阅读说明：本技术 马齿苋多糖提取物及制备方法及其应用 (Purslane polysaccharide extract, preparation method and application thereof ) 是由 王振辉 牛超 张真真 黄梅 *** 杨兰萍 李胜楠 孔继川 周德军 冯思颉 刘志 于 2019-11-06 设计创作，主要内容包括：本发明涉及天然药物领域,具体来说是马齿苋多糖提取物的制备方法及其应用,包括预处理、脱脂、酶解及提纯的过程,本发明制备经由预处理、脱脂、酶解的操作得到了马齿苋多糖提取物粗品,然后本发明通过高压超临界二氧化碳萃取的方式对马齿苋多糖提取物粗品进行了提纯,得到了纯度较高的马齿苋多糖提取物,并将制备得到的马齿苋多糖提取物在抗氧化及抗衰老药物中得到应用。(The invention relates to the field of natural medicines, in particular to a preparation method and application of a purslane polysaccharide extract, and the preparation method comprises the processes of pretreatment, degreasing, enzymolysis and purification.)

1. The preparation method of the purslane polysaccharide extract is characterized by comprising the following steps:

(1) pretreatment: selecting fresh purslane, removing residual roots and impurities, cleaning, scalding the purslane in boiling water for 30s-1min, drying in the air, and crushing to obtain purslane powder;

(2) degreasing: adding the purslane powder prepared in the step (1) into anhydrous ether, refluxing for 6-8h in a soxhlet extractor, taking out residues, washing with water, and drying at low temperature to obtain defatted purslane powder;

(3) enzymolysis: adding cellulose C into the defatted purslane powder prepared in the step (2)₁Performing first enzymolysis treatment on the enzyme, then filtering, adding papain, performing second enzymolysis treatment, filtering, adding papain again into filter residues, performing third enzymolysis treatment, filtering, combining filtrate, performing centrifugal separation, retaining supernatant solution, and drying to obtain a crude product of the purslane polysaccharide extract;

(4) and (3) purification: performing high-pressure supercritical carbon dioxide extraction on the crude product of the purslane polysaccharide extract prepared in the step (3), and filling gasified CO into an extractor₂And ethanol, and after extraction, CO is added₂Separating under reduced pressure with the mixture of herba Portulacae polysaccharide extract and ethanol to obtain mixture of herba Portulacae polysaccharide extract and ethanol and impurities, repeating the above operation for 3-5 times, mixing the obtained mixture of herba Portulacae polysaccharide extract and ethanol, and vacuum drying to obtain herba Portulacae polysaccharide extract.

2. The method for preparing the purslane polysaccharide extract according to claim 1, wherein the cellulose C in the step (2)₁The enzyme is endoglucosidase, and the mass ratio of the volume of the ether to the purslane is 1 mL: 1-2 g.

3. The method of claim 1, wherein the purslane polysaccharide extract is prepared by the methodCharacterized in that in the step (3), the purslane is mixed with the cellulose C₁The mass ratio of the total amount of the enzyme to the total amount of the papain is 100: 0.1-0.3: 0.4-0.6, and the papain in the second enzymolysis treatment accounts for 40% of the total amount of the papain.

4. The method for preparing the purslane polysaccharide extract according to claim 1, wherein the first enzymatic treatment in the step (3) is: adjusting pH to 4-5, soaking in 5 times volume of warm water at 50-70 deg.C for 1-2 hr; the second enzymolysis treatment method comprises the following steps: adjusting pH to 6-7, soaking in 5 times volume of 45-55 deg.C warm water for 1-2 hr; the third enzymolysis treatment method comprises the following steps: adjusting pH to 6-7, and soaking in 10 times volume of 45-55 deg.C warm water for 12-24 hr.

5. The method for preparing the purslane polysaccharide extract according to claim 1, wherein the centrifugation process in the step (3) is as follows: centrifuging at 12000-20000rpm/min for 10-15 min.

6. The method of claim 1, wherein the purslane polysaccharide extract is obtained by a method comprising the steps of, the pressure in the extraction kettle of the supercritical extraction in the step (4) is 15-25MPa, the extraction temperature is 25-40 ℃, introducing carbon dioxide at a flow rate of 15-25L/h, introducing the carbon dioxide at a pressure of 6-15Mpa for 2-3h, separating the carbon dioxide fluid sequentially in a primary separator and a secondary separator after extraction is finished, wherein the flow rate of the carbon dioxide is 10-20L/h, the pressure in the primary separator is 8-10 MPa, the temperature is 10-25 ℃, the pressure in the secondary separator is 6-8 MPa, the temperature is 35-45 ℃, discharging impurities from the primary separator, and discharging the purslane polysaccharide extract from the secondary separator.

7. A purslane polysaccharide extract prepared by the method of any one of claims 1-5.

8. Use of the purslane polysaccharide extract of claim 7 in the preparation of an anti-aging medicament.

9. Use of the purslane polysaccharide extract of claim 7 in the preparation of an antioxidant medicament.

Technical Field

The invention relates to the field of natural medicines, in particular to a purslane polysaccharide extract, a preparation method and application thereof.

Background

The purslane is a medicine and food dual-purpose purslane plant, belongs to an annual fleshy herbaceous plant, contains various chemical components, wherein purslane polysaccharide is one of main components of the purslane, and the highest content can reach 11 percent, so the purslane polysaccharide is a natural functional food with great development potential and development value, and has wide application prospect in the aspects of preventing the occurrence of various diseases and delaying the development of complications.

