CN103665181A - Simple porphyra yezoensis crude polysaccharide preparation method - Google Patents
Simple porphyra yezoensis crude polysaccharide preparation method Download PDFInfo
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- CN103665181A CN103665181A CN201310667095.8A CN201310667095A CN103665181A CN 103665181 A CN103665181 A CN 103665181A CN 201310667095 A CN201310667095 A CN 201310667095A CN 103665181 A CN103665181 A CN 103665181A
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Abstract
The invention discloses a simple porphyra yezoensis crude polysaccharide preparation method. The simple porphyra yezoensis crude polysaccharide preparation method comprises the following steps: (1) pounding porphyra yezoensis fronds to pieces by using a dry method; (2) adding an ethanol solution with the volume concentration of above 75%, soaking for 12-24 hours, and filtering to obtain filter residues; (3) boiling the filter residues for 1-2 hours at the temperature of 90-95 DEG C according to a mass ratio of water to materials being (20-40): 1, filtering to obtain filtrate, repeatedly adding water to the filter residues so as to boil the filter residues again, and combining filtrate; (4) heating and concentrating the filtrate, adding ethanol till the final volume concentration of ethanol is 75-80%, standing for 5-10 days, and filtering so as to obtain crude polysaccharide precipitates. Compared with the prior art, the simple porphyra yezoensis crude polysaccharide preparation method has the advantages of high efficiency and low cost which are not reported before application. Compared with a conventional technology, the simple porphyra yezoensis crude polysaccharide preparation method has the advantages that the cost of centrifugal equipment is saved, large-scale production is facilitated, and the cost of decoloring and degreasing technologies is reduced than before.
Description
Technical field
The present invention relates to the simple and easy preparation of Crude Polysaccharide from Porphyra yezoensis, belong to field of processing of aquatic products.
Background technology
Laver belongs to rhodophyta (Rhodophyta), former Rhodophyceae (Protoflorideophyceae), Bangiales section (Bangiaceae), Porphyra (Porphyra), at China coast, all has distribution.The common laver kind of China has more than 10 kinds, and Main Cultivars has two kinds of yezoensis laver (Porphyra yezoensis Ueda) and porphyra haitanensis (Porphyra haitanensis T.J.Chang et B.F.Zheng).Yezoensis laver frond is light violet redness or brown, is the common kind of northern China bank.
Yezoensis laver is a kind of large-scale economical alga, it has high nutritive value, contains abundant containing 18 seed amino acids (wherein with the content of L-Ala, L-glutamic acid, aspartic acid for the highest), cholesterol galactoside, cholesterol half mannoside, palmityl cholesterol galactoside, palmityl cholesterol mannoside and carotene, VITMAIN B1, B2, C1, nicotinic acid, protein and calcium, phosphorus, iron etc.Traditional Chinese medicine is thought, yezoensis laver taste is sweet, salty, cold in nature, have that softening and eliminating sputum, heat-clearing nourish heart, the function of invigorating kidney to promote diuresis, tradition is used for the treatment of the diseases such as dysphoria and insomnia due to cough and asthma, swelling and pain in the throat, high fever, palpitation with fear palpitation, vertigo, edema due to deficiency of the kidney, dysuria.
In laver, contain polysaccharide, and be laver important component part, carried out laver amylose research the fifties.
The conventional preliminary extracting method of polysaccharide in laboratory has at present: by individual character employing water (cold water or hot water) extraction method, neutral salt solution extraction method, alkali extraction method, the lytic enzyme of homopolysaccharide do not digest extraction method, proteinase hydrolization method etc., the different solubility in different concns spirituous solution according to polysaccharide component again, progressively alcohol is analysed and is obtained thick laver amylose precipitation.In the preliminary leaching process of polysaccharide, temperature, pH value, the control of the key factors such as time is very important, and the structure of polysaccharide, activity will be played to very large impact.
