CN105273093B - A kind of preparation method of seaweed polysaccharide sulfate - Google Patents
A kind of preparation method of seaweed polysaccharide sulfate Download PDFInfo
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- CN105273093B CN105273093B CN201410328156.2A CN201410328156A CN105273093B CN 105273093 B CN105273093 B CN 105273093B CN 201410328156 A CN201410328156 A CN 201410328156A CN 105273093 B CN105273093 B CN 105273093B
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Abstract
The invention discloses a kind of preparation method of seaweed polysaccharide sulfate, this method is using sea-tangle as raw material, preprocessed, two step water extraction, the analysis of two step alcohol, a step alcohol is molten and dries obtained seaweed polysaccharide sulfate, seaweed polysaccharide sulfate in sea-tangle can effectively be extracted, and pass through the well-designed of each technological parameter, it is possible to increase the efficiency and yield of seaweed polysaccharide sulfate.
Description
Technical field
The present invention relates to the extracting method of medicine, more particularly to a kind of preparation method of seaweed polysaccharide sulfate.
Background technology
Different from the similar variety production technique through chemical modification or chemical process, seaweed polysaccharide sulfate preparation method uses
Natural bioactive technology.During extraction separation and purification, lead hydroxide, calcium chloride, quaternary amine, ether, acetone are not added
, also should not soda acid processing or sulphonation chemistry modification, it is only necessary to two kinds of systems of drinkable water and medical ethanol Deng precipitating reagent and organic reagent
Standby medium.This technique neither destroys the natural chemical composition of active component, turn avoid extraction and the pollution that purge process is brought
With the harmful effect of strong acid, highly basic to medicine.
Extraction seaweed polysaccharide sulfate active principle simultaneously, other non-active ingredients, as iodine, algin (alginic acid and its
Salt), mannitol etc. needs effectively to remove.
Sea-tangle is larger in northern China marine site yield, can be planted with large area.Sea-tangle is embodied in People's Republic of China's medicine
Allusion quotation one (page 195, version in 2010) kelp, it is the drying of Laminariaceae plant sea-tangle (Laminaria japonica Aresch)
Thallus.Its character is that curling is folded into bulk, or is tangled into handle, and entirety is in dark brown or green and brown color, and surface has bloom.With
Water logging is soft, is expanded into flat long ribbon shape, and long 50-150 centimetres, wide 10-40 centimetres, middle part is thicker, and edge is relatively thin and is in wavy.
Function is with curing mainly as softening and resolving hard mass, dissolving phlegm, Li Shui.For goiter and tumor, scrofula, testiclar gall, phlegm and retained fluid oedema.And marine alga is sargassum
The drying frond of section's plant Sargassum or sargassum fusifome, practise again and claim big leaflet marine alga, marine alga character shrinkage curling, dark brown, have
It is long 30-60 centimetres by bloom.
General in the prior art all to use marine alga to extract seaweed polysaccharide sulfate for raw material, this method one is can not be fully sharp
With the sea-tangle that northern China marine site yield is larger, second, its extracting method is not suitable for sea-tangle.
The content of the invention
It is an object of the invention to provide a kind of preparation method of seaweed polysaccharide sulfate, to prepare marine alga by raw material of sea-tangle
Sulfated polysaccharide.
The preparation method of the seaweed polysaccharide sulfate of the present invention, comprises the following steps：
S10, by sea-tangle be placed in the water of 38-42 times of its own quality soak 35-45 minutes after, for its own matter
The 38-42 times of water measured of amount is rinsed well；
S20, sea-tangle is pre-processed, reject the unqualified sea-tangle that leaf is yellow, leaf is broken and mouldy, qualified sea-tangle is cut into face
The fragment of product 7-10 square centimeters；Because sea-tangle is in long ribbon shape, being adapted to cut into pieces using chopping processes is advantageous to improve efficiency
And yield.
