CN115624959B - Urokinase adsorption material and preparation method and application thereof - Google Patents
Urokinase adsorption material and preparation method and application thereof Download PDFInfo
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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
- 238000001179 sorption measurement Methods 0.000 title claims abstract description 46
- 239000000463 material Substances 0.000 title claims abstract description 35
- 238000002360 preparation method Methods 0.000 title claims abstract description 22
- 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
- 210000002700 urine Anatomy 0.000 claims abstract description 23
- 238000001035 drying Methods 0.000 claims abstract description 22
- 239000002243 precursor Substances 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
- 238000000034 method Methods 0.000 claims abstract description 16
- 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
- 239000002244 precipitate Substances 0.000 claims abstract description 11
- 238000006243 chemical reaction Methods 0.000 claims description 22
- 239000003463 adsorbent Substances 0.000 claims description 12
- 230000001376 precipitating effect Effects 0.000 claims description 9
- 238000010992 reflux Methods 0.000 claims description 9
- 230000035484 reaction time Effects 0.000 claims 1
- 238000003756 stirring Methods 0.000 abstract description 11
- 238000004519 manufacturing process Methods 0.000 abstract description 7
- 238000004132 cross linking Methods 0.000 abstract description 4
- 239000012043 crude product Substances 0.000 abstract description 3
- 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
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 description 3
- 230000020764 fibrinolysis Effects 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
- SRSXLGNVWSONIS-UHFFFAOYSA-N benzenesulfonic acid Chemical compound OS(=O)(=O)C1=CC=CC=C1 SRSXLGNVWSONIS-UHFFFAOYSA-N 0.000 description 2
- 229940092714 benzenesulfonic acid Drugs 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 238000006467 substitution reaction Methods 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
- 241000282412 Homo Species 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
- 230000000740 bleeding effect Effects 0.000 description 1
- 230000007012 clinical effect Effects 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
- 238000007654 immersion Methods 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 230000001766 physiological effect Effects 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
- 230000002537 thrombolytic effect Effects 0.000 description 1
- 238000009777 vacuum freeze-drying Methods 0.000 description 1
Classifications
-
- 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
-
- 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
-
- 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
Abstract
The invention discloses a urokinase rapid adsorption material, a preparation method and application thereof, wherein the preparation method of the urokinase rapid adsorption 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, washing the precipitate, and drying to obtain a precursor; and (3) reacting the precursor with concentrated sulfuric acid, filtering, and washing the precipitate to obtain the catalyst. The application is the application of urokinase adsorption material in adsorbing urokinase crude 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 the urokinase in urine, does not need additional stirring or acidification treatment, simplifies the enrichment process of the urokinase, and has the advantages of simple preparation method, low cost and suitability for mass production.
Description
Technical Field
The invention belongs to the technical field of separation and extraction, and particularly relates to a urokinase rapid adsorption material and a preparation method and application thereof.
Background
Urokinase is an enzyme protein isolated from urine of healthy humans or obtained from tissue culture of human kidneys. In the human body, it is only necessary to exist in two forms of small molecular weight urokinase (molecular weight 33000) and large molecular weight urokinase (molecular weight 54000).
Research shows that the large molecular weight urokinase is the type of human physiological activity, and proves that the large molecular weight urokinase has the advantages of quick effect, strong fibrinolysis, long delay time and the like on new thrombosis (good response to treatment in the early stage of disease), the clinical effect is better than twice that of small molecular weight urokinase, and the large molecular weight urokinase does not have the side effect that the small molecular weight urokinase possibly causes fibrinolysis hyperfunction to cause bleeding. It is because of the fact that large molecular weight urokinase exhibits its excellent thrombolytic effect of effectively activating plasminogen to participate in fibrinolysis process, and urokinase is used as the first choice for treating cerebral thrombosis, venous embolism, acute myocardial infarction and other diseases.
Urokinase has important medical value, and market demand increases year by year, but raw materials of urokinase are in a tension state all the time. At present, the traditional urine collection mode is replaced by the pipeline type urine collection mode, and how to efficiently and rapidly collect urokinase from urine is a difficult problem facing the industry at present.
CN114736892a discloses a process for extracting urokinase by using modified silica gel, which is characterized in that the modified silica gel is mixed with male urine, and urokinase is collected by obtaining a urokinase-containing silica gel substance after uniform stirring. The invention can increase the macromolecular urokinase content in urokinase products and has high activity yield. However, the modified silica gel of the invention needs stirring when adsorbing urokinase, and is not matched with the existing pipeline urine collection mode.
