CN111560081A

CN111560081A - Method for extracting and purifying burdock tea polysaccharide

Info

Publication number: CN111560081A
Application number: CN202010636071.6A
Authority: CN
Inventors: 杨建昊; 毛猛
Original assignee: Huarong Shengfeng Tea Industry Co ltd
Current assignee: Huarong Shengfeng Tea Industry Co ltd
Priority date: 2020-07-03
Filing date: 2020-07-03
Publication date: 2020-08-21

Abstract

The invention discloses an extraction and purification method of burdock tea polysaccharide, which mainly comprises the following steps: extracting polysaccharide by using a water extraction and alcohol precipitation method: weighing the prepared burdock tea, crushing, putting into a container, adding water, uniformly mixing, and concentrating by using a rotary evaporator; adding ethanol into the concentrated solution, and dissolving overnight; the precipitate was collected by filtration through a centrifuge as crude polysaccharide. Deproteinizing the crude polysaccharide by a Sevag method; placing the crude polysaccharide solution in a separating funnel, adding Sevag reagent, performing shaking centrifugation to obtain supernatant, dialyzing with distilled water to obtain solvent, adding hydrogen peroxide for heat preservation and decolorization, concentrating again, and vacuum freeze-drying to obtain pure polysaccharide. The total content of the burdock tea polysaccharide purified by the method is improved by 1.5 times compared with that of the raw material burdock root. And the polysaccharide has better in-vitro antioxidant activity, improves the economic value of the burdock tea and has good market development prospect.

Description

Method for extracting and purifying burdock tea polysaccharide

Technical Field

The invention belongs to the field of polysaccharide purification, and particularly relates to an extraction and purification method of burdock tea polysaccharide.

Background

Polysaccharide is a substance widely existing in organisms, is a natural high-molecular polymer, is an important macromolecule in organisms, and is one of basic substances for maintaining normal operation of life activities. Nowadays, the research on plant polysaccharides is receiving increasing attention, the international science even proposes the 21 st century which is the polysaccharide century, and through scientific verification, many plant polysaccharides have the health-care effects of regulating immunity, resisting tumors, reducing blood sugar and blood fat and the like, and the research on the plant polysaccharides is paid attention from the catering industry to the medical industry.

Arctium lappaL, also known as Eupatorium adenophorum, belongs to the order Campanulales, family Compositae, and is distributed mainly in China, Western Europe, the region of Kaishimir, Europe, etc. The burdock can be used as a cooking food material, and fruits, leaves and roots of the burdock can also be used as a medicine. The radix Arctii contains active substances such as polysaccharide, arctiin and lignan, such as arginine, aspartic acid, dietary fiber, mineral elements and vitamin C, B6, and has effects of lowering blood sugar, reducing cholesterol, resisting oxidation, clearing heat and diminishing inflammation. However, no polysaccharide purification mode aiming at burdock exists in the market at present, and the traditional purification mode has extremely low extraction efficiency of burdock tea polysaccharide and lacks of economic value.

Disclosure of Invention

The present invention aims to provide a method for extracting and purifying burdock tea polysaccharide, which can respond to the problems involved in the background technology.

The purpose of the invention can be realized by the following technical scheme: weighing the prepared burdock tea, crushing, putting into a beaker, adding ten times of tea powder mass of water, uniformly mixing, carrying out water bath at 65 ℃ for 2.5h, setting the rotating speed at 70r/min and the bath temperature at 65 ℃ for 25min by using a rotary evaporator, and concentrating to 0.2 time of the original volume. The concentrated solution is added with 95 percent ethanol with 5 times volume and dissolved for 12 hours. Centrifuging at 5000r/min for 10min, and filtering to obtain crude polysaccharide. Deproteinizing crude polysaccharide by Sevag method, preparing Sevag reagent with chloroform-n-butanol volume ratio of 4:1, placing crude polysaccharide solution in separating funnel, adding Sevag reagent with 0.2 times solution volume, oscillating for 18min, centrifuging at 5000r/min for 8min to obtain supernatant, repeating the previous operation until no milky denatured protein is precipitated between chloroform layer and n-butanol layer, mixing the supernatants, pouring into semi-permeable membrane bag with cutoff relative molecular mass of 7000, dialyzing with distilled water for 2.5d, and changing water every 2 h. Preparing the dialyzed polysaccharide into a solution of 5mg/mL, adding 1 part of 30% hydrogen peroxide solvent into each 50 parts of the dialyzed polysaccharide, keeping the temperature at 50 ℃ for decoloring for 3 hours until the color value is not reduced any more, and performing vacuum freeze-drying after concentration to obtain a polysaccharide pure product.

The invention has the beneficial effects that: the total content of the burdock tea polysaccharide purified by the method is improved by 1.5 times compared with that of the raw material burdock root. And the polysaccharide has better in-vitro antioxidant activity, improves the economic value of the burdock tea and has good market development prospect.

