CN108329397B - Method for improving alcohol precipitation efficiency of sorbus hupehensis leaf polysaccharide by adding inorganic salt - Google Patents
Method for improving alcohol precipitation efficiency of sorbus hupehensis leaf polysaccharide by adding inorganic salt Download PDFInfo
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- CN108329397B CN108329397B CN201810191681.2A CN201810191681A CN108329397B CN 108329397 B CN108329397 B CN 108329397B CN 201810191681 A CN201810191681 A CN 201810191681A CN 108329397 B CN108329397 B CN 108329397B
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- polysaccharide
- alcohol precipitation
- inorganic salt
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
Abstract
The invention relates to a method for improving alcohol precipitation efficiency of sorbus hupehensis leaf polysaccharide by adding inorganic salt, which is characterized by comprising the following steps of: the method comprises the following steps: ultrasonic-assisted extraction is carried out on the Hubei mountain ash leaves under certain conditions; step two: carrying out solid-liquid separation on the extracting solution in the step one to obtain supernatant containing polysaccharide; step three: concentrating the supernatant of the polysaccharide obtained in the step two; step four: adding ethanol with the volume four times that of the concentrated solution and a certain amount of inorganic salt into the concentrated solution in the third step, and then carrying out alcohol precipitation; step five: and (4) filtering the alcohol precipitation solution in the fourth step, and freeze-drying filter residues to obtain the extracted polysaccharide. The method can effectively improve the yield of polysaccharide alcohol precipitation and reduce the waste of the extracting solution; the method has the advantages of simple process, convenient operation and low cost, and is suitable for large-scale application.
Description
Technical Field
The invention belongs to the technical field of polysaccharide extraction, and particularly relates to a method for improving the alcohol precipitation efficiency of sorbus hupehensis leaf polysaccharide by adding inorganic salt.
Background
Polysaccharides are natural compounds enriched in plants, animals and microorganisms, have various pharmacological activities such as antioxidant, immunomodulatory and antitumor activities, and are widely used in medical, functional foods and pharmaceuticals.
The fructus Sorbi Pohuashanensis is rich in vitamins and sugar, and can be used as jam, fruit cake and wine, and the seed contains fat and amygdalin for preparing soap and for medical industry; the branch bark contains tannin, and can be used in tanning industry, and the twig and leaf can be used as feed. The determination shows that the leaves of the plant have a small amount of polysaccharide, and the plant has high antioxidant activity and high potential utilization value. Most of the traditional methods for extracting polysaccharide are an extraction method, a hot water reflux method and the like, which generally require longer extraction time and extraction temperature and consume more energy, but the extraction rate is not high, and a large amount of polysaccharide is lost particularly in the alcohol precipitation process of an extracting solution. The alcohol precipitation process is therefore a critical step in reducing polysaccharide loss.
Disclosure of Invention
The invention aims to provide a method for improving the alcohol precipitation efficiency of sorbus hupehensis leaf polysaccharide by adding inorganic salt aiming at the problem of low yield of the traditional alcohol precipitation polysaccharide so as to improve the extraction rate of the polysaccharide as a raw material.
The technical solution of the invention is as follows: a method for improving alcohol precipitation efficiency of sorbus hupehensis leaf polysaccharide by adding inorganic salt is characterized by comprising the following steps:
the method comprises the following steps: ultrasonic-assisted extraction is carried out on the Hubei mountain ash leaves under certain conditions;
step two: carrying out solid-liquid separation on the extracting solution in the step one to obtain supernatant containing polysaccharide;
step three: concentrating the supernatant of the polysaccharide obtained in the step two;
step four: adding ethanol with the volume four times that of the concentrated solution and a certain amount of inorganic salt into the concentrated solution in the third step, and then carrying out alcohol precipitation;
step five: and (4) filtering the alcohol precipitation solution in the fourth step, and freeze-drying filter residues to obtain the extracted polysaccharide.
The first step is as follows: the method comprises the following specific steps of carrying out ultrasonic-assisted extraction on the sorbus hupehensis leaves under certain conditions: pulverizing folium Sorbi Pohuashanensis, and sieving with 60 mesh sieve to obtain folium Sorbi Pohuashanensis powder; adding distilled water into the powder of the kalopanax hupehensis leaf according to the material-liquid ratio of 1:30, uniformly mixing, and performing ultrasonic assisted extraction at 60 ℃ under 350 watts for 40 minutes.