S30, sea-tangle be added to decocted three times in the water of 9-11 times of its own quality, respectively 1.9-2.1 hours,
1.4-1.6 hours, 1.4-1.6 hours；The seaweed polysaccharide sulfate extracted by the way of decoction in sea-tangle can be more fully
Seaweed polysaccharide sulfate in sea-tangle is extracted.
S40, decoction liquor it will be concentrated under reduced pressure into relative density 1.05-1.15 after merging three times and obtain concentrate I；Due to decocting
Boil liquid is concentrated under reduced pressure so that the concentration increase of seaweed polysaccharide sulfate in concentrate, can more easily extract marine alga sulphur
Acid polysaccharide, ethanol is not only saved, and the yield extracted is high.
S50, ethanol is added to ethanol volumetric concentration 29-31% into concentrate I, supernatant is centrifuged to obtain after standing overnight
And sediment；
S60, sediment is detected, if SO in sediment4 2-Weight/mass percentage composition be more than 5%, ethanol is added into sediment
To ethanol volumetric concentration 29-31%, stand and centrifuge to obtain supernatant to after being layered；If SO in sediment4 2-Weight/mass percentage composition it is small
In equal to 5%, sediment is abandoned；The seaweed polysaccharide sulfate in sediment can be extracted by this step, make marine alga in the sediment of discarding
The content of sulfated polysaccharide is relatively low.
S70, by step S50 and step S60 supernatant merge, being concentrated under reduced pressure into relative density 1.1-1.2 must concentrate
Liquid II；
S80, ethanol is added to ethanol volumetric concentration 83-86% into concentrate II, after standing overnight, abandon supernatant, sink
Shallow lake centrifuges after being washed with percentage by volume 74-76% ethanol；
S90, carry out being spray-dried to obtain seaweed polysaccharide sulfate after the precipitation after centrifugation is dissolved with the 7-9 times of water measured.
Extracted by above-mentioned two step water, the analysis of two step alcohol and the molten safety of a step alcohol, science, succinct extraction, separation and
The production line of purifying, the seaweed polysaccharide sulfate in sea-tangle can effectively be extracted, and setting meticulously by each technological parameter
Meter, it is possible to increase the efficiency and yield of seaweed polysaccharide sulfate.
Preferably, sea-tangle is placed in the step S10 after being soaked 40 minutes in the water of 40 times of its own quality, be used for
The water of 40 times of amounts of its own quality is rinsed well.
Preferably, in the step S20, qualified sea-tangle is cut into the fragment of 9 square centimeters of area.
Preferably, in the step S30, sea-tangle is added to and decocted three times in the water of its own quality 10, respectively 2
Hour, 1.5 hours, 1.5 hours.
Preferably, in the step S40, will three times decoction liquor merge after be concentrated under reduced pressure into relative density 1.1 concentrate
I。
Preferably, in the step S50, ethanol is added into concentrate I to ethanol volumetric concentration 30%, after standing overnight
Centrifuge to obtain supernatant and sediment.
Preferably, in the step S60, if SO in sediment4 2-Weight/mass percentage composition be more than 5%, second is added into sediment
Alcohol stands to ethanol volumetric concentration 30% and centrifuges to obtain supernatant to after being layered.
Preferably, in the step S70, the supernatant in step S50 and step S60 is merged, is concentrated under reduced pressure into relative
Density 1.17 obtains concentrate II.
Preferably, in the step S80, ethanol is added into concentrate II to ethanol volumetric concentration 85%, is stood overnight
Afterwards, supernatant is abandoned, precipitation centrifuges after being washed with the ethanol of percentage by volume 75%.
Preferably, in the step S90, carry out being spray-dried extra large after the precipitation after centrifugation is dissolved with the water of 8 times of amounts
Algae sulfated polysaccharide.
The preparation method of the seaweed polysaccharide sulfate of the present invention can effectively extract the seaweed polysaccharide sulfate in sea-tangle, have peace
Entirely, science, it is succinct the advantages of.
