CN104788586A - Strontium chondroitin sulfate and preparation method thereof - Google Patents
Strontium chondroitin sulfate and preparation method thereof Download PDFInfo
- Publication number
- CN104788586A CN104788586A CN201510150350.0A CN201510150350A CN104788586A CN 104788586 A CN104788586 A CN 104788586A CN 201510150350 A CN201510150350 A CN 201510150350A CN 104788586 A CN104788586 A CN 104788586A
- Authority
- CN
- China
- Prior art keywords
- chondroitin sulfate
- strontium
- hours
- cartilage
- preparation
- 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.)
- Granted
Links
Landscapes
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention relates to strontium chondroitin sulfate and a preparation method thereof. The molecular formula of the strontium chondroitin sulfate is H2O(C14H19NSrO14S)x, wherein x is an integer of 60-100. According to the strontium chondroitin sulfate, the problems of petechiae and hematoma caused by calcium deposition of blood vessel parts and blood capillary parts can be avoided, and osteoporosis and osteoarthritis can be better treated in an aid way.
Description
Technical field
The invention belongs to biological technical field, particularly relate to a kind of chondroitin sulfate strontium and preparation method thereof.
Background technology
Chondroitin sulfate is a kind of mucopolysaccharide class material extracted from animal cartilage, forms primarily of D-Glucose aldehydic acid and N-acetyl-D-amino semi-lactosi.It has provide protection to bone, can prevent that bone is hardening to become fragile, and stops further calcium to harden, the snappiness making bone keep certain, contributes to the recovery of hyperosteogeny, have important effect in Bone Defect Repari, arthropathic control etc.
The method of tradition extraction chondroitin sulfate mainly contains alkali and puies forward enzymolysis process, alkaline extraction, ultrasonic assistant method and acetic acid extraction method etc., the chondroitin sulfate that these traditional methods are prepared mainly exists with the form of Sodium chondroitin sulfate A, and the affinity of chondroitin sulfate to calcium is better than sodium, after using, the calcium at blood vessel and capillary vessel position is easily caused to stockpile and cause petechiae and hemotoncus, poor to the preventing effectiveness of osteoporosis and osteoarthritis.
Summary of the invention
Given this, be necessary to provide a kind of chondroitin sulfate strontium, the problem of the petechiae that this chondroitin sulfate strontium can be avoided the calcium at blood vessel and capillary vessel position to stockpile and cause and hemotoncus, and can assisting therapy osteoporosis and osteoarthritis preferably.
In addition, a kind of preparation method of chondroitin sulfate strontium is also provided.
A kind of chondroitin sulfate strontium, the molecular formula of described chondroitin sulfate strontium is H
2o (C
14h
19nSrO
14s)
x, wherein, x is the integer of 60 ~ 100.
A preparation method for chondroitin sulfate strontium, comprises the steps:
The aqueous solution of Sodium chondroitin sulfate A is crossed cation exchange resin column, when the pH value of effluent liquid collects effluent liquid from neutrality becomes 4.5 ~ 5, until the pH value of effluent liquid becomes 6.5 ~ 7, stops collecting;
Regulate pH value to 6 ~ 6.5 of the described effluent liquid collected, then add soluble strontium salt, stirring and dissolving 20 minutes ~ 60 minutes, through standing and reacting 3 hours ~ 5 hours, obtain reaction solution, wherein, the mass ratio of described water-soluble strontium salt and described Sodium chondroitin sulfate A is 0.4:1 ~ 0.8:1; And
In described reaction solution, add mass percentage is that the aqueous solution of the ethanol of 90% ~ 95% obtains mixed solution, and the mass percentage of ethanol in described mixed solution is 60% ~ 65%, and left standstill by described mixed solution, then suction filtration, obtaining molecular formula is H
2o (C
14h
19nSrO
14s)
xchondroitin sulfate strontium, wherein, x is the integer of 60 ~ 100.
Wherein in an embodiment, the mass ratio of the Zeo-karb in described cation exchange resin column and described Sodium chondroitin sulfate A is 3:1 ~ 4:1.
Wherein in an embodiment, in the aqueous solution of described Sodium chondroitin sulfate A, the mass percentage of described Sodium chondroitin sulfate A is 5% ~ 10%.
Wherein in an embodiment, described water-soluble strontium salt is strontium chloride or Strontium Ranelate.
Wherein in an embodiment, the time that described mixed solution leaves standstill is 12 hours ~ 15 hours.
Wherein in an embodiment, the suction filtration thing also comprised suction filtration obtains carries out cleaning and dry step, and described cleaning and dry step are: use dehydrated alcohol to clean described suction filtration thing, then in 60 DEG C ~ 80 DEG C vacuum-dryings 2 hours ~ 4 hours.
