CN108753872B - High-yield extraction method of lotus seed polysaccharide - Google Patents
High-yield extraction method of lotus seed polysaccharide Download PDFInfo
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- NINOYEKNLLERTK-UHFFFAOYSA-M COC(C)(O)OC.[Cl-].[Li+] Chemical compound COC(C)(O)OC.[Cl-].[Li+] NINOYEKNLLERTK-UHFFFAOYSA-M 0.000 description 1
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- 229920001491 Lentinan Polymers 0.000 description 1
- XCUCMLUTCAKSOZ-FIRIVFDPSA-N Liensinine Chemical compound C([C@@H]1C=2C=C(C(=CC=2CCN1C)OC)OC=1C(O)=CC=C(C=1)C[C@H]1N(C)CCC=2C=C(C(=CC=21)OC)OC)C1=CC=C(O)C=C1 XCUCMLUTCAKSOZ-FIRIVFDPSA-N 0.000 description 1
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- 150000002605 large molecules Chemical class 0.000 description 1
- 229940115286 lentinan Drugs 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 235000019341 magnesium sulphate Nutrition 0.000 description 1
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- -1 polysaccharide alkali metal salt Chemical class 0.000 description 1
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- MUPFEKGTMRGPLJ-ZQSKZDJDSA-N raffinose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO[C@@H]2[C@@H]([C@@H](O)[C@@H](O)[C@@H](CO)O2)O)O1 MUPFEKGTMRGPLJ-ZQSKZDJDSA-N 0.000 description 1
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Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/14—Preparation of compounds containing saccharide radicals produced by the action of a carbohydrase (EC 3.2.x), e.g. by alpha-amylase, e.g. by cellulase, hemicellulase
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/04—Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds
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- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
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- Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- General Health & Medical Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Biochemistry (AREA)
- General Chemical & Material Sciences (AREA)
- Microbiology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Genetics & Genomics (AREA)
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- Medicines Containing Plant Substances (AREA)
Abstract
The invention discloses a high-yield extraction method of lotus seed polysaccharide, relating to the technical field of extraction of lotus seed polysaccharide, and comprising the following steps: the lotus seed polysaccharide is prepared by cleaning lotus seeds, adding microcrystalline cellulose, carrying out coarse crushing, superfine crushing, emulsifying and filtering to obtain filtrate and filter residues, carrying out centrifugal demulsification on the filtrate to obtain supernatant and precipitate, and carrying out different extraction modes on the filter residues, the supernatant and the precipitate. The invention utilizes the microcrystalline cellulose to improve the extraction rate of the lotus seed polysaccharide, and different extraction modes are carried out according to different parts of the lotus seeds after crushing, filtering and centrifugal demulsification, so that the yield and the yield of the polysaccharide are improved, and the purity of the polysaccharide is improved.
Description
Technical Field
The invention relates to the technical field of extraction of lotus seed polysaccharide, in particular to a high-yield extraction method of lotus seed polysaccharide.
Background
Polysaccharides (polysaccharides) are formed by condensation and dehydration of a plurality of monosaccharide molecules, and are carbohydrate substances with complex and huge molecular mechanisms. Carbohydrates and their derivatives, which meet the concept of high molecular weight compounds, are called polysaccharides. Polysaccharides are also important in their very wide distribution in nature. Some are components constituting animal and plant cell walls, such as peptidoglycan and cellulose; some are nutrients for animal and plant storage, such as glycogen and starch; some have special biological activity, such as heparin in human body has anticoagulant effect, and polysaccharide in pneumococcal cell wall has antigen effect. The structural units of polysaccharides are monosaccharides, with the relative molecular masses of polysaccharides ranging from tens of thousands to tens of millions.
More and more researches show that: human life processes are almost exclusively associated with sugar chains: such as cell-cell communication, recognition and interaction, cellular motility and adhesion, pathogen interaction with host cells, and the like. This is because the sugar chain carries biological information. It plays a crucial role in the molecular recognition process on the cell surface.
Four methods are commonly used for extraction of polysaccharides: A. hot water extraction method: according to most polysaccharidesThe extraction is carried out with the property of greater solubility and stability in hot water-this method causes minimal damage to the extracted polysaccharides. B. Dilute alkali water solution extraction method: acidic polysaccharides or polysaccharides with relatively high molecular weight have low solubility in hot water, so 5-15% NaOH or Na is usually used2CO3Extracting with water solution. C. An enzymolysis method comprises the following steps: the method is used in the preparation of polysaccharide health product (such as lentinan health product). However, most of them adopt a method combining hot water extraction and enzymatic extraction, i.e., extraction with hot water is performed first, and then extraction with enzymatic method is performed on the residue, so that the yield of polysaccharide can be greatly improved. D. Other methods are as follows: the method for extracting polysaccharide from polysaccharide alkali metal salt by using aprotic solvent such as dimethyl sulfoxide as solvent comprises extracting polysaccharide with acidic aqueous solution such as lithium chloride-dimethoxy ethanol, and is mainly not used due to high cost and low yield.