Brief description of the drawings
Fig. 1 is that the seaweed polysaccharide sulfate monose obtained by embodiment 1 forms gas chromatogram.
Embodiment
The preparation method of the seaweed polysaccharide sulfate of the present invention, comprises the following steps：
S10, by sea-tangle be placed in the water of 38-42 times of its own quality soak 35-45 minutes after, for its own matter
The 38-42 times of water measured of amount is rinsed well；
S20, sea-tangle is pre-processed, reject the unqualified sea-tangle that leaf is yellow, leaf is broken and mouldy, qualified sea-tangle is cut into face
The fragment of product 7-10 square centimeters；Because sea-tangle is in long ribbon shape, being adapted to cut into pieces using chopping processes is advantageous to improve efficiency
And yield.
S30, sea-tangle be added to decocted three times in the water of 9-11 times of its own quality, respectively 1.9-2.1 hours,
1.4-1.6 hours, 1.4-1.6 hours；The seaweed polysaccharide sulfate extracted by the way of decoction in sea-tangle can be more fully
Seaweed polysaccharide sulfate in sea-tangle is extracted.
S40, decoction liquor it will be concentrated under reduced pressure into relative density 1.05-1.15 after merging three times and obtain concentrate I；Due to decocting
Boil liquid is concentrated under reduced pressure so that the concentration increase of seaweed polysaccharide sulfate in concentrate, can more easily extract marine alga sulphur
Acid polysaccharide, ethanol is not only saved, and the yield extracted is high.
S50, ethanol is added to ethanol volumetric concentration 29-31% into concentrate I, supernatant is centrifuged to obtain after standing overnight
And sediment；
S60, sediment is detected, if SO in sediment4 2-Weight/mass percentage composition be more than 5%, ethanol is added into sediment
To ethanol volumetric concentration 29-31%, stand and centrifuge to obtain supernatant to after being layered；If SO in sediment4 2-Weight/mass percentage composition it is small
In equal to 5%, sediment is abandoned；The seaweed polysaccharide sulfate in sediment can be extracted by this step, make marine alga in the sediment of discarding
The content of sulfated polysaccharide is relatively low.
S70, by step S50 and step S60 supernatant merge, being concentrated under reduced pressure into relative density 1.1-1.2 must concentrate
Liquid II；
S80, ethanol is added to ethanol volumetric concentration 83-86% into concentrate II, after standing overnight, abandon supernatant, sink
Shallow lake centrifuges after being washed with percentage by volume 74-76% ethanol；
S90, carry out being spray-dried to obtain seaweed polysaccharide sulfate after the precipitation after centrifugation is dissolved with the 7-9 times of water measured.
Technical scheme is further elaborated with reference to embodiment.
Embodiment 1
1. seaweed polysaccharide sulfate processing parameter
1.1 sea-tangles select and identification
The place of production：North of China marine site.
Form：Dry bundling.
Identification：Identified one by one by seaweed polysaccharide sulfate quality standard by professional, reject leaf Huang, leaf
The unqualified sea-tangle such as broken, mouldy.
1.2 immersion processes
Feed intake：200 kilograms of sea-tangles.
Immersion way：In four batches, 50 kilograms every batch are put into stainless steel pond, sea-tangle is totally immersed into 40 up and down after unclamping
Again in amount water.
Soak time：40 minutes.
1.3 cleanings and chopping process
Cleaning way：Sea-tangles are rinsed to clean with 40 times of amount water.
Scavenging period：Every batch 10 minutes.
Shred size：3 centimetres of big fractionlets of particle diameter.
1.4 water extraction process
Take sea-tangle fragment to be put into extractor, by successively boiling extracts three times, add the water of 10 times of amounts every time, boil for the first time
Boiling 2 hours, second and third time is respectively boiled 1.5 hours.
1.5 concentrations one
Extract concentrates on concentration tank and carries out first time concentration three times.
Mode：It is concentrated under reduced pressure.
