CN115624959A - Urokinase adsorbing material and preparation method and application thereof - Google Patents
Urokinase adsorbing material and preparation method and application thereof Download PDFInfo
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- CN115624959A CN115624959A CN202211211533.5A CN202211211533A CN115624959A CN 115624959 A CN115624959 A CN 115624959A CN 202211211533 A CN202211211533 A CN 202211211533A CN 115624959 A CN115624959 A CN 115624959A
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- 102000003990 Urokinase-type plasminogen activator Human genes 0.000 title claims abstract description 85
- 108090000435 Urokinase-type plasminogen activator Proteins 0.000 title claims abstract description 85
- 229960005356 urokinase Drugs 0.000 title claims abstract description 85
- 239000000463 material Substances 0.000 title claims abstract description 45
- 238000002360 preparation method Methods 0.000 title claims abstract description 31
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims abstract description 48
- 238000005406 washing Methods 0.000 claims abstract description 30
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 claims abstract description 28
- 238000001035 drying Methods 0.000 claims abstract description 22
- 239000002243 precursor Substances 0.000 claims abstract description 22
- 210000002700 urine Anatomy 0.000 claims abstract description 22
- 238000001914 filtration Methods 0.000 claims abstract description 20
- 239000000741 silica gel Substances 0.000 claims abstract description 17
- 229910002027 silica gel Inorganic materials 0.000 claims abstract description 17
- PJANXHGTPQOBST-VAWYXSNFSA-N Stilbene Natural products C=1C=CC=CC=1/C=C/C1=CC=CC=C1 PJANXHGTPQOBST-VAWYXSNFSA-N 0.000 claims abstract description 15
- PJANXHGTPQOBST-UHFFFAOYSA-N stilbene Chemical compound C=1C=CC=CC=1C=CC1=CC=CC=C1 PJANXHGTPQOBST-UHFFFAOYSA-N 0.000 claims abstract description 15
- 235000021286 stilbenes Nutrition 0.000 claims abstract description 15
- 229910000147 aluminium phosphate Inorganic materials 0.000 claims abstract description 14
- QAOWNCQODCNURD-UHFFFAOYSA-N sulfuric acid Substances OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 claims abstract description 14
- YUYCVXFAYWRXLS-UHFFFAOYSA-N trimethoxysilane Chemical compound CO[SiH](OC)OC YUYCVXFAYWRXLS-UHFFFAOYSA-N 0.000 claims abstract description 14
- 238000000034 method Methods 0.000 claims abstract description 13
- 230000001376 precipitating effect Effects 0.000 claims abstract description 13
- 238000006243 chemical reaction Methods 0.000 claims description 20
- 238000010992 reflux Methods 0.000 claims description 9
- 239000003463 adsorbent Substances 0.000 claims description 3
- 230000035484 reaction time Effects 0.000 claims description 2
- 238000001179 sorption measurement Methods 0.000 abstract description 21
- 238000003756 stirring Methods 0.000 abstract description 9
- 238000004132 cross linking Methods 0.000 abstract description 4
- 238000011031 large-scale manufacturing process Methods 0.000 abstract description 2
- 230000020477 pH reduction Effects 0.000 abstract description 2
- 239000004593 Epoxy Substances 0.000 abstract 1
- BLRPTPMANUNPDV-UHFFFAOYSA-N Silane Chemical compound [SiH4] BLRPTPMANUNPDV-UHFFFAOYSA-N 0.000 abstract 1
- 239000003054 catalyst Substances 0.000 abstract 1
- 229910000077 silane Inorganic materials 0.000 abstract 1
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 description 21
- 238000002156 mixing Methods 0.000 description 14
- 230000000694 effects Effects 0.000 description 8
- 239000008213 purified water Substances 0.000 description 8
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 8
- 230000000052 comparative effect Effects 0.000 description 7
- 239000002244 precipitate Substances 0.000 description 7
- SRSXLGNVWSONIS-UHFFFAOYSA-N benzenesulfonic acid Chemical compound OS(=O)(=O)C1=CC=CC=C1 SRSXLGNVWSONIS-UHFFFAOYSA-N 0.000 description 3
- 229940092714 benzenesulfonic acid Drugs 0.000 description 3
- 239000000843 powder Substances 0.000 description 3
- 239000002994 raw material Substances 0.000 description 3
- 239000011347 resin Substances 0.000 description 3
