CN1267456C - Process for preparing glucose-reducing sand sagebrush polysaccharide and its use - Google Patents
Process for preparing glucose-reducing sand sagebrush polysaccharide and its use Download PDFInfo
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- CN1267456C CN1267456C CN 200510002228 CN200510002228A CN1267456C CN 1267456 C CN1267456 C CN 1267456C CN 200510002228 CN200510002228 CN 200510002228 CN 200510002228 A CN200510002228 A CN 200510002228A CN 1267456 C CN1267456 C CN 1267456C
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Abstract
The present invention relates to sand sagebrush polysaccharide, and a preparation method and an application thereof. Waste slag formed by degreasing the seeds of the sand sagebrush is used as a raw material which is processed by extraction, separation, enzymolysis and column chromatography is prepared into the sand sagebrush polysaccharide. The present invention is characterized in that 50 to 80 % of alcohol is used for extracting, chloroform and normal butyl alcohol mixed liquor is used for extracting, papain is used for carrying out enzymolysis, and sephadex G-200 is used for carrying out column chromatography; thus, the sand sagebrush polysaccharide is prepared. The sand sagebrush polysaccharide is used for treating chemical diabetes caused by alloxan, reducing the damage of diabetes to the liver and kidney, decreasing content of glutamic oxalacetic transaminase enzyme, serum glutamic pyruvic transaminase and urea nitrogen in blood, and inhibiting the y weight reduction caused by diabetes. The sand sagebrush polysaccharide has the advantages of simple production process, good blood sugar lowering effect, no toxic side effect, easy use and good treating effect on diabetes.
Description
Technical field
The present invention relates to a kind of waste residue of sloughing behind the oil with natural sand sagebrush (Artemisia filifolia) seed is raw material, through the artemisia desertorum polysaccharide preparation method and the application thereof with blood sugar reducing function of extraction, separation, enzymolysis, column chromatography preparation.
Background technology
Diabetes are that a kind of what be associated with inherited genetic factors and multiple environmental factors is the metabolism disturbance syndrome of feature with the chronic hyperglycemia, are the common diseases of the serious harm human health in a kind of global range.In recent years, global diabetes prevalence and mortality ratio all present a progressively ascendant trend, become that world today's medical circle needs to be resolved hurrily and one of the emphasis problem of capturing.Antidiabetic drug commonly used is the preparation based on synthetic drugs, and the liver kidney is had certain damage, and is not easy for patients to accept.
Patent JP95/00560 has applied for having hypoglycemic activity new pyridine type thiazolidine compound and salt thereof; Patent CN86104075 (86104075) has applied for hypoglycemic drug--the preparation method of thiazolidinediones, they treatment in the hyperglycemia animal application method and the formula of medicine that contains them; Patent CN1252273 (99121819.1) has applied for the preparation and the application of sulfourea hypoglycemic agents-seleno-methionine chrome yellow urea as hypoglycemic of new generation; Patent CN1465342 (02112414.0) has applied for a kind of thiazolidine salt derivative hypoglycemic medicine; CN1113491 (95102055.2) has the new hydroxylamine compound of hypoglycemic activity by suitable 1-(2-, 3-or 4-oil of mirbane sulphonyl) the derivative preparation of selective reduction.
Patent US20030072822 has applied for a kind of method of tarragon ethanol extraction treatment diabetes; Patent US20040151783 has applied for a kind of forest Gymnema latifolium Wall. ex Wight polysaccharide component with hypoglycemic activity; But patent US20030054015 has applied for a kind of composition of essence of exciting sympathetic nerves, and wherein Oil of grapefruit has blood sugar decreasing effect to the diabetic subject.Patent US20030198697 has applied for a kind of fatty acid component that extracts from Job's tears, have hypoglycemic activity.The health-caring product capable of reducing blood sugar or the medicine of a kind of high-efficient pure natural that contains hypoglycemic effective constituent that patent CN1188651 (97100285.1) extracts from wild Graptopelaum Paraguayense; Patent CN1151303 (95117499.1) has applied for that a kind of is that raw material extracts novel process hypoglycemic and nutritive substance with plants such as Crassulaceae Crassula argentea Thunb; Patent CN1137411 (95105954.8) has applied for a kind of by the 12 kinds of hypoglycemic Chinese medicine of rehabilitation pancreas islet " rehabilitation pancreas islet Hypolycemic pellet " and making methods that Chinese medicine is formed such as genseng, gekkos; Patent CN1401318 (02138749.4) extracts, separates efficient part and effective constituent with hypoglycemic activity from the Chinese medicine Winged Euonymus Twig, comprise terpene, phenolic acids etc., has the blood glucose in diabetic mice of falling effect, influence mda and superoxide-dismutase simultaneously, can alleviate or suppress the generation and the development of diabetic complication; Patent CN1456337 (03128089.7) has applied for a kind of balsam pear goods and preparation method who contains a plurality of blood-sugar decreasing actives; Patent CN1306752 (00101247.9) discloses a kind of prescription and production technique with edible and healthy biscuit one Baile biscuit with dietary fiber of hypoglycemic activity.This biscuit mainly contains flour, food fibre, active polysaccharide, Chinese yam, balsam pear etc., develops through scientific approach.
