CN1908019A - Chitosan oligosaccharide sulphate and preparation method thereof - Google Patents
Chitosan oligosaccharide sulphate and preparation method thereof Download PDFInfo
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- CN1908019A CN1908019A CN 200610020011 CN200610020011A CN1908019A CN 1908019 A CN1908019 A CN 1908019A CN 200610020011 CN200610020011 CN 200610020011 CN 200610020011 A CN200610020011 A CN 200610020011A CN 1908019 A CN1908019 A CN 1908019A
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- chitosan oligosaccharide
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Abstract
The invention discloses a Chitosan oligosaccharide sulfate and preparing method, which is reacted by Chitosan oligosaccharide and sulfuric acid, wherein the polymerization degree of Chitosan oligosaccharide is 2-30. The preparing method comprises the following steps: dissolving chitosan in the weakly-acidic solution; degrading the solution into Chitosan oligosaccharide polymerization degree at 2-30 through enzyme; adding sulfuric acid to produce the Chitosan oligosaccharide sulfate; separating and purifying to obtain the solid product. The invention is stable to reserve, which can be raw material of drug and health food to resist tumour and adjust immunity.
Description
Technical field
The present invention relates to chitosan oligosaccharide sulphate and its preparation method, the product that makes is easy to preserve, the raw material that this product can be used as is antitumor, regulate immunizing power and arthritic medicine and protective foods.
Background technology
From the intravital alkaline polysaccharide of animal and plant, nontoxic, harmless, the source is abundant, and many unique physiologically actives are arranged, and is considered to " life the 6th key element " as a kind of for chitin.Oligosaccharides in the Mammals milk contains the acetylglucosamine unit.Yet along with the raising of living condition, people absorb chitin from the low-diet thing chance reduces, and therefore, need carry out necessary replenishing, and but, chitin is water insoluble and sour.Chitosan is the derivative behind the chitin deacetylase base, dissolves in some acid solution, but is insoluble to neutral medium, influences its digestion and absorption in vivo, has a strong impact on its physiological activity.
Glucosamine monose has had the history of nearly half a century in Europe as the medicine of protection and treatment of arthritis.Oligochitosan is the little chitosan of the polymerization degree, and the chitosan of this specification sheets middle finger 2 to 30 is solvable under the physiological pH condition.Last century, the eighties scientist discovered that crust six sugar and shell six sugar have the obvious suppression effect to mouse transplantability cancer cells.Recently studies show that the polymerization degree has better anti-tumor activity and immunologic enhancement greater than 6 water-soluble chitosan oligosaccharide.
At present, the method for preparation of industrialization oligochitosan mainly is to adopt enzymatic degradation, generally the chitosan raw material is dissolved in the weakly acid solns such as acetic acid, lactic acid, citric acid earlier, degrade then, products therefrom behind membrane sepn, lyophilize, product mainly is the salt of weak acid of oligochitosan.But these product shelf-lifes are short, and variable brownish black in air and in the higher temperature loses water-soluble and active.We discover that the oligochitosan hydrochloride is more stable than oligochitosan salt of weak acid, and product is easy to preserve.But introduce chlorion in the oligochitosan hydrochloride, be unwell to cardiovascular diseases and nephrotic,, overcome the weak point of oligochitosan salt of weak acid and hydrochloride so we are prepared into chitosan oligosaccharide sulphate with oligochitosan.
Summary of the invention
Problem to be solved by this invention provides a kind of suitable crowd oligochitosan salt more widely, i.e. chitosan oligosaccharide sulphate and its preparation method.That the chitosan oligosaccharide sulphate that makes can be used as is antitumor, regulate the raw material of immunizing power and arthritic medicine and protective foods.
Technical scheme provided by the invention is: chitosan oligosaccharide sulphate is that the salt product that 2 to 30 oligochitosan and sulfuric acid reaction form is chitosan oligosaccharide sulphate by the polymerization degree.