- 229920005989 resin Polymers 0.000 description 3
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 230000020764 fibrinolysis Effects 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 description 1
- 206010008132 Cerebral thrombosis Diseases 0.000 description 1
- 208000005189 Embolism Diseases 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 208000032843 Hemorrhage Diseases 0.000 description 1
- 201000001429 Intracranial Thrombosis Diseases 0.000 description 1
- 102000013566 Plasminogen Human genes 0.000 description 1
- 108010051456 Plasminogen Proteins 0.000 description 1
- 208000007536 Thrombosis Diseases 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 206010000891 acute myocardial infarction Diseases 0.000 description 1
- 239000000908 ammonium hydroxide Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000007012 clinical effect Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 239000003480 eluent Substances 0.000 description 1
- 239000002158 endotoxin Substances 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- 210000005084 renal tissue Anatomy 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 230000002537 thrombolytic effect Effects 0.000 description 1
- 238000009777 vacuum freeze-drying Methods 0.000 description 1
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
- B01J20/22—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof comprising organic material
- B01J20/26—Synthetic macromolecular compounds
- B01J20/262—Synthetic macromolecular compounds obtained otherwise than by reactions only involving carbon to carbon unsaturated bonds, e.g. obtained by polycondensation
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
- C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
- C12N9/64—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue
- C12N9/6421—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue from mammals
- C12N9/6424—Serine endopeptidases (3.4.21)
- C12N9/6456—Plasminogen activators
- C12N9/6462—Plasminogen activators u-Plasminogen activator (3.4.21.73), i.e. urokinase
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y304/00—Hydrolases acting on peptide bonds, i.e. peptidases (3.4)
- C12Y304/21—Serine endopeptidases (3.4.21)
- C12Y304/21073—Serine endopeptidases (3.4.21) u-Plasminogen activator (3.4.21.73), i.e. urokinase
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Abstract
The invention discloses a rapid urokinase adsorbing material and a preparation method and application thereof, wherein the preparation method of the rapid urokinase adsorbing material comprises the following steps: reacting silica gel with phosphoric acid, washing and drying to obtain activated silica gel; reacting the obtained activated silica gel with trimethoxy epoxy silane and stilbene, filtering, precipitating, washing and drying to obtain a precursor; and (3) reacting the precursor with concentrated sulfuric acid, filtering, precipitating and washing to obtain the catalyst. The application is the application of the urokinase adsorbing material in adsorbing a crude urokinase product from human urine. The urokinase adsorption material is a silica gel modified material, is prepared by crosslinking and sulfonating trimethoxy silane and stilbene with silica gel, has the advantages of high adsorption speed, large adsorption capacity and the like compared with the prior art, can be directly placed in a pipeline to adsorb urokinase in urine, does not need additional stirring or acidification treatment, simplifies the enrichment process of urokinase, and is simple in preparation method, low in cost and suitable for large-scale production.
Description
Technical Field
The invention belongs to the technical field of separation and extraction, and particularly relates to a rapid urokinase adsorbing material as well as a preparation method and application thereof.
Background
Urokinase is an enzyme protein isolated from healthy human urine, or obtained from human kidney tissue culture. It exists in human body in the form of small molecular weight urokinase (molecular weight 33000) and large molecular weight urokinase (molecular weight 54000).