Detailed Description

The technical solutions of the present invention will be described clearly and completely with reference to the following embodiments, and it should be understood that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

Examples

Weighing the prepared burdock tea, crushing, putting into a beaker, adding ten times of tea powder mass of water, uniformly mixing, carrying out water bath at 65 ℃ for 2.5h, setting the rotating speed at 70r/min and the bath temperature at 65 ℃ for 25min by using a rotary evaporator, and concentrating to 0.2 time of the original volume. The concentrated solution is added with 95 percent ethanol with 5 times volume and dissolved for 12 hours. Centrifuging at 5000r/min for 10min, and filtering to obtain crude polysaccharide. Deproteinizing crude polysaccharide by Sevag method, preparing Sevag reagent with chloroform-n-butanol volume ratio of 4:1, placing crude polysaccharide solution in separating funnel, adding Sevag reagent with 0.2 times solution volume, oscillating for 18min, centrifuging at 5000r/min for 8min to obtain supernatant, repeating the previous operation until no milky denatured protein is precipitated between chloroform layer and n-butanol layer, mixing the supernatants, pouring into semi-permeable membrane bag with cutoff relative molecular mass of 7000, dialyzing with distilled water for 2.5d, and changing water every 2 h. Preparing the dialyzed polysaccharide into a solution of 5mg/mL, adding 1 part of 30% hydrogen peroxide solvent into 50 parts of the dialyzed polysaccharide, keeping the temperature and decoloring for 3 hours at 50 ℃ until the color value is not reduced any more, and performing vacuum freeze-drying after concentration to obtain a pure polysaccharide product. And sealing and packaging the prepared pure polysaccharide, and storing the polysaccharide in a ventilated and dry place.

The preferred embodiments of the invention disclosed above are intended to be illustrative only. The preferred embodiments are not intended to be exhaustive or to limit the invention to the precise embodiments disclosed. Obviously, many modifications and variations are possible in light of the above teaching. The embodiments were chosen and described in order to best explain the principles of the invention and the practical application, to thereby enable others skilled in the art to best utilize the invention. The invention is limited only by the claims and their full scope and equivalents.

Claims

1. The method for extracting and purifying the burdock tea polysaccharide is characterized by comprising the following specific steps of: weighing the prepared burdock tea, crushing, putting into a beaker, adding ten times of tea powder mass of water, uniformly mixing, carrying out water bath at 65 ℃ for 2.5h, setting the rotating speed at 70r/min and the bath temperature at 65 ℃ for 25min by using a rotary evaporator, and concentrating to 0.2 time of the original volume. The concentrated solution is added with 95 percent ethanol with 5 times volume and dissolved for 12 hours. Centrifuging at 5000r/min for 10min, and filtering to obtain crude polysaccharide. Deproteinizing crude polysaccharide by Sevag method, preparing Sevag reagent with chloroform-n-butanol volume ratio of 4:1, placing crude polysaccharide solution in separating funnel, adding Sevag reagent with 0.2 times solution volume, oscillating for 18min, centrifuging at 5000r/min for 8min to obtain supernatant, repeating the previous operation until no milky denatured protein is precipitated between chloroform layer and n-butanol layer, mixing the supernatants, pouring into semi-permeable membrane bag with cutoff relative molecular mass of 7000, dialyzing with distilled water for 2.5d, and changing water every 2 h. Preparing the dialyzed polysaccharide into a solution of 5mg/mL, adding 1 part of 30% hydrogen peroxide solvent into each 50 parts of the dialyzed polysaccharide, keeping the temperature at 50 ℃ for decoloring for 3 hours until the color value is not reduced any more, and performing vacuum freeze-drying after concentration to obtain a polysaccharide pure product.

2. The extraction and purification method of burdock tea polysaccharide as claimed in claim 1, wherein the burdock tea raw material is ground, added with water in a ratio of 1:10, mixed uniformly, and subjected to water bath at 65 ℃ for 2.5 h.

3. The extraction and purification method of burdock tea polysaccharide as claimed in claim 1, wherein a rotary evaporator is used, the rotation speed is set to 70r/min, the bath temperature is 65 ℃, the time is 25min, and the burdock tea polysaccharide is concentrated to 0.2 time of the original volume.

4. The extraction and purification method of burdock tea polysaccharide according to claim 1, wherein the burdock tea powder solution is dissolved by 95% ethanol with 5 times volume, and the dissolving time is 12 hours.

5. The method for extracting and purifying burdock tea polysaccharide as claimed in claim 1, wherein Sevag method is adopted to deproteinize the crude polysaccharide, and Sevag reagent is prepared with chloroform-n-butanol volume ratio of 4: 1.

6. The method for extracting and purifying burdock tea polysaccharide as claimed in claim 1, wherein the Sevag reagent is added, the oscillation is carried out for 18min, the centrifugation is carried out for 8min at 5000r/min, the supernatant is obtained, and the steps are repeated for many times, so that the polysaccharide in the tea powder is extracted to the maximum extent.

7. The method for extracting and purifying burdock tea polysaccharide as claimed in claim 1, wherein the supernatant obtained by purification is distilled and dialyzed for half two days, water is replaced every 2 hours, and decolorization treatment is carried out again after dialysis.