Efficient ionizing radiation extraction process of polygonatum polysaccharide

Abstract

The invention provides an efficient ionizing radiation extraction process of polygonatum polysaccharide, and belongs to the field of extraction processes of main functional components of traditional Chinese medicinal materials. The rhizoma polygonati is cleaned, drained, cut into slices, freeze-dried, crushed and sieved. The preparation method comprises the following steps: mixing the raw materials, uniformly stirring, then subpackaging in PE bags, sealing, carrying out electron beam radiation extraction, selecting a dose rate of 20kGy / pass, and a radiation absorbed dose of 0-320kGy. Water with the solid-to-liquid ratio of 1: 10-1: 30 is added into the irradiated rhizoma polygonati powder, reflux extraction is carried out 1-5 times and 1-3 h each time, extraction is carried out at the extraction temperature of 70-110 DEG C, crude rhizoma polygonati polysaccharide obtained through extraction is separated and purified through a DEAE-52 chromatographic column, and the refined rhizoma polygonati polysaccharide is obtained. Through the process, the extraction rate of the polygonatum polysaccharide extraction process can be effectively improved.

Description

Technical Field

[0001] The invention belongs to the field of extraction of main functional components of traditional Chinese medicine, and relates to a high-efficiency ionizing radiation extraction process of polygonatum sibiricum polysaccharide, a main functional component in the natural plant polygonatum, and particularly to a new method for extracting polygonatum sibiricum polysaccharide, a main functional component in polygonatum sibiricum, by using ionizing radiation technology as an extraction process, which is environmentally friendly and high-efficiency. Background Art

[0002] Polygonatum sibiricum is a traditional Chinese medicine in my country. It is both a medicine and food. Its rhizome is used as a medicinal part. It has the effects of nourishing qi and yin, moistening the lungs, strengthening the spleen, and benefiting the kidneys. It has been used in my country for hundreds of years. People in southern my country like to cook Polygonatum sibiricum with meat and eat it as a health food. In clinical Chinese medicine, it is often used to treat symptoms such as weak spleen and stomach, fatigue and weakness. There are currently three known species of Polygonatum sibiricum, namely, Polygonatum sibiricum, Polygonatum yunnanensis, and Polygonatum multiflorum.

[0003] Polygonatum has pharmacological effects such as enhancing immune system function, delaying aging, lowering blood sugar and blood lipids, antibacterial and anti-inflammatory, antiviral and anti-tumor. Polygonatum contains a variety of active ingredients that are beneficial to the human body, such as polysaccharides, saponins, lignans, cardiac glycosides, alkaloids, vitamins, and anthraquinones. Polygonatum polysaccharide, one of the main medicinal ingredients in Polygonatum, generally has a content of more than 10.00% in Polygonatum. Due to its high content and wide range of effects, it is considered to be the most important chemical component in Polygonatum that exerts its pharmacological effects. Polygonatum polysaccharide has good medicinal effects in antibacterial and anti-inflammatory, antioxidant, hypoglycemic, anti-tumor and immune regulation. Therefore, how to efficiently extract Polygonatum polysaccharide from Polygonatum is the key process control point.

[0004] The extraction of polygonatum polysaccharide is generally carried out in three steps: removing impurities, breaking cell walls and extracting polysaccharide. Polygonatum polysaccharide is mostly concentrated in the root of polygonatum, and the pharmacopoeia also stipulates that the root can be used as medicine, so the root of polygonatum needs to be washed, sliced, dried and crushed before it can be used for extraction.

[0005] Polygonatum cells have a cell wall that maintains the cell structure. It is mainly composed of pectin, cellulose and hemicellulose, which can effectively protect the homeostatic environment in the cell. To release polysaccharides from the cytoplasm, the cell wall must be broken. Different methods of breaking the cell wall determine different extraction methods.

[0006] Chinese patent CN104059157A provides an ultrasonic extraction process of polygonatum polysaccharide, selects suitable polygonatum rhizome, screens and cleans, and then puts it into a pulverizer to crush it into particles; puts the polygonatum particles into a closed container, evacuates, and then puts the container containing the polygonatum particles into a heating device for heating; takes out the heated polygonatum particles, cools to room temperature, and then sends the heated polygonatum particles into a grinder to grind into powder; puts the polygonatum powder into an oven, sets the time to 30 seconds, and the temperature to 110°C for baking; puts the baked polygonatum powder into 40-60°C purified water and soaks it for 30 minutes, acts under the condition of ultrasonic power of 440W / L for 5-10min, ultrasonically extracts 3 times, filters the filtrate, concentrates it, adds ethanol, stands for 24 hours, removes the supernatant, decompresses and filters, and dries to obtain polygonatum polysaccharide. However, the operation process of this method is too complicated, and the container requirements are too high, which is not conducive to mass production.