Final concentration：Relative density 1.10 (50 DEG C of heat are surveyed).
1.6 alcohol precipitations one
Concentrate adds ethanol to volumetric concentration 30%, carries out first time precipitation, stands overnight.
1.7 centrifugations one
Collected after centrifugation supernatant, and sediment sample is taken, carry out assay.【Sediment detects】By seaweed polysaccharide sulfate matter
Amount standard is carried out.
1.8 sediments weight is molten
If sediment SO4 2->When 5%, with 30% ethanol molten sediment again, stand overnight.
1.9 centrifuge again
The SO into sediment4 2-<When 5%, sediment is removed in centrifugation, collects supernatant.
1.10 concentration two
Centrifuged supernatant concentrates on concentration tank.
Mode：It is concentrated under reduced pressure.
Final concentration：Relative density 1.17 (50 DEG C of heat are surveyed).
1.11 alcohol precipitation two
Concentrate adds ethanol to final concentration 85%, stands overnight.
1.12 wash precipitation
Incline supernatant, adds ethanol to wash precipitation to volumetric concentration 75%, stand overnight.
1.13 centrifugation two
Deposit sample is taken to carry out assay after centrifugation.
1.14 spray drying
After centrifugation, add 8 times of amount water dissolvings, spray drying, produce seaweed polysaccharide sulfate.
Total amount is made：200 kilograms of sea-tangles obtain 5~6 kilograms of extracts.3~5 kilograms are obtained after spray drying process to do
Powder.Yield 1.5~2.5%.The seaweed polysaccharide sulfate monose composition gas chromatogram of gained is shown in Fig. 1.
The packaging of 1.15 seaweed polysaccharide sulfates
For lucifuge, protection against the tide, the seaweed polysaccharide sulfate after spray drying answers aluminium-plastic sealing to pack.
2. quality testing
Determined by above-mentioned technological process, identification, sediment detection, initial survey, the matter of Zhong Jian tetra- must be passed through in process of production
Examine the outpost of the tax office, identified by seaweed polysaccharide sulfate quality standard and assay, this technique may insure product quality stabilization,
Controllable and uniformity.
Sea-tangle dissolution first during freshwater soaking is non-active component iodine and mannitol, substantially molten in 1 hour
Go out；Active component in neutral hot water also can dissolution quickly, while also can a small amount of non-effective position algin of dissolution.For
This, we devise the extraction of two step water, the analysis of two step alcohol and the molten safety of a step alcohol, science, succinct extraction, separation and purifying
Reparation technology route.
Emphasis is presented below：
Water logging extracts the determination of iodine and mannitol parameter
First by soaking and washing the process of sea-tangle, by non-effective position iodine and mannitol dissolution.From with a collection of cigarette
500 grams of Taiwan Straits domain production complete stool sea-tangle, every part of 3 samples, with 40 times of edible originally water washings and soaks, soaks different by 5 parts
Time, water once (10 minutes do not change water with 20 minutes persons) is changed within 20 minutes, (3 centimetres) are shredded after leaching, extracts seaweed polysaccharide sulfate
Condition it is identical, with SO4 2-Content as monitoring seaweed polysaccharide sulfate quality index, be shown in Table 1.
Influence (n=3) of the kelp soaking wash time of table 1 to seaweed polysaccharide sulfate quality
Hereafter, it is final to determine to the further adjustment of above-mentioned parameter by test of many times：For Shandong Yantai production complete stool sea
Band is soaked 40~50 minutes with 30~40 times of edible clear water, and water is changed once in centre.
Extracting in boiling water
Laboratory test process choice boiling water at atmospheric pressure 6 hours, is extracted in four times, adds the water of 10 times of amounts every time, for the first time
Decoct 2 hours；Second 1.5 hours；1.5 hours for the third time；4th time 1 hour.In pilot scale, decoct points for 3 times totally 5 hours with
Dividing 4 decoctions, the product quality of totally 6 hours and yield have no significant difference.It is final we determined that the extractions of 5 hours 3 times extractions
Technique.