However, in the prior art, the purslane polysaccharide extract is extracted from purslane natural raw materials, and the purslane polysaccharide extract obtained by extraction contains a large amount of inorganic impurities, so that the purity of the purslane polysaccharide extract is low, and the application of the purslane polysaccharide extract in medicines is limited.

Disclosure of Invention

Aiming at the defects of the prior art, the invention aims to provide a purslane polysaccharide extract, a preparation method and application thereof.

In order to achieve the purpose, the invention adopts the technical scheme that:

the preparation method of the purslane polysaccharide extract comprises the following steps:

(1) pretreatment: selecting fresh purslane, removing residual roots and impurities, cleaning, scalding the purslane in boiling water for 30s-1min, drying in the air, and crushing to obtain purslane powder;

(2) degreasing: adding the purslane powder prepared in the step (1) into anhydrous ether, refluxing for 6-8h in a soxhlet extractor, taking out residues, washing with water, and drying at low temperature to obtain defatted purslane powder;

(3) enzymolysis: adding cellulose C into the defatted purslane powder prepared in the step (2)₁Performing first enzymolysis treatment on the enzyme, then filtering, adding papain, performing second enzymolysis treatment, filtering, adding papain again into filter residues, performing third enzymolysis treatment, filtering, combining filtrate, performing centrifugal separation, retaining supernatant solution, and drying to obtain a crude product of the purslane polysaccharide extract;

(4) and (3) purification: performing high-pressure supercritical carbon dioxide extraction on the crude product of the purslane polysaccharide extract prepared in the step (3), and filling gasified CO into an extractor₂And ethanol, and after extraction, CO is added₂Separating under reduced pressure with the mixture of herba Portulacae polysaccharide extract and ethanol to obtain mixture of herba Portulacae polysaccharide extract and ethanol and impurities, repeating the above operation for 3-5 times, mixing the obtained mixture of herba Portulacae polysaccharide extract and ethanol, and vacuum drying to obtain herba Portulacae polysaccharide extract.

Preferably, the cellulose C in the step (2)₁The enzyme is endoglucosidase, and the mass ratio of the volume of the ether to the purslane is 1 mL: 2-4 g.

Preferably, in the step (3), the purslane is mixed with the cellulose C₁The mass ratio of the enzyme to the papain is 100: 0.1-0.3: 0.2-0.3.

Preferably, the first enzymolysis treatment in step (3) is performed by: adjusting pH to 4-5, soaking in 5 times volume of warm water at 50-70 deg.C for 1-2 hr; the first enzymolysis treatment method comprises the following steps: adjusting pH to 6-7, soaking in 5 times volume of 45-55 deg.C warm water for 1-2 hr; the third enzymolysis treatment method comprises the following steps: adjusting pH to 6-7, and soaking in 10 times volume of 45-55 deg.C warm water for 12-24 hr.

Preferably, the centrifugation process of step (3) is: centrifuging at 12000-20000rpm/min for 10-15 min.

Preferably, the pressure in the supercritical extraction kettle in the step (4) is 15-25MPa, the extraction temperature is 25-40 ℃, the flow rate of carbon dioxide is 15-25L/h, the pressure of carbon dioxide is 6-15MPa, the extraction time is 2-3h, after the extraction is finished, the carbon dioxide fluid sequentially enters a primary separator and a secondary separator for separation, the flow rate of carbon dioxide is 10-20L/h, the pressure in the primary separator is 8MPa-10MPa, the temperature is 10-25 ℃, the pressure in the secondary separator is 6MPa-8MPa, the temperature is 35-45 ℃, impurities are discharged from the primary separator, and the purslane polysaccharide extract is discharged from the secondary separator.

The invention also protects the purslane polysaccharide extract prepared by the method.

Preferably, the purslane polysaccharide extract is applied to the preparation of anti-aging drugs.

Preferably, the purslane polysaccharide extract is applied to the preparation of anti-oxidation medicaments.

Compared with the prior art, the invention has the beneficial effects that:

1. in the invention, the residual roots and impurities of fresh purslane are removed in a pretreatment stage, then the purslane is scalded in boiling water for 30s-1min, so that the cell wall of the purslane is damaged in the boiling water, cells on the surface of the leaf are aged, purslane polysaccharide is easy to separate out from the cell wall, then degreasing is carried out, fat in the purslane is removed under the action of anhydrous ether, the influence of the fat in the purslane on the following enzymolysis operation is avoided, and then the enzymolysis process is carried out, wherein in the enzymolysis process, the residual roots and the impurities are firstly removed through cellulose C₁The method comprises the steps of breaking a crystalline structure of a cellulose chain by enzyme, performing enzymolysis by papain to hydrolyze protein in purslane to generate a large amount of purslane polysaccharide, performing enzymolysis for the third time to ensure that the enzymolysis is more thorough to obtain enough purslane polysaccharide, performing centrifugal separation and retaining supernatant solution to obtain a crude product of the purslane polysaccharide extract, purifying the crude product of the purslane polysaccharide extract by a high-pressure supercritical carbon dioxide extraction mode,the purslane polysaccharide extract with higher purity is obtained.

2. The purslane polysaccharide extract prepared by the invention has high purity, and the purslane polysaccharide extract has oxidation resistance, anti-tumor activity and anti-aging effect, so that the purslane polysaccharide extract with high purity prepared by the invention can be widely applied to preparation of anti-aging drugs and anti-oxidation drugs.

Detailed Description

In order to make the technical solutions of the present invention better understood and enable those skilled in the art to practice the present invention, the following embodiments are further described, but the present invention is not limited to the following embodiments.