The lotus seeds are also called Shuiziandan, and are important special economic resources and export agricultural products in China. Polysaccharides are important components of all living organisms, are widely present in cell walls of animals, plants and microorganisms, are another important class of biomolecules in organisms besides nucleic acids and proteins, and scientific research has confirmed that the polysaccharides have many biological activities including anti-tumor, immunity, blood sugar reduction, and virus resistance, and have almost no toxic side effects on organisms. Therefore, basic research on the lotus seeds breaks through the technical bottleneck, and the method has important significance for promoting the development of the current lotus seed deep processing industry. The existing lotus seed extraction methods comprise a water decoction method, ultrasonic extraction, microwave extraction and the like, and literature reports about amylase and glucoamylase dual-enzyme degradation are reported, but the extraction rate is low.
The functions of the lotus seeds comprise: 1. and (3) cancer prevention and resistance: the lotus seed is good at tonifying the five internal organs, promoting the circulation of qi and blood of twelve meridians, and smoothing qi and blood without rotting, and the oxycephalonine contained in the lotus seed has an inhibiting effect on nasopharyngeal carcinoma, so that the nutrition and health-care functions of cancer prevention and anticancer of the lotus seed are achieved. 2. Blood pressure reduction: the non-crystalline alkaloid N-9 contained in semen Nelumbinis has blood pressure lowering effect. 3. Strengthening heart and soothing nerves: alkaloid contained in the lotus plumule has a remarkable cardiotonic effect, and the liensinine has strong calcium and arrhythmia resisting effects. 4. Nourishing and tonifying deficiency, and arresting seminal emission: the raffinose contained in the lotus seeds is a tonic suitable for both the old and the young, and is a common nutritional product for people with chronic diseases, postpartum diseases or old and weak people; the aloperine has the function of inhibiting libido, and has good effects of stopping spermatorrhea and arresting seminal emission when being taken by young people with dreaminess, frequent spermatorrhea or spermatorrhea. 5. Clearing away heart-fire and removing freckles: lotus seed with plumula Nelumbinis can clear heart fire and dispel freckle, but it can not be decocted for a long time.
The extraction method of the lotus seed polysaccharide disclosed in the Chinese invention patent with the application number of 201410292488.X comprises the following steps: taking lotus seeds, adding water and pulping to obtain pulp; adding magnesium sulfate into the slurry, stirring, steaming with pressure cooker, cooling, and sieving to obtain filtrate; diluting the filtrate with water, and filtering with a ceramic microfiltration membrane to obtain a ceramic membrane clear solution; and adsorbing the clear liquid of the ceramic membrane by using ion exchange resin, eluting by using a sodium chloride solution, concentrating the eluent by using a nanofiltration membrane, and drying the concentrated solution to obtain the lotus seed polysaccharide. However, the yield, yield and purity of the polysaccharide extracted by the method are not high.
Disclosure of Invention
The invention aims to provide a method for extracting lotus seed polysaccharide with high yield aiming at the defects in the prior art.
The invention solves the technical problem, adopts the technical scheme that a high-yield extraction method of lotus seed polysaccharide is provided, and comprises the following steps:
A. pretreatment: cleaning semen Nelumbinis, adding microcrystalline cellulose, pulverizing, and micronizing to obtain semen Nelumbinis fine powder;
B. emulsification: putting the lotus seed fine powder into water, adding an emulsifier, and fully emulsifying to obtain a lotus seed emulsion containing microcrystalline cellulose;
C. and (3) filtering: filtering the lotus seed emulsion to obtain filtrate and filter residue;
D. and (3) filter residue treatment: 1) alkali soaking: c, soaking the filter residue in the step C in alkali liquor to obtain a soaking liquid, and 2) neutralizing: neutralizing the soaking liquid with acid, adjusting pH to 6.5-7.5, and 3) filtering for one time: filtering the neutralized soaking liquid to obtain filtrate and filter residue, and 4) carrying out secondary filtration: repeating the operations 1), 2) and 3) on the filter residue in 3) to obtain filtrate and filter residue, and 5) concentrating: combining the filtrates in 3) and 4), concentrating to obtain a concentrated solution, and 6) precipitating with ethanol: adding ethanol into the concentrated solution for alcohol precipitation to obtain crude lotus seed polysaccharide and drying, 7): nanofiltration drying: dissolving the dried crude lotus seed polysaccharide in distilled water to obtain a polysaccharide solution, concentrating the polysaccharide solution by using a nanofiltration membrane to obtain a concentrated solution, and drying the concentrated solution to obtain lotus seed polysaccharide;
E. and (3) centrifugal demulsification: c, centrifuging and demulsifying the filtrate obtained in the step C to obtain supernatant and precipitate;
F. and (3) supernatant treatment: e, adsorbing the supernatant obtained in the step E by using ion exchange resin, eluting by using 3-5wt.% sodium chloride solution to obtain eluent, concentrating the eluent by using a nanofiltration membrane to obtain concentrated solution, and drying the concentrated solution to obtain lotus seed polysaccharide;
G. and (3) precipitation treatment: 1) and (3) extraction: adding enzyme liquid into the precipitate obtained in the step E, performing enzymolysis for 2-3h at 25-35 ℃, performing ultrasonic extraction to obtain a leaching solution, and 2) centrifuging and concentrating: centrifuging the leaching solution to obtain supernatant, concentrating the supernatant to obtain a concentrated solution, and 3) precipitating with ethanol: adding ethanol into the concentrated solution for alcohol precipitation to obtain crude lotus seed polysaccharide and drying, and 4) nano-filtering and drying: dissolving the dried crude lotus seed polysaccharide in distilled water to obtain a polysaccharide solution, concentrating the polysaccharide solution by using a nanofiltration membrane to obtain a concentrated solution, and drying the concentrated solution to obtain the lotus seed polysaccharide.
