CN109293801A - A method of heparin sodium is prepared by raw material of chitterlings - Google Patents
A method of heparin sodium is prepared by raw material of chitterlings Download PDFInfo
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- CN109293801A CN109293801A CN201811096277.3A CN201811096277A CN109293801A CN 109293801 A CN109293801 A CN 109293801A CN 201811096277 A CN201811096277 A CN 201811096277A CN 109293801 A CN109293801 A CN 109293801A
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- resin
- chitterlings
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- heparin sodium
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- 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
- C08B37/0075—Heparin; Heparan sulfate; Derivatives thereof, e.g. heparosan; Purification or extraction methods 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
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Abstract
The invention discloses using chitterlings as raw material, pass through the collection of pig intestinal mucosa liquid, it digests (salt solution), filter residue, resin adsorption, butane equipment extraction element under liquid gas biphase equilibrium state is passed through to the resin for having adsorbed heparin sodium, using butane as solvent, principle using heparin sodium insoluble in butane, washing removes and is attached to the lipid material and impurity that the heparin sodium being adsorbed in resin surface layer and resin wraps, ensure heparin sodium from the elution in resin, improve the consummate degree of heparin sodium, improve the yield of heparin sodium, and this addition product of casing can be obtained, propose super added value of product, it reduces costs.
Description
Technical field
The invention belongs to utilize chitterlings heparin sodium extracting technique field, one kind is concretely related to chitterlings as original
The method that material prepares heparin sodium.
Background technique
The current main source of heparin is chitterlings, and demand of the international market to heparin bulk pharmaceutical chemicals is very powerful, drug
Function is main are as follows: treats various diseases concurrent sowing property intravascular coagulation early stage, prevents artery and vein thrombus and pulmonary embolism, be
The most effective in the world and maximum anticoagulation medicine of quantity.Traditional heparin sodium is main from the extraction preparation method in chitterlings
It wants are as follows: the technical process such as chitterlings knifing, mucous membrane collection, enzymatic hydrolysis, filter residue, resin adsorption, elution dry, alcohol precipitates, drying are deposited
Its surface layer makes heparin sodium be difficult to elute by lipid or impurity package after because of resin adsorption heparin sodium, and contains a certain amount of lipid
Substance and impurity, affect quality and efficiency.This technology carries out degreasing class and miscellaneous using the butane under liquid gas biphase equilibrium state
The preparation method of matter improves the consummate degree of heparin sodium, improves the yield of heparin sodium.
Summary of the invention
The object of the present invention is to provide a kind of method for preparing heparin sodium as raw material using chitterlings, it is intended to solve chitterlings
The problem of high efficiency extraction heparin sodium.
In order to solve the above technical problems, the object of the present invention is achieved like this:
A method of heparin sodium is prepared by raw material of chitterlings, it is characterised in that: is included the following steps
S1: the preparation of intestinal mucosa: pouring into clear water in chitterlings, chitterlings is separated into intestinal mucosa, intestines skin and casing, by intestines
Mucous membrane is sent to mucous membrane pond, and intestines skin is crushed after extruding obtains intestines skin paste and is sent to mucus pool；
S2: enzymatic hydrolysis: being pumped with mucous membrane and squeeze into intestinal mucosa and intestines skin paste in enzymatic vessel from mucous membrane pond, is added into enzymatic vessel
The weight ratio of water, mucous membrane and water is 1:7-9, and being stirring evenly and then adding into 2709 basic protein enzyme adjustment pH values is 7.5-7.8, mucous membrane
Weight ratio with 2709 alkali proteases is 500:1, and heating makes temperature reach 55 DEG C -60 DEG C, and adding weight percent is 3%-
5% casing special-purpose salt surveys its salinity and reaches 2%-2.5%, keeps the temperature 2-3 hours, and continuing heating makes temperature reach 80 DEG C -85 DEG C, will
PH value adjustment is filtered after heat preservation 50-65 minutes with 90-100 mesh nylon cloth to 8-8.5, collects filtered fluid；
S3: absorption: above-mentioned filtered fluid is squeezed into adsorption tanks, stirs, resin, resin are added when temperature drops to 55 DEG C -65 DEG C
Dosage is the 20% of filtrate, mixing speed be 60-70 turn/when, stir 6-7 hour, continue to be cooled to 40 DEG C -45 DEG C, release and give up
Liquid collects filtering resin with 90-100 mesh nylon leaching net；
S4: elution: resin being drained, is transferred in liquid gas two-phase degreasing class device, and the butane for being passed through liquefied carries out resin
Washing is 1-1.5 hour to the control of resin wash time, by washing after, butane under liquid gas biphase equilibrium state takes off
Lipid device is in the state that butane drains and takes out resin, and resin is washed with 1.2 times of 5% dilute sodium chloride aqueous solution,
Time about 10-20 minutes, drained after washing, will be washed after resin carried out with the sodium-chloride water solution of 18%-20% concentration
Elution, elution carry out in three times: for the first time with 1.2-2 times of amount of resin volume submergence resin, the time about 4 hours；It is for the second time
Resin is submerged with 1.2-1.5 times of amount of resin volume, the time about 2 hours；Third time is to submerge resins with 1.2 times of amounts of resin volume,
Time about 1 hour, then waste liquid is outwelled, the salt water that mass fraction is 20%-24% is added, temperature is maintained at 50 DEG C -55 DEG C, stirs
After mixing 3-4 hours, eluent is collected；
S5: sedimentation desalination: above-mentioned eluent being filtered and is moved into settling tank, and it is 80%-85%'s that mass fraction is added while stirring
Precooled ethyl alcohol adjusts pH value to 7-8, stirs evenly sealing precipitating 23-25 hours, siphon supernatant liquor is collected for recycling ethyl alcohol
Sediment is stand-by；
S6: dehydration and drying: being filtered dry dehydration for above-mentioned sediment, is put into after oven drying up to heparin sodium.