The present invention also provides the preparation method of above-mentioned chitosan oligosaccharide sulphate: stir in the weak organic acid solution with chitosan adding 0.5～5wt%, the concentration of control chitosan is 1～10wt%, press the enzyme-added reagent of 0.1～5wt% of chitosan weight then, control reaction mixture pH value is 4.5 to 6.5, stirred 2～24 hours at 40～60 ℃, when the reaction principal product polymerization degree 2 to 30 the time, cooling is filtered.Filtrate can be selected following method to handle and obtain the chitosan oligosaccharide sulphate product.Method one: press amino and H in the chitosan
2SO
4The ratio of amount of substance be 1: the ratio of 0.4-1.0 was boiled 5～15 minutes to adding sulfuric acid, filtered and dezymotized back (can adopt activated carbon decolorizing) concentrating under reduced pressure, adds organic solvent sedimentation and washing, and is dry then.Method two: filtrate is dezymotized with the ultra-filtration membrane ultrafiltration of 5-20kDa and is obtained ultrafiltrated, the ultrafiltrated after the classification (can adopt activated carbon decolorizing) concentrate with nanofiltration membrane or concentrating under reduced pressure after, press amino and H in the chitosan
2SO
4The ratio of amount of substance be 1: the ratio of 0.4-1.0 adopts organic solvent sedimentation and washing to adding sulfuric acid, and is dry then.
Described organic solvent is ethanol, methyl alcohol, propyl alcohol, Virahol, butanols, primary isoamyl alcohol, acetone and/or butanone.
Described enzyme reagent is chitoanase, chitin enzyme, cellulase, hemicellulase, amylase, proteolytic enzyme and/or N,O-Diacetylmuramidase; Described weak organic acid is acetic acid, lactic acid or citric acid.
Yield of the present invention is higher, cost is lower; Gained chitosan oligosaccharide sulphate product is stable, water-soluble, is easy to preserve, and can be used as antitumor, as to regulate immunizing power and arthritic medicine and protective foods raw material.
Embodiment
Embodiment 1: get small lobsters shell chitosan (deacetylation 88%) 10 grams, with 200 milliliters of 2wt% dilute acetic acid dissolvings, add α-Dian Fenmei 0.3 gram, in 50-55 ℃ of water-bath, react, when the major constituent polymerization degree of chitosan product drops to 30 when following, naturally cooling adopts the ultrafiltration membrance filter of 10kDa, filtrate to add concentrating under reduced pressure behind the 2.5 gram sulfuric acid, add ethanol sedimentation and washing, filtration is drained, and solids by using vacuum-drying obtains the chitosan oligosaccharide sulphate product.
Embodiment 2: get commercially available crab shell chitosan (deacetylation 86%) 50 grams, add 2000 milliliters of 1wt% dilute acetic acid, stirred 4 hours, add R-10 cellulase 3 grams, 45 ℃ of reactions, when the major constituent polymerization degree of chitosan product drops to 30 when following, cooling is filtered, add 25 gram sulfuric acid after, boil about 10 minutes, filtration is dezymotized, the filtrate decolorizing with activated carbon, concentrating under reduced pressure adds acetone sedimentation and washing, centrifugal after, 50 ℃ of oven dry of solid product obtain the chitosan oligosaccharide sulphate product.
Embodiment 3: get commercially available shrimp shell chitosan (deacetylation 93%) 20 grams, add and stir 2 milliliters of enzyme liquid of adding (produce in Wuhan, contains half cellulase and chitoanase) after 2 hours in 2000 milliliters of 1wt% lactic acid, 48 ℃ of reactions, when the major constituent polymerization degree of chitosan product drops to about 30 the time, cooling is filtered, and filtrate adds concentrating under reduced pressure behind the 5 gram sulfuric acid, with methyl alcohol sedimentation and washing, filtration is drained, and solid product is dried below 70 ℃, obtains the chitosan oligosaccharide sulphate product.
Embodiment 4: get chitosan (deacetylation 73%) 10 grams, add in 200 milliliters of 4wt% acetic acid, stir and add N,O-Diacetylmuramidase 0.5 gram after 2 hours, in 50 ℃ of water-baths, react, when the major constituent polymerization degree of chitosan product drops to about 4-8, cooling, filter, filtrate adds concentrating under reduced pressure behind the 3 gram sulfuric acid, and with ethanol sedimentation and washing, filtration is drained.Solid product adds in 200 ml waters and dissolves, decolorizing with activated carbon, concentrating under reduced pressure adds primary isoamyl alcohol sedimentation and washing, centrifugal after, 50 ℃ of oven dry of solid product obtain the chitosan oligosaccharide sulphate product.