Wherein in an embodiment, also comprise the preparation process of described Sodium chondroitin sulfate A:
Pulverize after cartilage degrease, obtain cartilage powder;
Described cartilage powder is mixed with deionized water, then adds basifier, be stirred to described cartilage powder in 30 DEG C ~ 40 DEG C and dissolve, obtain Cartilage solution, wherein, the mass ratio of described basifier and described cartilage is 0.01:1 ~ 0.5:1, and described basifier is sodium hydroxide or sodium bicarbonate;
Regulate pH value to 8 ~ 9 of described Cartilage solution, add pancreatin, in 40 DEG C ~ 50 DEG C enzymolysis 4 hours ~ 5 hours, be warming up to 80 DEG C ~ 100 DEG C insulation reaction 2 hours ~ 4 hours again, after filtration, obtain settled solution, the mass ratio of described pancreatin and described cartilage is 0.0045:1 ~ 0.0055:1; And
Regulate pH value to 6 ~ 6.5 of described settled solution, add dehydrated alcohol and obtain mixture, and the mass percentage of ethanol in described mixture is 70% ~ 75%, then suction filtration, obtains Sodium chondroitin sulfate A.
Wherein in an embodiment, in the step mix described cartilage powder with described deionized water, the quality of described deionized water is 5 times ~ 30 times of the quality of cartilage.
Wherein in an embodiment, by the step of described cartilage degrease be: by 80 DEG C ~ 100 DEG C boilings 2 hours ~ 3 hours in deionized water of described cartilage, after removing described grease, then 80 DEG C ~ 100 DEG C boilings 2 hours ~ 4 hours in deionized water, dry after cooling.
Above-mentioned chondroitin sulfate strontium is a kind of new chondroitin sulfate, strontium in this chondroitin sulfate strontium enters cell by calcium channel, combine with relevant calcium binding site in cell, have and promote that skeleton development and osteoid are formed and regulate the multiple action of bone metabolism, and strontium has the effect promoting Oesteoblast growth and suppress osteoclast formation, on a cellular level, strontium reduces bone by osteoclast and heavily to absorb and by scleroblast promoting bone growing, and the affinity of chondroitin sulfate to strontium and calcium is suitable, can avoid and use traditional chondroitin sulfate existed with the form of Sodium chondroitin sulfate A easily to cause the calcium at blood vessel and capillary vessel position to stockpile, and cause the problem of petechiae and hemotoncus, can also assisting therapy osteoporosis and osteoarthritis preferably.
Accompanying drawing explanation
Fig. 1 is the schema of the preparation method of the chondroitin sulfate strontium of an embodiment;
The relative expression quantity of interleukin (IL-1 β) mRNA when concentration of what Fig. 2 represented the is chondroitin sulfate strontium of embodiment 1 is respectively 0mmol/L, 1mmol/L, 3mmol/L and 5mmol/L;
The relative expression quantity of collagen protein (CollagenI) mRNA when concentration of what Fig. 3 represented the is chondroitin sulfate strontium of embodiment 1 is respectively 0mmol/L, 1mmol/L, 3mmol/L and 5mmol/L;
The relative expression quantity of membranin (RANKL) mRNA when concentration of what Fig. 4 represented the is chondroitin sulfate strontium of embodiment 1 is respectively 0mmol/L, 1mmol/L, 3mmol/L and 5mmol/L;
The relative expression quantity of interleukin (IL-1 β) mRNA when concentration of what Fig. 5 represented is Sodium chondroitin sulfate A is respectively 0mmol/L, 1mmol/L, 3mmol/L and 5mmol/L;
The relative expression quantity of collagen protein (CollagenI) mRNA when concentration of what Fig. 6 represented is Sodium chondroitin sulfate A is respectively 0mmol/L, 1mmol/L, 3mmol/L and 5mmol/L;
The relative expression quantity of membranin (RANKL) mRNA when concentration of what Fig. 7 represented is Sodium chondroitin sulfate A is respectively 0mmol/L, 1mmol/L, 3mmol/L and 5mmol/L.
Embodiment
Mainly in conjunction with the drawings and the specific embodiments chondroitin sulfate strontium and preparation method thereof is described in further detail below.
The chondroitin sulfate strontium of one embodiment, its molecular formula is H
2o (C
14h
19nSrO
14s)
x, wherein, x is the integer of 60 ~ 100.
This chondroitin sulfate strontium can be applied to Bone Defect Repari, the arthropathic aspect such as to prevent.
Above-mentioned chondroitin sulfate strontium is a kind of new chondroitin sulfate, strontium in this chondroitin sulfate strontium enters cell by calcium channel, combine with relevant calcium binding site in cell, have and promote that skeleton development and osteoid are formed and regulate the multiple action of bone metabolism, and the affinity of chondroitin sulfate to strontium and calcium is suitable, avoid and use traditional chondroitin sulfate existed with the form of Sodium chondroitin sulfate A easily to cause the calcium at blood vessel and capillary vessel position to stockpile, and cause the problem of petechiae and hemotoncus.
As shown in Figure 1, the preparation method of the chondroitin sulfate strontium of an embodiment, can be used for preparing above-mentioned chondroitin sulfate strontium, this preparation method comprises the steps:
Step S110: the aqueous solution of Sodium chondroitin sulfate A is crossed cation exchange resin column, when the pH value of effluent liquid collects effluent liquid from neutrality becomes 4.5 ~ 5, until the pH value of effluent liquid becomes 6.5 ~ 7, stops collecting.
Wherein, Sodium chondroitin sulfate A can be bought by market; Also can by preparing voluntarily.