Preferred embodiment on the basis of above scheme and as above scheme: step S1 includes to squeeze chitterlings
It handles and cleans the chitterlings after extrusion process and obtain the chitterlings after small intestine cleaning waste liquid and cleaning, then use archenteron-scrapping machine pair
Chitterlings after the cleaning carry out isolated intestinal mucosa, casing and intestines skin.
Preferred embodiment on the basis of above scheme and as above scheme: step S1 also includes that the cleaning casing obtains
To casing cleaning waste liquid and cleaning after casing, by the casing after cleaning around processing formed casing the step of.
The present invention is prominent and beneficial compared with prior art to be had the technical effect that using casing, intestines skin and the intestines in chitterlings
Mucous membrane, which is combined, produces heparin sodium, improves the utilization rate of chitterlings, increases the added value of chitterlings, produces 100,000,000 international units
Heparin sodium crude only needs 1200~1300 chitterlings, and the potency of heparin sodium is 90U/ mg~110U/mg；Extracting heparin
During sodium, while this byproduct of casing is also made, has thus greatly reduced costs of production and operation, realize chitterlings
Comprehensive utilization, reduce waste.
Specific embodiment
To keep the purposes, technical schemes and advantages of the application clearer, below in conjunction with embodiment, to technical solution into
Row clearly and completely describes,
Embodiment one
A method of heparin sodium is prepared by raw material of chitterlings, it is characterised in that: is included the following steps
S1: the preparation of intestinal mucosa: pouring into clear water in chitterlings, chitterlings is separated into intestinal mucosa, intestines skin and casing, by intestines
Mucous membrane is sent to mucous membrane pond, and intestines skin is crushed after extruding obtains intestines skin paste and is sent to mucus pool；
S2: enzymatic hydrolysis: being pumped with mucous membrane and squeeze into intestinal mucosa and intestines skin paste in enzymatic vessel from mucous membrane pond, is added into enzymatic vessel
The weight ratio of water, mucous membrane and water be 1:9, be stirring evenly and then adding into 2709 basic protein enzyme adjustment pH values be 7.8, mucous membrane with
The weight ratio of 2709 alkali proteases is 500:1, and heating makes temperature reach 60 DEG C, and it is special to add the casing that weight percent is 5%
It with salt, surveys its salinity and reaches 2.5%, keep the temperature 3 hours, continuing heating makes temperature reach 85 DEG C, by pH value adjustment to 8.5, heat preservation 65
It after minute, is filtered with 100 mesh nylon cloths, collects filtered fluid；
S3: absorption: above-mentioned filtered fluid is squeezed into adsorption tanks, is stirred, resin is added when temperature drops to 65 DEG C, resin dosage is
The 20% of filtrate, mixing speed is 70 turns/when, it stirs 7 hours, continues to be cooled to 45 DEG C, waste liquid is released, with 100 mesh nylon leaching nets
Collect filtering resin；
S4: elution: resin being drained, is transferred in liquid gas two-phase degreasing class device, and the butane for being passed through liquefied carries out resin
Washing, to resin wash time control be 1.5 hours, by washing after, the butane degreasing under liquid gas biphase equilibrium state
Class device is in the state that butane drains and takes out resin, and resin is washed with 1.2 times of 5% dilute sodium chloride aqueous solution, when
Between about 20 minutes, drained after washing, will be washed after resin eluted with the sodium-chloride water solution of 20% concentration, elute
It carries out in three times: for the first time with 2 times of amounts of resin volume submergence resin, the time about 4 hours；Second is with resin volume 1.5
Amount submergence resin again, the time about 2 hours；Third time is that resins are submerged with 1.2 times of amounts of resin volume, the time about 1 hour, then
Waste liquid is outwelled, the salt water that mass fraction is 24% is added and collects eluent after temperature is maintained at 55 DEG C, stirring 3-4 hours；
S5: sedimentation desalination: eluent is filtered (or separated with supercentrifuge, 3600 revs/min -4000 revs/min, 20 points
Clock), filtrate is transferred in settling tank after removing impurity, and the precooled ethyl alcohol that mass fraction is 85% is added while stirring, adjusts
PH value stirs evenly sealing precipitating 25 hours, it is stand-by to collect sediment for recycling ethyl alcohol for siphon supernatant liquor to 8；
S6: dehydration and drying: being filtered dry dehydration for above-mentioned sediment, is put into after oven drying up to heparin sodium.