Embodiment 5: water intaking soluble chitosan (deacetylation 52%) 10 grams, with 400 milliliters of rare citric acid dissolvings of 1wt%, add stomach en-0.5 gram, in 40-50 ℃ of water-bath, react, when the major constituent polymerization degree of chitosan product drops to 20 when following, cooling adopts the ultrafiltration membrance filter of 10kDa, filtrate to add concentrating under reduced pressure behind the 3 gram sulfuric acid, add butanone sedimentation and washing, filtration is drained, and 50 ℃ of oven dry of solids obtain the chitosan oligosaccharide sulphate product.
Embodiment 5: get chitosan (deacetylation 85%) 5 grams, with 500 milliliters of 1wt% dilute acetic acid dissolvings, add 2 milliliters of chitoanase solution, in 45 ℃ of water-baths, react, when the major constituent polymerization degree of chitosan product drops to 10 when following, cooling, filter, add 2 gram sulfuric acid after, boil about 10 minutes, filtration is dezymotized, filtrate decolorizing with activated carbon, concentrating under reduced pressure, add ethanol/acetone (v/v) sedimentation in 1: 1 and washing, after centrifugal, 50 ℃ of oven dry of solid product obtain the chitosan oligosaccharide sulphate product.
Claims (7)
1. chitosan oligosaccharide sulphate is that the salt product that 2 to 30 oligochitosan and sulfuric acid reaction form is chitosan oligosaccharide sulphate by the polymerization degree.
2. the preparation method of chitosan oligosaccharide sulphate, it is characterized in that: stir in the weak organic acid solution with chitosan adding 0.8～5wt%, the concentration of control chitosan is 1～10wt%, press the enzyme-added reagent of 0.1～5wt% of chitosan weight then, control reaction mixture pH value stirred 2～24 hours at 40～60 ℃ 4.5 to 6.5, when reacting the principal product polymerization degree 2 to 30 the time, cooling filters to get filtrate; Press amino and H in the chitosan
2SO
4The ratio of amount of substance be 1: the ratio of 0.4-1.0 adds sulfuric acid, boils 5～15 minutes, filters and dezymotizes the back concentrating under reduced pressure, adds organic solvent sedimentation and washing, and drying obtains chitosan oligosaccharide sulphate then.
3. preparation method according to claim 2 is characterized in that: filter and dezymotize back elder generation decolorizing with activated carbon and then concentrating under reduced pressure.
4. the preparation method of chitosan oligosaccharide sulphate, it is characterized in that: stir in the weak organic acid solution with chitosan adding 0.8～5wt%, the concentration of control chitosan is 1～10wt%, press the enzyme-added reagent of 0.1～5wt% of chitosan weight then, control reaction mixture pH value stirred 2～24 hours at 40～60 ℃ 4.5 to 6.5, when reacting the principal product polymerization degree 2 to 30 the time, cooling filters to get filtrate; Filtrate is dezymotized with the ultra-filtration membrane ultrafiltration of 5-20kDa and is obtained ultrafiltrated, the gained ultrafiltrated concentrate with nanofiltration membrane or concentrating under reduced pressure after, press amino and H in the chitosan
2SO
4The ratio of amount of substance be 1: the ratio of 0.4-1.0 adds sulfuric acid, adopts organic solvent sedimentation and washing to obtain chitosan oligosaccharide sulphate.
5. preparation method according to claim 4 is characterized in that: the gained ultrafiltrated is used earlier decolorizing with activated carbon, and then concentrates or concentrating under reduced pressure with nanofiltration membrane.
6. according to claim 2 or 3 or 4 or 5 described preparation methods, it is characterized in that: organic solvent is ethanol, methyl alcohol, propyl alcohol, Virahol, butanols, primary isoamyl alcohol, acetone and/or butanone.
7. according to claim 2 or 3 or 4 or 5 described preparation methods, it is characterized in that: described enzyme reagent is chitoanase, chitin enzyme, cellulase, hemicellulase, amylase, proteolytic enzyme and/or N,O-Diacetylmuramidase; Described weak organic acid is acetic acid, lactic acid or citric acid.
