Method for detecting uric acid by using mesoporous organic silicon hollow nanosphere fixed uricase

A technology of mesoporous silicone and nanospheres, which is applied in the field of mesoporous silicone hollow nanospheres immobilizing uricase to detect uric acid, can solve the problems of unfavorable uricase reuse, weak fixation and easy inhibition of uricase, etc. Fixed amount, good linearity, and the effect of expanding the detection range

Inactive Publication Date: 2019-08-09
JILIN UNIV
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

However, these methods all involve a series of complex processes such as cladding-corrosion. These multi-step processes require more energy and time, and often require the use of toxic reagents. At the same time, the resulting PMO hollow shell The mesoporous channels are disordered and the orientation is arbitrary, which limits the penetration of guest molecules to some extent. Therefore, it is still a big challenge to develop a simple and effective one-step strategy to synthesize HPMOs with ordered vertical mesoporous channels. challenge
[0007] The traditional carrier material immobilized uricase usually has a small amount of adsorption, the immobilization is not firm, and it is easy to inhibit the activity of uricase, which is not conducive to the repeated use of uricase in the detection of uric acid. Therefore, the carrier material with a large amount of immobilization and does not destroy the structure of uricase has always been research focus

Method used

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  • Method for detecting uric acid by using mesoporous organic silicon hollow nanosphere fixed uricase
  • Method for detecting uric acid by using mesoporous organic silicon hollow nanosphere fixed uricase
  • Method for detecting uric acid by using mesoporous organic silicon hollow nanosphere fixed uricase

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Experimental program
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Effect test

Embodiment 1

[0029] (1) Accurately measure 37mL of ethanol and 5mL of ultrapure water with a graduated cylinder, and pour them into a 100mL beaker.

[0030] (2) Add 1.6 mL of ammonia water (mass fraction 28%), and then quickly add 3 mL of tetraethyl orthosilicate.

[0031] (3) After stirring for 2 hours at room temperature, centrifuge at 12000 rpm for 3 minutes in a high-speed centrifuge.

[0032] (4) Wash twice with ultrapure water and then wash twice with ethanol, centrifuge and dry naturally at room temperature to obtain monodisperse SiO with a mass of about 300 mg 2 nanospheres.

Embodiment 2

[0034] (1) Accurately measure 22mL of ultrapure water and 11mL of ethanol in a 100mL beaker.

[0035] (2) get the 100mg monodisperse SiO that embodiment 1 synthesizes 2 Nanospheres were added thereto, and the mixture was sonicated for 30 minutes.

[0036] (3) Add 120mg cetyltrimethylammonium bromide and 2mL ammonia water (mass fraction 28%), stir at room temperature until cetyltrimethylammonium bromide is completely dissolved, then add 0.22mL drop by drop 1,2-bis(triethoxysilyl)ethane, and continued to stir at room temperature for 2h.

[0037] (3) Put the above mixture into a reactor, put the reactor in an oven at 100°C for 24 hours, take out the reactor and cool it to room temperature, and centrifuge the mixture in the reactor at 12000 rpm for 3 minutes.

[0038] (4) Wash twice with ultrapure water and ethanol respectively, and put it into an oven at 80°C to dry.

[0039] (5) Take out the dried sample and add a little ethanol for ultrasonication, then add 65mL ethanol and ...

Embodiment 3

[0042] (1) Take 1mg / mL uricase stock solution, dilute it with glycine buffer solution with pH=8.5 to prepare 0.1mg / mL, 0.2mg / mL, 0.3mg / mL, 0.4mg / mL, 0.5mg / mL uricase dilution solution, measure the absorbance of the uricase dilution solution at 293nm, and establish the standard curve of the uricase absorbance (such as image 3 ), so as to calculate the immobilized amount of uricase.

[0043] (2) Take 2 mL of the uricase stock solution with a concentration of 1 mg / mL and dilute it 5 times with a glycine buffer solution with a pH of 8.5 to obtain 10 mL of a uricase solution with a concentration of 0.2 mg / mL. 10 mg of synthesized mesoporous silicone hollow nanospheres were added thereto, and stirred at 4° C. for 8 h.

[0044] (3) centrifuge 5min with high-speed centrifuge 12000rpm, get supernatant and measure its absorbance at 293nm place, according to the standard curve of step (1) ( image 3 ) to calculate the concentration of uricase in the supernatant, which is 0.0372mg / mL; ...

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Abstract

The invention discloses a method for detecting uric acid by using mesoporous organic silicon hollow nanosphere fixed uricase, and belongs to the technical field of uric acid detection. The experimenttakes monodispersed SiO2 nanospheres as a hard template, and adopts a one-step growth-induced corrosion method to prepare mesoporous organic silicon hollow nanospheres. The method is simple and easy to operate, is low in requirement and highly controllable in synthesis process, and can synthesize hollow-structured mesoporous organic silicon nanospheres with different functional groups by using different precursors. The synthesized mesoporous organic silicon hollow nanospheres have ordered pore channels and uniform pore diameters, and internally contain hollow cavities. The synthesized mesoporous organic silicon hollow nanosphere fixed uricase is used to detect serum uric acid, so that the method for detecting uric acid by using the mesoporous organic silicon hollow nanosphere fixed uricaseis established, and an application prospect in rapid detection of serum uric acid is achieved.

Description

technical field [0001] The invention belongs to the technical field of uric acid detection, and in particular relates to a method for detecting uric acid by immobilizing uricase with mesoporous organosilicon hollow nanospheres. Background technique [0002] The content of serum uric acid in the body is closely related to the health status of many human systems, and is related to the occurrence and progression of various diseases. Therefore, timely detection and monitoring of serum uric acid content is of great significance to the health of the human body and the prevention and treatment of diseases. [0003] At present, the commonly used methods for detecting uric acid mainly include high performance liquid chromatography, isotope dilution mass spectrometry, and enzymatic method. The first two detection methods have disadvantages such as complicated operation, expensive equipment, and long detection cycle, which are not suitable for rapid detection of uric acid. The enzymat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/33
CPCG01N21/33
Inventor 王春艳王润伟苗婷婷段秉怡王欢田牧晓于惠源
Owner JILIN UNIV
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