugation disappear at the pot edge. That is, the appropriate rate is 90 RPM.

[0069] (6) Placing positive and negative electrode plates in the reaction tank and connecting them to a direct current. By adjusting the distance between the positive and negative electrode plates and the direct current voltage, the current is maintained between 0.5 A and 1.0 A, and the continuous treatment is 9 hours. The sample A9 of the example is obtained.

[0070] The samples of Comparative Example 1 and Examples 1-4 are tested:

[0071] The detection method is as follows:

[0072] (1) The detection method of Aloe vera polysaccharide content in Aloe vera gel is executed according to the polysaccharide detection method in the Light Industry Standard of the People's Republic of China QB / T 2489-2018.

[0073] (2) The dry weight of alcohol-sedimented polysaccharide is obtained by taking 100 ml of Aloe vera gel sample, adding 200 ml of 95% ethanol, standing for 12 hours, and then taking the precipitate part to dry in a 40°C oven and weighing.

[0074] (3) The sugar degree is detected by a refractometer.

[0075] Among them, the detection results of Aloe vera polysaccharide content in Aloe vera gel obtained by using and not using electric field treatment in Example 1 and Comparative Example 1 are shown in Table 1.

[0076] The detection results of Aloe vera polysaccharide content in Aloe vera gel obtained by Example 1 and Examples 2-4 are shown in Table 2. From the above Tables 1 and 2, it can be seen that:

[0077] (1) The polysaccharide content of the sample treated by electric field is significantly higher than that of the sample without electric field treatment.

[0078] (2) The polysaccharide content increases significantly with the increase of electric field treatment time, and the increase is fastest at the beginning of treatment, and slows down in the period of 9 hours to 12 hours.

[0079] (3) The dry weight of polysaccharide precipitated by alcohol increases significantly with the increase of electric field treatment time, and the increase is fastest at the beginning of treatment, and slows down in the period of 9 hours to 12 hours. The trend is consistent with the polysaccharide content result.

[0080] (4) The sugar content decreases significantly with the increase of electric field treatment time, because the monosaccharides in aloe gel polymerize to form polysaccharides under the action of electric field, resulting in a decrease in monosaccharide content and a decrease in sugar content.

[0081] Liquid chromatography-mass spectrometry analysis:

[0082] (1) Take 100 ml of aloe gel from samples A0, A6 and A12, respectively, and precipitate with alcohol of 95% concentration, and then freeze-dry the obtained polymer.

[0083] (2) Use different molecular weight dextran as standard, adopt high performance gel permeation chromatography (HPGPC), use high performance gel permeation chromatography column and differential detector for detection, and use GPC software to analyze the results. The logarithmic value of the relative molecular weight of the standard is used as the vertical coordinate, and the retention time of the corresponding chromatographic peak is used as the horizontal coordinate. The correction curve equation is drawn to determine the molecular weight and molecular weight distribution of polysaccharide.

[0084] (3) The experimental samples are shown in Table 3. (4) The standard substances are shown in Table 4. (5) The experimental instruments are shown in Table 5. (6) Experimental method.

[0085] a. Establishment of molecular weight correction curve.

[0086] Take 5 dextran control samples with known molecular weight, dissolve and dilute to prepare a solution containing 2 mg per 1 ml.

[0087] b. Preparation of test solution.

[0088] Take an appropriate amount of the product, accurately weigh and dissolve in water, and dilute to prepare a solution containing about 1 mg-2 mg per 1 ml, shake well, filter, and take the filtrate, i.e. the product is obtained.

[0089] Note: When the sample is not water-soluble, it is dissolved by ultrasonic or heating. The preparation information of the test sample solution is shown in Table 6. c. Chromatographic conditions.

[0090] Instrument: High performance liquid chromatograph.

[0091] Detector: Differential detector RID-20A.

[0092] Chromatographic column: TSK GPWXL chromatographic column (7.8 mm*300 mm, particle size 13 μm).

[0093] Mobile phase: water.

[0094] Flow rate: 0.6 ml / min.

[0095] Column temperature: 30°C.

[0096] Detector temperature: 4°C.

[0097] Injection volume: 20 μ.

[0098] (7) Experimental results.

[0099] The chromatograms of the test samples A0, A6 and A12 are shown in Figure 1 , Figure 2 , Figure 3 From the chromatograms, it can be seen that the polysaccharide peaks with a retention time of 15-20 min gradually and significantly increase from A0, A6 to A12 (the black box part). It can be seen that under the action of the electric field, the main polysaccharides that increase are those with a molecular weight less than D4, and the polysaccharides with a retention time of 15-20 min.

[0100] From the liquid chromatography-mass spectrometry detection of 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 number of monosaccharides decreases (the sugar degree decreases), and the amount of polysaccharides increases, because monosaccharides are synthesized into polysaccharides through polymerization. It is shown that the method can significantly increase the content of polysaccharides, and the content of polysaccharides can be controlled by adjusting the time of electric field treatment. The increased aloe polysaccharides are those with a molecular weight less than the D4 standard (weight average molecular weight 13.05 kDa).

[0101] The above only describes preferred embodiments of the present application and is not intended to limit the present application. Any modification, equivalent replacement and improvement made within the spirit and principle of the present application shall be included in the protection scope of the present application.

Claims

1. A method for increasing the content of aloe polysaccharide in an aloe product using an electric field, characterized by, The method comprises the following steps: The treated aloe leaves are planted in pure water; The planted aloe leaves are peeled and pulped to obtain aloe gel; The obtained aloe gel is placed in a reaction tank, the temperature of the aloe gel in the reaction tank is maintained at 4-12℃, and the aloe gel is stirred at the same time; The positive and negative electrode plates are placed in the reaction tank, and the aloe gel is treated by direct current, and the current is maintained at 0.5A-1A, and the aloe product with increased aloe polysaccharide content is obtained after treatment.

2. The method of claim 1, wherein, In the step of taking the treated aloe leaves, the tip and tail of the aloe leaves are removed, and the aloe leaves are reserved.

3. The method of claim 2, wherein, In the step of taking the treated aloe leaves, the length of the removed tip of the aloe leaves is 15-20cm.

4. The method according to any one of claims 1 to 3, characterized in that, The selected aloe is Aloe barbadensis Miller, and the growth time of the selected aloe is more than 10 months.

5. The method of claim 1, wherein, In the step of planting the aloe in pure water, the planting time of the aloe in pure water is 24 hours.

6. The method of claim 1, wherein, In the step of peeling and pulping the planted aloe leaves to obtain aloe gel, the pulping amount is not more than 40L each time, and the pulping time is 20-30 minutes.

7. The method of claim 1, wherein, The volume of the reaction tank is 300L, the diameter of the reaction tank is 85cm, the depth of the reaction tank is 75cm, the reaction tank is internally provided with a circulating water jacket, the circulating water jacket is connected with an external cold water circulating unit, and the circulating water temperature is set to 4-12℃.

8. The method of claim 7, wherein, In the step of stirring the aloe gel, the stirring rate is configured to make the wave brought by stirring disappear just to the edge of the reaction tank.

9. The method of claim 1, wherein, In the step of treating the aloe gel by direct current, the direct current treatment time is 3-12 hours.

10. The method of claim 1, wherein, The electrode of the positive and negative electrode plates is a ruthenium-iridium-titanium electrode.

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