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| CNB2006100200111A CN100441692C (en) | 2006-08-17 | 2006-08-17 | Chitosan oligosaccharide sulphate and preparation method thereof |
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| CNB2006100200111A CN100441692C (en) | 2006-08-17 | 2006-08-17 | Chitosan oligosaccharide sulphate and preparation method thereof |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102475714A (en) * | 2010-11-29 | 2012-05-30 | 中国科学院大连化学物理研究所 | Application of chitosan oligosaccharide single compound in immunopotentiating drugs |
| CN102558387A (en) * | 2010-12-13 | 2012-07-11 | 青岛中仁药业有限公司 | Method for extracting chitin and antibacterial peptide from fly larvae |
| CN103041377A (en) * | 2012-09-20 | 2013-04-17 | 杭州师范大学 | Preparation method of lysozyme loaded chitosan/gamma polyglutamic acid composite nanoparticles |
| CN109535280A (en) * | 2018-12-03 | 2019-03-29 | 青岛博智汇力生物科技有限公司 | A kind of ascorbic acid chitosan oligosaccharide compound salt and its preparation method and application |
| CN109576329A (en) * | 2018-12-29 | 2019-04-05 | 广东医科大学 | A kind of preparation method, product and its application of the non-agar oligosaccharides of the asparagus of anti-melanin genesis |
| CN112626147A (en) * | 2021-01-12 | 2021-04-09 | 中国科学院海洋研究所 | Preparation method of chitosan oligosaccharide salt |
| CN117229429A (en) * | 2023-08-17 | 2023-12-15 | 江西师范大学 | Chitosan oligosaccharide sulfate and preparation method thereof |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1177856C (en) * | 2001-08-14 | 2004-12-01 | 华东理工大学 | Process for industrial production of oligochitose and chitooligose |
| CN1184237C (en) * | 2003-01-07 | 2005-01-12 | 江南大学 | Preparation of crust oligosaccharide and use |
| CN1228350C (en) * | 2003-11-25 | 2005-11-23 | 武汉大学 | Water-soluble Chitooligosaccharide preparation method |
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2006
- 2006-08-17 CN CNB2006100200111A patent/CN100441692C/en not_active Expired - Fee Related
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102475714A (en) * | 2010-11-29 | 2012-05-30 | 中国科学院大连化学物理研究所 | Application of chitosan oligosaccharide single compound in immunopotentiating drugs |
| CN102558387A (en) * | 2010-12-13 | 2012-07-11 | 青岛中仁药业有限公司 | Method for extracting chitin and antibacterial peptide from fly larvae |
| CN103041377A (en) * | 2012-09-20 | 2013-04-17 | 杭州师范大学 | Preparation method of lysozyme loaded chitosan/gamma polyglutamic acid composite nanoparticles |
| CN103041377B (en) * | 2012-09-20 | 2014-11-05 | 杭州师范大学 | Preparation method of lysozyme loaded chitosan/gamma polyglutamic acid composite nanoparticles |
| CN109535280A (en) * | 2018-12-03 | 2019-03-29 | 青岛博智汇力生物科技有限公司 | A kind of ascorbic acid chitosan oligosaccharide compound salt and its preparation method and application |
| CN109576329A (en) * | 2018-12-29 | 2019-04-05 | 广东医科大学 | A kind of preparation method, product and its application of the non-agar oligosaccharides of the asparagus of anti-melanin genesis |
| CN109576329B (en) * | 2018-12-29 | 2021-05-18 | 广东医科大学 | Preparation method, product and application of anti-melanin synthesized asparagus non-agar oligosaccharide |
| CN112626147A (en) * | 2021-01-12 | 2021-04-09 | 中国科学院海洋研究所 | Preparation method of chitosan oligosaccharide salt |
| CN117229429A (en) * | 2023-08-17 | 2023-12-15 | 江西师范大学 | Chitosan oligosaccharide sulfate and preparation method thereof |
| CN117229429B (en) * | 2023-08-17 | 2024-03-15 | 江西师范大学 | Chitosan oligosaccharide sulfate and preparation method thereof |
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| CN100441692C (en) | 2008-12-10 |
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