Further, step S1 include extrusion process is carried out to chitterlings and the chitterlings that clean after extrusion process obtain it is small
Intestines clean the chitterlings after waste liquid and cleaning, and it is viscous then to carry out isolated intestines to the chitterlings after the cleaning using archenteron-scrapping machine
Film, casing and intestines skin.
Further, step S1 also includes that the cleaning casing obtains the casing after casing cleaning waste liquid and cleaning, will be clear
Casing after washing around processing formed casing the step of.
Embodiment two
A method of heparin sodium is prepared by raw material of chitterlings, it is characterised in that: is included the following steps
S1: the preparation of intestinal mucosa: pouring into clear water in chitterlings, chitterlings is separated into intestinal mucosa, intestines skin and casing, by intestines
Mucous membrane is sent to mucous membrane pond, and intestines skin is crushed after extruding obtains intestines skin paste and is sent to mucus pool；
S2: enzymatic hydrolysis: being pumped with mucous membrane and squeeze into intestinal mucosa and intestines skin paste in enzymatic vessel from mucous membrane pond, is added into enzymatic vessel
The weight ratio of water, mucous membrane and water is 1:7, and being stirring evenly and then adding into 2709 basic protein enzyme adjustment pH values is 7.5, mucous membrane and 2709
The weight ratio of alkali protease is 500:1, and heating makes temperature reach 55 DEG C, and it is dedicated to add the casing that weight percent is 3%
Salt surveys its salinity and reaches 2%, keeps the temperature 2 hours, and continuing heating makes temperature reach 80 DEG C, by pH value adjustment to 8, keeps the temperature 50 minutes
Afterwards, it is filtered with 90 mesh nylon cloths, collects filtered fluid；
S3: absorption: above-mentioned filtered fluid is squeezed into adsorption tanks, is stirred, resin is added when temperature drops to 55 DEG C, resin dosage is
The 20% of filtrate, mixing speed is 60 turns/when, it stirs 6 hours, continues to be cooled to 40 DEG C, waste liquid is released, with 90 mesh nylon leaching nets
Collect filtering resin；
S4: elution: resin being drained, is transferred in liquid gas two-phase degreasing class device, and the butane for being passed through liquefied carries out resin
Washing, to resin wash time control be 1 hour, by washing after, the butane degreasing class under liquid gas biphase equilibrium state
Device is in the state that butane drains and takes out resin, resin is washed with 1.2 times of 5% dilute sodium chloride aqueous solution, the time
About 10 minutes, drained after washing, will be washed after resin eluted with the sodium-chloride water solution of 18% concentration, elution point
It carries out three times: for the first time with 1.2 times of amounts of resin volume submergence resin, the time about 4 hours；Second is with resin volume 1.2
Amount submergence resin again, the time about 2 hours；Third time is that resins are submerged with 1.2 times of amounts of resin volume, the time about 1 hour, then
Waste liquid is outwelled, the salt water that mass fraction is 20% is added and collects eluent after temperature is maintained at 50 DEG C, stirring 3 hours；
S5: sedimentation desalination: eluent is filtered (or separated with supercentrifuge, 3600 revs/min -4000 revs/min, 20 points
Clock), filtrate is transferred in settling tank after removing impurity, and the precooled ethyl alcohol that mass fraction is 80% is added while stirring, adjusts
PH value stirs evenly sealing precipitating 23 hours, it is stand-by to collect sediment for recycling ethyl alcohol for siphon supernatant liquor to 7；
S6: dehydration and drying: being filtered dry dehydration for above-mentioned sediment, is put into after oven drying up to heparin sodium.