Wu Qiongying etc. adopt microwave auxiliary extraction technology, research microwave power, treatment time and the impact of solid-liquid ratio on Porphyra yezoensis Polysaccharides extraction yield.On the basis of single factor experiment, by orthogonal test, determine that optimal extract process is that microwave power 180W, microwave extraction time 8min, solid-liquid ratio are 1:40, with this understanding. Porphyra yezoensis Polysaccharides extraction yield is 5.358%.The optimum process condition that He Ronghai etc. extract through the definite Crude Polysaccharide from Porphyra yezoensis of orthogonal test is: solid-liquid ratio is that the dry laver powder of 15g adds 2L water, extracting solution circulation rotating speed is 12r/s, extracting solution temperature is 40～C, ultrasonic power is 800W, supersound process time/ultrasonic gap is that 3s/2s and ultrasonic omnidistance time are 2h, and corresponding yield is 26.29%.
Polysaccharide has stronger polarity, adds the organic solvents such as ethanol, acetone or methyl alcohol in its aqueous solution, can produce precipitation.The contained polar group of homopolysaccharide and molecular weight are not different yet.The solubleness that molecular weight is little is large, branch many compared with the good water solubility of straight chain, adopt the organic solvent of different concns, can obtain the polysaccharide of different components.Conventional method is: 1. ethanol precipitation; 2. the saline solns precipitator method.Precipitation agent conventional in the precipitator method has: methyl alcohol, ethanol, acetone, barium salt or zinc salt (for dermatan sulfate), potassium acetate etc.
In polysaccharide molecule, ionogen has hydroxyl and sulfate, PK value, and the connected group of three-dimensional arrangement position and sulfate differs, and the PK value of different hydroxyls and sulfate has again small difference.Meanwhile, have hydroxyl and sulfate quantity also to have difference in various polysaccharide, these difference become the important evidence of separation of polysaccharides.Conventional method is: 1. quaternary ammonium salt complexometry; 2. electrophoretic separation.
Except above-mentioned several method can carry out separation polysaccharide, also has some other method.As: the method for column chromatography, conventional has: ion exchange chromatography (being suitable for separated various acidic polysaccharose and neutral polysaccharide), gel-filtration, have another name called gel chromatography or molecular sieve chromatography and (utilize the difference of molecular size range, polysaccharide fraction is carried out to separation and purification), affinity chromatography etc., active carbon column chromatography etc., first two method is widely used in the polysaccharide purification of laboratory stage at present.
Also have in addition some technology, as plate technique method, enzyme process, ultracentrifuge method, high performance liquid chromatography (HPLC) method etc. also have general application in the purifying of polysaccharide.
Summary of the invention
The object of this invention is to provide a kind of simple and easy method of preparing Crude Polysaccharide from Porphyra yezoensis.
In order to achieve the above object, the present invention is achieved through the following technical solutions:
A simple and easy method of preparing Crude Polysaccharide from Porphyra yezoensis, comprises the following steps:
(1) yezoensis laver frond dry method is smashed to pieces;
(2) add the more than 75% alcohol solution dipping 12-24h of volumetric concentration, filter to obtain filter residue;
(3) according to water material mass ratio 20-40:1, filter residue is boiled to 1-2h at 90-95 ℃, cross leaching filtrate, filter residue repeats to add poach once, merging filtrate;
(4) heating concentrated filtrate, adding ethanol is 75-80% to ethanol final volume concentration, filters to obtain Crude polysaccharides precipitation after standing 5-10 days.
Preferably, step (1) by laver frond smash to pieces to particle diameter be 3-5mm.
Preferably, step (2), (3) and (4) adopt 200 order to 800 order filtered through gauze.
The present invention above-mentionedly prepares Crude Polysaccharide from Porphyra yezoensis method simply owing to having adopted, thereby has compared to existing technology cost-effective advantage, before this aspect application, has no report; In addition, technique of the present invention is compared to the long-time standing rear filtration of existing process using and replaces routine centrifugal, has saved centrifugation apparatus cost, is conducive to large-scale production simultaneously.
Embodiment
For technique means, creation characteristic that the present invention is realized, reach object and effect is easy to understand, below further set forth the present invention.
Embodiment 1:
By 600 grams of yezoensis laver frond dry method smash to pieces to particle diameter be 3-5mm, the alcohol solution dipping 12h that adds volumetric concentration 75%, 200 order filtered through gauze obtain filter residue, filter residue water material mass ratio be 20: 1 times 90 ℃ boil 1h, 600 order filtered through gauze are got filtrate, filter residue repeats to add poach once, merging filtrate, heating concentrated filtrate is to 6L, and adding ethanol to ethanol final volume concentration is 75%, and after standing 5 days, 800 order filtered through gauze obtain Crude polysaccharides precipitation.Precipitation obtains 56.3 grams of Crude polysaccharides after vacuum lyophilization, and yield is 9.4%, and it is 43.1% that Crude polysaccharides adopts phenol sulfuric acid process to record wherein total sugar content.
