CN1158308C - Seaweed polyose product and its preparing method and use - Google Patents

Seaweed polyose product and its preparing method and use Download PDF

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CN1158308C
CN1158308C CNB011146036A CN01114603A CN1158308C CN 1158308 C CN1158308 C CN 1158308C CN B011146036 A CNB011146036 A CN B011146036A CN 01114603 A CN01114603 A CN 01114603A CN 1158308 C CN1158308 C CN 1158308C
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seaweed
preparation
product
polysaccharide
scpg
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CN1377896A (en
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钟红茂
赵迪
刘锡金
于嘉陵
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South China Sea Institute of Oceanology of CAS
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South China Sea Institute of Oceanology of CAS
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Abstract

The present invention provides a seaweed polysaccharide product and a preparation method thereof. The product is obtained by separation and extraction of usual seaweed of microsporaceae of oscillatoriaceae of dictyosiphoniales of phaeophyta in marine plants. The molecular weight of the product is from 5000 to 20000 daltons. The solid product is brown or brownish black. Monosaccharide of the product mainly contains 90 to 95% of rhamnose and fucose and 10 to 5% of arabinose and glucose. The product can be developed into novel antiarrhythmic medicine. A preparation method is reliable and is easily operated, the present invention is suitable for large-scale preparation, the environment is not polluted, and the cost is low.