Further, step S1 include extrusion process is carried out to chitterlings and the chitterlings that clean after extrusion process obtain it is small
Intestines clean the chitterlings after waste liquid and cleaning, and it is viscous then to carry out isolated intestines to the chitterlings after the cleaning using archenteron-scrapping machine
Film, casing and intestines skin.
Further, step S1 also includes that the cleaning casing obtains the casing after casing cleaning waste liquid and cleaning, will be clear
Casing after washing around processing formed casing the step of
Embodiment three
A method of heparin sodium is prepared by raw material of chitterlings, it is characterised in that: is included the following steps
S1: the preparation of intestinal mucosa: pouring into clear water in chitterlings, chitterlings is separated into intestinal mucosa, intestines skin and casing, by intestines
Mucous membrane is sent to mucous membrane pond, and intestines skin is crushed after extruding obtains intestines skin paste and is sent to mucus pool；
S2: enzymatic hydrolysis: being pumped with mucous membrane and squeeze into intestinal mucosa and intestines skin paste in enzymatic vessel from mucous membrane pond, is added into enzymatic vessel
The weight ratio of water, mucous membrane and water is 1:8, and being stirring evenly and then adding into 2709 basic protein enzyme adjustment pH values is 7.6, mucous membrane and 2709
The weight ratio of alkali protease is 500:1, and heating makes temperature reach 58 DEG C, and it is dedicated to add the casing that weight percent is 4%
Salt surveys its salinity and reaches 2.25%, keeps the temperature 2.5 hours, and continuing heating makes temperature reach 82.5 DEG C, and pH value adjustment to 8.25 is protected
Temperature after sixty minutes, is filtered with 90-100 mesh nylon cloth, collects filtered fluid；
S3: absorption: above-mentioned filtered fluid is squeezed into adsorption tanks, is stirred, resin is added when temperature drops to 60 DEG C, resin dosage is
The 20% of filtrate, mixing speed be 60-70 turn/when, stir 6.5 hours, continue to be cooled to 42.5 DEG C, release waste liquid, use 90-100
Mesh nylon leaching net collects filtering resin；
S4: elution: resin being drained, is transferred in liquid gas two-phase degreasing class device, and the butane for being passed through liquefied carries out resin
Washing, to resin wash time control be 1.25 hours, by washing after, the butane degreasing under liquid gas biphase equilibrium state
Class device is in the state that butane drains and takes out resin, and resin is washed with 1.2 times of 5% dilute sodium chloride aqueous solution, when
Between about 15 minutes, drained after washing, will be washed after resin eluted with the sodium-chloride water solution of 19% concentration, elute
It carries out in three times: for the first time with 1.6 times of amounts of resin volume submergence resin, the time about 4 hours；Second is with resin volume
1.35 times of amounts submergence resins, the time about 2 hours；Third time is that resins are submerged with 1.2 times of amounts of resin volume, the time about 1 hour,
Then waste liquid is outwelled, the salt water that mass fraction is 22% is added, after temperature is maintained at 52.5 DEG C, stirring 3.5 hours, collection is washed
De- liquid；
S5: sedimentation desalination: eluent is filtered (or separated with supercentrifuge, 3600 revs/min -4000 revs/min, 20 points
Clock), filtrate is transferred in settling tank after removing impurity, the precooled ethyl alcohol that mass fraction is 82.5% is added while stirring,
It adjusts pH value to 7.5, stirs evenly sealing precipitating 24 hours, it is stand-by to collect sediment for recycling ethyl alcohol for siphon supernatant liquor；
S6: dehydration and drying: being filtered dry dehydration for above-mentioned sediment, is put into after oven drying up to heparin sodium.
Further, step S1 include extrusion process is carried out to chitterlings and the chitterlings that clean after extrusion process obtain it is small
Intestines clean the chitterlings after waste liquid and cleaning, and it is viscous then to carry out isolated intestines to the chitterlings after the cleaning using archenteron-scrapping machine
Film, casing and intestines skin.
Further, step S1 also includes that the cleaning casing obtains the casing after casing cleaning waste liquid and cleaning, will be clear
Casing after washing around processing formed casing the step of
Above-described embodiment is only presently preferred embodiments of the present invention, is not limited the scope of the invention according to this, therefore: Fan Yibenfa
The equivalence changes that bright structure, shape, principle are done, should all be covered by within protection scope of the present invention.