Research shows that the high molecular urokinase is a human body physiological activity type, and proves that the high molecular urokinase has the advantages of quick response, strong fibrinolysis effect, long delay time and the like on new-onset thrombus (good response to treatment in the early disease onset period), the clinical effect of the high molecular urokinase is twice of that of the low molecular urokinase, and the high molecular urokinase does not have the side effect that the low molecular urokinase can possibly cause hyperfibrino causing hemorrhage. Since large molecular weight urokinase exhibits its excellent thrombolytic effect of activating plasminogen to participate in fibrinolysis process, urokinase is used as the first choice drug for treating cerebral thrombosis, venous embolism and acute myocardial infarction.
Urokinase has an important medical value, and the market demand is increasing year by year, but the raw materials thereof are in a state of tension. At present, the traditional urine collection mode is replaced by a pipeline urine collection mode, and how to efficiently and quickly collect urokinase from urine is a problem in the industry at present.
CN114736892A discloses a process for extracting urokinase by using modified silica gel, which utilizes the modified silica gel to be mixed with male urine, and the mixture is stirred uniformly to obtain a urokinase-containing silica gel substance, thereby realizing the collection of urokinase. The invention can improve the content of macromolecular urokinase in urokinase products and has high activity yield. However, the modified silica gel needs to be stirred when adsorbing urokinase, and is not matched with the existing pipeline urine collection mode.
CN111979216A discloses a method for extracting urokinase from benzenesulfonic acid modified resin, which is to put surface benzenesulfonic acid modified resin in urine to adsorb urine for 24-120 hours so as to fully absorb urokinase in urine. The invention can improve the purity of the extracted urokinase, but the invention has poor adsorption speed, needs surface benzene sulfonic acid resin to be immersed and contacted with urine for a long time, and is not matched with the prior pipeline urine collection mode.
CN105238772A discloses a method for purifying urokinase powder, which needs to regulate the pH value of male urine to 5.0-5.5 to obtain acidified urine, and then the acidified urine is absorbed by a diatomite column. The yield of the urokinase separated by the invention is more than 97 percent, the content of endotoxin is low, but the urokinase can be adsorbed only after being acidified, and the method is not matched with the existing collection mode of the pipeline urine.
Therefore, a urokinase rapid adsorption material is developed, so that urokinase can be rapidly and efficiently adsorbed from urine, the urokinase rapid adsorption material is matched with the existing pipeline urine collection mode, and the urokinase rapid adsorption material has important significance for production of urokinase.
Disclosure of Invention
In order to solve the technical problems, the invention provides a urokinase adsorbing material and a preparation method and application thereof.
In order to achieve the purpose, the invention adopts the following technical scheme:
a preparation method of a urokinase adsorbing material comprises the following steps:
(4) Reacting silica gel with phosphoric acid, washing and drying to obtain activated silica gel;
(5) Reacting the activated silica gel obtained in the step (1) with trimethoxy silane and stilbene, filtering, precipitating, washing and drying to obtain a precursor;
(6) And (3) reacting the precursor obtained in the step (2) with concentrated sulfuric acid, filtering, precipitating and washing to obtain the urokinase adsorbing material.
Preferably, the mass ratio of the silica gel to the phosphoric acid in the step (1) is 1:2-3; more preferably 1:2.5.
preferably, the phosphoric acid in step (1) is 2.5-3.5M; more preferably 3M.
Preferably, the reaction time in step (1) is: 2-5h; further preferably 3 hours.
Preferably, the drying temperature in step (1) is: 160-180 ℃.
Preferably, the mass ratio of the activated silica gel, the trimethoxy silane and the stilbene in the step (2) is 1:0.5-0.8:4-8; more preferably 1:0.55-0.75:5-7; optimally 1:0.6:6.
preferably, the reaction in step (2) is: carrying out reflux reaction for 4-8h; further preferably 5-7h; most preferably 6h.
Preferably, the mass percentage of the concentrated sulfuric acid in the step (3) is 50-70%.
Preferably, the mass ratio of the precursor to the concentrated sulfuric acid in the step (3) is 1:2-3.