Description

A kind of seaweed polyose product and its production and use
The present invention relates to a kind of seaweed polyose product that extracts from the marine plant algae and its production and use.
The common marine alga of Chnoosporaceae (Chnoospora) (Chnoospora spp.) extensively is distributed in the coastal marine site of China, aboundresources, relevant is that the pharmacology of the raw material polysaccharose substance that separates, extract and the research of chemical aspect have not yet to see report with this marine alga.Thereby explore the preparation method who from this marine alga raw material, extracts its effective medicinal components and be suitable for industrial application, so that this marine algae resource is fully utilized, have important value.
The objective of the invention is to propose a kind of from the marine alga raw material new product and preparation method thereof of the Sargassum polysaccharides of institute's separation and Extraction, and the new purposes of this material as antiarrhythmic drug proposed.
The seaweed polyose product that the present invention produced is the phaeophyta (Phaeophyta) from marine plant, Dictyosiphoniales (Dictgosiphonales), Scytosiphonales (Scytosiphonaceae), separation and Extraction gained in the common marine alga of Chnoosporaceae (Chnoospora) (Chnoospora spp.), its molecular weight is between the 5000-20000 dalton, solid-stately be brown or brownish black, chromatograph, its monose composition is mainly: rhamnosyl and the Fucose of 90-95% (weight), pectinose and the glucose of 10-5% (weight).
The step of preparation process of these goods is as follows:
(1) will extract with water boiling and extraction or with the direct boiling of dry seaweed behind the dry seaweed weeding of grease soluble substance;
(2) will go up step decoction thing and cross leaching filtrate;
(3) filtrate of obtaining is passed through ion exchange column or the desalination of ionodialysis method;
(4) more earlier after anionite-exchange resin and Zeo-karb intercept to such an extent that molecular weight is the daltonian polysaccharide of 1000-20000, separate, purify to such an extent that molecular weight is the daltonian polysaccharide of 5000-20000 with high pressure liquid chromatography at last;
(5) above-mentioned gains are concentrated after, oven dry is pulverized, or directly with the enriched material spraying drying, is promptly got described seaweed polyose product.
Said the method for extracting behind the dry seaweed weeding of grease soluble substance can be adopted the technology that may further comprise the steps:
(1) with the aqueous solution soaking of dry seaweed with non-fat-soluble solvent;
(2) soak is filtered separately slag and filtrate;
(3) the algae-residue boiling is extracted repeatedly.
Said non-fat-soluble solvent can be methyl alcohol, ethanol, Virahol, butanols or acetone.
The said anionite-exchange resin of the present invention can adopt 701 or weak base 330.
The said Zeo-karb of the present invention can adopt 724 or weak acid 101.
Result according to pharmacology, pharmacodynamic study shows that the prepared Sargassum polysaccharides product of the present invention has tangible effect to cardiovascular systems, especially can resist multiple drug-induced irregular pulse animal model, can be developed into the novel anti antiarrhythmic medicament.
The present invention adopts modern scientific method, and the medicinal ingredients in the described marine alga raw material is separated, purifies, and prepared this Sargassum polysaccharides product safety is nontoxic, the reliable easily row of its preparation method, and suitable for mass production, free from environmental pollution, cost is low.
The antiarrhythmic effect that the present invention is good proves through experimentation on animals, below is its main pharmacodynamics experimental result:
Test of pesticide effectiveness result
I. sample source: the Sargassum polysaccharides that Chinese Academy of Science Nanhai Ocean Research Institute provides (being called for short SCPG) product.
II. laboratory animal: NIH pure lines small white mouse and SD rat are provided by Guangdong Province's medical animal feedlot.Cavy is provided by the military region, Guangzhou military medicine institute.
III. test method and result:
(1) chloroform is brought out the antagonistic action of quivering in the mouse chamber
The NIH mouse is some, and body weight is 18-22g, male and female half and half, 20 every group.Give the SPA polysaccharide soln of animal ip or iv doses, drop into behind the 5min in the inverted culture dish of 500ml beaker (in the rayon balls that contains the 2ml chloroform is arranged, whenever change an animal later on and add the 0.5ml chloroform), after treating that animal breath stops, chest is cutd open in taking-up immediately and whether visual inspection ventricular fibrillation takes place.Blank group ip or iv equal-volume physiological saline, positive controls adopts the verapamil of 5mg/kg dosage and the lignocaine of 15mg/kg dosage.Calculate direct probability with " the accurate test of significance table of four fold table ", judge significance.Test-results sees Table 1, table 2.
Table 1.SCPG polysaccharide (ip) brings out the antagonistic action of quivering in the mouse chamber to chloroform
Several chambers percentage that quivers quivers in dosage number of mice chamber
Mg/kg (only) (only) %
The physiological saline group--20 18 90
SCPG organizes 17.05 20 10 50*
SCPG organizes 34.10 20 8 40*
SCPG organizes 51.10 20 6 30*
Verapamil group 5.00 20 2 20*
* compare p＜0.01 with the blank group
Table 2.SCPG polysaccharide (iv) brings out the antagonistic action of quivering in the mouse chamber to chloroform
Several chambers percentage that quivers quivers in dosage number of mice chamber
Mg/kg (only) (only) %
The physiological saline group--20 18 90
SCPG organizes 2.50 20 8 40*
SCPG organizes 3.75 20 6 30*
SCPG organizes 5.00 20 4 20*
Lignocaine group 15.00 20 6 30*
* compare p＜0.01 with the blank group
Test-results shows, the blank group chamber incidence of quivering is 90%, and incidence and blank group are quivered relatively in the chamber of the various dose group of two kinds of different way of administration ip and iv, p＜0.01 illustrates that mouse chamber that the SCPG polysaccharide of these two kinds of route of administration various dose brings out for chloroform quivers and have significant antagonistic action.In addition, SCPG polysaccharide and its result of positive controls comparison also show, SCPG polysaccharide group than the effect of verapamil group a little less than, but the effect of many cacaines of Billy group is strong.
(2) the SCPG polysaccharide brings out the antagonistic action of quivering in the mouse chamber to bariumchloride
It is some to get mouse, body weight 18-22g, male and female half and half, random packet, 20 every group.The SPA polysaccharide soln of tail vein injection 2.50mg/kg, 3.75mg/kg, 5.00mg/kg dosage, control group is injected isopyknic physiological saline, the BaCl of tail vein injection 0.75mg/kg behind the 2min 2Solution draws vertebra to put to death immediately after the administration, cut open brisket and observe to examine whether ventricular fibrillation takes place.Calculate direct probability with " the accurate test of significance table of four fold table ", judge its significance.The results are shown in Table 3.
Table 3.SCPG polysaccharide is to BaCl 2Bring out the antagonistic action of quivering in the mouse chamber