Preferably, the alcohol precipitation in step D and the alcohol precipitation in step G can be carried out in combination. The operation is convenient, the production efficiency is improved, the two concentrated solutions are simultaneously subjected to alcohol precipitation, the possibility of collision among particles can be increased, and the alcohol precipitation time is shortened.
Preferably, ethanol is added in the ethanol precipitation in the step D and the step G, and the ethanol concentration is 85-90%. On one hand, the ethanol with the concentration can reduce the usage amount of the ethanol, and on the other hand, the ethanol with the concentration is simple to recover, so that the cost is saved.
Preferably, the alcohol content of the solution is 70-80% when alcohol precipitation occurs.
Preferably, in step G, the enzyme solution is prepared from distilled water, amylase, pectinase and acid cellulase.
Preferably, the weight of the enzyme solution is 5-10 times of the weight of the precipitate, the amylase accounts for 0.003-0.008% of the weight of the precipitate, and the weight ratio of the amylase, the pectinase and the acid cellulase is 2-5:3-10: 1-2. The enzymolysis reaction of various enzymes in the proportion is coordinated without inhibiting effect, so that the yield, yield and purity of the final polysaccharide product are improved.
Preferably, in the step A, before the microcrystalline cellulose is added, the lotus seeds are dried at the temperature of 30-40 ℃, and the moisture content of the dried lotus seeds is 5-10%. Thus, after the lotus seeds are crushed, the exposed surface of the lotus seeds is provided with a plurality of pores, and the microcrystalline cellulose can enter the pores by stirring, so that the contact area is increased, the superfine crushing efficiency is improved, and the crushing is more uniform; if the moisture content is too low, the lotus seeds are easily damaged by friction heat generated by the lotus seeds and the microcrystalline cellulose in the superfine grinding, and the 5-10% moisture can protect the lotus seeds and cannot influence the grinding effect.
Preferably, in step a, the coarsely pulverized lotus seed and microcrystalline cellulose are frozen before the coarsely pulverized lotus seed and the coarsely pulverized microcrystalline cellulose. The cell wall of the frozen lotus seed loses elasticity and is easy to break when being crushed, and meanwhile, the frozen lotus seed is subjected to superfine crushing to avoid friction heat generation to damage the components of the lotus seed.
Preferably, the freezing temperature is below-5 ℃ and the freezing time is 1-3 h.
Preferably, the emulsifier is tween-80.
The invention has the following beneficial effects:
1. the microcrystalline cellulose is added, so that the lotus seed can be more thoroughly crushed, the subsequent extraction of polysaccharide is facilitated, and the microcrystalline cellulose is insoluble in an emulsifier, so that the subsequent removal of the microcrystalline cellulose is very convenient and can be recycled.
2. And (3) separating the emulsified filtrate, the filter residue and the supernatant and the precipitate after centrifugal demulsification of the emulsified filtrate to extract the polysaccharide, and adopting different extraction modes according to the conditions of all parts, so that on one hand, the yield and the yield of the polysaccharide are improved, and on the other hand, the purity of the polysaccharide is also improved.
Detailed Description
The following are specific examples of the present invention and further describe the technical solutions of the present invention, but the present invention is not limited to these examples.
