阅读说明：本技术 一种分离纯化白刺果多糖的方法 (Method for separating and purifying nitraria tangutorum bobr polysaccharide ) 是由 赵建强 贾坤 张大兵 马伟 于 2020-07-08 设计创作，主要内容包括：本发明公开了一种分离纯化白刺果多糖的方法。本发明利用膜分离纯化白刺果多糖,该方法包括如下步骤：对大孔树脂进行预处理后装入层析柱中,将白刺果果浆加入去离子水稀释,过滤去除不容物,将得到的白刺果多糖水提取液通过大孔树脂进行脱色,得白刺果多糖脱色液,用超滤设备对白刺果多糖脱色液进行分离纯化,得白刺果多糖超滤流出液,再用纳滤设备对白刺果多糖超滤流出液进行分离纯化,得白刺果多糖纳滤浓缩液,将白刺果多糖纳滤液浓缩液进行浓缩,干燥,获得含量大于80%的白刺果多糖。该工艺操作简单,总收率高,绿色环保,工艺占地少,成本低,环境污染小,适宜连续性、规模化生产。(The invention discloses a method for separating and purifying nitraria tangutorum bobr polysaccharide. The invention utilizes membrane separation to purify nitraria tangutorum bobr polysaccharide, and the method comprises the following steps: pretreating macroporous resin, then loading into a chromatographic column, adding deionized water into the Nitraria sibirica pall pulp for dilution, filtering to remove insoluble substances, decoloring the obtained Nitraria sibirica pall polysaccharide water extract through the macroporous resin to obtain Nitraria sibirica polysaccharide decoloring solution, separating and purifying the Nitraria sibirica polysaccharide decoloring solution by using ultrafiltration equipment to obtain Nitraria sibirica polysaccharide ultrafiltration effluent, separating and purifying the Nitraria sibirica polysaccharide ultrafiltration effluent by using nanofiltration equipment to obtain Nitraria sibirica polysaccharide nanofiltration concentrated solution, concentrating the Nitraria sibirica polysaccharide nanofiltration concentrated solution, and drying to obtain Nitraria sibirica polysaccharide with the content of more than 80%. The process has the advantages of simple operation, high total yield, environmental protection, small process occupation area, low cost, small environmental pollution and suitability for continuous and large-scale production.)

1. A method for separating and purifying Nitraria sibirica pall polysaccharide is characterized by comprising the following steps:

(1) pretreating macroporous resin;

(2) preparing a nitraria tangutorum polysaccharide water extracting solution;

(3) decolorizing the nitraria tangutorum polysaccharide water extract to obtain a nitraria tangutorum polysaccharide decolorized solution;

(4) separating and purifying the nitraria tangutorum bobr polysaccharide decolorization solution by using ultrafiltration equipment to obtain nitraria tangutorum bobr polysaccharide ultrafiltration effluent;

(5) separating and purifying the nitraria tangutorum bobr polysaccharide ultrafiltration effluent by using nanofiltration equipment to obtain nitraria tangutorum bobr polysaccharide nanofiltration concentrated solution;

(6) and (4) carrying out reduced pressure concentration and drying on the nitraria tangutorum bobr polysaccharide nanofiltration concentrated solution to obtain a nitraria tangutorum bobr polysaccharide finished product.

2. The method according to claim 1, wherein the macroporous resin in step (1) is a nonpolar resin having a resin skeleton of styrene-divinylbenzene, including DM130, D312, HZ-818, HZ-816, HZ-835, HZ-20SS, XDA-8, XDA-6, LSA-21.

3. The method according to claim 1, characterized in that the macroporous resin pretreatment method of step (1) is specifically: soaking macroporous resin in ethanol, stirring for 6 hr, loading into chromatographic column, eluting with ethanol until the effluent is dropped into water and no turbidity occurs, and eluting with deionized water until no alcohol smell exists.

4. The method according to claim 1, wherein the method for preparing the aqueous extract of Nitraria tangutorum bobr polysaccharide in step (2) comprises: diluting the Nitraria sibirica pall fruit pulp by 4-5 times with deionized water, stirring well, adding acid to adjust pH to 4-5, heating in water bath, filtering to remove insoluble substances to obtain Nitraria sibirica pall polysaccharide water extract.

