CN102153672B - Method for extracting chondroitin sulfate - Google Patents
Method for extracting chondroitin sulfate Download PDFInfo
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- CN102153672B CN102153672B CN 201010609853 CN201010609853A CN102153672B CN 102153672 B CN102153672 B CN 102153672B CN 201010609853 CN201010609853 CN 201010609853 CN 201010609853 A CN201010609853 A CN 201010609853A CN 102153672 B CN102153672 B CN 102153672B
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- enzymolysis
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Abstract
The invention discloses a method for extracting chondroitin sulfate. Pig nose cartilage is used as a main raw material in the method; and fine chondroitin sulfate is obtained by digestion, impurity removal, water distribution, enzymolysis, decolorization, sedimentation, enzymolysis, nano-filtration, sedimentation and drying. The product produced by using the method has high purity, any toxic or harmful substance is not produced, and the obtained product has high safety and can be widely applied to the fields of medicaments, health-care products and the like.
Description
Technical field
The inventive method relates to pig cartilage deep process technology field, is specifically related to a kind of process for extracting of CHS, be a kind of be raw material with the hog snout cartilage, through biological enzymolysis technology, thereby extract the method for CHS.
Background technology
CHS is the acidic mucopolysaccharide that from animal tissues, extracts preparation; Be a kind of of TGSS C3; By D-glucuronic acid and N-acetylamino galactosamine with β-1; The polysaccharide that repetition disaccharide unit that the 4-glycosidic link is formed by connecting is formed, and sulphating takes place on the C-4 position of N-acetylamino galactosamine or C-6 position hydroxyl.CHS corneal collegen filament have provide protection, can promote the growth of fiber in the matrix, strengthen permeability, improve blood circulation, quicken metabolism, promote the absorption of penetrating fluid and the elimination of inflammation; Its polyanion has strong water-retentivity, can improve the water metabolism of cornea tissue, and corneal has stronger avidity, can form the ventilative water-retaining film of one deck at anterior corneal surface, promotes the healing of corneal wound and improves the eye dryness symptom.In addition, it brings glad tidings for the arthralgia patient, mainly shows the pad effect that provides on the one hand, and impact and friction when relaxing action supply important oxygen on the other hand and nutrient substance is delivered to the joint, help to remove IA refuse.
At present, extract the method that CHS generally adopts diluted alkaline or concentrated base to combine with enzymolysis both at home and abroad.In the diluted alkaline enzymolysis process, the dilute alkaline soln consumption is big, makes when alcohol precipitation, and consumption is bigger; In the concentrated base enzymolysis process, the alkali consumption is big, for purification at the back brings difficulty.
Summary of the invention
To the deficiency that exists in the prior art, the object of the present invention is to provide a kind of process for extracting of CHS, be a kind of to improve product purity, energy-conservation and to improve Product Safety be the process for extracting of purpose, this method helps industrialized production.
In order to realize above-mentioned technical purpose, the step of the process for extracting of a kind of CHS of the present invention is following:
1, takes by weighing commercially available hog snout cartilage as raw material, cut off reticular tissue, add water boil after cleaning; Remove degrease, will from water, pull out except that the hog snout cartilage behind the degrease then, rub with mincer; Add the water of 3 times of weight of raw material hog snout cartilage, adjust pH is warming up to 45-50 ℃ to 8.5-9.0; Add the bovine trypsin (enzyme activity is 3.3 ten thousand U/g) that accounts for raw material pig cartilage weight 2%, stir enzymolysis 4-5h down at 45-50 ℃, the pH value stabilization that keeps reaction system in the enzymolysis process is at 8.5-9.0;
2, with the enzymolysis solution adjust pH to 6.8-7.0, add the white bole of raw material hog snout cartilage weight 1%, 1% active carbon powder successively, 45 ℃ of whip attachment 1h remove by filter white bole and gac then, must filtrate;
3, pH value of filtrate is transferred to 6.0, add the long-pending 95wt% ethanol of its triploid then, leave standstill after fully stirring, remove supernatant, must precipitate, be the CHS bullion;
4, the pure water that adds 9 times of bullion weight is with dissolving crude product; The sodium-chlor that adds bullion weight 1% more successively, 1% bovine trypsin (enzyme activity is 3.3 ten thousand U/g), 45-50 ℃ is stirred enzymolysis 3-4h down; The pH value stabilization of control reaction system is at 8.5-9.0 in the enzymolysis process; Be warming up to 90-95 ℃ then, keep 10-15min, cross then and filter filtered liq with the enzyme that goes out;
The pH value of 5, regulating filtered liq is used the filter filtration of filter membrane aperture as 5nm, to remove inorganic salt and small molecular weight impurity then to 2.0-3.0;
6, regulate gained pH value of filtrate to 6.5, write down the liquor capacity of this moment, the 90wt% ethanol that adds 2 times of this volumes then precipitates, and isolates post precipitation, adds and the isopyknic absolute ethyl alcohol dehydration of deposition, and last vacuum-drying gets the CHS elaboration.