The technique (including seaweed polysaccharide sulfate) for the extracting in boiling water polysaccharide that the traditional handicraft and document of traditional Chinese medicine extraction are recorded, temperature
80 DEG C of degree condition, 90 DEG C, 100 DEG C of decocting times are 4h, 6h, 8h.[referring to Du Bin, Chinese Pharmaceutical Journal volume 2002,37 (9)
p657]。
Be concentrated under reduced pressure parameter
It is concentrated under reduced pressure and (50~60 DEG C, 60~70 DEG C, 70~80 DEG C) is concentrated by triple effect, starts to during relative density 1.10
Alcohol precipitation.
First time alcohol precipitation parameter
Japan's English one that exports to the west removes algin impurity with 20% ethanol, and Anno (1966), Wang Zuoyun etc. are then with 30%
Ethanol, it has been found that do not contain active ingredient with the precipitation obtained by the ethanol of two kinds of concentration, and 30% ethanol is separated out
Precipitation capacity it is more one times, be further added by the concentration of ethanol to 35%, the amount of precipitation has increased slightly, and starts marine alga sulfuric acid occur more
The phenomenon of the co-precipitation of sugar.The final concentration of alcohol that first time alcohol precipitation is determined is 30%.
Second of concentration parameter
Condition is identical with first time, and to save ethanol consumption, this is concentrated into relative density as 1.17.
Second of alcohol precipitation parameter
Extract starts seaweed polysaccharide sulfate precipitation occur in 65% concentration of alcohol.To have precipitated seaweed polysaccharide sulfate
Entirely, experiment determines the concentration of alcohol using 85%, to obtain the crude product of the maximum amount of seaweed polysaccharide sulfate, can save ethanol again
Dosage.
Purify parameter
Crude product seaweed polysaccharide sulfate is washed with the ethanol of low concentration.Determined after experiment, with 75% ethanol clean result
Most preferably, 2 are shown in Table.
The ethanol purification seaweed polysaccharide sulfate of table 2 is tested
The rate of transform
Total reducing sugar, total protein and the sulfate content of sea-tangle and corresponding finished product seaweed polysaccharide sulfate to scale up test are carried out
Detection, is as a result listed in table 3.
The sea-tangle of table 3 and seaweed polysaccharide sulfate (%) result containing survey and the rate of transform (n=3)
Finally it should be noted that：The above embodiments are merely illustrative of the technical solutions of the present invention, rather than its limitations；Although
The present invention is described in detail with reference to the foregoing embodiments, it will be understood by those within the art that：It still may be used
To be modified to the technical scheme described in foregoing embodiments, or equivalent substitution is carried out to which part technical characteristic；
And these modification or replace, do not make appropriate technical solution essence depart from various embodiments of the present invention technical scheme spirit and
Scope.
Claims (10)
1. a kind of preparation method of seaweed polysaccharide sulfate, it is characterised in that comprise the following steps：
S10, by sea-tangle be placed in the water of 38-42 times of its own quality soak 35-45 minutes after, for its own quality
The 38-42 times of water measured is rinsed well；
S20, sea-tangle is pre-processed, reject the unqualified sea-tangle that leaf is yellow, leaf is broken and mouldy, qualified sea-tangle is cut into area
The fragment of 7-10 square centimeters；
S30, sea-tangle be added to decocted three times in the water of 9-11 times of its own quality, respectively 1.9-2.1 hours,
1.4-1.6 hours, 1.4-1.6 hours；
S40, decoction liquor it will be concentrated under reduced pressure into relative density 1.05-1.15 after merging three times and obtain concentrate I；
S50, ethanol is added to ethanol volumetric concentration 29-31% into concentrate I, supernatant and heavy is centrifuged to obtain after standing overnight
Slag；
S60, sediment is detected, if SO in sediment4 2-Weight/mass percentage composition be more than 5%, ethanol is added into sediment to second
Alcohol volumetric concentration 29-31%, stand and centrifuge to obtain supernatant to after being layered；If SO in sediment4 2-Weight/mass percentage composition be less than etc.