Patent PCT/KR98/00325 applied for a kind of as Hypoylycemic agents as the Vitamin P complex of molecular formula (I) or contain the purposes of reduction blood glucose level mammals of the plant milk extract of same Vitamin P complex; Patent CN1348762 (00116167.9) is with the medicament of kuh-seng flavones preparation treatment diabetes and treat the combination drug of diabetes with at least a preparation in kuh-seng flavones and astragalus polysaccharides, panaxan, genseng general glycoside, Radix Ophiopogonis polysaccharide, THFPS, the mulberry leaf total polysaccharides effective constituent; Patent CN1389146 (02138749.4) is with the Cyclocarya paliurus extract, be equipped with flavour agent, auxiliary substance, and add in sodium alginate, carboxymethyl cellulose, agar, the xanthan gum two or more material as stablizer, hypoglycemic, hypotensive, the blood fat reducing healthcare food that make; Patent CN1245060 (98110398.7) is (1) L-carnitine, (2) chitosan, and (3) rich chromium microbial polysaccharide part is by the preparation method of certain formula combination at a kind of high-efficient fat reducing and hypoglycemic medicine or protective foods; Patent CN1398630 (02136740.X) extracts saccharide complex, its production method and the purposes with hypoglycemic activity from the Chinese medicinal materials bighead atractylodes rhizome.Patent does not all relate to preparation method and the application that artemisia desertorum polysaccharide is the hypoglycemic natural product of raw material both at home and abroad.
Summary of the invention
The waste residue that the objective of the invention is to slough behind the oil with sand sagebrush (Artemisia filifolia) is a raw material, a kind of natural energy lowering blood glucose through preparations such as extraction, separation, enzymolysis, column chromatographies, and can reduce that diabetes cause the damage of liver, kidney is promptly reduced the content of glutamic-oxal(o)acetic transaminase in the blood, gpt, blood urea nitrogen, and suppress diabetes and cause natural biological polysaccharide artemisia desertorum polysaccharide that loses weight and its production and application.
Step of the present invention is as follows,
Step 1: the sand sagebrush (Artemisia filifolia) seed is through CO
2After supercritical extraction was sloughed grease, water fully dissolved, and the ratio of raw material and water is 1: 30-50, in the aqueous solution, add ethanol, and that ethanol content reaches is static behind the 50-80%, spend the night, centrifugal, collecting precipitation, vacuum-drying gets substance A;
Step 2: the substance A that takes by weighing certainweight, water dissolution with 80-100 times of weight, in the aqueous solution, add papoid and carry out enzymolysis, use 1 again: 2-4 chloroform and propyl carbinol mixed solution extract, collect that the water layer solution decompression concentrates, dry substance B, the A aqueous solution and extraction liquid ratio by volume are that 4: 1, papoid consumption are the 24.00U/2000ml aqueous solution;
Step 3: substance B is passed through sephadex G-200 column chromatography, elutriant is: 0.05molNacl solution, flow velocity: 8ml/h, every part of 1.5ml of elutriant behind the collection chromatography, put into the dialysis tubing 36-48h that dialyses behind the elutriant concentrating under reduced pressure, dialyzate add 2-3 doubly measure ethanol static, spend the night, centrifugal, collecting precipitation, vacuum-drying gets pure sand sagebrush (Artemisia filifolia) natural seed biological polyoses.