5. The method according to claim 1, wherein the decolorization treatment of the aqueous extract of Nitraria tangutorum polysaccharide in step (3) is specifically: and (2) introducing the nitraria tangutorum bobr polysaccharide water extracting solution into a chromatographic column filled with macroporous resin at a flow rate of 0.5-3.0 times of the volume of the resin per hour, stopping feeding when the effluent liquid at the lower end of the chromatographic column is detected to have the transmittance (610 nm) of less than or equal to 50% by using a visible spectrophotometer, introducing deionized water at the same flow rate, and collecting the nitraria tangutorum bobr polysaccharide decolorizing solution.

6. The method according to claim 1, characterized in that the method for separating and purifying the nitraria tangutorum bobr polysaccharide decolorization solution by using the ultrafiltration device in the step (4) is specifically as follows: introducing the nitraria tangutorum bobr polysaccharide decolorization solution into an ultrafiltration device, controlling the frequency of the ultrafiltration device to be 36.5-38.0 Hz, the inlet pressure to be 1.2-1.3 MPa and the outlet pressure to be 0.1-1.2 MPa during ultrafiltration, repeatedly adding deionized water for washing when the nitraria tangutorum bobr polysaccharide decolorization solution in the ultrafiltration device is 1/3 in original volume, collecting ultrafiltration effluent liquid, and stopping ultrafiltration and collecting the ultrafiltration effluent liquid when a visible spectrophotometer detects and calculates that the sugar content of the liquid in the ultrafiltration device is 0.

7. The method according to claim 1, wherein the step (5) of separating and purifying the polysaccharide ultrafiltration effluent of the nitraria tangutorum bobr by using a nanofiltration device comprises: introducing the collected nitraria tangutorum bobr polysaccharide ultrafiltration effluent into a nanofiltration device, controlling the frequency of the nanofiltration device to be 36.5-38.0 Hz, the inlet pressure to be 12.0-12.3 MPa and the outlet pressure to be 10.0-10.2 MPa during nanofiltration, adding deionized water for washing when the sugar content of the nitraria tangutorum bobr polysaccharide concentrated solution in the nanofiltration device is detected and calculated to be 5% by using a spectrophotometer, and collecting the nitraria tangutorum bobr polysaccharide nanofiltration concentrated solution.

8. The method according to claim 1, wherein the concentration under reduced pressure and the drying of the nitraria tangutorum bobr polysaccharide nanofiltration concentrated solution in the step (6) are specifically as follows: controlling the temperature of a water bath kettle to be 60 ℃ and the vacuum degree to be-0.09 MPa, carrying out reduced pressure concentration, collecting reduced pressure concentrated solution, stopping concentration when the sugar content of the reduced pressure concentrated solution is 15% through detection and calculation of a visible spectrophotometer, carrying out microfiltration on the collected reduced pressure concentrated solution by using a 0.22 micron membrane, and carrying out spray drying to obtain the nitraria tangutorum polysaccharide finished product.

9. A method according to claim 4, wherein the acid is an organic edible acid, including citric acid, acetic acid, tartaric acid, malic acid.

10. The method according to claim 5, wherein the deionized water is introduced in an amount of 1.0 to 2.0 times by volume of the resin.

11. The preparation method according to claim 6, wherein the ultrafiltration membrane used by the ultrafiltration equipment is made of ceramic, polyethersulfone, polyvinylidene fluoride, cellulose acetate or composite material; the module types are tubular, hollow fiber, roll type and plate frame type; the molecular weight cutoff is 8-12 ten thousand.

12. The preparation method according to claim 7, characterized in that the nanofiltration membrane used by the nanofiltration equipment is made of polyethersulfone, polyvinylidene fluoride and composite materials; the component types are tubular, rolled and plate-frame; the molecular weight cutoff is 600-700.

Technical Field

The invention relates to a method for separating and purifying nitraria tangutorum bobr polysaccharide, and belongs to the technical field of polysaccharide purification.