Embodiment 2:
By 600 grams of yezoensis laver frond dry method smash to pieces to particle diameter be 3-5mm, the alcohol solution dipping 20h that adds volumetric concentration 80%, 200 order filtered through gauze obtain filter residue, filter residue water material mass ratio be 30: 1 times 90 ℃ boil 1h, 400 order filtered through gauze are got filtrate, filter residue repeats to add poach once, merging filtrate, heating concentrated filtrate is to 6L, and adding ethanol to ethanol final volume concentration is 80%, and after standing 10 days, 600 order filtered through gauze obtain Crude polysaccharides precipitation.Precipitation obtains 57.5 grams of Crude polysaccharides after vacuum lyophilization, and yield is 9.6%, and it is 43.8% that Crude polysaccharides adopts phenol sulfuric acid process to record wherein total sugar content.
Embodiment 3:
By 600 grams of yezoensis laver frond dry method smash to pieces to particle diameter be 3-5mm, the alcohol solution dipping 24h that adds volumetric concentration 85%, 300 order filtered through gauze obtain filter residue, filter residue water material mass ratio be 40: 1 times 95 ℃ boil 1h, 500 order filtered through gauze are got filtrate, filter residue repeats to add poach once, merging filtrate, heating concentrated filtrate is to 6L, and adding ethanol to ethanol final volume concentration is 75%, and after standing 10 days, 700 order filtered through gauze obtain Crude polysaccharides precipitation.Precipitation obtains 58.0 grams of Crude polysaccharides after vacuum lyophilization, and yield is 9.7%, and it is 44.4% that Crude polysaccharides adopts phenol sulfuric acid process to record wherein total sugar content.
The above is preferred embodiment of the present invention, but the present invention should not be confined to the disclosed content of this embodiment.So every, do not depart from the equivalence completing under principles of this disclosure or revise, all falling into the scope of protection of the invention.
Claims (3)
1. a simple and easy method of preparing Crude Polysaccharide from Porphyra yezoensis, is characterized in that, comprises the following steps:
(1) yezoensis laver frond dry method is smashed to pieces;
(2) add the more than 75% alcohol solution dipping 12-24h of volumetric concentration, filter to obtain filter residue;
(3) according to water material mass ratio 20-40:1, filter residue is boiled to 1-2h at 90-95 ℃, cross leaching filtrate, filter residue repeats to add poach once, merging filtrate;
(4) heating concentrated filtrate, adding ethanol is 75-80% to ethanol final volume concentration, filters to obtain Crude polysaccharides precipitation after standing 5-10 days.
2. the simple and easy method of preparing Crude Polysaccharide from Porphyra yezoensis according to claim 1, is characterized in that, step (1) by laver frond smash to pieces to particle diameter be 3-5mm.
3. the simple and easy method of preparing Crude Polysaccharide from Porphyra yezoensis according to claim 1, is characterized in that, step (2), (3) and (4) adopt 200 order to 800 order filtered through gauze.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109400745A (en) * | 2018-11-21 | 2019-03-01 | 温州大学苍南研究院 | A kind of preparation of low molecular weight Porphyra yezoensis Polysaccharides and its application in anti-human cervical cancer cell tumour |
| CN111494505A (en) * | 2020-04-27 | 2020-08-07 | 乌林奇 | Active molecule composition for target regulation and prevention of thrombus |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109400745A (en) * | 2018-11-21 | 2019-03-01 | 温州大学苍南研究院 | A kind of preparation of low molecular weight Porphyra yezoensis Polysaccharides and its application in anti-human cervical cancer cell tumour |
| CN109400745B (en) * | 2018-11-21 | 2021-01-05 | 温州大学苍南研究院 | Preparation of low molecular weight porphyra yezoensis polysaccharide and application of low molecular weight porphyra yezoensis polysaccharide in resisting human cervical cancer cell tumor |
| CN111494505A (en) * | 2020-04-27 | 2020-08-07 | 乌林奇 | Active molecule composition for target regulation and prevention of thrombus |
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Application publication date: 20140326 |