And the solid-liquid separation mode in the second step is suction filtration.
And concentrating the supernatant obtained in the third step until the volume of the supernatant is one-fifteenth of the volume of the added distilled water.
Adding a certain amount of inorganic salt into the step four, wherein the inorganic salt is sodium sulfate, ammonium sulfate or sodium chloride; the addition amount of the inorganic salt is 1.2-2.8% of the mass fraction of the alcohol precipitation system.
And the alcohol precipitation time in the fourth step is 12 hours.
The invention has the advantages that:
1. the method for improving the alcohol precipitation efficiency of the sorbus hupehensis leaf polysaccharide by adding the inorganic salt can effectively improve the alcohol precipitation yield of the polysaccharide and reduce the waste of the extracting solution;
2. according to the method, a certain amount of inorganic salt is added in the alcohol precipitation process of the extracting solution, so that the alcohol precipitation yield is improved, the alcohol precipitation time is reduced, and the alcohol precipitation efficiency is greatly improved;
3. the method has the advantages of simple process, convenient operation, low cost and suitability for large-scale application.
Detailed Description
The invention will now be further described with reference to specific examples, but the invention is not limited thereto.
Example 1
A method for improving alcohol precipitation efficiency of sorbus hupehensis leaf polysaccharide by adding inorganic salt comprises the following steps:
the method comprises the following steps: ultrasonic-assisted extraction is carried out on the Hubei mountain ash leaves under certain conditions;
step two: carrying out solid-liquid separation on the extracting solution in the step one to obtain supernatant containing polysaccharide;
step three: concentrating the supernatant of the polysaccharide obtained in the step two;
step four: adding ethanol with the volume four times that of the concentrated solution and a certain amount of inorganic salt into the concentrated solution in the third step, and then carrying out alcohol precipitation;
step five: and (4) filtering the alcohol precipitation solution in the fourth step, and freeze-drying filter residues to obtain the extracted polysaccharide.
The first step is as follows: the method comprises the following specific steps of carrying out ultrasonic-assisted extraction on the sorbus hupehensis leaves under certain conditions: pulverizing folium Sorbi Pohuashanensis, and sieving with 60 mesh sieve to obtain folium Sorbi Pohuashanensis powder; adding distilled water into the powder of the kalopanax hupehensis leaf according to the material-liquid ratio of 1:30, uniformly mixing, and performing ultrasonic assisted extraction at 60 ℃ under 350 watts for 40 minutes.
And the solid-liquid separation mode in the second step is suction filtration. The suction filtration refers to a method for reducing the pressure in a suction filtration bottle by using an air pump to achieve the purpose of solid-liquid separation.
And concentrating the supernatant obtained in the third step until the volume of the supernatant is one-fifteenth of the volume of the added distilled water.
Adding a certain amount of inorganic salt into the step four, wherein the inorganic salt is sodium sulfate, ammonium sulfate or sodium chloride; the addition amount of the inorganic salt is 1.2-2.8% of the mass fraction of the alcohol precipitation system.
And the alcohol precipitation time in the fourth step is 12 hours.
Example 2
Pulverizing folium Sorbi Pohuashanensis, and sieving with 60 mesh sieve to obtain folium Sorbi Pohuashanensis powder; weighing 5 g of 60-mesh kalopanax hupehensis leaf powder, adding 150mL of distilled water according to the material-liquid ratio of 1:30, carrying out ultrasonic assisted extraction at 60 ℃ under 350 watts for 40 minutes, after the extraction is finished, filtering to obtain a supernatant, concentrating the supernatant to 10mL, adding ethanol with four times of volume of a concentrated solution, adding 1.2% by mass of sodium chloride of an ethanol precipitation system, carrying out ethanol precipitation for 12 hours, carrying out suction filtration to obtain a polysaccharide filter cake, and carrying out freeze drying to obtain the polysaccharide, wherein the polysaccharide yield is 3.47%.