Molecular weight, chemical constitution: molecular weight Mn is about 1.42 * 10
5G/mol, the sharp coefficient of second dimension is 3.64 * 10
-5Cm
3Mol/g
2Be made up of six kinds of monose such as L-arabinose, D-wood sugar, D-lyxose, D-seminose, D-glucose, D-semi-lactosis, mol ratio is 0.14: 0.09: 1.51: 0.17: 0.07: 1.45, and white powder.
This natural biological polysaccharide is mainly used in the therapeutical chemistry diabetes, and can reduce diabetes the warps such as damage of liver, kidney are shown the rat pharmacological evaluation: feeding said preparations in 30 days makes the blood sugar of hyperglycemia animal reduce by 13.98%, and gpt (GPT) reduces by 25.48%, glutamic-oxal(o)acetic transaminase (GOT) reduces by 28.7%, urea nitrogen content reduces by 51.27%, significantly slow down the weight loss that diabetes cause.
Characteristics of the present invention: the waste residue that the present invention sloughs behind the oil with sand sagebrush (Artemisia filifolia) is a raw material, a kind of natural energy lowering blood glucose through preparations such as extraction, separation, enzymolysis, column chromatographies, and can reduce diabetes the damage of liver, kidney is promptly reduced the content of glutamic-oxal(o)acetic transaminase in the blood, gpt, blood urea nitrogen, and can suppress diabetes and cause the preparation that loses weight.Compare with other antidiabetic drug that to have production technique simple, hypoglycemic effect is good, has no side effect, and is easy to use, and I type, type ii diabetes etc. are all had curative effect preferably.
Embodiment
Embodiment 1:
The preparation method of hypoglycemic natural product and application thereof in the sand sagebrush (Artemisia filifolia) seed
Step 1: the sand sagebrush (Artemisia filifolia) seed is through CO
2The part water that supercritical extraction is sloughed behind the grease fully dissolves (ratio of raw material and water is 1: 30), in the aqueous solution, add ethanol (ethanol content reaches 50%) static, spend the night, centrifugal, collecting precipitation vacuum-drying gets substance A;
Step 2: take by weighing the water dissolution of the substance A of certainweight with 80 times of weight, in water dissolution, add papoid and carry out enzymolysis, use chloroform and propyl carbinol (1: 2) mixed solution to extract again, collect concentrated, the dry substance B that gets of water layer solution decompression, the A aqueous solution and extraction liquid ratio by volume are 4: 1, and the papoid consumption is the 24.00U/2000ml aqueous solution;
Step 3: (elutriant is: 0.05molNacl solution by sephadex G-200 column chromatography with substance B, flow velocity: 8ml/h, every part of 1.5ml of elutriant behind the collection chromatography), put into the dialysis tubing 36h that dialyses behind the elutriant concentrating under reduced pressure, dialyzate add 2 times of amount ethanol static, spend the night, centrifugal, collecting precipitation vacuum-drying gets pure hypoglycemic natural product.
Blood sugar reducing function: the chemotype diabetes rat that tetraoxypyrimidine causes takes after this product that blood sugar, gpt, glutamic-oxal(o)acetic transaminase, blood urea nitrogen all reduce result such as following table in the blood:
| Control group | The hyperglycemia group | Hyperglycemia group+this product | ||
| Blood sugar (m mol/L) | 3.77 | 5.71 | 4.48 | Reduce by 13.98% |
| Gpt (I μ/L) | 27.0375 | 77.025 | 51.5136 | Reduce by 25.48% |
| Glutamic-oxal(o)acetic transaminase (I μ/L) | 146.375 | 230 | 165 | Reduce by 28.7% |
| Blood urea nitrogen (m mol/L) | 7.0225 | 26.7425 | 13.0136 | Reduce by 51.27% |
Body weight change: every group of weight average value seen (table 1), the control group weight increase, hyperglycemia group body weight obviously reduces, reduced rate is 7.69%, and the diabetes rat whose body weight of feeding pure natural artemisia desertorum polysaccharide group slightly alleviates, reduced rate is 1.56%, illustrates that it has certain restraining effect to alleviating of hyperglycemia whose body weight.
Rat body weight (g/ only) is respectively organized in table 1 experiment
| Fate | 1 | 30 days | Growth tendency |
| The hyperglycemia group | 260.1 | 240.1 | -7.69％ |
| Hyperglycemia+artemisia desertorum polysaccharide | 269.2 | 265.5 | -1.56％ |
| Control group | 266.2 | 289 | +8.56％ |
Utilize sound attitude light scattering apparatus, gas-chromatography, infrared spectra, nucleus magnetic resonance etc. to characterize the molecular weight and the chemical constitution of pure hypoglycemic natural product.