Example 1:
a high-yield extraction method of lotus seed polysaccharide comprises the following steps:
A. pretreatment: cleaning semen Nelumbinis, adding microcrystalline cellulose, pulverizing, and micronizing to obtain semen Nelumbinis fine powder;
B. emulsification: putting the lotus seed fine powder into water, adding an emulsifier Tween-80, and fully emulsifying to obtain a lotus seed emulsion containing microcrystalline cellulose;
C. and (3) filtering: filtering the lotus seed emulsion to obtain filtrate and filter residue;
D. and (3) filter residue treatment: 1) alkali soaking: c, soaking the filter residue in the step C in alkali liquor to obtain a soaking liquid, and 2) neutralizing: neutralizing the soaking liquid with acid, adjusting the pH to 7.5, and 3) filtering for one time: filtering the neutralized soaking liquid to obtain filtrate and filter residue, and 4) carrying out secondary filtration: repeating the operations 1), 2) and 3) on the filter residue in 3) to obtain filtrate and filter residue, and 5) concentrating: combining the filtrates in 3) and 4), concentrating to obtain a concentrated solution, and 6) precipitating with ethanol: adding 90% ethanol into the concentrated solution, adjusting the ethanol content of the concentrated solution to 70%, performing alcohol precipitation to obtain crude lotus seed polysaccharide, and drying, 7): nanofiltration drying: dissolving the dried crude lotus seed polysaccharide in distilled water to obtain a polysaccharide solution, concentrating the polysaccharide solution by using a nanofiltration membrane to obtain a concentrated solution, and drying the concentrated solution to obtain lotus seed polysaccharide;
E. and (3) centrifugal demulsification: c, centrifuging and demulsifying the filtrate obtained in the step C to obtain supernatant and precipitate;
F. and (3) supernatant treatment: e, adsorbing the supernatant obtained in the step E by using ion exchange resin, eluting by using a 5wt.% sodium chloride solution to obtain an eluent, concentrating the eluent by using a nanofiltration membrane to obtain a concentrated solution, and drying the concentrated solution to obtain lotus seed polysaccharide;
G. and (3) precipitation treatment: 1) and (3) extraction: adding enzyme liquid containing water, amylase, pectinase and acid cellulase into the precipitate in the step E, performing enzymolysis for 2 hours at 35 ℃, performing ultrasonic extraction to obtain a leaching solution, wherein the weight of the enzyme liquid is 10 times that of the precipitate, the amylase accounts for 0.003 percent of the precipitate, the weight ratio of the amylase to the pectinase to the acid cellulase is 2:10:1, 2) centrifuging and concentrating: centrifuging the leaching solution to obtain supernatant, concentrating the supernatant to obtain a concentrated solution, and 3) precipitating with ethanol: adding 90% ethanol into the concentrated solution, adjusting the ethanol content of the concentrated solution to 70%, performing alcohol precipitation to obtain crude lotus seed polysaccharide, and drying, 4) performing nanofiltration drying: dissolving the dried crude lotus seed polysaccharide in distilled water to obtain a polysaccharide solution, concentrating the polysaccharide solution by using a nanofiltration membrane to obtain a concentrated solution, and drying the concentrated solution to obtain the lotus seed polysaccharide.
The yield of the lotus seed polysaccharide is 3.9%, the yield is 95.2%, and the purity reaches 45%.
Example 2:
a high-yield extraction method of lotus seed polysaccharide comprises the following steps:
A. pretreatment: cleaning semen Nelumbinis, oven drying at 30 deg.C until the water content of the semen Nelumbinis is 10%, adding microcrystalline cellulose into the dried semen Nelumbinis, coarse pulverizing, and micronizing in micronizer to obtain semen Nelumbinis fine powder;
B. emulsification: putting the lotus seed fine powder into water, adding an emulsifier Tween-80, and fully emulsifying to obtain a lotus seed emulsion containing microcrystalline cellulose;
C. and (3) filtering: filtering the lotus seed emulsion to obtain filtrate and filter residue;
D. and (3) filter residue treatment: 1) alkali soaking: c, soaking the filter residue in the step C in alkali liquor to obtain a soaking liquid, and 2) neutralizing: neutralizing the soaking liquid with acid, adjusting the pH to 6.5, and 3) filtering for one time: filtering the neutralized soaking liquid to obtain filtrate and filter residue, and 4) carrying out secondary filtration: repeating the operations 1), 2) and 3) on the filter residue in 3) to obtain filtrate and filter residue, and 5) concentrating: combining the filtrates in 3) and 4), and concentrating to obtain a concentrated solution for later use;
E. and (3) centrifugal demulsification: c, centrifuging and demulsifying the filtrate obtained in the step C to obtain supernatant and precipitate;
F. and (3) supernatant treatment: e, adsorbing the supernatant obtained in the step E by using ion exchange resin, eluting by using a 4wt.% sodium chloride solution to obtain an eluent, concentrating the eluent by using a nanofiltration membrane to obtain a concentrated solution, and drying the concentrated solution to obtain lotus seed polysaccharide;
G. and (3) precipitation treatment: 1) and (3) extraction: adding enzyme liquid containing water, amylase, pectinase and acid cellulase into the precipitate in the step E, performing enzymolysis for 3 hours at 25 ℃, performing ultrasonic extraction to obtain a leaching solution, wherein the weight of the enzyme liquid is 5 times that of the precipitate, the amylase accounts for 0.008 percent of the precipitate, and the weight ratio of the amylase, the pectinase and the acid cellulase is 5:3:3, 2) centrifuging and concentrating: centrifuging the leaching solution to obtain supernatant, concentrating the supernatant to obtain a concentrated solution, and 3) precipitating with ethanol: mixing the standby concentrated solution obtained in the step D) and the concentrated solution obtained in the step G) in the step 2), adding ethanol with the concentration of 85%, adjusting the ethanol content of the mixed solution to 75%, performing alcohol precipitation to obtain crude lotus seed polysaccharide, and drying, 4) performing nanofiltration drying: dissolving the dried crude lotus seed polysaccharide in distilled water to obtain a polysaccharide solution, concentrating the polysaccharide solution by using a nanofiltration membrane to obtain a concentrated solution, and drying the concentrated solution to obtain the lotus seed polysaccharide.