[0007] Chinese patent CN109354630A provides a high-temperature extraction method of polygonatum polysaccharide, comprising the following steps: (1) extracting a crude polysaccharide extract: vacuum drying polygonatum, crushing, mixing with water, leaching at 70-80°C for 30-40min, then ultrasonic extraction, centrifugation, rotary evaporation of the supernatant obtained by centrifugation, then adding an ethanol solution and stirring, standing, filtering, and freeze-drying the obtained precipitate to obtain a crude polysaccharide extract; (2) extraction: adding the crude polysaccharide extract to a chloroform-n-butanol mixed solution according to a material ratio of 1g:10mL, repeating the extraction 3 times, combining the aqueous phases obtained by the 3 extractions, and then adding an ethanol solution to the aqueous phase until the mass concentration of ethanol is 80%, standing for 12-24h, filtering, washing the obtained solid, and drying to obtain polygonatum polysaccharide. However, the extraction rate of polygonatum polysaccharide obtained by this method is not high, and the industrial production efficiency is too low.

[0008] Chinese patent CN105418782A provides a microwave-assisted extraction method for polygonatum polysaccharide, comprising the following steps: (1) raw material preparation; (2) crushing; (3) defatting; (4) microwave-assisted extraction; (5) concentration; (6) protein removal and decolorization; (7) preparation of polygonatum polysaccharide. Although this patent uses microwave-assisted extraction technology to extract polygonatum polysaccharide from ultrafine polygonatum particles, the extraction mechanism is unclear and it is difficult to scale up to industrial production levels.

[0009] Industrial methods of destroying the cell wall of Polygonatum sibiricum include enzymolysis, high temperature, acid and alkali, ultrasound, microwave, etc. Among them, although enzymolysis technology helps to improve the extraction rate and protect the active ingredients of drugs in the process of traditional Chinese medicine extraction, the disadvantages of enzymolysis technology in the process of traditional Chinese medicine extraction are also prominent: first, the enzyme has very strict requirements on temperature and pH value during the reaction. However, at this stage, due to the limitations of science and technology, the reaction equipment still has certain deficiencies in ensuring the reaction conditions; second, although enzymolysis technology can well ensure the extraction efficiency and medicinal properties of the active ingredients of drugs in the process of traditional Chinese medicine extraction, there are also some problems. These problems are mainly manifested in that enzymolysis technology can cause chemical changes in some components of the drug, destroying the original structure of the drug components. The destruction of the structure of the active ingredients greatly reduces the purity of the active ingredients of the drug. High-temperature extraction is simple and easy to operate, but there are too many impurities, which is not conducive to precise control in a certain molecular weight area. The advantage of the acid-base method is that the acid-base medium can significantly improve the extraction rate of polysaccharides. The use of dilute alkali solution for extraction can save time and reduce the consumption of raw materials and reagents. And the extracted polysaccharide content is high. However, the disadvantages are also obvious: when acid is used as a medium, it has a certain destructive effect on glycosidic bonds, reduces the yield of polysaccharides, and also corrodes the container. It is not suitable for use except for weak acid; the concentration of the solution extracted by alkaline method increases, making filtration difficult. The principle of ultrasonic extraction is ultrasonic cavitation, which accelerates the leaching of plant active ingredients. The advantages are short extraction time and simple extraction method, and it can also effectively avoid the destruction of active ingredients by high temperature and high pressure. However, the disadvantages are also obvious: high requirements for the container, and there are problems with noise and equipment amplification.

[0010] In order to overcome the problem that the extraction of polygonatum polysaccharide is difficult to industrialize and to achieve efficient extraction of polygonatum polysaccharide, an efficient and rapid ionizing radiation extraction process of polygonatum polysaccharide has emerged. In the ionizing radiation extraction process, the reaction process does not require the addition of any substances toxic to the human body, is pollution-free, has mild reaction conditions, can be carried out at room temperature, and has high industrial production efficiency, extremely low cost, and good production controllability.