Preferably, the reaction in step (3) is: reacting for 1-2h at 70-100 ℃.
The invention also provides the urokinase adsorbing material prepared by the preparation method.
The invention also provides application of the urokinase adsorbing material prepared by the preparation method in adsorbing a crude urokinase product from human urine.
A method for rapidly adsorbing urokinase, which comprises the following steps: the urokinase adsorbing material prepared by the preparation method is used for adsorption.
The beneficial effects of the invention are as follows:
(1) The urokinase adsorption material prepared by the invention is a silica gel modified material, is prepared by crosslinking and sulfonating trimethoxy silane, stilbene and silica gel, has the advantages of high adsorption speed, large adsorption capacity and the like compared with the prior art, can be directly placed in a pipeline to adsorb urokinase in urine, does not need additional stirring or acidification treatment, and simplifies the urokinase enrichment process.
(2) The preparation method is simple, low in cost and suitable for large-scale production.
Detailed Description
The embodiments of the present invention are described below with reference to specific embodiments, and other advantages and effects of the present invention will be easily understood by those skilled in the art from the disclosure of the present specification. The invention is capable of other and different embodiments and of being practiced or of being carried out in various ways, and its several details are capable of modification in various respects, all without departing from the spirit and scope of the present invention.
Before the present embodiments are further described, it is to be understood that the scope of the invention is not to be limited to the specific embodiments described below; it is also to be understood that the terminology used in the examples herein is for the purpose of describing particular embodiments only, and is not intended to limit the scope of the present invention.
When numerical ranges are given in the examples, it is understood that both endpoints of each of the numerical ranges and any number between the two endpoints are optional unless otherwise specified in the invention. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.
The sources of the raw materials used in the present invention are not limited, and the raw materials used in the present invention are all those commonly available in the art unless otherwise specified.
Example 1 urokinase adsorbing material and preparation method thereof
The preparation method comprises the following steps:
(1) According to the mass ratio of 1:2.5, adding 3M phosphoric acid into the silica gel, mixing and stirring for reaction for 3 hours, washing for 3-5 times by using purified water, and drying for 2.5 hours at 170 ℃ to obtain activated silica gel;
(2) According to the mass ratio of 1:0.6:6, adding trimethoxy silane and stilbene into 200g of activated silica gel, mixing, adding 300ml of toluene, carrying out reflux reaction for 6 hours, filtering, washing precipitate, and drying to obtain a precursor;
(3) According to the mass ratio of 1:2.5, adding 60 percent concentrated sulfuric acid by mass into the precursor, reacting for 1.5 hours at 90 ℃, filtering, precipitating and washing to obtain the urokinase adsorbing material. Example 2 urokinase adsorbing material and preparation method thereof
The preparation method comprises the following steps:
(1) According to the mass ratio of 1:2, adding 3.5M phosphoric acid into silica gel, mixing and stirring for reaction for 5 hours, washing for 3-5 times by using purified water, and drying for 2.5 hours at 170 ℃ to obtain activated silica gel;
(2) According to the mass ratio of 1:0.5:8, adding trimethoxy silane and stilbene into 200g of activated silica gel, mixing, adding 300ml of toluene, carrying out reflux reaction for 8 hours, filtering, washing precipitate, and drying to obtain a precursor;
(3) According to the mass ratio of 1:2, adding concentrated sulfuric acid with the mass percent of 70% into the precursor, reacting for 1 hour at 70 ℃, filtering, precipitating and washing to obtain the urokinase adsorbing material.
Example 3 urokinase adsorbing material and preparation method thereof
The preparation method comprises the following steps:
(1) According to the mass ratio of 1:3, adding 2.5M phosphoric acid into silica gel, mixing and stirring for reaction for 2 hours, washing for 3-5 times by using purified water, and drying for 2.5 hours at 170 ℃ to obtain activated silica gel;
(2) According to the mass ratio of 1:0.8:4, adding trimethoxy silane and stilbene into 200g of activated silica gel, mixing, adding 300ml of toluene, carrying out reflux reaction for 4 hours, filtering, washing precipitate, and drying to obtain a precursor;
(3) According to the mass ratio of 1: and 3, adding 50% concentrated sulfuric acid by mass into the precursor, reacting for 2 hours at the temperature of 100 ℃, filtering, precipitating and washing to obtain the urokinase adsorbing material.