Wherein, the preparation process of Sodium chondroitin sulfate A is as follows: pulverize after cartilage degrease, obtain cartilage powder; Cartilage powder is mixed with deionized water, then adds basifier, be stirred to cartilage powder in 30 DEG C ~ 40 DEG C and dissolve, obtain Cartilage solution; Wherein, the mass ratio of basifier and cartilage is 0.01:1 ~ 0.5:1; Basifier is sodium hydroxide or sodium bicarbonate; Regulate pH value to 8 ~ 9 of Cartilage solution, add pancreatin, in 40 DEG C ~ 50 DEG C enzymolysis 4 hours ~ 5 hours, be warming up to 80 DEG C ~ 100 DEG C insulation reaction 2 hours ~ 4 hours again, after filtration, obtain settled solution, the mass ratio of pancreatin and cartilage is 0.0045:1 ~ 0.0055:1; Regulate pH value to 6 ~ 6.5 of settled solution, add dehydrated alcohol and obtain mixture, and the mass percentage of ethanol in mixture is 70% ~ 75%, then suction filtration, obtains Sodium chondroitin sulfate A.
Wherein, regulate in the step of pH value to 8 ~ 9 of Cartilage solution, use reagent for concentration be the salt aqueous acid of 3 ~ 4mol/L.
Wherein, in the step mix cartilage powder with deionized water, the quality of deionized water is 5 times ~ 30 times of the quality of cartilage.
Wherein, regulate in the step of pH value to 6 ~ 6.5 of settled solution, working concentration is pH value to 6 ~ 6.5 of the salt aqueous acid adjustment settled solution of 3 ~ 4mol/L.
Wherein, by the step of cartilage degrease be: by cartilage 80 DEG C ~ 100 DEG C boilings 2 hours ~ 3 hours in deionized water, after removing grease, then 80 DEG C ~ 100 DEG C boilings 2 hours ~ 4 hours in deionized water, dry after cooling.To guarantee that the grease in cartilage is removed as much as possible.
In step S110, in the aqueous solution of Sodium chondroitin sulfate A, the mass percentage of Sodium chondroitin sulfate A is 5% ~ 10%.
In step S110, the Zeo-karb in cation exchange resin column and the mass ratio of Sodium chondroitin sulfate A are 3:1 ~ 4:1.To make in Sodium chondroitin sulfate A sodium by complete exchange.
Wherein, Zeo-karb is with vinylbenzene and divinylbenzene polymerization, the polymkeric substance obtained through sulfuric acid sulfonation.Zeo-karb is met water and certain electron ion in the activated ion of a certain tool of itself and water can be exchanged mutually, namely replacement(metathesis)reaction occurs, and removes soluble ion in water.
Step S120: regulate pH value to 6 ~ 6.5 of effluent liquid of collecting, then add water-soluble strontium salt, stirring and dissolving 20 minutes ~ 60 minutes, through standing and reacting 3 hours ~ 5 hours, obtains reaction solution.Wherein, the mass ratio of water-soluble strontium salt and Sodium chondroitin sulfate A is 0.4:1 ~ 0.8:1.
Wherein, in step S120, regulate pH value to 6 ~ 6.5 of the effluent liquid collected, the reagent of use is the aqueous solution of the strontium hydroxide of 10% ~ 20% for mass percentage.The purity that the pH value of the effluent liquid using the adjustment of the aqueous solution of strontium hydroxide to collect can be avoided introducing extraneous ion in preparation process and affect product.
Wherein, water-soluble strontium salt is strontium chloride or Strontium Ranelate.Strontium, as one of important trace element in skeleton composition, enters cell by calcium channel, combines in cell with relevant calcium binding site, has the formation promoting skeleton development and osteoid and the multiple action regulating bone metabolism.
Step S130: add the aqueous solution that mass percentage is the ethanol of 90% ~ 95% in reaction solution, obtain mixed solution, and the mass percentage of ethanol in mixed solution is 60% ~ 65%, left standstill by mixed solution, then suction filtration, obtaining molecular formula is H
2o (C
14h
19nSrO
14s)
xchondroitin sulfate strontium, wherein, x is the integer of 60 ~ 100.
By making the mass percentage of the ethanol in mixed solution be 60% ~ 65%, precipitate to make chondroitin sulfate strontium.
Wherein, in step S130, the time that mixed solution leaves standstill is 12 hours ~ 15 hours.
Wherein, the suction filtration thing also comprised step S130 suction filtration obtains carries out cleaning and dry step, and cleaning and dry step are: use dehydrated alcohol to clean suction filtration thing, then in 60 DEG C ~ 80 DEG C vacuum-dryings 2 hours ~ 4 hours.Concrete, use dehydrated alcohol to clean 3 times to suction filtration thing.
The preparation method of above-mentioned chondroitin sulfate strontium is simple, and by using Sodium chondroitin sulfate A to be raw material, and by adopting above-mentioned steps, can prepare chondroitin sulfate strontium, be a kind of new chondroitin sulfate, strontium in this chondroitin sulfate strontium enters cell by calcium channel, combine with relevant calcium binding site in cell, have and promote that skeleton development and osteoid are formed and regulate the multiple action of bone metabolism, and strontium has the effect promoting Oesteoblast growth and suppress osteoclast formation, on a cellular level, strontium reduces bone by osteoclast and heavily to absorb and by scleroblast promoting bone growing, and the affinity of chondroitin sulfate to strontium and calcium is suitable, chondroitin sulfate strontium be not only avoid and use traditional chondroitin sulfate existed with the form of Sodium chondroitin sulfate A easily to cause the calcium at blood vessel and capillary vessel position to stockpile, and cause the problem of petechiae and hemotoncus, and can assisting therapy sacroiliitis and osteoporosis etc. preferably.And the high purity more than 95% of chondroitin sulfate strontium that the preparation method of above-mentioned chondroitin sulfate strontium prepares.And the raw material used in the preparation method of sulfuric acid ossein strontium is cartilage, wide material sources.