The bronsted lowry acids and bases bronsted lowry that is used to regulate the pH value in the inventive method is respectively hydrochloric acid and the solid sodium hydroxide of 35～37wt%.
Compared with prior art, advantage of the inventive method and beneficial effect are following:
1, adopt white bole and activated carbon decolorizing, both avoided objectionable impurities residual in product, can guarantee the natural whiteness of product again, products obtained therefrom is white uniform powder;
2, adopt nanofiltration of the present invention, the liquid towards material separates, concentrates and purifying, all operates, does not have phase-state change, energy-efficient at normal temperatures, and do not produce pollution in the production process;
3, the purity of this law products obtained therefrom is high, does not produce any hazardous and noxious substances, and safety has no side effect, and can be widely used in fields such as medicine, healthcare products.
Embodiment
Below in conjunction with concrete embodiment the inventive method is done further detailed description.
The bronsted lowry acids and bases bronsted lowry that is used to regulate the pH value in following examples is respectively hydrochloric acid and the solid sodium hydroxide of 35～37wt%.
Embodiment 1:
A kind of process for extracting of CHS, its step is following:
1, takes by weighing commercially available hog snout cartilage 1kg, cut off reticular tissue, add water boil after cleaning; Remove degrease, will from water, pull out except that the hog snout cartilage behind the degrease then, rub with mincer; Add the 3kg pure water, adjust pH to 8.5 is warming up to 45 ℃; Add 20g bovine trypsin (enzyme activity is 3.3 ten thousand U/g), stir enzymolysis 4h down at 45 ℃, the pH value stabilization that keeps reaction system in the enzymolysis process is 8.5;
2, with enzymolysis solution adjust pH to 6.8, add 10g white bole, 10g active carbon powder successively, 45 ℃ of whip attachment 1h remove by filter white bole and gac then, must filtrate;
3, pH value of filtrate is transferred to 6.0, add the long-pending 95wt% ethanol of its triploid then, leave standstill after fully stirring, remove supernatant, must precipitate 400g, be the CHS bullion;
4, the pure water that adds 3.6kg is with dissolving crude product; Add 4g sodium-chlor more successively, 4g bovine trypsin (enzyme activity is 3.3 ten thousand U/g), 45 ℃ are stirred enzymolysis 4h down; The pH value stabilization of control reaction system is 8.5 in the enzymolysis process; Be warming up to 90 ℃ then, keep 15min, cross then and filter filtered liq with the enzyme that goes out;
5, regulate the pH value to 2.0 of filtered liq, use of the filter filtration of filter membrane aperture then, to remove inorganic salt and small molecular weight impurity as 5nm;
6, regulate gained pH value of filtrate to 6.5; Write down the liquor capacity of this moment, the 90wt% ethanol that adds 2 times of this volumes then precipitates, and isolates post precipitation; Add and the isopyknic absolute ethyl alcohol dehydration of deposition, last vacuum-drying gets CHS elaboration 250g.