Example 4 urokinase adsorbing material and preparation method thereof
The preparation method comprises the following steps:
(1) According to the mass ratio of 1:2.5, adding 3M phosphoric acid into silica gel, mixing and stirring for reaction for 3 hours, washing for 3-5 times by using purified water, and drying for 2.5 hours at 170 ℃ to obtain activated silica gel;
(2) According to the mass ratio of 1:0.55:5, adding trimethoxy silane and stilbene into 200g of activated silica gel, mixing, adding 300ml of toluene, carrying out reflux reaction for 6 hours, filtering, washing precipitate, and drying to obtain a precursor;
(3) According to the mass ratio of 1:2.5, adding 60 percent by mass of concentrated sulfuric acid into the precursor, reacting for 1.5h at 90 ℃, filtering, precipitating and washing to obtain the urokinase adsorbing material.
Example 5 urokinase adsorbing material and preparation method thereof
The preparation method comprises the following steps:
(1) According to the mass ratio of 1:2.5, adding 3M phosphoric acid into silica gel, mixing and stirring for reaction for 3 hours, washing for 3-5 times by using purified water, and drying for 2.5 hours at 170 ℃ to obtain activated silica gel;
(2) According to the mass ratio of 1:0.75:7, adding trimethoxy silane and stilbene into 200g of activated silica gel, mixing, adding 300ml of toluene, carrying out reflux reaction for 6 hours, filtering, washing precipitate, and drying to obtain a precursor;
(3) According to the mass ratio of 1:2.5, adding 60 percent by mass of concentrated sulfuric acid into the precursor, reacting for 1.5h at 90 ℃, filtering, precipitating and washing to obtain the urokinase adsorbing material.
Comparative example 1 urokinase adsorbing material and preparation method thereof
The preparation method comprises the following steps:
(1) According to the mass ratio of 1:2.5, adding 3M phosphoric acid into silica gel, mixing and stirring for reaction for 3 hours, washing for 3-5 times by using purified water, and drying for 2.5 hours at 170 ℃ to obtain activated silica gel;
(2) According to the mass ratio of 1:0.4:8, adding trimethoxy silane and stilbene into 200g of activated silica gel, mixing, adding 300ml of toluene, carrying out reflux reaction for 9 hours, filtering, washing precipitate, and drying to obtain a precursor;
(3) According to the mass ratio of 1:2.5, adding 60 percent by mass of concentrated sulfuric acid into the precursor, reacting for 1.5h at 90 ℃, filtering, precipitating and washing to obtain the urokinase adsorbing material.
Comparative example 2 urokinase adsorbing material and preparation method thereof
The preparation method comprises the following steps:
(1) According to the mass ratio of 1:2.5, adding 3M phosphoric acid into the silica gel, mixing and stirring for reaction for 3 hours, washing for 3-5 times by using purified water, and drying for 2.5 hours at 170 ℃ to obtain activated silica gel;
(2) According to the mass ratio of 1:0.8: adding trimethoxy silane and stilbene into 200g of activated silica gel, mixing, adding 600ml of toluene, carrying out reflux reaction for 3 hours, filtering, washing precipitate, and drying to obtain a precursor;
(3) According to the mass ratio of 1:2.5, adding 60 percent concentrated sulfuric acid by mass into the precursor, reacting for 1.5 hours at 90 ℃, filtering, precipitating and washing to obtain the urokinase adsorbing material.
5mg of the crude urokinase was added to purified water to prepare a urokinase solution having a concentration of 0.001%, 500ml of the urokinase solution was passed through a modified silica gel column (the volume ratio of the modified silica gel urokinase-adsorbing material to the column was 0.5.