CN111979216a discloses a method for extracting urokinase by using benzenesulfonic acid modified resin, which comprises placing the surface benzenesulfonic acid modified resin in urine to adsorb urine for 24-120 hours, and fully absorbing urokinase in urine. The invention can improve the purity of the extracted urokinase, but the invention has poor adsorption speed, needs long-term immersion contact of the surface benzene sulfonated resin and urine, and is not matched with the existing pipeline urine collection mode.
CN105238772A discloses a urokinase powder purifying method, which comprises the steps of adjusting pH of male urine to 5.0-5.5 to obtain acidified urine, and adsorbing the acidified urine by a diatomite column. The yield of urokinase separated by the invention is more than 97%, the endotoxin content is low, but the urokinase can be adsorbed after the urine is acidified, and the urokinase is not matched with the existing pipeline urine collecting mode.
Therefore, the urokinase rapid adsorption material is developed, so that urokinase can be rapidly and efficiently adsorbed from urine, and is matched with the existing pipeline urine collection mode, and has important significance for urokinase production.
Disclosure of Invention
In order to solve the technical problems, the invention provides a urokinase adsorption material, and a preparation method and application thereof.
In order to achieve the above purpose, the present invention adopts the following technical scheme:
a preparation method of urokinase adsorption material, comprising 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, washing the precipitate, 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 adsorption material.
Preferably, in the step (1), the mass ratio of the silica gel to the phosphoric acid is 1:2-3; further preferably 1:2.5.
preferably, the phosphoric acid in step (1) is 2.5-3.5M; further preferably 3M.
Preferably, the reaction in step (1) is for a time of: 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, trimethoxysilane and stilbene in the step (2) is 1:0.5-0.8:4-8; further preferably 1:0.55-0.75:5-7; most preferably 1:0.6:6.
preferably, the reaction in step (2) is: reflux reaction for 4-8h; further preferably 5 to 7 hours; 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 at 70-100 deg.c for 1-2 hr.
The invention also provides the urokinase adsorption material prepared by the preparation method.
The invention also provides application of the urokinase adsorption material prepared by the preparation method in adsorbing crude urokinase from human urine.
A method for rapid adsorption of urokinase comprising the steps of: the urokinase adsorption material prepared by the preparation method is used for adsorption.
The beneficial effects of the invention are as follows:
(1) 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 the urokinase in urine, does not need additional stirring or acidification treatment, and simplifies the enrichment process of the urokinase.
(2) The preparation method of the invention is simple, has low cost and is suitable for mass production.
Detailed Description
Other advantages and effects of the present invention will become apparent to those skilled in the art from the following disclosure, which describes the embodiments of the present invention with reference to specific examples. The invention may be practiced or carried out in other embodiments that depart from the specific details, and the details of the present description may be modified or varied from the spirit and scope of the present invention.
Before the embodiments of the invention are explained in further detail, it is to be understood that the invention is not limited in its scope to the particular embodiments described below; it is also to be understood that the terminology used in the examples of the invention is for the purpose of describing particular embodiments only, and is not intended to limit the scope of the invention.
Where numerical ranges are provided in the examples, it is understood that unless otherwise stated herein, both endpoints of each numerical range and any number between the two endpoints are significant both in the numerical range. 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 invention does not limit the sources of the adopted raw materials, and if no special description exists, the adopted raw materials are all common commercial products in the technical field.
Example 1 urokinase adsorbent and method for producing the same
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: 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 the precipitate, and drying to obtain a precursor;
(3) According to the mass ratio of 1:2.5, adding 60% concentrated sulfuric acid into the precursor, reacting for 1.5h at 90 ℃, filtering, precipitating and washing to obtain the urokinase adsorption material. Example 2 urokinase adsorbent and method for producing the same
The preparation method comprises the following steps:
(1) According to the mass ratio of 1:2, adding 3.5M phosphoric acid into the 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 the precipitate, and drying to obtain a precursor;
(3) According to the mass ratio of 1:2, adding 70% concentrated sulfuric acid into the precursor, reacting for 1h at 70 ℃, filtering, precipitating and washing to obtain the urokinase adsorption material.
Example 3 urokinase adsorbent and method for producing the same
The preparation method comprises the following steps:
(1) According to the mass ratio of 1:3, adding 2.5M phosphoric acid into the 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: 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 the precipitate, and drying to obtain a precursor;
(3) According to the mass ratio of 1: and 3, adding 50% concentrated sulfuric acid into the precursor, reacting for 2 hours at 100 ℃, filtering, precipitating and washing to obtain the urokinase adsorption material.