Background

The white thorn is shrub of the genus nitraria of the family Zygophyllaceae, named 1759 years by Linnah, has a large amount of distribution in Qinghai firewood wood basin, has the characteristics of strong stress resistance, drought resistance and saline-alkali resistance, can adapt to severe and severe environments, and is a typical desert plant. The nitraria tangutorum bobr has good medicinal value besides ecological effects of fixing sand and the like, such as nourishing yin and moisturizing lung, relieving exterior syndrome and soothing nerves, and is a traditional authentic medicinal material for minority nationalities such as Mongolian, Tibetan and Hui nationality. As the Chelidonium majus basin of Qinghai province is less polluted by modern industrialization, the population is sparse, the environment is very clean, and the Nitraria tangutorum bobr has the characteristics of pure nature, green, no pollution and the like, is a raw and auxiliary material source of modern food, medicine and health care products, can be eaten raw or cooked, has quite rich nutrient components of Nitraria tangutorum bobr through detection, and has very high development value.

Extracting water-soluble polysaccharide from nitraria tangutorum bobr, so that the resource of the nitraria tangutorum bobr is fully utilized; the polysaccharide is macromolecule in natural organic compound, and is formed by connecting monosaccharide through glycosidic bond; more and more domestic and foreign researches prove that the active polysaccharide has the effects of resisting tumors, enhancing immunity, resisting oxidation, reducing blood sugar and the like.

The traditional nitraria tangutorum bobr polysaccharide purification method comprises technological methods such as a water extraction method, an alcohol precipitation method, a Sevag method, an ultrasonic method, a biological enzyme method, a dialysis method, an active carbon decoloration method and the like, but the methods have various defects, for example, the Sevag method needs to consume a large amount of organic solvent, the operation is complicated, the boiling point of the organic solvent is low, the organic solvent is volatile and is not suitable for being used in a large amount, and the organic solvent can cause polysaccharide degradation, so that the physiological activity of the polysaccharide is influenced; the biological enzyme method is expensive and is not suitable for industrial production; the dialysis method is slow in speed and high in environmental requirement, and can be only carried out in a laboratory; the ultrasonic method has huge energy consumption, and the active carbon can adsorb the polysaccharide to cause the loss of the polysaccharide.

Disclosure of Invention

Aiming at the technical problems, the invention provides a novel method for separating and purifying nitraria tangutorum bobr polysaccharide by using a membrane.

A method for separating and purifying Nitraria sibirica pall polysaccharide is characterized by comprising the following steps:

(1) pretreating macroporous resin;

(2) preparing a nitraria tangutorum polysaccharide water extracting solution;

(3) decolorizing the nitraria tangutorum polysaccharide water extract to obtain a nitraria tangutorum polysaccharide decolorized solution;

(4) separating and purifying the nitraria tangutorum bobr polysaccharide decolorization solution by using ultrafiltration equipment to obtain nitraria tangutorum bobr polysaccharide ultrafiltration effluent;

(5) separating and purifying the nitraria tangutorum bobr polysaccharide ultrafiltration effluent by using nanofiltration equipment to obtain nitraria tangutorum bobr polysaccharide nanofiltration concentrated solution;

(6) and (4) carrying out reduced pressure concentration and drying on the nitraria tangutorum bobr polysaccharide nanofiltration concentrated solution to obtain a nitraria tangutorum bobr polysaccharide finished product.

The macroporous resin in the step (1) is nonpolar resin, and the resin skeleton of the macroporous resin is styrene-divinylbenzene, which comprises DM130, D312, HZ-818, HZ-816, HZ-835, HZ-20SS, XDA-8, XDA-6 and LSA-21.

The method for pretreating the macroporous resin in the step (1) comprises the following steps: soaking macroporous resin in ethanol, stirring for 6 hr, loading into chromatographic column, eluting with ethanol until the effluent is dropped into water and no turbidity occurs, and eluting with deionized water until no alcohol smell exists.

The method for preparing the nitraria tangutorum bobr polysaccharide water extracting solution in the step (2) specifically comprises the following steps of: diluting the Nitraria sibirica pall fruit pulp by 4-5 times with deionized water, stirring well, adding acid to adjust pH to 4-5, heating in water bath, filtering to remove insoluble substances to obtain Nitraria sibirica pall polysaccharide water extract.