Eluting the modified silica gel column with eluent (2% ammonium hydroxide and 1M sodium chloride) at the speed of 5 times of column volume/h, stopping collecting when the ultraviolet absorption of the collected liquid is less than 50mAU, freezing the collected liquid at-45 ℃ for 3 hours, then heating to 20 ℃ for vacuum freeze drying, keeping for 2 hours, and taking out to obtain crude urokinase powder.
The adsorption performance of the modified silica gel urokinase adsorption materials prepared in each example and comparative example on crude urokinase was calculated, and the results are shown in table 1. The urokinase adsorbing material prepared in the embodiment 1 has the adsorption rate as high as 89.4% and the best adsorption effect, and the urokinase adsorbing materials prepared in the embodiments 2 to 4 have the better adsorption effect and the adsorption rates of more than 80%, so that the process production requirements are met. Comparative examples 1 and 2 changed the amounts of trimethoxysilane and stilbene and the time of the cross-linking reaction, and the urokinase adsorbing materials prepared therefrom had poor adsorption effects, with adsorption rates of 60.6% and 63.2%, indicating that the degree of the cross-linking reaction all had an important effect on the urokinase adsorbing materials of the present invention.
TABLE 1 adsorption Performance of modified silica gel urokinase adsorbent materials prepared in examples 1-5 and comparative examples 1-2
| Crude mg of initial urokinase | Crude urokinase powder mg | Adsorption Rate% | |
| Example 1 | 5.00 | 4.47 | 89.4 |
| Example 2 | 5.00 | 4.08 | 81.6 |
| Example 3 | 5.00 | 4.2 | 84.0 |
| Example 4 | 5.00 | 4.14 | 82.8 |
| Example 5 | 5.00 | 4.26 | 85.2 |
| Comparative example 1 | 5.00 | 3.03 | 60.6 |
| Comparative example 2 | 5.00 | 3.16 | 63.2 |
The present invention has been further described with reference to specific embodiments, which are only exemplary and do not limit the scope of the present invention. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention, and that such changes and substitutions are intended to be within the scope of the invention.
Claims (10)
1. A preparation method of a urokinase adsorbing material is characterized by comprising the following steps:
(1) Reacting silica gel with phosphoric acid, washing and drying to obtain activated silica gel;
(2) Reacting the activated silica gel obtained in the step (1) with trimethoxy silane and stilbene, filtering, precipitating, washing and drying to obtain a precursor;
(3) And (3) reacting the precursor obtained in the step (2) with concentrated sulfuric acid, filtering, precipitating and washing to obtain the urokinase adsorbing material.
2. The preparation method according to claim 1, wherein the mass ratio of the silica gel to the phosphoric acid in the step (1) is 1:2-3.
3. The method according to claim 1, wherein the phosphoric acid in the step (1) is 2.5 to 3.5M.
4. The method according to claim 1, wherein the reaction time in the step (1) is: 2-5h; the drying temperature is as follows: 160-180 ℃.
5. The preparation method according to claim 1, wherein the mass ratio of the activated silica gel, the trimethoxysilane and the stilbene in the step (2) is 1:0.5-0.8:4-8.
6. The method according to claim 5, wherein the reaction in the step (2) is: and refluxing and reacting for 4-8h.
7. The preparation method according to claim 1, wherein the mass percent of the concentrated sulfuric acid in the step (3) is 50-70%; the mass ratio of the precursor to concentrated sulfuric acid is 1:2-3.
8. A urokinase adsorbent material prepared by the preparation method according to any one of claims 1 to 7.
9. Use of the urokinase adsorbent material according to claim 8 for adsorbing crude urokinase from human urine.
10. A method for rapidly adsorbing urokinase, which comprises adsorbing urokinase with the urokinase-adsorbing material according to claim 7.
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| Publication number | Priority date | Publication date | Assignee | Title |
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