In 5%, sediment is abandoned；
S70, by step S50 and step S60 supernatant merge, be concentrated under reduced pressure into relative density 1.1-1.2 and obtain concentrate
II；
S80, ethanol is added to ethanol volumetric concentration 83-86% into concentrate II, after standing overnight, abandon supernatant, precipitation is used
Centrifuged after percentage by volume 74-76% ethanol washing；
S90, carry out being spray-dried to obtain seaweed polysaccharide sulfate after the precipitation after centrifugation is dissolved with the 7-9 times of water measured.
2. the preparation method of seaweed polysaccharide sulfate according to claim 1, it is characterised in that put sea-tangle in step S10
Enter after being soaked 40 minutes in the water of 40 times of its own quality, the water for 40 times of amounts of its own quality is rinsed well.
3. the preparation method of seaweed polysaccharide sulfate according to claim 1, it is characterised in that in step S20, by qualified sea
Band is cut into the fragment of 9 square centimeters of area.
4. the preparation method of seaweed polysaccharide sulfate according to claim 1, it is characterised in that in step S30, by sea-tangle plus
Enter into the water in its own quality 10 and decoct three times, respectively 2 hours, 1.5 hours, 1.5 hours.
5. the preparation method of seaweed polysaccharide sulfate according to claim 1, it is characterised in that in step S40, will decoct three times
Boil liquid is concentrated under reduced pressure into relative density 1.1 and obtains concentrate I after merging.
6. the preparation method of seaweed polysaccharide sulfate according to claim 1, it is characterised in that in step S50, to concentrate
Ethanol is added in I to ethanol volumetric concentration 30%, supernatant and sediment are centrifuged to obtain after standing overnight.
7. the preparation method of seaweed polysaccharide sulfate according to claim 1, it is characterised in that in step S60, if in sediment
SO4 2-Weight/mass percentage composition be more than 5%, ethanol is added into sediment to ethanol volumetric concentration 30%, stands to after being layered and centrifuges
Obtain supernatant.
8. the preparation method of seaweed polysaccharide sulfate according to claim 1, it is characterised in that in step S70, by step
Supernatant in S50 and step S60 merges, and is concentrated under reduced pressure into relative density 1.17 and obtains concentrate II.
9. the preparation method of seaweed polysaccharide sulfate according to claim 1, it is characterised in that in step S80, to concentrate
Ethanol is added in II to ethanol volumetric concentration 85%, after standing overnight, abandons supernatant, the ethanol of precipitation percentage by volume 75%
Centrifuged after washing.
10. the preparation method of seaweed polysaccharide sulfate according to claim 1, it is characterised in that in step S90, will centrifuge
Precipitation afterwards carries out being spray-dried to obtain seaweed polysaccharide sulfate after being dissolved with the water of 8 times of amounts.
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| CN105963325A (en) * | 2016-05-06 | 2016-09-28 | 中国人民解放军军事医学科学院放射与辐射医学研究所 | Application of sulfated polysaccharides from seaweed |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1279953A (en) * | 2000-08-04 | 2001-01-17 | 中国人民解放军军事医学科学院放射医学研究所 | Application of seeweed polyose sulphate |
| CN103239472A (en) * | 2013-05-10 | 2013-08-14 | 首都医科大学 | Novel medicinal application of fucoidan polysaccharide sulfate |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1279953A (en) * | 2000-08-04 | 2001-01-17 | 中国人民解放军军事医学科学院放射医学研究所 | Application of seeweed polyose sulphate |
| CN103239472A (en) * | 2013-05-10 | 2013-08-14 | 首都医科大学 | Novel medicinal application of fucoidan polysaccharide sulfate |
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