Be below specific embodiment part:
Embodiment 1
The preparation process of the chondroitin sulfate strontium of the present embodiment is as follows:
(1) preparation of Sodium chondroitin sulfate A: by animal cartilage 80 DEG C of boilings 3 hours in deionized water, after removing grease, then 80 DEG C of boilings 4 hours in deionized water, dry after cooling.
Pulverize after cartilage degrease, obtain cartilage powder, cartilage powder is placed in retort, add the deionized water mixing of cartilage 5 times, be then 0.01:1 according to the mass ratio of sodium hydroxide and cartilage, add sodium hydroxide, be stirred to cartilage powder in 40 DEG C to dissolve, obtain Cartilage solution; Working concentration is that the salt aqueous acid of 3mol/L regulates the pH value to 8 of Cartilage solution, is 0.0045:1, adds pancreatin according to the mass ratio of pancreatin and cartilage, in 40 DEG C of enzymolysis 4 hours, then is warming up to 80 DEG C of insulation reaction 4 hours, after filtration, obtains settled solution; Working concentration is that the salt aqueous acid of 3mol/L regulates the pH value to 6 of settled solution, adds dehydrated alcohol and obtains mixture, and the mass percentage of ethanol in mixture is 70%, and then suction filtration, obtains Sodium chondroitin sulfate A.
(2) by mass percentage be 8% the aqueous solution of Sodium chondroitin sulfate A cross cation exchange resin column, when the pH value of effluent liquid collects effluent liquid from neutrality becomes 4.5, until the pH value of effluent liquid becomes 7, stop collecting.Wherein, the Zeo-karb in cation exchange resin column and the mass ratio of Sodium chondroitin sulfate A are 3:1.
(3) functional quality percentage composition is the pH value to 6 of the effluent liquid of the aqueous solution adjustment collection of the strontium hydroxide of 10%, then adds strontium chloride, stirring and dissolving 20 minutes, through standing and reacting 3 hours, obtain reaction solution, wherein, the mass ratio of strontium chloride and Sodium chondroitin sulfate A is 0.4:1.
(4) in reaction solution, add mass percentage is that the aqueous solution of the ethanol of 90% obtains mixed solution, and the mass percentage of ethanol in mixed solution is 60%, mixed solution is left standstill 12 hours, then suction filtration, dehydrated alcohol is used to clean 3 times to suction filtration thing, then in 60 DEG C of vacuum-dryings 4 hours, obtaining molecular formula is H
2o (C
14h
19nSrO
14s)
60chondroitin sulfate strontium.
The purity of the chondroitin sulfate strontium of the present embodiment is 95%.
Embodiment 2
The preparation process of the chondroitin sulfate strontium of the present embodiment is as follows:
(1) preparation of Sodium chondroitin sulfate A: by animal cartilage 100 DEG C of boilings 2 hours in deionized water, after removing grease, then 100 DEG C of boilings 2 hours in deionized water, dry after cooling.
Pulverize after cartilage degrease, obtain cartilage powder, cartilage powder is placed in retort, add the deionized water mixing of cartilage 30 times, be then 0.5:1 according to the mass ratio of sodium bicarbonate and cartilage, add sodium bicarbonate, be stirred to cartilage powder in 30 DEG C to dissolve, obtain Cartilage solution; Working concentration is that the salt aqueous acid of 4mol/L regulates the pH value to 9 of Cartilage solution, is 0.0055:1, adds pancreatin according to the mass ratio of pancreatin and cartilage, in 50 DEG C of enzymolysis 5 hours, then is warming up to 100 DEG C of insulation reaction 2 hours, after filtration, obtains settled solution; Working concentration is that the salt aqueous acid of 4mol/L regulates the pH value to 6.5 of settled solution, adds dehydrated alcohol and obtains mixture, and the mass percentage of ethanol in mixture is 75%, and then suction filtration, obtains Sodium chondroitin sulfate A.
(2) by mass percentage be 5% the aqueous solution of Sodium chondroitin sulfate A cross cation exchange resin column, when the pH value of effluent liquid collects effluent liquid from neutrality becomes 5, until the pH value of effluent liquid becomes 6.5, stop collecting.Wherein, the Zeo-karb in cation exchange resin column and the mass ratio of Sodium chondroitin sulfate A are 4:1.
(3) functional quality percentage composition is the pH value to 6.5 of the effluent liquid of the aqueous solution adjustment collection of the strontium hydroxide of 20%, then adds strontium chloride, stirring and dissolving 60 minutes, through standing and reacting 5 hours, obtain reaction solution, wherein, the mass ratio of strontium chloride and Sodium chondroitin sulfate A is 0.8:1.