Several chambers percentage that quivers quivers in dosage number of mice chamber
Group
Mg/kg (only) (only) %
The physiological saline group--20 20 100
SCPG organizes 2.50 20 10 50*
SCPG organizes 3.75 20 9 45*
SCPG organizes 5.00 20 8 40*
Lignocaine group 15.00 20 6 30*
* compare p＜0.01 with the blank group
Test-results shows, the physiological saline control group chamber incidence of quivering is 100%, and the administration group is respectively 50%; 45%; 40%, compare with the blank group, its p value is all less than 0.01.Illustrate that three SCPG polysaccharide dosages are for BaCl 2Quivering in the mouse chamber of bringing out has tangible antagonistic action.
(3) the SCPG polysaccharide brings out the antagonistic action of big white mouse ventricular arrhythmia to bariumchloride
Get some of body weight 200-300g SD big white mouse, be divided into four groups at random, 10 every group.Fixing with urethane intraperitoneal anesthesia (1.2g/kg) layback position, after tracing normal ECG II and leading, control group is injected the 10%BaCl of 6mg/kg by femoral vein 2Solution is observed electrocardioscope, and ARR time and time length appear in record.The medication group is injected 2.50mg/kg by femoral vein immediately after obvious irregular pulse occurring, the SCPG polysaccharide antagonism of 3.75mg/kg or 5.00mg/kg dosage, and observe and can recover sinus rhythm and time of recovery, relatively obtain the P value with control group with t value method.The results are shown in Table 4.
Table 4.SCPG polysaccharide is (iv) to BaCl 2Bring out the ARR effect of big white mouse (x ± SD)
When dosage number of animals irregular pulse continues
(mg/kg) (min) between (only)
The blank group--10 176 ± 31
SCPG organizes 2.50 10 123 ± 32*
SCPG organizes 3.75 10 76 ± 37**
SCPG organizes 5.00 10 58 ± 33***
Lignocaine group 15.00 10 71 ± 35**
Compare * p＞0.2, * * p＜0.05, * * * p＜0.02 with the blank group
Test-results shows that three dosage 2.50mg/kg of marine alga SCPG polysaccharide, 3.75mg/kg, 5.00mg/kg compared with the blank group the ARR time length, and the p value of first dosage is greater than 0.05, and illustrating does not have significant difference.The p value that the back is two groups is less than 0.05, and the prompting dosage is that 3.75mg/kg and 5.00mg/kg bring out the antagonistic action that the rat ventricular arrhythmia has highly significant to bariumchloride.Test-results shows that also the effect of the many cacaines of SCPG polysaccharide Billy is strong.
(4) the SCPG polysaccharide brings out the ARR antagonistic action of cavy to ouabain
Get some of 350-400g healthy guinea pigs, be divided into three groups at random, 5 every group, with urethane intraperitoneal anesthesia (1.2g/kg), anesthesia layback position is fixing, cuts right leg and skin of neck.The right lateral thigh venous cannula connects the constant speed syringe that fills 0.03% ouabain.Normal ECG II leads before tracing experiment.The administration group is injected 3.12mg/kg by jugular vein, 6.25mg/kg or the SCPG polysaccharide of 12.50mg/kg dosage, the blank group is injected the physiological saline of equivalent, then with 3 μ g/min speed instillation ouabains, use the electrocardioscope watch-keeping, and the ouabain dosage when record ventricular premature contraction (VP), ventricular tachycardia (VT), ventricular fibrillation (VF) and heartbeat occur and stops (CA) respectively, relatively obtain the P value with control group with t value method.The results are shown in Table 5 and accompanying drawing 1.
Table 5.SCPG polysaccharide to ouabain bring out the ARR antagonistic action of cavy (x ± SD, n=5)
Required ouabain dosage (μ g/kg) appears in all kinds of irregular pulse
Chamber property early chamber property is crossed ventricle and is quivered to jump and stop
(VP) speed (VT) of fighting moving (VF) (CA)
Physiology salt 147 ± 25 180 ± 24 221 ± 22 272 ± 26
The water control group
12.50mg 225±12** 260±20*** 316±37*** 381±20***
/ kg dosage group
6.25mg/ 183±33* 224±33* 292±39*** 348±26***
Kg dosage group
3.12mg/ 157±24* 206±16* 266±26* 332±23**
Kg dosage group
Compare * p＜0.1, * * p＜0.05, * * * p＜0.01 with the blank group.
Test-results shows, 2 effects that dosage is 12.50mg/kg and 6.25mg/kg of marine alga SCPG polysaccharide, compare with the blank group, the P value is all less than 0.05 or 0.01, illustrate that cavy irregular pulse that the SCPG polysaccharide of this dosage brings out ouabain all has significantly or antagonistic action, the especially ouabain of highly significant bring out cavy ventricular fibrillation and heartbeat are stopped to have clearly antagonistic action.Its effect is strengthened along with the increase of dosage, presents tangible dose-effect relationship.
Conclusion
From test-results, marine alga SCPG polysaccharide can resist multiple drug-induced irregular pulse, for example to chloroform, BaCl 2Quiver in the mouse chamber of bringing out, to BaCl 2The cavy ventricular premature contraction (VP) that rat ventricular arrhythmia that brings out and ouabain bring out, chamber property is overrun (VT), and ventricular fibrillation (VF) and heartbeat stop (CA) all significant antagonistic action.Can affirm the drug effect of its antiarrhythmic effect according to top test-results, prompting marine alga SCPG polysaccharide has the prospect that well is developed to Antiarrhythmic Agent.
Fig. 1 brings out the ARR interactively of cavy for the SCPG polysaccharide antagonism ouabain of various dose.
Embodiment one:
Get dry seaweed (phaeophyta Phaeophyta, Dictyosiphoniales Dictgosiphonales, Scytosiphonales Scytosiphonaceae, the common marine alga Chnoospora spp. of Chnoosporaceae Chnoospora) 2 kilogram, pulverize the back or do not pulverize, 2 times of amounts of ethanol of adding 95%, soaked 24 hours, after the filtration, the algae-residue boiling is extracted 5 times, filters out filtrate, filtrate is by the ion exchange column desalination, be the daltonian polysaccharide of 1000-20000 with 701 anionite-exchange resin and 724 Zeo-karbs intercepting molecular weight again, separate with high pressure liquid chromatography at last, purify to such an extent that molecular weight is the daltonian polysaccharide of 5000-20000, then in oven dry below 70 ℃, pulverize the oven dry thing, promptly get brown Sargassum polysaccharides.
Embodiment two:
With the former algae of described dry seaweed or after pulverizing, claim 2 kilograms, add the about 2 times of amounts of 95% methyl alcohol, soak 24 times, after the filtration, the filter residue boiling, extract 3 times, filter out filtrate, filtrate is the daltonian polysaccharide of 1000-20000 with 701 anionite-exchange resin and 724 Zeo-karbs intercepting molecular weight with the desalination of ionodialysis method again, separate, purify to such an extent that molecular weight is the daltonian polysaccharide of 2000-20000 with high pressure liquid chromatography at last, in oven dry below 70 ℃, pulverize the oven dry thing then, promptly get brown Sargassum polysaccharides.
Embodiment three:
Claim 2 kilograms with described former dry seaweed, directly boiling, extract 5 times, filter out filtrate, filtrate is the daltonian polysaccharide of 1000-20000 with weak base 330 anionite-exchange resin and weak acid 101 Zeo-karbs intercepting molecular weight by the ion exchange column desalination again, separate, purify to such an extent that molecular weight is the daltonian polysaccharide of 5000-20000 with high pressure liquid chromatography at last, in oven dry below 70 ℃, pulverize the oven dry thing then, promptly get brown Sargassum polysaccharides.