Sound attitude light scattering apparatus, gas-chromatography confirm: pure hypoglycemic natural product weight-average molecular weight Mn is about 1.42 * 10
5G/mol, the sharp coefficient of second dimension is 3.64 * 10
-5Cm
3Mol/g
2Be made up of six kinds of monose such as L-arabinose, D-wood sugar, D-lyxose, D-seminose, D-glucose, D-semi-lactosis, mol ratio is 0.14: 0.09: 1.51: 0.17: 0.07: 1.45, and white powder.
Glucose (Glu) is measured with the continuous detecting method respectively with glutamic-oxal(o)acetic transaminase (GOT) with determination of glucose oxidase, gpt (GPT), and blood urea nitrogen (BuN) uses indexs such as end-point method mensuration.Instrument uses automatic clinical chemistry analyzer, OLYcupus AU400 type (Japan).Tetraoxypyrimidine is available from sigma company product.
Take blood sugar, gpt, glutamic-oxal(o)acetic transaminase, the urea nitrogen content of the chemotype diabetic animal that tetraoxypyrimidine causes are all reduced, show that hypoglycemic effect is better.
Embodiment 2:
The preparation method of hypoglycemic natural product and application thereof in the sand sagebrush (Artemisia filifolia) seed
Step 1: the sand sagebrush (Artemisia filifolia) seed is through CO
2The part water that supercritical extraction is sloughed behind the grease fully dissolves (ratio of raw material and water is 1: 50), in the aqueous solution, add ethanol (ethanol content reaches 80%) static, spend the night, centrifugal, collecting precipitation vacuum-drying gets substance A;
Step 2: take by weighing the water dissolution of the substance A of certainweight with 100 times of weight, in water dissolution, add papoid and carry out enzymolysis, use chloroform and propyl carbinol (1: 4) mixed solution to extract again, collect that the water layer solution decompression concentrates, dry substance B, the A aqueous solution and extraction liquid ratio by volume are that 4: 1, papoid consumption are the 24.00U/2000ml aqueous solution;
Step 3: (elutriant is: 0.05molNacl solution by sephadex G-200 column chromatography with substance B, flow velocity: 8ml/h, every part of 1.5ml of elutriant behind the collection chromatography), put into the dialysis tubing 48h that dialyses behind the elutriant concentrating under reduced pressure, dialyzate add 3 times of amount ethanol static, spend the night, centrifugal, collecting precipitation vacuum-drying gets pure hypoglycemic natural product.
Rat test result and embodiment 1 with, it is same with embodiment 1 to utilize sound attitude light scattering apparatus, gas-chromatography, infrared spectra, nucleus magnetic resonance etc. to characterize the molecular weight and the chemical constitution of pure hypoglycemic natural products.
Embodiment 3:
The preparation method of hypoglycemic natural product and application thereof in the sand sagebrush (Artemisia filifolia) seed
Step 1: the sand sagebrush (Artemisia filifolia) seed is through CO
2The part water that supercritical extraction is sloughed behind the grease fully dissolves (ratio of raw material and water is 1: 40), in the aqueous solution, add ethanol (ethanol content reaches 70%) static, spend the night, centrifugal, collecting precipitation vacuum-drying gets substance A;
Step 2: take by weighing the water dissolution of the substance A of certainweight with 90 times of weight, in water dissolution, add papoid and carry out enzymolysis, use chloroform and propyl carbinol (1: 3) mixed solution to extract again, collect that the water layer solution decompression concentrates, dry substance B, the A aqueous solution and extraction liquid ratio by volume are that 4: 1, papoid consumption are the 24.00U/2000ml aqueous solution;
Step 3: (elutriant is: 0.05molNacl solution by sephadex G-200 column chromatography with substance B, flow velocity: 8ml/h, every part of 1.5ml of elutriant behind the collection chromatography), put into the dialysis tubing 36-48h that dialyses behind the elutriant concentrating under reduced pressure, dialyzate add 2.5 times of amount ethanol static, spend the night, centrifugal, collecting precipitation vacuum-drying gets pure hypoglycemic natural product.