(4) in reaction solution, add mass percentage is that the aqueous solution of the ethanol of 95% obtains mixed solution, and the mass percentage of ethanol in mixed solution is 65%, mixed solution is left standstill 15 hours, then suction filtration, dehydrated alcohol is used to clean 3 times to suction filtration thing, then in 80 DEG C of vacuum-dryings 2 hours, obtaining molecular formula is H
2o (C
14h
19nSrO
14s)
80chondroitin sulfate strontium.
The purity of the chondroitin sulfate strontium of the present embodiment is 98%.
Embodiment 3
The preparation process of the chondroitin sulfate strontium of the present embodiment is as follows:
(1) preparation of Sodium chondroitin sulfate A: by animal cartilage 90 DEG C of boilings 2.5 hours in deionized water, after removing grease, then 90 DEG C of boilings 3.5 hours in deionized water, dry after cooling.
Pulverize after cartilage degrease, obtain cartilage powder, cartilage powder is placed in retort, add the deionized water mixing of cartilage 20 times, be then 0.2:1 according to the mass ratio of sodium hydroxide and cartilage, add sodium hydroxide, be stirred to cartilage powder in 35 DEG C to dissolve, obtain Cartilage solution; Working concentration is that the salt aqueous acid of 4mol/L regulates the pH value to 8 of Cartilage solution, is 0.0050:1, adds pancreatin according to the mass ratio of pancreatin and cartilage, in 50 DEG C of enzymolysis 4 hours, then is warming up to 90 DEG C of insulation reaction 3 hours, after filtration, obtains settled solution; Working concentration is that the salt aqueous acid of 4mol/L regulates the pH value to 6 of settled solution, adds dehydrated alcohol and obtains mixture, and the mass percentage of ethanol in mixture is 72%, and then suction filtration, obtains Sodium chondroitin sulfate A.
(2) by mass percentage be 10% the aqueous solution of Sodium chondroitin sulfate A cross cation exchange resin column, when the pH value of effluent liquid collects effluent liquid from neutrality becomes 4.5, until the pH value of effluent liquid becomes 6.5, stop collecting.Wherein, the Zeo-karb in cation exchange resin column and the mass ratio of Sodium chondroitin sulfate A are 3.5:1.
(3) functional quality percentage composition is the pH value to 6 of the effluent liquid of the aqueous solution adjustment collection of the strontium hydroxide of 15%, then adds Strontium Ranelate, stirring and dissolving 30 minutes, through standing and reacting 4 hours, obtain reaction solution, wherein, the mass ratio of Strontium Ranelate and Sodium chondroitin sulfate A is 0.6:1.
(4) in reaction solution, add mass percentage is that the aqueous solution of the ethanol of 95% obtains mixed solution, and the mass percentage of ethanol in mixed solution is 60%, mixed solution is left standstill 14 hours, then suction filtration, dehydrated alcohol is used to clean 3 times to suction filtration thing, then in 70 DEG C of vacuum-dryings 3 hours, obtaining molecular formula is H
2o (C
14h
19nSrO
14s)
100chondroitin sulfate strontium.
The purity of the chondroitin sulfate strontium of the present embodiment is 96%.
Embodiment 4
The preparation process of the chondroitin sulfate strontium of the present embodiment is as follows:
(1) preparation of Sodium chondroitin sulfate A: by animal cartilage 80 DEG C of boilings 3 hours in deionized water, after removing grease, then 100 DEG C of boilings 2 hours in deionized water, dry after cooling.
Pulverize after cartilage degrease, obtain cartilage powder, cartilage powder is placed in retort, add the deionized water mixing of cartilage 5 times, be then 0.4:1 according to the mass ratio of sodium bicarbonate and cartilage, add sodium bicarbonate, be stirred to cartilage powder in 40 DEG C to dissolve, obtain Cartilage solution; Working concentration is that the salt aqueous acid of 3mol/L regulates the pH value of Cartilage solution to be 9, is 0.0050:1, adds pancreatin according to the mass ratio of pancreatin and cartilage, in 40 DEG C of enzymolysis 4 hours, then is warming up to 80 DEG C of insulation reaction 4 hours, after filtration, obtains settled solution; Working concentration is that the salt aqueous acid of 3mol/L regulates the pH value to 6 of settled solution, adds dehydrated alcohol and obtains mixture, and the mass percentage of ethanol in mixture is 75%, and then suction filtration, obtains Sodium chondroitin sulfate A.
(2) by mass percentage be 6% the aqueous solution of Sodium chondroitin sulfate A cross cation exchange resin column, when the pH value of effluent liquid collects effluent liquid from neutrality becomes 4.8, until the pH value of effluent liquid becomes 6.8, stop collecting.Wherein, the Zeo-karb in cation exchange resin column and the mass ratio of Sodium chondroitin sulfate A are 3:1.
(3) functional quality percentage composition is the pH value to 6.5 of the effluent liquid of the aqueous solution adjustment collection of the strontium hydroxide of 10%, then Strontium Ranelate is added, stirring and dissolving 50 minutes, through standing and reacting 2.5 hours, obtain reaction solution, wherein, the mass ratio of Strontium Ranelate and Sodium chondroitin sulfate A is 0.7:1.