Example 4 urokinase adsorbent and method for preparing the same
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 1:0.55:5, 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 the precipitate, and drying to obtain a precursor;
(3) According to the mass ratio of 1:2.5, adding 60% concentrated sulfuric acid into the precursor, reacting for 1.5h at 90 ℃, filtering, precipitating and washing to obtain the urokinase adsorption material.
Example 5 urokinase adsorbent and method for preparing the same
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 1:0.75:7, 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 the precipitate, and drying to obtain a precursor;
(3) According to the mass ratio of 1:2.5, adding 60% concentrated sulfuric acid into the precursor, reacting for 1.5h at 90 ℃, filtering, precipitating and washing to obtain the urokinase adsorption material.
Comparative example 1 urokinase adsorbent 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 1:0.4:8, 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 the precipitate, and drying to obtain a precursor;
(3) According to the mass ratio of 1:2.5, adding 60% concentrated sulfuric acid into the precursor, reacting for 1.5h at 90 ℃, filtering, precipitating and washing to obtain the urokinase adsorption material.
Comparative example 2 urokinase adsorbent 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 1:0.8:3, 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 the precipitate, and drying to obtain a precursor;
(3) According to the mass ratio of 1:2.5, adding 60% concentrated sulfuric acid into the precursor, reacting for 1.5h at 90 ℃, filtering, precipitating and washing to obtain the urokinase adsorption material.
Adding 5mg urokinase crude product into purified water to obtain urokinase solution with concentration of 0.001%, taking 500ml urokinase solution, passing through modified silica gel column (modified silica gel urokinase adsorption material and column volume ratio of 0.5:1) at flow rate of 10 times column volume/h, and discarding effluent.
Eluting the modified silica gel column with eluent (2% ammonium hydroxide and 1M sodium chloride) at a rate of 5 times 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 urokinase crude product powder.
The adsorption properties of the modified silica gel urokinase adsorbent materials prepared in each example and comparative example on crude urokinase were calculated, and the results are shown in Table 1. The urokinase adsorption material prepared in the embodiment 1 has the adsorption rate up to 89.4%, the adsorption effect is optimal, the urokinase adsorption materials prepared in the embodiments 2 to 4 have good adsorption effect, the adsorption rate is more than 80%, and the process production requirements are met. Comparative examples 1 and 2 changed the amounts of trimethoxysilane and stilbene and the time of the crosslinking reaction, and the prepared urokinase adsorption materials had poor adsorption effect and adsorption rates of 60.6% and 63.2%, indicating that the degree of the crosslinking reaction has an important effect on the urokinase adsorption materials of the present invention.
TABLE 1 adsorption Properties of modified silica gel urokinase adsorbent materials prepared in examples 1-5 and comparative examples 1-2
| Crude urokinase of initial stage mg | Urokinase crude 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 invention has been further described above in connection with specific embodiments, which are exemplary only and do not limit the scope of the invention in any way. It will be understood by those skilled in the art that various changes and substitutions of details and forms of the technical solution of the present invention may be made without departing from the spirit and scope of the present invention, but these changes and substitutions fall within the scope of the present invention.
Claims (8)
1. The preparation method of the urokinase adsorption material is characterized by comprising the following steps of:
(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, washing the precipitate, and drying to obtain a precursor;
(3) Reacting the precursor obtained in the step (2) with concentrated sulfuric acid, filtering, precipitating and washing to obtain the urokinase adsorption material;
the mass ratio of the activated silica gel to the trimethoxysilane to the stilbene in the step (2) is 1:0.5-0.8:4-8;
the reaction in the step (2) is as follows: reflux reaction for 4-8h.
2. The method according to claim 1, wherein the mass ratio of silica gel to phosphoric acid in 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 step (1) is: 2-5h; the drying temperature is as follows: 160-180 ℃.
5. The method according to claim 1, wherein the mass percentage of the concentrated sulfuric acid in the step (3) is 50-70%; the mass ratio of the precursor to the concentrated sulfuric acid is 1:2-3.
6. A urokinase adsorbent material prepared by the method of any one of claims 1 to 5.
7. Use of the urokinase adsorbent material according to claim 6 for adsorbing crude urokinase from human urine.
8. A method for rapidly adsorbing urokinase, characterized in that the urokinase adsorbent according to claim 6 is used for the adsorption.
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| WO1989012099A1 (en) * | 1988-06-10 | 1989-12-14 | Damon Biotech, Inc. | Recovery of urokinase compounds |
| EP0484057A2 (en) * | 1990-11-02 | 1992-05-06 | The Dow Chemical Company | Antithrombogenic surfaces, their preparation, and materials therefore |
| CN101056846A (en) * | 2004-09-03 | 2007-10-17 | 细胞基因公司 | Diphenylethylene compounds and uses thereof |
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