The method for decoloring the polysaccharide water extract of the nitraria tangutorum bobr specifically comprises the following steps: and (2) introducing the nitraria tangutorum bobr polysaccharide water extracting solution into a chromatographic column filled with macroporous resin at a flow rate of 0.5-3.0 times of the volume of the resin per hour, stopping feeding when the effluent liquid at the lower end of the chromatographic column is detected to have the transmittance (610 nm) of less than or equal to 50% by using a visible spectrophotometer, introducing deionized water at the same flow rate, and collecting the nitraria tangutorum bobr polysaccharide decolorizing solution.

The method for separating and purifying the nitraria tangutorum bobr polysaccharide decolorized solution by using ultrafiltration equipment specifically comprises the following steps: introducing the nitraria tangutorum bobr polysaccharide decolorization solution into an ultrafiltration device, controlling the frequency of the ultrafiltration device to be 36.5-38.0 Hz, the inlet pressure to be 1.2-1.3 MPa and the outlet pressure to be 0.1-1.2 MPa during ultrafiltration, repeatedly adding deionized water for washing when the nitraria tangutorum bobr polysaccharide decolorization solution in the ultrafiltration device is 1/3 in original volume, collecting ultrafiltration effluent liquid, and stopping ultrafiltration and collecting the ultrafiltration effluent liquid when a visible spectrophotometer detects and calculates that the sugar content of the liquid in the ultrafiltration device is 0.

The method for separating and purifying the nitraria tangutorum bobr polysaccharide ultrafiltration effluent by using nanofiltration equipment specifically comprises the following steps: introducing the collected nitraria tangutorum bobr polysaccharide ultrafiltration effluent into a nanofiltration device, controlling the frequency of the nanofiltration device to be 36.5-38.0 Hz, the inlet pressure to be 12.0-12.3 MPa and the outlet pressure to be 10.0-10.2 MPa during nanofiltration, adding deionized water for washing when the sugar content of the nitraria tangutorum bobr polysaccharide concentrated solution in the nanofiltration device is detected and calculated to be 5% by using a spectrophotometer, and collecting the nitraria tangutorum bobr polysaccharide nanofiltration concentrated solution.

The method for carrying out reduced pressure concentration and drying on the nitraria tangutorum bobr polysaccharide nanofiltration concentrated solution in the step (6) specifically comprises the following steps: controlling the temperature of a water bath kettle to be 60 ℃ and the vacuum degree to be-0.09 MPa, carrying out reduced pressure concentration, collecting reduced pressure concentrated solution, stopping concentration when the sugar content of the reduced pressure concentrated solution is 15% through detection and calculation of a visible spectrophotometer, carrying out microfiltration on the collected reduced pressure concentrated solution by using a 0.22 micron membrane, and carrying out spray drying to obtain the nitraria tangutorum polysaccharide finished product.

In the step (2), the acid is organic edible acid, including citric acid, acetic acid, tartaric acid and malic acid.

And (3) in the method, the amount of the deionized water is 1.0-2.0 times of the volume of the resin.

The ultrafiltration membrane used by the ultrafiltration equipment in the method of the step (4) is made of ceramics, polyethersulfone, polyvinylidene fluoride, cellulose acetate and composite materials; the module types are tubular, hollow fiber, roll type and plate frame type; the molecular weight cutoff is 8-12 ten thousand.

The nanofiltration membrane used by the nanofiltration equipment in the step (5) is made of polyether sulfone, polyvinylidene fluoride and composite materials; the component types are tubular, rolled and plate-frame; the molecular weight cutoff is 600-700.

The method has the advantages of utilizing the membrane to separate and purify the nitraria tangutorum bobr polysaccharide, being simpler in operation, low in labor intensity, simple in process, small in process occupation area, low in cost, small in environmental pollution, high in content of the nitraria tangutorum bobr polysaccharide and suitable for industrial production.

Detailed Description

The following examples are intended to illustrate the invention but are not intended to limit the scope of the invention.