Claims (7)

1. seaweed polyose product, it is characterized in that the phaeophyta from marine plant, Dictyosiphoniales, Scytosiphonales, separation and Extraction gained in the marine alga of Chnoosporaceae, its molecular weight are between the 5000-20000 dalton, solid-stately are brown or brownish black, its monose composition is mainly: rhamnosyl and the Fucose of 90-95% (weight), pectinose and the glucose of 10-5% (weight).
2. the preparation method of the described Sargassum polysaccharides of claim 1 is characterized in that comprising the steps:
(1) will extract with water boiling and extraction or with the direct boiling of dry seaweed behind the dry seaweed weeding of grease soluble substance;
(2) will go up step decoction thing and cross leaching filtrate;
(3) filtrate of obtaining is passed through ion exchange column or the desalination of ionodialysis method;
(4) more earlier after anionite-exchange resin and Zeo-karb intercept to such an extent that molecular weight is the daltonian polysaccharide of 1000-20000, separate, purify to such an extent that molecular weight is the daltonian polysaccharide of 5000-20000 with high pressure liquid chromatography at last;
(5) above-mentioned gains are concentrated after, oven dry is pulverized, or directly with the enriched material spraying drying, is promptly got described seaweed polyose product.
3. the preparation method of Sargassum polysaccharides according to claim 2 is characterized in that said step with water boiling and extraction behind the dry seaweed weeding of grease soluble substance being comprised:
(1) with the aqueous solution soaking of dry seaweed with non-fat-soluble solvent;
(2) soak is filtered separately slag and filtrate;
(3) the algae-residue boiling is extracted repeatedly.
4. the preparation method of Sargassum polysaccharides according to claim 3 is characterized in that said non-fat-soluble solvent is methyl alcohol, ethanol, Virahol, butanols or acetone.
5. the preparation method of Sargassum polysaccharides according to claim 2 is characterized in that said anionite-exchange resin is 701 or weak base 330.
6. the preparation method of Sargassum polysaccharides according to claim 2 is characterized in that said Zeo-karb is 724 or weak acid 101.
7. the described seaweed polyose product of claim 1 is as the application of preparation antiarrhythmic drug.
CNB011146036A 2001-04-05 2001-04-05 Seaweed polyose product and its preparing method and use Expired - Fee Related CN1158308C (en)

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Publication number Priority date Publication date Assignee Title
CN1314711C (en) * 2004-03-27 2007-05-09 中国科学院水生生物研究所 Method for extracting Gexianmi amylose from nostoc
CN1322015C (en) * 2004-03-27 2007-06-20 中国科学院水生生物研究所 Method for extracting amylose of desert algae
CN1304428C (en) * 2004-03-27 2007-03-14 中国科学院水生生物研究所 Method for extracting polysaccharide by using blooms blue algae
CN1313499C (en) * 2004-09-21 2007-05-02 珠江医院 Sargassum polysaccharide, its preparation method and use
CN105647229B (en) * 2016-01-21 2017-08-29 王君华 A kind of state of aggregation natural dye based on marine alga gel and preparation method thereof

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