Rat test result and embodiment 1 with, it is same with embodiment 1 to utilize sound attitude light scattering apparatus, gas-chromatography, infrared spectra, nucleus magnetic resonance etc. to characterize the molecular weight and the chemical constitution of pure hypoglycemic natural products.
Claims (3)
1. the artemisia desertorum polysaccharide preparation method of a hypoglycemic is characterized in that: the waste residue that utilizes the natural biological sand sagebrush (Artemisia filifolia) to slough behind the oil obtains through the following step for raw material:
Step 1: the sand sagebrush (Artemisia filifolia) seed is through CO
2After supercritical extraction was sloughed grease, water fully dissolved, and the ratio of raw material and water is 1: 30-50, in the aqueous solution, add ethanol, and that ethanol content reaches is static behind the 50-80%, spend the night, centrifugal, collecting precipitation, vacuum-drying gets substance A;
Step 2: the substance A that takes by weighing certainweight, water dissolution with 80-100 times of weight, in water dissolution, add papoid and carry out enzymolysis, use 1 again: 2-4 chloroform and propyl carbinol mixed solution extract, collect that the water layer solution decompression concentrates, dry substance B, the A aqueous solution and extraction liquid ratio by volume are that 4: 1, papoid consumption are the 24.00U/2000ml aqueous solution;
Step 3: substance B is passed through sephadex G-200 column chromatography, elutriant is: 0.05molNacl solution, flow velocity: 8ml/h, every part of 1.5ml of elutriant behind the collection chromatography, put into the dialysis tubing 36-48h that dialyses behind the elutriant concentrating under reduced pressure, dialyzate add 2-3 doubly measure ethanol static, spend the night, centrifugal, collecting precipitation, vacuum-drying gets pure sand sagebrush (Artemisia filifolia) natural seed biological polyoses.
2. the artemisia desertorum polysaccharide of the preparation method of artemisia desertorum polysaccharide according to claim 1 preparation, it is characterized in that: it is made up of L-arabinose, D-wood sugar, D-lyxose, D-seminose, D-glucose, six kinds of monose of D-semi-lactosi, mol ratio was followed successively by 0.14: 0.09: 1.51: 0.17: 0.07: 1.45, weight-average molecular weight Mn was 1.42 * 10
5G/mol.
3. the application of artemisia desertorum polysaccharide according to claim 2 is characterized in that: be used for the treatment of the chemotype diabetes that tetraoxypyrimidine causes.
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| CN 200510002228 CN1267456C (en) | 2005-01-18 | 2005-01-18 | Process for preparing glucose-reducing sand sagebrush polysaccharide and its use |
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| CN1267456C true CN1267456C (en) | 2006-08-02 |
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Families Citing this family (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100553496C (en) * | 2006-06-02 | 2009-10-28 | 中国科学院长春应用化学研究所 | A kind of preparation process of sand sage gelatin diet |
| CN101219157B (en) * | 2007-11-16 | 2010-12-08 | 西北师范大学 | method for preparing product for reducing blood sugar |
| CN101240036B (en) | 2008-02-26 | 2010-06-09 | 西北师范大学 | Microwave auxiliary extraction method for artemisia seed polysaccharide |
| CN100572397C (en) * | 2008-05-07 | 2009-12-23 | 西北师范大学 | A kind of preparation method of lily polysaccharide of strengthening immunity |
| CN101412762B (en) * | 2008-12-10 | 2011-01-05 | 西北师范大学 | Method for catalytically synthesizing sand sagebrush polysaccharide sulfate |
| CN102268101A (en) * | 2011-08-31 | 2011-12-07 | 南京财经大学 | Enzyme-assisted extraction method of high-purity porphyra polysaccharide |
| CN102987206A (en) * | 2012-12-27 | 2013-03-27 | 上海普维健康食品有限公司 | Composition capable of reducing blood glucose |
| CN103156865B (en) * | 2013-03-14 | 2015-12-23 | 广西壮族自治区中国科学院广西植物研究所 | L-arabinose is preparing the application in medicine or health product |
| CN106397622B (en) * | 2016-10-11 | 2018-10-02 | 重庆三峡医药高等专科学校 | The extraction of bulbus fritillariae cirrhosae polysaccharide and separation purifying technique |
| CN114644719A (en) * | 2022-03-24 | 2022-06-21 | 珠海中科先进技术研究院有限公司 | Hai antler polysaccharide extracting solution and preparation method and application thereof |
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