The examining report of table 1 embodiment 1 products obtained therefrom
| Test item | Standard code | Detected result |
| Product purity | ≥76％ | 98.5％ |
| Proterties | White powder | White powder |
| PH value (the 1wt% aqueous solution) | 4.0-6.0 | 5.5 |
| Weight loss on drying | ≤5％ | 1.4％ |
| Nitrogen content | 2.2-3.8％ | 3.0％ |
| Heavy metal (in the Pb element) | ＜2.0ppm | 0.5ppm |
Embodiment 2:
A kind of process for extracting of CHS, its step is following:
1, takes by weighing commercially available hog snout cartilage 2kg, cut off reticular tissue, add water boil after cleaning; Remove degrease, will from water, pull out except that the hog snout cartilage behind the degrease then, rub with mincer; Add the 6kg pure water, adjust pH to 8.6 is warming up to 46 ℃; Add 40g bovine trypsin (enzyme activity is 3.3 ten thousand U/g), stir enzymolysis 4.2h down at 46 ℃, the pH value stabilization that keeps reaction system in the enzymolysis process is 8.6;
2, with enzymolysis solution adjust pH to 6.9, add 20g white bole, 20g active carbon powder successively, 45 ℃ of whip attachment 1h remove by filter white bole and gac then, must filtrate;
3, pH value of filtrate is transferred to 6.0, add the long-pending 95wt% ethanol of its triploid then, leave standstill after fully stirring, remove supernatant, must precipitate 820g, be the CHS bullion;
4, the pure water that adds 7.4kg is with dissolving crude product; Add 8g sodium-chlor more successively, 8g bovine trypsin (enzyme activity is 3.3 ten thousand U/g), 46 ℃ are stirred enzymolysis 4h down; The pH value stabilization of control reaction system is 8.6 in the enzymolysis process; Be warming up to 91 ℃ then, keep 14min, cross then and filter filtered liq with the enzyme that goes out;
5, regulate the pH value to 2.3 of filtered liq, use of the filter filtration of filter membrane aperture then, to remove inorganic salt and small molecular weight impurity as 5nm;
6, regulate gained pH value of filtrate to 6.5; Write down the liquor capacity of this moment, the 90wt% ethanol that adds 2 times of this volumes then precipitates, and isolates post precipitation; Add and the isopyknic absolute ethyl alcohol dehydration of deposition, last vacuum-drying gets CHS elaboration 505g.
The examining report of table 2 embodiment 2 products obtained therefroms
| Test item | Standard code | Detected result |
| Product purity | ≥76％ | 97.9％ |
| Proterties | White powder | White powder |
| PH value (the 1wt% aqueous solution) | 4.0-6.0 | 5.2 |
| Weight loss on drying | ≤5％ | 1.9％ |
| Nitrogen content | 2.2-3.8％ | 3.2％ |
| Heavy metal (in the Pb element) | ＜2.0ppm | 0.8ppm |
Embodiment 3:
A kind of process for extracting of CHS, its step is following:
1, takes by weighing commercially available hog snout cartilage 10kg, cut off reticular tissue, add water boil after cleaning; Remove degrease, will from water, pull out except that the hog snout cartilage behind the degrease then, rub with mincer; Add the 30kg pure water, adjust pH to 8.8 is warming up to 48 ℃; Add 200g bovine trypsin (enzyme activity is 3.3 ten thousand U/g), stir enzymolysis 4.5h down at 48 ℃, the pH value stabilization that keeps reaction system in the enzymolysis process is 8.8;
2, with enzymolysis solution adjust pH to 6.9, add 100g white bole, 100g active carbon powder successively, 45 ℃ of whip attachment 1h remove by filter white bole and gac then, must filtrate;
3, pH value of filtrate is transferred to 6.0, add the long-pending 95wt% ethanol of its triploid then, leave standstill after fully stirring, remove supernatant, must precipitate 4.1kg, be the CHS bullion;
4, the pure water that adds 36.9kg is with dissolving crude product; Add 40g sodium-chlor more successively, 40g bovine trypsin (enzyme activity is 3.3 ten thousand U/g), 48 ℃ are stirred enzymolysis 3.5h down; The pH value stabilization of control reaction system is 8.8 in the enzymolysis process; Be warming up to 93 ℃ then, keep 12min, cross then and filter filtered liq with the enzyme that goes out;
5, regulate the pH value to 2.5 of filtered liq, use of the filter filtration of filter membrane aperture then, to remove inorganic salt and small molecular weight impurity as 5nm;
6, regulate gained pH value of filtrate to 6.5; Write down the liquor capacity of this moment, the 90wt% ethanol that adds 2 times of this volumes then precipitates, and isolates post precipitation; Add and the isopyknic absolute ethyl alcohol dehydration of deposition, last vacuum-drying gets CHS elaboration 2.55kg.