The yield of the lotus seed polysaccharide is 4.1%, the yield is 95.5%, and the purity reaches 47%.
Example 3:
a high-yield extraction method of lotus seed polysaccharide comprises the following steps:
A. pretreatment: cleaning semen Nelumbinis, oven drying at 40 deg.C until the water content of the dried semen Nelumbinis is 5%, adding microcrystalline cellulose into the dried semen Nelumbinis, coarse pulverizing, freezing at-8 deg.C for 2 hr, and micronizing in micronizer to obtain semen Nelumbinis fine powder;
B. emulsification: putting the lotus seed fine powder into water, adding an emulsifier Tween-80, and fully emulsifying to obtain a lotus seed emulsion containing microcrystalline cellulose;
C. and (3) filtering: filtering the lotus seed emulsion to obtain filtrate and filter residue;
D. and (3) filter residue treatment: 1) alkali soaking: c, soaking the filter residue in the step C in alkali liquor to obtain a soaking liquid, and 2) neutralizing: neutralizing the soaking liquid with acid, adjusting the pH to 7, and 3) filtering for one time: filtering the neutralized soaking liquid to obtain filtrate and filter residue, and 4) carrying out secondary filtration: repeating the operations 1), 2) and 3) on the filter residue in 3) to obtain filtrate and filter residue, and 5) concentrating: combining the filtrates in 3) and 4), and concentrating to obtain a concentrated solution for later use;
E. and (3) centrifugal demulsification: c, centrifuging and demulsifying the filtrate obtained in the step C to obtain supernatant and precipitate;
F. and (3) supernatant treatment: e, adsorbing the supernatant obtained in the step E by using ion exchange resin, eluting by using 3wt.% sodium chloride solution to obtain an eluent, concentrating the eluent by using a nanofiltration membrane to obtain a concentrated solution, and drying the concentrated solution to obtain lotus seed polysaccharide;
G. and (3) precipitation treatment: 1) and (3) extraction: adding enzyme liquid containing water, amylase, pectinase and acid cellulase into the precipitate in the step E, performing enzymolysis for 2-3h at 30 ℃, performing ultrasonic extraction to obtain a leaching solution, wherein the weight of the enzyme liquid is 7 times that of the precipitate, the amylase accounts for 0.005% of the precipitate, the weight ratio of the amylase to the pectinase to the acid cellulase is 4:7:1, 2) centrifuging and concentrating: centrifuging the leaching solution to obtain supernatant, concentrating the supernatant to obtain a concentrated solution, and 3) precipitating with ethanol: mixing the standby concentrated solution obtained in the step D) and the concentrated solution obtained in the step G) in the step 2), adding 88% ethanol, adjusting the ethanol content of the mixed solution to 80%, performing alcohol precipitation to obtain crude lotus seed polysaccharide, and drying, 4) performing nanofiltration drying: dissolving the dried crude lotus seed polysaccharide in distilled water to obtain a polysaccharide solution, concentrating the polysaccharide solution by using a nanofiltration membrane to obtain a concentrated solution, and drying the concentrated solution to obtain the lotus seed polysaccharide.
The yield of the lotus seed polysaccharide is 4.6%, the yield is 96.0%, and the purity reaches 49%.