(4) in reaction solution, add mass percentage is that the aqueous solution of the ethanol of 95% obtains mixed solution, and the mass percentage of ethanol in mixed solution is 65%, mixed solution is left standstill 13 hours, then suction filtration, dehydrated alcohol is used to clean 3 times to suction filtration thing, then in 60 DEG C of vacuum-dryings 4 hours, obtaining molecular formula is H
2o (C
14h
19nSrO
14s)
90chondroitin sulfate strontium.
The purity of the chondroitin sulfate strontium of the present embodiment is 95%.
Result detects:
The chondroitin sulfate strontium of Sodium chondroitin sulfate A and embodiment 1 is carried out mrna expression detection.
The chronic joint disease of osteoarthritis to be articular cartilage degeneration be feature, osteoarthritis cartilage cell epimatrix synthetics and degradation is unbalance is one of major reason causing cartilage degeneration.Wherein, matrix metalloproteinase has Degradation to cartilage cell epimatrix, makes arthroncus, and external force resistance effect declines, and finally causes articular cartilage damage.And due to interleukin (IL-1 β) can inducer substance metalloprotein expression of enzymes, and impel it to activate, thus cause cartilage matrix to degrade.Membranin (RANKL) is the membranin on scleroblast film, can activate osteoclast, prevent osteoclast apoptosis.And collagen protein (CollagenI) is the important component part of joint cartilage.Therefore, the effect of Sodium chondroitin sulfate A and chondroitin sulfate strontium is obtained by the relative expression quantity of the mRNA detecting interleukin (IL-1 β), collagen protein (CollagenI) and membranin (RANKL).
Adopt quantitative real time PCR Instrument (ABI, Vii A7) mRNA relative expression quantity is detected, detecting the reagent used is RNAiso Plus (TaKaRa, D9108A) and SYBR Green dye method mRNA detection kit (Geneup).Concrete grammar is: joined by material to be detected (chondroitin sulfate strontium or Sodium chondroitin sulfate A) in the DMEM substratum containing 10%FBS and be mixed with the culture solution that concentration is the material to be detected of 1mmol/L, 3mmol/L and 5mmol/L respectively, the DMEM substratum (containing 10%FBS) of sulfur acid chrondroitin strontium as a control group, is not often organized in triplicate; Cultivate after 48 hours respectively, extract total serum IgE with Trizol, reverse transcription obtains 4 cDNA samples; Then the mRNA relative expression quantity of interleukin (IL-1 β), collagen protein (CollagenI) and membranin (RANKL) in 4 cDNA is detected respectively.
The relative expression quantity of interleukin (IL-1 β) mRNA when concentration of what Fig. 2 represented the is chondroitin sulfate strontium of embodiment 1 is respectively 0mmol/L, 1mmol/L, 3mmol/L and 5mmol/L; The relative expression quantity of collagen protein (CollagenI) mRNA when concentration of what Fig. 3 represented the is chondroitin sulfate strontium of embodiment 1 is respectively 0mmol/L, 1mmol/L, 3mmol/L and 5mmol/L; The relative expression quantity of membranin (RANKL) mRNA when concentration of what Fig. 4 represented the is chondroitin sulfate strontium of embodiment 1 is respectively 0mmol/L, 1mmol/L, 3mmol/L and 5mmol/L.The relative expression quantity of interleukin (IL-1 β) mRNA when concentration of what Fig. 5 represented is Sodium chondroitin sulfate A is respectively 0mmol/L, 1mmol/L, 3mmol/L and 5mmol/L; The relative expression quantity of collagen protein (CollagenI) mRNA when concentration of what Fig. 6 represented is Sodium chondroitin sulfate A is respectively 0mmol/L, 1mmol/L, 3mmol/L and 5mmol/L; The relative expression quantity of membranin (RANKL) mRNA when concentration of what Fig. 7 represented is Sodium chondroitin sulfate A is respectively 0mmol/L, 1mmol/L, 3mmol/L and 5mmol/L.
As can be seen from Fig. 2 ~ 7, along with the increase of the concentration of chondroitin sulfate strontium or Sodium chondroitin sulfate A, the relative expression quantity of interleukin (IL-1 β) mRNA reduces gradually, the relative expression quantity of collagen protein (CollagenI) mRNA increases gradually, and the relative expression quantity of membranin (RANKL) mRNA reduces gradually.And compared with the Sodium chondroitin sulfate A of same concentrations, although the relative expression quantity of the interleukin of chondroitin sulfate strontium (IL-1 β) is more or less the same, but the relative expression quantity of collagen protein (CollagenI) mRNA significantly increases, and the relative expression quantity of membranin (RANKL) also considerably reduces, obviously, the performance of chondroitin sulfate strontium is better than Sodium chondroitin sulfate A, and chondroitin sulfate strontium can assisting therapy sacroiliitis and osteoporosis better.
The above embodiment only have expressed several embodiment of the present invention, and it describes comparatively concrete and detailed, but therefore can not be interpreted as the restriction to the scope of the claims of the present invention.It should be pointed out that for the person of ordinary skill of the art, without departing from the inventive concept of the premise, can also make some distortion and improvement, these all belong to protection scope of the present invention.Therefore, the protection domain of patent of the present invention should be as the criterion with claims.