The examining report of table 3 embodiment 3 products obtained therefroms
| Test item | Standard code | Detected result |
| Product purity | ≥76％ | 97.9％ |
| Proterties | White powder | White powder |
| PH value (the 1wt% aqueous solution) | 4.0-6.0 | 4.5 |
| Weight loss on drying | ≤5％ | 2.0％ |
| Nitrogen content | 2.2-3.8％ | 2.8％ |
| Heavy metal (in the Pb element) | ＜2.0ppm | 0.9ppm |
Embodiment 4:
A kind of process for extracting of CHS, its step is following:
1, takes by weighing commercially available hog snout cartilage 20kg, cut off reticular tissue, add water boil after cleaning; Remove degrease, will from water, pull out except that the hog snout cartilage behind the degrease then, rub with mincer; Add the 60kg pure water, adjust pH to 9.0 is warming up to 50 ℃; Add 400g bovine trypsin (enzyme activity is 3.3 ten thousand U/g), stir enzymolysis 5h down at 50 ℃, the pH value stabilization that keeps reaction system in the enzymolysis process is 9.0;
2, with enzymolysis solution adjust pH to 7.0, add 200g white bole, 200g active carbon powder successively, 45 ℃ of whip attachment 1h remove by filter white bole and gac then, must filtrate;
3, pH value of filtrate is transferred to 6.0, add the long-pending 95wt% ethanol of its triploid then, leave standstill after fully stirring, remove supernatant, must precipitate 8.1kg, be the CHS bullion;
4, the pure water that adds 72.9kg is with dissolving crude product; Add 80g sodium-chlor more successively, 80g bovine trypsin (enzyme activity is 3.3 ten thousand U/g), 50 ℃ are stirred enzymolysis 3h down; The pH value stabilization of control reaction system is 9.0 in the enzymolysis process; Be warming up to 95 ℃ then, keep 10min, cross then and filter filtered liq with the enzyme that goes out;
5, regulate the pH value to 3.0 of filtered liq, use of the filter filtration of filter membrane aperture then, to remove inorganic salt and small molecular weight impurity as 5nm;
6, regulate gained pH value of filtrate to 6.5; Write down the liquor capacity of this moment, the 90wt% ethanol that adds 2 times of this volumes then precipitates, and isolates post precipitation; Add and the isopyknic absolute ethyl alcohol dehydration of deposition, last vacuum-drying gets CHS elaboration 5.2kg.
The examining report of table 4 embodiment 4 products obtained therefroms
| Test item | Standard code | Detected result |
| Product purity | ≥76％ | 98.0％ |
| Proterties | White powder | White powder |
| PH value (the 1wt% aqueous solution) | 4.0-6.0 | 4.7 |
| Weight loss on drying | ≤5％ | 1.9％ |
| Nitrogen content | 2.2-3.8％ | 3.1％ |
| Heavy metal (in the Pb element) | ＜2.0ppm | 1.0ppm |
Claims (1)
1. the process for extracting of a CHS, step is following:
1), take by weighing the hog snout cartilage as raw material, cut off reticular tissue, add water boil after cleaning; Remove degrease, will from water, pull out except that the hog snout cartilage behind the degrease then, rub with mincer; Add the water of 3 times of weight of raw material hog snout cartilage, adjust pH is warming up to 45-50 ℃ to 8.5-9.0; The bovine trypsin that adds 3.3 ten thousand U/g that account for raw material pig cartilage weight 2% stirs enzymolysis 4-5h down at 45-50 ℃, and the pH value stabilization that keeps reaction system in the enzymolysis process is at 8.5-9.0;
2), after enzymolysis accomplishes, the enzymolysis solution adjust pH to 6.8-7.0, is added the white bole of raw material hog snout cartilage weight 1%, 1% active carbon powder successively, 45 ℃ of whip attachment 1h remove by filter white bole and gac then, must filtrate;