[0011] The problem to be solved by the present invention is to disclose a high-efficiency, environmentally friendly, pollution-free and extremely low-cost efficient and rapid ionizing radiation extraction process for extracting polygonatum polysaccharide, the main functional component in polygonatum, and its application, so as to overcome the above-mentioned defects of the prior art.

[0012] After searching, there is no patent application in China that is identical to the present invention. Summary of the invention

[0013] The purpose of the present invention is to provide a high-efficiency ionizing radiation extraction process for polygonatum polysaccharide in view of various problems existing in the prior art. The technical problem to be solved by the present invention is how to efficiently extract polygonatum polysaccharide, the main functional component in polygonatum.

[0014] The purpose of the present invention can be achieved by the following technical scheme: a highly efficient ionizing radiation extraction process of polygonatum polysaccharide, characterized in that the polygonatum is irradiated by electron beam technology to break the cell wall of polygonatum cells that maintains the cell structure, thereby releasing polysaccharide from the cytoplasm, thereby achieving the purpose of improving the extraction rate of polygonatum polysaccharide. A irradiation extraction process of polygonatum polysaccharide and its application, comprising the following steps:

[0015] ① Weigh a certain amount of Polygonatum sibiricum, wash and drain, cut into thin slices, and chop the fibrous root samples.

[0016] Freeze-dry for 1-2 days, crush and sieve to obtain polygonatum powder.

[0017] ② Take the crushed and sieved Polygonatum sibiricum powder, pack it into PE bags and seal them, and perform electron beam radiation extraction. The selected dose rate is 20kGy / pass and the radiation absorption dose is 0-320kGy.

[0018] ③ Add water with a solid-liquid ratio of 1:10-1:30 to the irradiated polygonatum powder, reflux extract 1-5 times, each time for 1-3 hours, the extraction temperature is 70-110°C, and filter.

[0019] ④ The filtrate was concentrated under reduced pressure, and after concentration, anhydrous ethanol solution was slowly added and refrigerated overnight.

[0020] ⑤ Centrifuge, discard the supernatant, wash the precipitate with 90% ethanol three times, freeze-dry,

[0021] Obtain crude polygonatum polysaccharide.

[0022] ⑥ The obtained crude polygonatum polysaccharide is separated and purified using a DEAE-52 chromatography column, and then concentrated, ethanol precipitated, centrifuged, freeze-dried, and dried to obtain polygonatum polysaccharide.

[0023] Furthermore, the raw material of Polygonatum sibiricum is crushed and sieved using 30 mesh before irradiation to increase the surface area of ​​Polygonatum sibiricum, thereby increasing its radiation contact surface and the contact area with water during extraction, which is beneficial to the dissolution of the effective ingredient Polygonatum sibiricum polysaccharide; at the same time, it also avoids excessive crushing that is not conducive to subsequent solid-liquid separation.

[0024] Furthermore, the ground and sieved polygonatum powder is put into a bag and then irradiated with an electron beam of 80-320 kGy to extract polygonatum polysaccharide.

[0025] Further, the filtrate was concentrated under reduced pressure at 60°C, and after concentration, 4 times the volume of anhydrous ethanol solution was slowly added, and refrigerated at 4°C overnight.

[0026] In order to improve the current situation of low extraction rate of Polygonatum sibiricum polysaccharide extraction process, we proposed an efficient and rapid ionizing radiation extraction process of Polygonatum sibiricum polysaccharide, which can effectively improve the extraction rate of Polygonatum sibiricum polysaccharide extraction process.

[0027] Compared with the prior art, this method has the following advantages:

[0028] 1. This method is easy to operate, and the reaction process does not have high requirements for temperature and pH value.

[0029] 2. The extraction mechanism is clear. The ionizing radiation extraction process is not likely to cause chemical changes in some components of the drug. Instead, under the radiation of the electron accelerator, the cell walls of the Polygonatum sibiricum tissue are destroyed, releasing more Polygonatum sibiricum polysaccharides, thereby greatly increasing the yield of polysaccharides.

[0030] 3. It has low requirements on production equipment, will not cause corrosion to the container, and is not easy to cause filtering difficulties.

[0031] 4. The extraction and sterilization process of polygonatum polysaccharide can be completed simultaneously, saving costs. From a comprehensive and long-term perspective, the economic cost of extracting polygonatum polysaccharide using irradiation extraction technology is lower. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] Figure 1 The effect of solid-liquid ratio on the extraction rate of Polygonatum sibiricum polysaccharide.

[0033] Figure 2 The effect of extraction temperature on the extraction rate of Polygonatum sibiricum polysaccharide.