Claims (10)
1. a chondroitin sulfate strontium, is characterized in that, the molecular formula of described chondroitin sulfate strontium is H
2o (C
14h
19nSrO
14s)
x, wherein, x is the integer of 60 ~ 100.
2. a preparation method for chondroitin sulfate strontium, is characterized in that, comprises the steps:
The aqueous solution of Sodium chondroitin sulfate A is crossed cation exchange resin column, when the pH value of effluent liquid collects effluent liquid from neutrality becomes 4.5 ~ 5, until the pH value of effluent liquid becomes 6.5 ~ 7, stops collecting;
Regulate pH value to 6 ~ 6.5 of the described effluent liquid collected, then add soluble strontium salt, stirring and dissolving 20 minutes ~ 60 minutes, through standing and reacting 3 hours ~ 5 hours, obtain reaction solution, wherein, the mass ratio of described water-soluble strontium salt and described Sodium chondroitin sulfate A is 0.4:1 ~ 0.8:1; And
In described reaction solution, add mass percentage is that the aqueous solution of the ethanol of 90% ~ 95% obtains mixed solution, and the mass percentage of ethanol in described mixed solution is 60% ~ 65%, and left standstill by described mixed solution, then suction filtration, obtaining molecular formula is H
2o (C
14h
19nSrO
14s)
xchondroitin sulfate strontium, wherein, x is the integer of 60 ~ 100.
3. the preparation method of chondroitin sulfate strontium according to claim 1, is characterized in that, the mass ratio of the Zeo-karb in described cation exchange resin column and described Sodium chondroitin sulfate A is 3:1 ~ 4:1.
4. the preparation method of chondroitin sulfate strontium according to claim 1, is characterized in that, in the aqueous solution of described Sodium chondroitin sulfate A, the mass percentage of described Sodium chondroitin sulfate A is 5% ~ 10%.
5. the preparation method of chondroitin sulfate strontium according to claim 1, is characterized in that, described water-soluble strontium salt is strontium chloride or Strontium Ranelate.
6. the preparation method of chondroitin sulfate strontium according to claim 1, is characterized in that, the time that described mixed solution leaves standstill is 12 hours ~ 15 hours.
7. the preparation method of chondroitin sulfate strontium according to claim 1, it is characterized in that, the suction filtration thing also comprised suction filtration obtains carries out cleaning and dry step, described cleaning and dry step are: use dehydrated alcohol to clean described suction filtration thing, then in 60 DEG C ~ 80 DEG C vacuum-dryings 2 hours ~ 4 hours.
8. the preparation method of chondroitin sulfate strontium according to claim 1, is characterized in that, also comprises the preparation process of described Sodium chondroitin sulfate A:
Pulverize after cartilage degrease, obtain cartilage powder;
Described cartilage powder is mixed with deionized water, then adds basifier, be stirred to described cartilage powder in 30 DEG C ~ 40 DEG C and dissolve, obtain Cartilage solution, wherein, the mass ratio of described basifier and described cartilage is 0.01:1 ~ 0.5:1, and described basifier is sodium hydroxide or sodium bicarbonate;
Regulate pH value to 8 ~ 9 of described Cartilage solution, add pancreatin, in 40 DEG C ~ 50 DEG C enzymolysis 4 hours ~ 5 hours, be warming up to 80 DEG C ~ 100 DEG C insulation reaction 2 hours ~ 4 hours again, after filtration, obtain settled solution, the mass ratio of described pancreatin and described cartilage is 0.0045:1 ~ 0.0055:1; And
Regulate pH value to 6 ~ 6.5 of described settled solution, add dehydrated alcohol and obtain mixture, and the mass percentage of ethanol in described mixture is 70% ~ 75%, then suction filtration, obtains Sodium chondroitin sulfate A.
9. the preparation method of chondroitin sulfate strontium according to claim 8, is characterized in that, in the step mix described cartilage powder with described deionized water, the quality of described deionized water is 5 times ~ 30 times of the quality of cartilage.