Comparative example 1:
a high-yield extraction method of lotus seed polysaccharide comprises the following steps:
A. pretreatment: cleaning semen Nelumbinis, pulverizing, and micronizing in micronizer to obtain semen Nelumbinis fine powder;
B. emulsification: putting the lotus seed fine powder into water, adding an emulsifier Tween-80, and fully emulsifying to obtain a lotus seed emulsion containing microcrystalline cellulose;
C. and (3) filtering: filtering the lotus seed emulsion to obtain filtrate and filter residue;
D. and (3) filter residue treatment: 1) alkali soaking: c, soaking the filter residue in the step C in alkali liquor to obtain a soaking liquid, and 2) neutralizing: neutralizing the soaking liquid with acid, adjusting the pH to 8, and 3) filtering for one time: filtering the neutralized soaking liquid to obtain filtrate and filter residue, and 4) carrying out secondary filtration: repeating the operations 1), 2) and 3) on the filter residue in 3) to obtain filtrate and filter residue, and 5) concentrating: combining the filtrates in 3) and 4), concentrating to obtain a concentrated solution, and 6) precipitating with ethanol: adding 90% ethanol into the concentrated solution, adjusting the ethanol content of the concentrated solution to 70%, performing alcohol precipitation to obtain crude lotus seed polysaccharide, and drying, 7): nanofiltration drying: dissolving the dried crude lotus seed polysaccharide in distilled water to obtain a polysaccharide solution, concentrating the polysaccharide solution by using a nanofiltration membrane to obtain a concentrated solution, and drying the concentrated solution to obtain lotus seed polysaccharide;
E. and (3) centrifugal demulsification: c, centrifuging and demulsifying the filtrate obtained in the step C to obtain supernatant and precipitate;
F. and (3) supernatant treatment: e, adsorbing the supernatant obtained in the step E by using ion exchange resin, eluting by using a 5wt.% sodium chloride solution to obtain an eluent, concentrating the eluent by using a nanofiltration membrane to obtain a concentrated solution, and drying the concentrated solution to obtain lotus seed polysaccharide;
G. and (3) precipitation treatment: 1) and (3) extraction: adding enzyme liquid containing water, amylase and pectinase into the precipitate in the step E, performing enzymolysis for 2 hours at 35 ℃, performing ultrasonic extraction to obtain a leaching solution, wherein the weight of the enzyme liquid is 10 times that of the precipitate, the amylase accounts for 0.003% of the precipitate, and the weight ratio of the amylase to the pectinase is 1:5, 2) centrifuging and concentrating: centrifuging the leaching solution to obtain supernatant, concentrating the supernatant to obtain a concentrated solution, and 3) precipitating with ethanol: adding 90% ethanol into the concentrated solution, adjusting the ethanol content of the concentrated solution to 70%, performing alcohol precipitation to obtain crude lotus seed polysaccharide, and drying, 4) performing nanofiltration drying: dissolving the dried crude lotus seed polysaccharide in distilled water to obtain a polysaccharide solution, concentrating the polysaccharide solution by using a nanofiltration membrane to obtain a concentrated solution, and drying the concentrated solution to obtain the lotus seed polysaccharide.
The yield of the lotus seed polysaccharide is 2.6%, the yield is 90.2%, and the purity is 30%.
Comparative example 2:
a high-yield extraction method of lotus seed polysaccharide comprises the following steps:
A. pretreatment: cleaning semen Nelumbinis, oven drying at 40 deg.C until the water content of the dried semen Nelumbinis is 5%, adding microcrystalline cellulose into the dried semen Nelumbinis, coarse pulverizing, freezing at-8 deg.C for 2 hr, and micronizing in micronizer to obtain semen Nelumbinis fine powder;
B. emulsification: putting the lotus seed fine powder into water, adding an emulsifier Tween-20, and fully emulsifying to obtain a lotus seed emulsion containing microcrystalline cellulose;
C. and (3) filtering: filtering the lotus seed emulsion to obtain filtrate and filter residue;
D. and (3) filter residue treatment: 1) alkali soaking: c, soaking the filter residue in the step C in alkali liquor to obtain a soaking liquid, and 2) neutralizing: neutralizing the soaking liquid with acid, adjusting the pH to 7, and 3) filtering for one time: filtering the neutralized soaking liquid to obtain filtrate and filter residue, and 4) carrying out secondary filtration: repeating the operations 1), 2) and 3) on the filter residue in 3) to obtain filtrate and filter residue, and 5) concentrating: combining the filtrates in 3) and 4), and concentrating to obtain a concentrated solution for later use;
E. and (3) centrifugal demulsification: c, centrifuging and demulsifying the filtrate obtained in the step C to obtain supernatant and precipitate;
F. and (3) supernatant treatment: e, adsorbing the supernatant obtained in the step E by using ion exchange resin, eluting by using 8wt.% sodium chloride solution to obtain an eluent, concentrating the eluent by using a nanofiltration membrane to obtain a concentrated solution, and drying the concentrated solution to obtain lotus seed polysaccharide;
G. and (3) precipitation treatment: 1) and (3) extraction: adding enzyme liquid containing water, amylase, pectinase and acid cellulase into the precipitate in the step E, performing enzymolysis for 2-3h at 30 ℃, performing ultrasonic extraction to obtain a leaching solution, wherein the weight of the enzyme liquid is 7 times that of the precipitate, the amylase accounts for 0.005% of the precipitate, the weight ratio of the amylase to the pectinase to the acid cellulase is 1:1:5, and 2) centrifuging and concentrating: centrifuging the leaching solution to obtain supernatant, concentrating the supernatant to obtain a concentrated solution, and 3) precipitating with ethanol: mixing the standby concentrated solution obtained in the step D) and the concentrated solution obtained in the step G) in the step 5), adding 60% ethanol, adjusting the ethanol content of the mixed solution to 50%, performing alcohol precipitation to obtain crude lotus seed polysaccharide, and drying, 4) performing nanofiltration drying: dissolving the dried crude lotus seed polysaccharide in distilled water to obtain a polysaccharide solution, concentrating the polysaccharide solution by using a nanofiltration membrane to obtain a concentrated solution, and drying the concentrated solution to obtain the lotus seed polysaccharide.