[0034] Figure 3 The effect of extraction time on the extraction rate of Polygonatum sibiricum polysaccharide.

[0035] Figure 4 The influence of extraction times on the extraction rate of Polygonatum sibiricum polysaccharide.

[0036] Figure 5 The effect of radiation absorption dose on the extraction rate of Polygonatum sibiricum polysaccharide.

[0037] Figure 6 This is a response surface experiment on radiation extraction of Polygonatum sibiricum polysaccharides. DETAILED DESCRIPTION

[0038] The following are specific embodiments of the present invention, which further describe the technical solutions of the present invention, but the present invention is not limited to these embodiments.

[0039] Example 1: Weigh 500g of Polygonatum sibiricum, wash and drain, cut into thin slices, cut the fibrous root sample into pieces, freeze-dry for 1-2 days, crush, and pass through a 30-mesh sieve to obtain Polygonatum sibiricum powder. Take 10g of the crushed and sieved Polygonatum sibiricum powder, pack it in PE bags and seal it, and irradiate it with electron beam. The irradiation dose is 80kGy and the dose rate is 20kGy / pass. The irradiated Polygonatum sibiricum powder is added with 10 times water and refluxed and extracted once, each time for 1h, and filtered. The filtrate is concentrated under reduced pressure at 60℃, and after concentration, 4 times the volume of anhydrous ethanol solution is slowly added, and refrigerated at 4℃ overnight. Centrifuge, discard the supernatant, wash the precipitate with 90% ethanol 3 times, and freeze-dry to obtain Polygonatum sibiricum crude polysaccharide. The obtained Polygonatum sibiricum crude polysaccharide is added to distilled water to prepare a crude polysaccharide solution, which is separated and purified using a DEAE-52 chromatography column, and then concentrated, precipitated with alcohol, centrifuged, freeze-dried, and dried to obtain Polygonatum sibiricum polysaccharide.

[0040] Example 2: Weigh 500g of Polygonatum sibiricum, wash and drain, cut into thin slices, cut the fibrous root sample into pieces, freeze-dry for 1-2 days, crush, and pass through a 30-mesh sieve to obtain Polygonatum sibiricum powder. Take 10g of the crushed and sieved Polygonatum sibiricum powder, pack it in PE bags and seal it, and irradiate it with electron beam. The irradiation dose is 240kGy and the dose rate is 20kGy / pass. The irradiated Polygonatum sibiricum powder is added with 20 times of water and refluxed and extracted twice, each time for 1.5h, and filtered. The filtrate is concentrated under reduced pressure at 60℃, and after concentration, 4 times the volume of anhydrous ethanol solution is slowly added, and refrigerated at 4℃ overnight. Centrifuge, discard the supernatant, wash the precipitate with 90% ethanol 3 times, and freeze-dry to obtain Polygonatum sibiricum crude polysaccharide. The obtained Polygonatum sibiricum crude polysaccharide is added to distilled water to prepare a crude polysaccharide solution, which is separated and purified using a DEAE-52 chromatography column, and then concentrated, precipitated with alcohol, centrifuged, freeze-dried, and dried to obtain Polygonatum sibiricum polysaccharide.

[0041] Example 3: Weigh 500g of Polygonatum sibiricum, wash and drain, cut into thin slices, cut the fibrous root sample into pieces, freeze-dry for 1-2 days, crush, and pass through a 30-mesh sieve to obtain Polygonatum sibiricum powder. Take 10g of the crushed and sieved Polygonatum sibiricum powder, pack it in PE bags and seal it, and irradiate it with electron beam. The irradiation dose is 320kGy and the dose rate is 20kGy / pass. The irradiated Polygonatum sibiricum powder is refluxed and extracted 5 times with 30 times water, each time for 3h, and filtered. The filtrate is concentrated under reduced pressure at 60℃, and after concentration, 4 times the volume of anhydrous ethanol solution is slowly added, and refrigerated at 4℃ overnight. Centrifuge, discard the supernatant, wash the precipitate with 90% ethanol 3 times, and freeze-dry to obtain Polygonatum sibiricum crude polysaccharide. The obtained Polygonatum sibiricum crude polysaccharide is added to distilled water to prepare a crude polysaccharide solution, which is separated and purified using a DEAE-52 chromatography column, and then concentrated, precipitated with alcohol, centrifuged, freeze-dried, and dried to obtain Polygonatum sibiricum polysaccharide.