Claims (3)
1. a kind of method for preparing heparin sodium as raw material using chitterlings, it is characterised in that: include the following steps
S1: the preparation of intestinal mucosa: pouring into clear water in chitterlings, chitterlings is separated into intestinal mucosa, intestines skin and casing, by intestines
Mucous membrane is sent to mucous membrane pond, and intestines skin is crushed after extruding obtains intestines skin paste and is sent to mucus pool；
S2: enzymatic hydrolysis: being pumped with mucous membrane and squeeze into intestinal mucosa and intestines skin paste in enzymatic vessel from mucous membrane pond, is added into enzymatic vessel
The weight ratio of water, mucous membrane and water is 1:7-9, and being stirring evenly and then adding into 2709 basic protein enzyme adjustment pH values is 7.5-7.8, mucous membrane
Weight ratio with 2709 alkali proteases is 500:1, and heating makes temperature reach 55 DEG C -60 DEG C, and adding weight percent is 3%-
5% casing special-purpose salt surveys its salinity and reaches 2%-2.5%, keeps the temperature 2-3 hours, and continuing heating makes temperature reach 80 DEG C -85 DEG C, will
PH value adjustment is filtered after heat preservation 50-65 minutes with 90-100 mesh nylon cloth to 8-8.5, collects filtered fluid；
S3: absorption: above-mentioned filtered fluid is squeezed into adsorption tanks, stirs, resin, resin are added when temperature drops to 55 DEG C -65 DEG C
Dosage is the 20% of filtrate, mixing speed be 60-70 turn/when, stir 6-7 hour, continue to be cooled to 40 DEG C -45 DEG C, release and give up
Liquid collects filtering resin with 90-100 mesh nylon leaching net；
S4: elution: resin being drained, is transferred in liquid gas two-phase degreasing class device, and the butane for being passed through liquefied carries out resin
Washing is 1-1.5 hour to the control of resin wash time, by washing after, butane under liquid gas biphase equilibrium state takes off
Lipid device is in the state that butane drains and takes out resin, and resin is washed with 1.2 times of 5% dilute sodium chloride aqueous solution,
Time about 10-20 minutes, drained after washing, will be washed after resin carried out with the sodium-chloride water solution of 18%-20% concentration
Elution, elution carry out in three times: for the first time with 1.2-2 times of amount of resin volume submergence resin, the time about 4 hours；It is for the second time
Resin is submerged with 1.2-1.5 times of amount of resin volume, the time about 2 hours；Third time is to submerge resins with 1.2 times of amounts of resin volume,
Time about 1 hour, then waste liquid is outwelled, the salt water that mass fraction is 20%-24% is added, temperature is maintained at 50 DEG C -55 DEG C, stirs
After mixing 3-4 hours, eluent is collected；
S5: sedimentation desalination: above-mentioned eluent being filtered and is moved into settling tank, and it is 80%-85%'s that mass fraction is added while stirring
Precooled ethyl alcohol adjusts pH value to 7-8, stirs evenly sealing precipitating 23-25 hours, siphon supernatant liquor is collected for recycling ethyl alcohol
Sediment is stand-by；
S6: dehydration and drying: being filtered dry dehydration for above-mentioned sediment, is put into after oven drying up to heparin sodium.
2. a kind of method for preparing heparin sodium as raw material using chitterlings according to claim 1, it is characterised in that: step S1
Comprising carrying out extrusion process to chitterlings and cleaning the pig after the chitterlings after extrusion process obtain small intestine cleaning waste liquid and cleaning
Then small intestine carries out isolated intestinal mucosa, casing and intestines skin to the chitterlings after the cleaning using archenteron-scrapping machine.
3. a kind of method for preparing heparin sodium as raw material using chitterlings according to claim 2, it is characterised in that: step S1
Also comprising cleaning the casing after the casing obtains casing cleaning waste liquid and cleaning, the casing after cleaning is formed into intestines around processing
Clothing the step of.
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| CN201811096277.3A CN109293801A (en) | 2018-09-19 | 2018-09-19 | A method of heparin sodium is prepared by raw material of chitterlings |
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| CN201811096277.3A CN109293801A (en) | 2018-09-19 | 2018-09-19 | A method of heparin sodium is prepared by raw material of chitterlings |
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| CN112812202A (en) * | 2020-12-28 | 2021-05-18 | 黄金时代股份有限公司 | Heparin crude production system of processing |
| CN115304689A (en) * | 2022-09-19 | 2022-11-08 | 巴彦淖尔海成生物科技有限公司 | Method for preparing crude heparin sodium by taking animal small intestine mucosa as raw material |
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Application publication date: 20190201 |