The yield of the lotus seed polysaccharide is 2.9%, the yield is 91.3%, and the purity is 28%.
Comparative example 3:
a high-yield extraction method of lotus seed polysaccharide comprises the following steps:
A. pretreatment: cleaning semen Nelumbinis, oven drying at 80 deg.C until the water content of the semen Nelumbinis is 10%, adding microcrystalline cellulose into the dried semen Nelumbinis, coarse pulverizing, and micronizing in micronizer to obtain semen Nelumbinis fine powder;
B. emulsification: putting the lotus seed fine powder into water, adding an emulsifier Tween-80, and fully emulsifying to obtain a lotus seed emulsion containing microcrystalline cellulose;
C. and (3) filtering: filtering the lotus seed emulsion to obtain filtrate and filter residue;
D. and (3) filter residue treatment: 1) alkali soaking: c, soaking the filter residue in the step C in alkali liquor to obtain a soaking liquid, and 2) neutralizing: neutralizing the soaking liquid with acid, adjusting the pH to 6.5, and 3) filtering for one time: filtering the neutralized soaking liquid to obtain filtrate and filter residue, and 4) carrying out secondary filtration: repeating the operations 1), 2) and 3) on the filter residue in 3) to obtain filtrate and filter residue, and 5) concentrating: combining the filtrates in 3) and 4), and concentrating to obtain a concentrated solution for later use;
E. and (3) centrifugal demulsification: c, centrifuging and demulsifying the filtrate obtained in the step C to obtain supernatant and precipitate;
F. and (3) supernatant treatment: e, adsorbing the supernatant obtained in the step E by using ion exchange resin, eluting by using a 4wt.% sodium chloride solution to obtain an eluent, concentrating the eluent by using a nanofiltration membrane to obtain a concentrated solution, and drying the concentrated solution to obtain lotus seed polysaccharide;
G. and (3) precipitation treatment: 1) and (3) extraction: and E, adding the precipitate in the step E into an enzyme solution containing water, amylase and acid cellulase, carrying out enzymolysis for 3 hours at 25 ℃, and carrying out ultrasonic extraction to obtain a leaching solution, wherein the weight of the enzyme solution is 2 times of the weight of the precipitate, the amylase accounts for 0.015 percent of the weight of the precipitate, and the weight ratio of the amylase to the acid cellulase is 5:3, 2) centrifuging and concentrating: centrifuging the leaching solution to obtain supernatant, concentrating the supernatant to obtain a concentrated solution, and 3) precipitating with ethanol: mixing the standby concentrated solution obtained in the step D) and the concentrated solution obtained in the step G) in the step 2), adding ethanol with the concentration of 85%, adjusting the ethanol content of the mixed solution to 75%, performing alcohol precipitation to obtain crude lotus seed polysaccharide, and drying, 4) performing nanofiltration drying: dissolving the dried crude lotus seed polysaccharide in distilled water to obtain a polysaccharide solution, concentrating the polysaccharide solution by using a nanofiltration membrane to obtain a concentrated solution, and drying the concentrated solution to obtain the lotus seed polysaccharide.
The yield of the lotus seed polysaccharide is 2.8%, the yield is 92.2%, and the purity is 31%.
The specific embodiments described herein are merely illustrative of the spirit of the invention. Various modifications or additions may be made to the described embodiments or alternatives may be employed by those skilled in the art without departing from the spirit or ambit of the invention as defined in the appended claims.