[0042] The selection of technical means is explained with reference to the accompanying drawings:

[0043] Figure 1-5 The effects of solid-liquid ratio, extraction temperature, extraction time, extraction times, and radiation absorption dose on the extraction rate of Polygonatum sibiricum polysaccharide are respectively selected. These five single-factor variables that have a greater impact on the PSCP extraction rate are selected to discuss the results of the PSCP extraction rate. It is determined that when the optimal solid-liquid ratio is 1:20, the PSCP extraction rate reaches the highest, which is 17.79%; when the optimal extraction temperature is 90℃, the PSCP extraction rate reaches the highest, which is 19.52%; when the optimal extraction time is 1.5h, the PSCP extraction rate reaches the highest, which is 15.19%; when the optimal number of extractions is 2 times, the PSCP extraction rate reaches the highest, which is 21.11%; when the optimal radiation dose is 240kGy, the PSCP extraction rate reaches the highest, which is 30.16%. Through the measurement results, three single factors that have a greater impact on radiation extraction of PSCP, namely extraction times, extraction temperature and radiation dose, are determined, and response surface experimental research is carried out.

[0044] Figure 6 This is a response surface experiment of radiation extraction of Polygonatum sibiricum polysaccharide. (a,b) Effects of extraction times and extraction temperature on PSCP extraction rate; (c,d) Effects of extraction temperature and radiation absorption dose on PSCP extraction rate; (e,f) Effects of extraction times and radiation absorption dose on PSCP extraction rate; (g) is the optimal process conditions calculated by response surface. (a,c,e: response surface diagram, b,d,f: contour diagram). Through Design-Expert 12 analysis, the optimal process conditions for radiation extraction of PSCP were determined to be 2 extraction times, extraction temperature 93℃, radiation absorption dose 240kGy, and the theoretical optimal PSCP yield was 32.40%. Its extraction rate is significantly higher than that of other extraction methods.

[0045] The specific embodiments described herein are merely examples of the spirit of the present invention. Those skilled in the art may make various modifications or additions to the specific embodiments described or replace them in similar ways, but they will not deviate from the spirit of the present invention or exceed the scope defined by the appended claims.

Claims

1. A highly efficient ionizing radiation extraction process for polygonatum polysaccharide, characterized in that: The following steps are involved: 1) Weigh a certain amount of Polygonatum sibiricum, wash and drain, cut into thin slices, cut the fibrous root sample into pieces, freeze-dry for 1-2 days, crush, sieve, and obtain Polygonatum sibiricum powder; 2) Take the crushed and sieved polygonatum powder, pack it into PE bags and seal them, and perform electron beam radiation extraction, with a selected dose rate of 20 kGy / pass and a radiation absorption dose of 0-320 kGy; 3) Add appropriate amount of water to the irradiated polygonatum powder, reflux extract 1-5 times, each time for 1-3 hours, the extraction temperature is 70-110°C, and filter; 4) The filtrate is concentrated under reduced pressure, and after concentration, anhydrous ethanol solution is slowly added and refrigerated overnight; 5) Centrifuge, discard the supernatant, wash the precipitate with 90% ethanol three times, and freeze-dry to obtain crude polygonatum polysaccharide; 6) The obtained crude polygonatum polysaccharide is separated and purified using a DEAE-52 chromatography column, and then concentrated, precipitated with alcohol, centrifuged, and freeze-dried to obtain polygonatum polysaccharide.

2. The high-efficiency ionizing radiation extraction process of polygonatum polysaccharide according to claim 1, characterized in that: In step 1), the mesh number of the sieve is 30 mesh, and the specific screening method is: the raw material of polygonatum is crushed and sieved with a 30-mesh sieve before irradiation to increase the radiation contact area of ​​the polygonatum and the contact area with the water extraction, thereby facilitating the dissolution of polygonatum polysaccharide.

3. The high-efficiency ionizing radiation extraction process of polygonatum polysaccharide according to claim 1, characterized in that: In step 2), the solid-to-liquid ratio of the polygonatum sibiricum powder mixed solution after adding water is 1:10-1:

30.

4. The high-efficiency ionizing radiation extraction process of polygonatum polysaccharide according to claim 1, characterized in that: In step 4), the alcohol precipitation method is: the filtrate is concentrated under reduced pressure at 60° C., and after concentration, 4 times the volume of anhydrous ethanol solution is slowly added, and refrigerated at 4° C. overnight.

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