10. the preparation method of chondroitin sulfate strontium according to claim 8, it is characterized in that, by the step of described cartilage degrease be: by 80 DEG C ~ 100 DEG C boilings 2 hours ~ 3 hours in deionized water of described cartilage, after removing described grease, 80 DEG C ~ 100 DEG C boilings 2 hours ~ 4 hours in deionized water again, dry after cooling.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510150350.0A CN104788586B (en) | 2015-03-31 | 2015-03-31 | Chondroitin sulfate strontium and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510150350.0A CN104788586B (en) | 2015-03-31 | 2015-03-31 | Chondroitin sulfate strontium and preparation method thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104788586A true CN104788586A (en) | 2015-07-22 |
| CN104788586B CN104788586B (en) | 2018-03-09 |
Family
ID=53553806
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510150350.0A Active CN104788586B (en) | 2015-03-31 | 2015-03-31 | Chondroitin sulfate strontium and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104788586B (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105440088A (en) * | 2015-11-20 | 2016-03-30 | 南方科技大学 | Glucosamine mixed strontium salt as well as preparation method and application thereof |
| CN108383927A (en) * | 2018-04-11 | 2018-08-10 | 南方科技大学 | chondroitin sulfate magnesium and preparation method thereof |
| CN110604742A (en) * | 2019-08-02 | 2019-12-24 | 南方科技大学 | Eucommia polysaccharide strontium complex and preparation method and application thereof |
| CN113583406A (en) * | 2021-08-03 | 2021-11-02 | 南方科技大学 | Composite polyether-ether-ketone material and preparation method and application thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2005123193A2 (en) * | 2004-06-17 | 2005-12-29 | Osteologix A/S | Treatments comprising strontium for rheumatic and arthritic diseases and pain |
| CN1974778A (en) * | 2006-12-05 | 2007-06-06 | 中国水产科学研究院南海水产研究所 | Sturgon chondrine extracting process |
| CN101831004A (en) * | 2010-03-24 | 2010-09-15 | 章云 | Hyaluronic acid strontium salt compounds, preparation method thereof and application in medicine |
| CN104451931A (en) * | 2014-12-10 | 2015-03-25 | 东华大学 | Method for preparing rare earth polysaccharide composite fibers |
-
2015
- 2015-03-31 CN CN201510150350.0A patent/CN104788586B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2005123193A2 (en) * | 2004-06-17 | 2005-12-29 | Osteologix A/S | Treatments comprising strontium for rheumatic and arthritic diseases and pain |
| CN1974778A (en) * | 2006-12-05 | 2007-06-06 | 中国水产科学研究院南海水产研究所 | Sturgon chondrine extracting process |
| CN101831004A (en) * | 2010-03-24 | 2010-09-15 | 章云 | Hyaluronic acid strontium salt compounds, preparation method thereof and application in medicine |
| CN104451931A (en) * | 2014-12-10 | 2015-03-25 | 东华大学 | Method for preparing rare earth polysaccharide composite fibers |
Non-Patent Citations (2)
| Title |
|---|
| HIDEMITSU UCHISAWA等: ""Binding between calcium ions and chondroitin sulfate chains of salmon nasal cartilage glycosaminoglycan"", 《INTERNATIONAL CONGRESS SERIES》 * |
| 叶勇等: "《制药工艺学》", 28 February 2014, 华南理工大学出版社 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105440088A (en) * | 2015-11-20 | 2016-03-30 | 南方科技大学 | Glucosamine mixed strontium salt as well as preparation method and application thereof |
| CN108383927A (en) * | 2018-04-11 | 2018-08-10 | 南方科技大学 | chondroitin sulfate magnesium and preparation method thereof |
| CN108383927B (en) * | 2018-04-11 | 2021-06-08 | 南方科技大学 | Chondroitin sulfate magnesium and preparation method thereof |
| CN110604742A (en) * | 2019-08-02 | 2019-12-24 | 南方科技大学 | Eucommia polysaccharide strontium complex and preparation method and application thereof |
| CN110604742B (en) * | 2019-08-02 | 2021-12-03 | 南方科技大学 | Eucommia polysaccharide strontium complex and preparation method and application thereof |
| CN113583406A (en) * | 2021-08-03 | 2021-11-02 | 南方科技大学 | Composite polyether-ether-ketone material and preparation method and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104788586B (en) | 2018-03-09 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104788586A (en) | Strontium chondroitin sulfate and preparation method thereof | |
| CN101182495B (en) | Joint production process for producing alkaline phosphatase and heparin sodium with pig small intestine as raw material | |
| CN102827550B (en) | Gelatin preparation process for acid process bone element gelatin | |
| CN102766225B (en) | A kind of piperylene gathers the preparation method of sodium sulfate and a kind of oligomeric xylose | |
| CN106031709B (en) | Milt DNA-NA, nucleoprotamine extract and preparation method thereof | |
| CN101851301A (en) | Method for extracting crude product of heparin sodium | |
| CN104513307A (en) | Method for recovery of Kunitz and Bowman-Birk trypsin inhibitors from soybean whey | |
| CN102286252A (en) | Process for producing gelatin by using acid method | |
| CN103627768A (en) | Fish skin collagen bioactive small peptide and preparation method thereof | |
| CN100591696C (en) | Method for separation purifying hyaluronic acid | |
| CN110407950A (en) | A method of mould intracellular polyse is extracted using green solvent | |
| CN100485002C (en) | Bone collagen gel liquid preparation method | |
| CN106032546A (en) | Method for extracting medical material-based collagen from fish scales | |
| CN102492166B (en) | Method for preparing feather keratin sponge by using feathers | |
| CN104829782A (en) | Preparation method of polymeric flocculant | |
| CN102586213B (en) | Method for extracting bromelain ananase through chitosan carboxylation derivative | |
| CN106011112A (en) | Special complex enzyme system for kelp comprehensive utilization and application thereof | |
| WO2020093893A1 (en) | Preparation method for alginic acid synergistic carrier for reducing urea conversion | |
| KR20100123514A (en) | The recovering method of a low molecular weight sericin from silk degumming process | |
| CN103804526A (en) | Method for purifying crude product of heparin sodium | |
| CN100467390C (en) | High efficiency sewage treating agent and its prepn process | |
| CN104177515B (en) | A kind of environment-friendly type processing method of Heveatex resource hierarchy time complete utilization | |
| CN102585034B (en) | Method for sulfonating pectin | |
| CN101851285B (en) | Method for preparing sulglycotide | |
| CN104628889A (en) | Extraction method of heparin sodium |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| EXSB | Decision made by sipo to initiate substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |