CN107987184A - A kind of preparation method of heparan - Google Patents
A kind of preparation method of heparan Download PDFInfo
- Publication number
- CN107987184A CN107987184A CN201711259994.9A CN201711259994A CN107987184A CN 107987184 A CN107987184 A CN 107987184A CN 201711259994 A CN201711259994 A CN 201711259994A CN 107987184 A CN107987184 A CN 107987184A
- Authority
- CN
- China
- Prior art keywords
- heparan
- preparation
- bone
- cartilage
- raw material
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/006—Heteroglycans, i.e. polysaccharides having more than one sugar residue in the main chain in either alternating or less regular sequence; Gellans; Succinoglycans; Arabinogalactans; Tragacanth or gum tragacanth or traganth from Astragalus; Gum Karaya from Sterculia urens; Gum Ghatti from Anogeissus latifolia; Derivatives thereof
- C08B37/0063—Glycosaminoglycans or mucopolysaccharides, e.g. keratan sulfate; Derivatives thereof, e.g. fucoidan
- C08B37/0069—Chondroitin-4-sulfate, i.e. chondroitin sulfate A; Dermatan sulfate, i.e. chondroitin sulfate B or beta-heparin; Chondroitin-6-sulfate, i.e. chondroitin sulfate C; Derivatives thereof
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
Landscapes
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Materials Engineering (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Medicinal Chemistry (AREA)
- Polymers & Plastics (AREA)
- Organic Chemistry (AREA)
- Dermatology (AREA)
- Sustainable Development (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The present invention provides a kind of preparation method of heparan, and this method concretely comprises the following steps：Using the bone of healthy animal and/or cartilage as raw material, raw material is crushed, after NaOH pretreatment add enzyme digested, it is filtered to obtain clear liquid, add ethanol precipitation, for sediment through water dissolving rear decoloring, filtering, filtrate is dry after ethanol precipitation, obtains heparan intermediate；Intermediate through poly-sulfated, ethanol precipitation, decoloration, be classified alcohol precipitation, be dried to obtain fine work heparan.The present invention provides a kind of method that can prepare the heparan to meet clinical needs, has simple production process, raw material sources are wide, cheap, and no waste discharge, meets environmental protection requirement, meet the requirement recycled.
Description
Technical field
The invention belongs to biological medicine class, is related to a kind of preparation method of heparan.
Background technology
Heparan is also known as poly-sulfated chondroitin sulfate (Heparinoid).1948, Jorpes and Gardell from liver and
When heparin is prepared in lung, a kind of polysaccharide being made of D-Glucose aldehydic acid and D-Glucose amine, these materials and heparin knot are obtained
Structure is similar, and simply degree is relatively low, and anticoagulant active is low, but after poly-sulfated, great changes will take place for its property.Pass through
Combined with blood vessel endothelium surface, promote the release of the endogenous glycosaminoglycan with antithrombotic acitivity, locally micro- follow can be improved
Ring, increases blood flow and blood flow velocity, so as to improve intermediate supersession, promotes the supply of skin-nourishing and the excretion of waste, play
Detumescence removes silt, anti-inflammatory analgesic, promotes connective tissue recovery and other effects；The inflammatory mediators such as histamine, serotonin, suppression can also be inactivated
Granulation tissue hyperplasia and smooth muscle cell breeding processed；Keratolytic, natural moisture preserving agent synergistic effect under, glue can be suppressed
The excessive growth of fibrillation, mitigates cicatrization；And skin surface moisture can be retained, soften collagenous fibres, elasticity enhancing is right
Hyperplastic scar has emollescence.
Meanwhile heparan has a broad antifungal spectrum, to for example suppurative bacterium (staphylococcus aureus) of many pathogenic bacteria, intestinal flora
(Escherichia coli), pathogenic fungus (Candida albicans), viral (poliovirus) etc. have good inhibitory action, can
Inflammation illness is efficiently controlled, improves affected part blood circulation, absorbs sepage, cures oedema and edema.
The content of the invention
The present invention provides a kind of method that can prepare the heparan to meet clinical needs.
A kind of preparation method of heparan, its preparation process are as follows：, will using the bone of healthy animal and/or cartilage as raw material
Raw material crushes, addition enzyme is digested after NaOH pretreatment, filtered to obtain clear liquid, adds ethanol precipitation, sediment
Through water dissolving rear decoloring, filtering, filtrate is dry after ethanol precipitation, obtains heparan intermediate；Intermediate is through poly-sulfated, second
Alcohol is precipitated, decolourized, is classified alcohol precipitation, is dried to obtain fine work heparan.
The bone and/or cartilage of the healthy animal, the including but not limited to tracheae of healthy animal, kneecap, each position
Bone, cartilage.
The pretreatment is：First bone and/or cartilage are put into reaction kettle after crushed, addition is no less than material quality
1.5 times of water, is pre-processed using the sodium hydroxide of 1%-10%, and the time is 6-24 hour, and temperature is 10 DEG C -80 DEG C.
Enzyme used in the enzymolysis include but is not limited to lipase, trypsase, papain, streptomyces proteinase,
Subtilopeptidase A.
The enzymatic hydrolysis condition is the inactivation condition of the reaction condition that enzymatic hydrolysis condition is above-mentioned enzyme and the enzyme.Such as pancreatin is anti-
When to answer condition be ph8-10,50-60 degrees Celsius of temperature, reaction time are 2-4 small, inactivation condition is more than 85 degrees Celsius, much
In 15 minutes.
The poly-sulfated step is：
(1) heparan intermediate, solvent volume are dissolved as solvent using n,N-Dimethylformamide and/or formamide：Class liver
Weight is 1-5L/g among plain；
(2) chlorosulfonic acid is dissolved in n,N-Dimethylformamide and/or formamide, chlorosulfonic acid and n,N-Dimethylformamide
And/or formamide volume ratio is 1:1-5 times；
(3) by two kinds of solution blendings in reaction kettle, condition is 18 DEG C -70 DEG C, when the time is 4~10 small.
The decolorising agent of the decoloration is activated carbon and/or hydrogen peroxide；When decolorising agent is activated carbon, activated carbon addition is
0.1-5%g/L (w/v), pH value 3-7, reaction temperature are 15 DEG C -100 DEG C, 30~180 minutes reaction time；When decolorising agent is
During hydrogen peroxide, it is 1%-10% (v/v) that hydrogen peroxide, which adds volume, and temperature is 15 DEG C -60 DEG C, pH value 8-12, the reaction time 2
~36 it is small when.
Heparan the key technical indexes prepared by the present invention includes：
Character：The odorless slight bitter powder of white；
PH：5.3-7.6；
Optical activity is：-11.7°——-14.7°；
Intrinsic viscosity 0.09 --- 0.18；
Free sulphur acidic group is below 13.0%；
D-Glucose aldehydic acid 19.0 --- 24.0%；
Nitrogen 1.6 --- 2.0%；
Organic sulfate 25.8 --- 37.3%；
High fever remaining 38.0 --- 48.0%；
Anticoagulant active：Below 80IU；
Hyaluronidase hinders activity 14%-26%.
The present invention provides a kind of method that can prepare the heparan to meet clinical needs, has simple production process,
Raw material sources are wide, cheap, and no waste discharge, meets environmental protection requirement, meet the requirement recycled.
Embodiment
Embodiment 1
Pig tracheae (band lung) 500kg is taken, cleaning, rub into rotten shape thing；8 times of amount purifying of raw material are added into retort
Water, opens mixer, adds NaOH to 2%g/L (w/V) concentration, rotten shape thing is added, when stirring 7 is small.With 1:1 hydrochloric acid is adjusted
Between PH8.5-9.0,4% pancreatin of material quality is added, temperature in tank is adjusted and, to 50 DEG C, pH value is adjusted with 40%NaOH solution,
It is maintained between PH8.5-9.0；Continue stirring 4 it is small when, solution is warming up to more than 85 DEG C, 15min is kept, with 1:1 hydrochloric acid
Between adjusting PH6.0-7.0.Filtering, filtrate adds 2% activated carbon of liquor capacity, 5% hydrogen peroxide is stirred decoloration, solution temperature
Degree rises to 50-60 DEG C, is maintained between PH10.0-11.0, when reaction 24 is small.PH is adjusted between 6.0-7.0 with hydrochloric acid.Cross
Filter, be deposited in 70 DEG C vacuum drying 10 it is small when, obtain heparan intermediate.
Heparan intermediate 50kg is taken to be slowly added in 150 liters of NN- dimethylformamides；Another take adds 100 liters of N, N- bis-
Methylformamide, is poured slowly into 50 liters of chlorosulfonic acids, merges two kinds of solution in retort, solution temperature is maintained at 40-60 DEG C
Between, when stirring 8 is small；Filtering, 2% activated carbon of addition liquor capacity, 5% hydrogen peroxide are stirred decoloration, and solution temperature adds to
50-60 DEG C, pH value is adjusted with 40%NaOH solution, is maintained between PH10.0-11.0, when reaction 24 is small.Adjusted with hydrochloric acid
Between PH6.0-7.0.Filter, 90% ethanol is added in filtrate, stablize to precipitation, sediment is dehydrated 30 points with soaked in absolute ethyl alcohol
Clock, filtering, repeatedly for three times, when 70 DEG C of vacuum drying 24 of precipitation are small, obtains fine work heparan.
Technical indicator meets following scope to the heparan of preparation after testing：
Character：The odorless slight bitter powder of white；
PH：5.3-7.6；
Optical activity is：-11.7°—-14.7°；
Intrinsic viscosity 0.09-0.18；
Free sulphur acidic group is below 13.0%；
D-Glucose aldehydic acid 19.0-24.0%；
Nitrogen 1.6-2.0%；
Organic sulfate 25.8-37.3%；
High fever remaining 38.0-48.0%；
Anticoagulant active：Below 80IU；
Hyaluronidase hinders activity 14%-26%.
Embodiment 2
Kneecap, larynx bone is taken to amount to 500kg, cleaning, rub into rotten shape thing；6 times of amount purified waters of raw material are added into retort,
Mixer is opened, adds NaOH to 4% (w/V) concentration, rotten shape thing is added, when stirring 6 is small.With 1:1 hydrochloric acid adjusts PH8.5-
Between 9.0,2% pancreatin of material quality is added, temperature in tank is adjusted and, to 55 DEG C, adjusts pH value with 40%NaOH solution, be maintained at
Between PH8.5-9.0；Continue stirring 6 it is small when, solution is warming up to more than 85 DEG C, 20min is kept, with 1:1 hydrochloric acid is adjusted
Between PH6.0-7.0.Filtering, filtrate adds 1% activated carbon of solution body, 3% hydrogen peroxide is stirred decoloration, and solution temperature adds to
50-60 DEG C, it is maintained between PH10.0-11.0, when reaction 24 is small.PH is adjusted between 6.0-7.0 with hydrochloric acid.Filtering, precipitation
When 70 DEG C of vacuum drying 10 are small, heparan intermediate is obtained.
Heparan intermediate 50kg is taken to be slowly added in 100 liters of NN- dimethylformamides；Another take adds 80 liters of N, N- diformazans
Base formamide, is poured slowly into 40 liters of chlorosulfonic acids, merge two kinds of solution in retort, solution temperature be maintained at 40-60 DEG C it
Between, when stirring 6 is small；Filtering, 1% activated carbon of addition liquor capacity, 3% hydrogen peroxide are stirred decoloration, and solution temperature adds to 50-
60 DEG C, pH value is adjusted with 40%NaOH solution, is maintained between PH10.0-11.0, when reaction 24 is small.PH6.0- is adjusted with hydrochloric acid
Between 7.0.Filter, 90% ethanol is added in filtrate, stablize to precipitation, sediment is dehydrated 30 minutes with soaked in absolute ethyl alcohol, mistake
Filter, repeatedly for three times, when 70 DEG C of vacuum drying 24 of precipitation are small, obtains fine work heparan.
Technical indicator meets following scope to the heparan of preparation after testing：
Character：The odorless slight bitter powder of white；
PH：5.3-7.6；
Optical activity is：-11.7°——-14.7°；
Intrinsic viscosity 0.09 --- 0.18；
Free sulphur acidic group is below 13.0%；
D-Glucose aldehydic acid 19.0 --- 24.0%；
Nitrogen 1.6 --- 2.0%；
Organic sulfate 25.8 --- 37.3%；
High fever remaining 38.0 --- 48.0%；
Anticoagulant active：Below 80IU；
Hyaluronidase hinders activity 14%-26%.
Claims (7)
1. a kind of preparation method of heparan, it is characterised in that this method concretely comprises the following steps：With the bone and/or cartilage of healthy animal
For raw material, raw material is crushed, enzyme is added after NaOH pretreatment is digested, it is filtered to obtain clear liquid, add ethanol and sink
Form sediment, for sediment through water dissolving rear decoloring, filtering, filtrate is dry after ethanol precipitation, obtains heparan intermediate；Among heparan
Body through poly-sulfated, ethanol precipitation, decoloration, be classified alcohol precipitation, be dried to obtain fine work heparan.
A kind of 2. preparation method of heparan according to claim 1, it is characterised in that the bone of the healthy animal and/
Or cartilage, including the tracheae of healthy animal, kneecap, the bone or cartilage at each position.
3. the preparation method of a kind of heparan according to claim 1, it is characterised in that the pretreatment is：First by bone
And/or cartilage is put into reaction kettle after crushed, the water no less than 1.5 times of material quality is added, uses concentration as 1%-10%
Sodium hydroxide solution pre-processed, the time for 6-24 it is small when, temperature be 10-80 DEG C.
4. the preparation method of a kind of heparan according to claim 1, it is characterised in that enzyme used in the enzymolysis is fat
Enzyme, pancreatin, papain, streptomyces proteinase or subtilopeptidase A.
5. the preparation method of a kind of heparan according to claim 1, it is characterised in that the enzymatic hydrolysis condition is enzyme used
Reaction condition and enzyme used inactivation condition.
6. the preparation method of a kind of heparan according to claim 1, it is characterised in that the poly-sulfated step is：
(1) heparan intermediate, solvent volume are dissolved as solvent using n,N-Dimethylformamide and/or formamide：In heparan
Between weight be 1-5L/g；
(2) chlorosulfonic acid is dissolved in n,N-Dimethylformamide and/or formamide, chlorosulfonic acid and n,N-Dimethylformamide and/
Or formamide volume ratio is 1:1-5；
(3) by two kinds of solution blendings in reaction kettle, condition is 18 DEG C -70 DEG C, when the time is 4~10 small.
7. the preparation method of a kind of heparan according to claim 1, it is characterised in that the decolorising agent of the decoloration is work
Property charcoal and/or hydrogen peroxide；When decolorising agent is activated carbon, activated carbon addition is 0.1-5%g/L, pH value 3-7, reaction temperature
It is 30~180 minutes to spend for 15 DEG C -100 DEG C, reaction time；When decolorising agent is hydrogen peroxide, it is 1%- that hydrogen peroxide, which adds volume,
10%, reaction temperature is 15 DEG C -60 DEG C, pH value 8-12, when the reaction time is 2~36 small.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711259994.9A CN107987184A (en) | 2017-12-04 | 2017-12-04 | A kind of preparation method of heparan |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711259994.9A CN107987184A (en) | 2017-12-04 | 2017-12-04 | A kind of preparation method of heparan |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN107987184A true CN107987184A (en) | 2018-05-04 |
Family
ID=62035516
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201711259994.9A Pending CN107987184A (en) | 2017-12-04 | 2017-12-04 | A kind of preparation method of heparan |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107987184A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111285943A (en) * | 2020-04-28 | 2020-06-16 | 山东冰文生物技术有限公司 | Preparation method of polysulfated chondroitin sulfate |
| CN111825777A (en) * | 2020-07-13 | 2020-10-27 | 山东众山生物科技有限公司 | Method for preparing heparinoids from chondroitin |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101033480A (en) * | 2007-03-16 | 2007-09-12 | 由守谊 | Preparation method for heparan |
| CN104877042A (en) * | 2015-06-10 | 2015-09-02 | 浙江三门恒康制药有限公司 | Preparation method of heparinoid |
-
2017
- 2017-12-04 CN CN201711259994.9A patent/CN107987184A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101033480A (en) * | 2007-03-16 | 2007-09-12 | 由守谊 | Preparation method for heparan |
| CN104877042A (en) * | 2015-06-10 | 2015-09-02 | 浙江三门恒康制药有限公司 | Preparation method of heparinoid |
Non-Patent Citations (2)
| Title |
|---|
| 于广利等著: "《糖药物学》", 31 October 2012, 中国海洋大学出版社 * |
| 朱洪法等著: "《精细化工产品配方与制造 第八册》", 30 June 2002, 金盾出版社 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111285943A (en) * | 2020-04-28 | 2020-06-16 | 山东冰文生物技术有限公司 | Preparation method of polysulfated chondroitin sulfate |
| CN111825777A (en) * | 2020-07-13 | 2020-10-27 | 山东众山生物科技有限公司 | Method for preparing heparinoids from chondroitin |
| CN111825777B (en) * | 2020-07-13 | 2022-05-27 | 山东众山生物科技有限公司 | Method for preparing heparinoids from chondroitin |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Salihu et al. | Citric acid: A green cross-linker of biomaterials for biomedical applications | |
| ES2662064T3 (en) | Method for producing crosslinked hyaluronic acid | |
| CN100523070C (en) | Method of preparing cellulose hydrogel | |
| EP1616019B1 (en) | Method for extracting collagen comprising microbial fermentation | |
| JPH0220292A (en) | Production of depolymerized chitosan | |
| Bayón et al. | Biopolymers from wastes to high-value products in biomedicine | |
| CN101161103A (en) | Method for preparing bean slags soluble food fibers by zymolysis method | |
| Moen et al. | Biological degradation of Ascophyllum nodosum | |
| CN104189944A (en) | High-purity natural collagenous fiber and preparation method thereof | |
| CN107987184A (en) | A kind of preparation method of heparan | |
| CN102134288B (en) | Process for extracting chondroitin sulfate from pigs | |
| CN1861640A (en) | Preparation process of chito oligosaccharace hydrochloride | |
| KR101841099B1 (en) | Cosmetic composition comprising fucoidan for mask pack and manufacturing method of the same that | |
| CN105326674A (en) | Preparation method of seaweed and silk fibroin moisturizing mask | |
| KR20100079362A (en) | Preparation of low molecular weight hualuronic acid by acid treatment | |
| CN100560730C (en) | A kind of preparation method of heparitin | |
| CN110922501A (en) | Process for extracting sodium alginate by using ultrasonic-assisted complex enzyme | |
| CN102321192B (en) | New preparation method of housefly larva chitosan | |
| CN101851259A (en) | Selenium chitosan oligosaccharide and preparation method thereof | |
| CN114949329A (en) | Preparation method of surface-modified silk fibroin-aloin double-layer fiber membrane | |
| US3761355A (en) | Comestible digestible protein from cellulose | |
| PT97565B (en) | PROCESS FOR THE PREPARATION OF XYLANASE | |
| CN101230377A (en) | Method for preparing antioxidant collagen peptide by spent of ocean alkaline protease 894 hydrolyzed aquatic products processing | |
| Zhang et al. | Hydrogels based on seafood chitin: From extraction to the development | |
| CN101307093A (en) | Method for separating and purifying collagen for biopharmaceutical use from sharkskin |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20180504 |