Example 3
Pulverizing folium Sorbi Pohuashanensis, and sieving with 60 mesh sieve to obtain folium Sorbi Pohuashanensis powder; weighing 5 g of 60-mesh kalopanax hupehensis leaf powder, adding 150mL of distilled water according to the material-liquid ratio of 1:30, carrying out ultrasonic assisted extraction at 60 ℃ under 350 watts for 40 minutes, after the extraction is finished, filtering to obtain a supernatant, concentrating the supernatant to 10mL, adding ethanol with four times of volume of a concentrated solution, adding 1.6% by mass of sodium chloride of an ethanol precipitation system, carrying out ethanol precipitation for 12 hours, carrying out suction filtration to obtain a polysaccharide filter cake, and carrying out freeze drying to obtain the polysaccharide, wherein the polysaccharide yield is 4.04%.
Example 4
Pulverizing folium Sorbi Pohuashanensis, and sieving with 60 mesh sieve to obtain folium Sorbi Pohuashanensis powder; weighing 5 g of 60-mesh kalopanax hupehensis leaf powder, adding 150mL of distilled water according to the material-liquid ratio of 1:30, carrying out ultrasonic assisted extraction at 60 ℃ under 350 watts for 40 minutes, after the extraction is finished, filtering to obtain a supernatant, concentrating the supernatant to 10mL, adding ethanol with four times of volume of the concentrated solution, adding ammonium sulfate with the mass fraction of 1.6% of an ethanol precipitation system, carrying out ethanol precipitation for 12 hours, carrying out suction filtration to obtain a polysaccharide filter cake, and carrying out freeze drying to obtain the polysaccharide, wherein the yield of the polysaccharide is 3.35%.
Example 5
Pulverizing folium Sorbi Pohuashanensis, and sieving with 60 mesh sieve to obtain folium Sorbi Pohuashanensis powder; weighing 5 g of 60-mesh kalopanax hupehensis leaf powder, adding 150mL of distilled water according to the material-liquid ratio of 1:30, carrying out ultrasonic assisted extraction at 60 ℃ under 350 watts for 40 minutes, after the extraction is finished, filtering to obtain a supernatant, concentrating the supernatant to 10mL, adding ethanol with four times of volume of a concentrated solution, adding ammonium sulfate with the mass fraction of 2% of an ethanol precipitation system, carrying out ethanol precipitation for 12 hours, carrying out suction filtration to obtain a polysaccharide filter cake, and carrying out freeze drying to obtain polysaccharide, wherein the polysaccharide yield is 3.44%.
Example 6
Pulverizing folium Sorbi Pohuashanensis, and sieving with 60 mesh sieve to obtain folium Sorbi Pohuashanensis powder; weighing 5 g of 60-mesh kalopanax hupehensis leaf powder, adding 150mL of distilled water according to the material-liquid ratio of 1:30, carrying out ultrasonic assisted extraction at 60 ℃ under 350 watts for 40 minutes, after the extraction is finished, filtering to obtain a supernatant, concentrating the supernatant to 10mL, adding ethanol with four times of volume of a concentrated solution, adding sodium sulfate with the mass fraction of 2% of an ethanol precipitation system, carrying out ethanol precipitation for 12 hours, carrying out suction filtration to obtain a polysaccharide filter cake, and carrying out freeze drying to obtain polysaccharide, wherein the polysaccharide yield is 6.7%. The effect of the embodiment is optimal.
Example 7
Pulverizing folium Sorbi Pohuashanensis, and sieving with 60 mesh sieve to obtain folium Sorbi Pohuashanensis powder; weighing 5 g of 60-mesh kalopanax hupehensis leaf powder, adding 150mL of distilled water according to the material-liquid ratio of 1:30, carrying out ultrasonic assisted extraction at 60 ℃ under 350 watts for 40 minutes, after the extraction is finished, filtering to obtain a supernatant, concentrating the supernatant to 10mL, adding four times of ethanol, adding sodium sulfate with the mass fraction of 2.4% of an ethanol precipitation system, carrying out ethanol precipitation for 12 hours, carrying out suction filtration to obtain a polysaccharide filter cake, and carrying out freeze drying to obtain polysaccharide, wherein the polysaccharide yield is 6.38%.
Comparative example
Pulverizing folium Sorbi Pohuashanensis, and sieving with 60 mesh sieve to obtain folium Sorbi Pohuashanensis powder; weighing 5 g of 60-mesh kalopanax hupehensis leaf powder, adding 150mL of distilled water according to the material-liquid ratio of 1:30, carrying out ultrasonic assisted extraction at 60 ℃ under 350 watts for 40 minutes, after the extraction is finished, filtering to obtain a supernatant, concentrating the supernatant to 10mL, adding ethanol with the volume four times that of the concentrated solution, carrying out alcohol precipitation for 24 hours, carrying out suction filtration to obtain a polysaccharide filter cake, and carrying out freeze drying to obtain the polysaccharide, wherein the polysaccharide yield is 2.54%.