3), pH value of filtrate is transferred to 6.0, add the long-pending 95wt% ethanol of its triploid then, leave standstill after the stirring, remove supernatant then, the CHS bullion;
4) pure water that, adds 9 times of bullion weight is with dissolving crude product; The sodium-chlor that adds bullion weight 1% more successively, the bovine trypsin of 1% 3.3 ten thousand U/g, 45-50 ℃ is stirred enzymolysis 3-4h down; The pH value stabilization of control reaction system is at 8.5-9.0 in the enzymolysis process; Be warming up to 90-95 ℃ then, keep 10-15min, cross then and filter filtered liq with the enzyme that goes out;
5), the pH value of regulating filtered liq is to 2.0-3.0, uses the filter membrane aperture to filter as the filter of 5nm then, to remove inorganic salt and small molecular weight impurity;
6), regulating step 5) gained pH value of filtrate to 6.5; Write down the liquor capacity of this moment, the 90wt% ethanol that adds 2 times of this volumes then precipitates, and isolates post precipitation; Add and the isopyknic absolute ethyl alcohol dehydration of deposition, last vacuum-drying gets the CHS elaboration.
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| CN 201010609853 CN102153672B (en) | 2010-12-28 | 2010-12-28 | Method for extracting chondroitin sulfate |
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| CN 201010609853 CN102153672B (en) | 2010-12-28 | 2010-12-28 | Method for extracting chondroitin sulfate |
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| CN102153672B true CN102153672B (en) | 2012-11-21 |
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Families Citing this family (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102965414B (en) * | 2012-11-27 | 2014-10-22 | 江南大学 | Method for extracting chondroitin sulfate from fermentation broth |
| CN104140472B (en) * | 2013-05-08 | 2017-11-28 | 清华大学 | Fine work chondroitin sulfate A (CSA) and C and prepare fine work chondroitin sulfate A (CSA) and C method |
| CN104413329A (en) * | 2013-08-22 | 2015-03-18 | 青岛蓝农谷农产品研究开发有限公司 | Chondroitin sulfate health product |
| CN103804517A (en) * | 2013-11-22 | 2014-05-21 | 青岛九龙生物医药有限公司 | Preparation method for increasing chondroitin sulfate yield |
| CN103788231B (en) * | 2014-02-24 | 2016-05-11 | 中国海洋大学 | A kind of method of preparing high-purity sulfuric acid Chondroitin A from rabbit ear cartilage |
| CN104004114A (en) * | 2014-06-06 | 2014-08-27 | 嘉兴纽迪康生物科技有限公司 | Preparation method of low-molecular-weight chondroitin sulfate |
| CN105504094B (en) * | 2016-01-20 | 2017-09-29 | 定陶县地元生化制品有限公司 | Liquid and preparation method thereof is digested without sodium ion chondroitin |
| CN109280093A (en) * | 2018-05-18 | 2019-01-29 | 山阳县恒桓生物科技有限公司 | A method of chondroitin sulfate is extracted using hog snout cartilage |
Citations (3)
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| CN1624004A (en) * | 2004-10-20 | 2005-06-08 | 天津科技大学 | Process for extracting chondroitin sulfate |
| CN101280027A (en) * | 2007-10-18 | 2008-10-08 | 日照众山生物科技有限公司 | Extracting method of chondroitin sulfate |
| CN101392035A (en) * | 2007-09-21 | 2009-03-25 | 哈尔滨奥霖药业有限公司 | Method for producing high-content chondroitin sulfate |
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Patent Citations (3)
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|---|---|---|---|---|
| CN1624004A (en) * | 2004-10-20 | 2005-06-08 | 天津科技大学 | Process for extracting chondroitin sulfate |
| CN101392035A (en) * | 2007-09-21 | 2009-03-25 | 哈尔滨奥霖药业有限公司 | Method for producing high-content chondroitin sulfate |
| CN101280027A (en) * | 2007-10-18 | 2008-10-08 | 日照众山生物科技有限公司 | Extracting method of chondroitin sulfate |
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