Claims (6)
1. The extraction method of the lotus seed polysaccharide with high yield is characterized by comprising the following steps:
A. pretreatment: cleaning semen Nelumbinis, adding microcrystalline cellulose, pulverizing, and micronizing to obtain semen Nelumbinis fine powder;
B. emulsification: putting the lotus seed fine powder into water, adding an emulsifier, and fully emulsifying to obtain a lotus seed emulsion containing microcrystalline cellulose, wherein the emulsifier is Tween-80;
C. and (3) filtering: filtering the lotus seed emulsion to obtain filtrate and filter residue;
D. and (3) filter residue treatment: 1) alkali soaking: c, soaking the filter residue in the step C in alkali liquor to obtain a soaking liquid, and 2) neutralizing: neutralizing the soaking liquid with acid, adjusting pH to 6.5-7.5, and 3) filtering for one time: filtering the neutralized soaking liquid to obtain filtrate and filter residue, and 4) carrying out secondary filtration: repeating the operations 1), 2) and 3) on the filter residue in 3) to obtain filtrate and filter residue, and 5) concentrating: combining the filtrates in 3) and 4), concentrating to obtain a concentrated solution, and 6) precipitating with ethanol: adding ethanol into the concentrated solution, performing ethanol precipitation to obtain crude lotus seed polysaccharide, and drying, wherein during the ethanol precipitation, the content of the ethanol in the solution is 70-80%, and the ratio of 7): nanofiltration drying: dissolving the dried crude lotus seed polysaccharide in distilled water to obtain a polysaccharide solution, concentrating the polysaccharide solution by using a nanofiltration membrane to obtain a concentrated solution, and drying the concentrated solution to obtain lotus seed polysaccharide;
E. and (3) centrifugal demulsification: c, centrifuging and demulsifying the filtrate obtained in the step C to obtain supernatant and precipitate;
F. and (3) supernatant treatment: e, adsorbing the supernatant obtained in the step E by using ion exchange resin, eluting by using 3-5wt.% sodium chloride solution to obtain eluent, concentrating the eluent by using a nanofiltration membrane to obtain concentrated solution, and drying the concentrated solution to obtain lotus seed polysaccharide;
G. and (3) precipitation treatment: 1) and (3) extraction: adding enzyme liquid into the precipitate in the step E, performing enzymolysis for 2-3h at 25-35 ℃, performing ultrasonic extraction to obtain a leaching solution, wherein the enzyme liquid is prepared by mixing distilled water, amylase, pectinase and acid cellulase, the weight of the enzyme liquid is 5-10 times of the weight of the precipitate, the amylase accounts for 0.003-0.008% of the weight of the precipitate, the weight ratio of the amylase to the pectinase to the acid cellulase is 2-5:3-10:1-2, and 2) centrifuging and concentrating: centrifuging the leaching solution to obtain supernatant, concentrating the supernatant to obtain a concentrated solution, and 3) precipitating with ethanol: adding ethanol into the concentrated solution for alcohol precipitation to obtain crude lotus seed polysaccharide, and drying, wherein in the alcohol precipitation, the alcohol content in the solution is 70-80%, and 4) carrying out nanofiltration drying: dissolving the dried crude lotus seed polysaccharide in distilled water to obtain a polysaccharide solution, concentrating the polysaccharide solution by using a nanofiltration membrane to obtain a concentrated solution, and drying the concentrated solution to obtain the lotus seed polysaccharide.
2. The method for extracting lotus seed polysaccharide with high yield according to claim 1, which is characterized in that: the alcohol precipitation in step D and the alcohol precipitation in step G can be combined.
3. The method for extracting lotus seed polysaccharide with high yield according to claim 1, which is characterized in that: and D, adding 85-90% ethanol into the ethanol precipitates in the step D and the step G.
4. The method for extracting lotus seed polysaccharide with high yield according to claim 1, which is characterized in that: in the step A, before microcrystalline cellulose is added, the lotus seeds are dried at the temperature of 30-40 ℃, and the moisture content of the dried lotus seeds is 5-10%.
5. The method for extracting lotus seed polysaccharide with high yield according to claim 1 or 4, which is characterized in that: in the step A, after coarse grinding and before superfine grinding, the coarsely ground lotus seeds and microcrystalline cellulose are frozen.
6. The method for extracting lotus seed polysaccharide with high yield according to claim 5, which is characterized in that: the freezing temperature is below-5 deg.C, and the freezing time is 1-3 hr.
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| CN102925497A (en) * | 2012-11-01 | 2013-02-13 | 恩施清江生物工程有限公司 | Method for preparing high-purity resveratrol from polygonum cuspidatum |
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| CN102925497A (en) * | 2012-11-01 | 2013-02-13 | 恩施清江生物工程有限公司 | Method for preparing high-purity resveratrol from polygonum cuspidatum |
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