Comparing examples 2 to 7 with the comparative examples, it is clear that the extraction effect with the addition of the inorganic salt is significantly higher than that of the comparative examples without the addition of the inorganic salt. Meanwhile, the polysaccharide yield of the sodium sulfate with the mass fraction of 2% in the alcohol precipitation system in the embodiment 6 is 6.7% which is the best scheme. The alcohol precipitation system related in the embodiment of the invention is the sum of four times of the volume of ethanol in the concentrated solution and the added concentrated solution.
The parts of the present embodiment not described in detail are common general knowledge in the industry, and are not described here.
Claims (4)
1. A method for improving the alcohol precipitation efficiency of sorbus hupehensis leaf polysaccharide by adding inorganic salt is characterized by comprising the following steps:
the method comprises the following steps: ultrasonic-assisted extraction is carried out on the Hubei mountain ash leaves under certain conditions;
step two: carrying out solid-liquid separation on the extracting solution in the step one to obtain supernatant containing polysaccharide;
step three: concentrating the supernatant of the polysaccharide obtained in the step two;
step four: adding ethanol with the volume four times that of the concentrated solution and a certain amount of inorganic salt into the concentrated solution in the third step, and then carrying out alcohol precipitation; the inorganic salt is sodium sulfate, ammonium sulfate or sodium chloride; the addition amount of the inorganic salt is 1.2-2.8% of the mass fraction of the alcohol precipitation system;
step five: and (4) filtering the alcohol precipitation solution in the fourth step, and freeze-drying filter residues to obtain the extracted polysaccharide.
2. The method for improving the alcohol precipitation efficiency of the sorbus hupehensis leaf polysaccharide by adding the inorganic salt as claimed in claim 1, wherein the first step is: the method comprises pulverizing folium Sorbi Pohuashanensis, and sieving with 60 mesh sieve to obtain folium Sorbi Pohuashanensis powder; adding distilled water into the powder of the kalopanax hupehensis leaf according to the material-liquid ratio of 1:30, uniformly mixing, and performing ultrasonic assisted extraction at 60 ℃ under 350 watts for 40 minutes.
3. The method for improving the alcohol precipitation efficiency of the sorbus hupehensis leaf polysaccharide as claimed in claim 2, wherein the supernatant obtained in the third step is concentrated to one-fifteenth of the volume of the added distilled water.
4. The method for improving the alcohol precipitation efficiency of the sorbus hupehensis leaf polysaccharide through the addition of the inorganic salt as claimed in claim 1, wherein the alcohol precipitation time in the fourth step is 12 hours.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0378486A1 (en) * | 1989-01-05 | 1990-07-18 | Societe De Developpement Des Utilisations Du Cuir Saduc | Process for the separation of sulphated glycosaminoglycans |
| CN105273094A (en) * | 2015-05-04 | 2016-01-27 | 西华大学 | Method for rapid separation of Ligusticum chuanqiong polysaccharide |
| CN106038737A (en) * | 2016-02-29 | 2016-10-26 | 西北农林科技大学 | Method for extracting flavones in Sorbus alnifolia through coupled supersonic wave-aqueous two-phase system |
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| CN103739735B (en) * | 2014-01-23 | 2016-02-17 | 李文东 | A kind of method extracting tea polysaccharide from black tea golden flower |
| CN105348406A (en) * | 2015-11-25 | 2016-02-24 | 江苏兴野食品有限公司 | Extraction and purification method for fistular onion stalk polysaccharide |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0378486A1 (en) * | 1989-01-05 | 1990-07-18 | Societe De Developpement Des Utilisations Du Cuir Saduc | Process for the separation of sulphated glycosaminoglycans |
| CN105273094A (en) * | 2015-05-04 | 2016-01-27 | 西华大学 | Method for rapid separation of Ligusticum chuanqiong polysaccharide |
| CN106038737A (en) * | 2016-02-29 | 2016-10-26 | 西北农林科技大学 | Method for extracting flavones in Sorbus alnifolia through coupled supersonic wave-aqueous two-phase system |
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