Enzyme-based magnetic nanoparticles, preparation method, application and detection method for glucose

A technology of magnetic nanoparticles and detection methods, applied in biochemical equipment and methods, determination/inspection of microorganisms, immobilization on or in inorganic carriers, etc., can solve problems such as poor effect and inactivation of biological enzymes

Inactive Publication Date: 2013-10-09
ANHUI NORMAL UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] The technical problem to be solved by the present invention is to overcome the defects of enzyme-based magnetic nanoparticles that cause certain damage to biological enzymes in the prior art, resulting in inactivation of biological enzymes and poor effects, and provide an enzyme-based magnetic nanoparticle and its preparation method, Application and glucose detection method, the enzyme-based magnetic nanoparticles can detect glucose, and can be reused, and the waste can be burned or treated at high temperature to achieve zero pollution

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  • Enzyme-based magnetic nanoparticles, preparation method, application and detection method for glucose

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Embodiment 1

[0058] The preparation method of enzyme-based magnetic nanoparticles, the steps comprising:

[0059] (1) Fe 3 o 4 Preparation of magnetic nanoparticles: take FeCl 3 ·6H 2 O, anhydrous sodium acetate, 1.0g polyethylene glycol was placed in ethylene glycol solution, stirred vigorously for 30 minutes, then transferred to an autoclave, reacted in an oven at 200°C for 8 hours, and cooled to room temperature; the synthesized After the product was washed with ethanol, it was dried in a vacuum oven for 6 hours to obtain Fe 3 o 4 magnetic nanoparticles;

[0060] (2) Polydopamine (PDA) wrapped Fe 3 o 4 Preparation: 50mg of the above Fe 3 o 4 Dissolve magnetic nanoparticles and 100 mg of dopamine in 100 mL of water, add 60 mg of tris hydrochloride buffer (Tris-HCl buffer, concentration 0.05 mg / l, pH 8.5), stir for 24 hours at room temperature Wash with 50 mL of water and dry for 6 hours to obtain polydopamine-coated Fe 3 o 4 (PDAFe 3 o 4 );

[0061] (3) 200 μL of horseradi...

Embodiment 2

[0064] (1) Fe 3 o 4 Preparation of magnetic nanoparticles: take FeCl 3 ·6H 2 O, anhydrous sodium acetate, 1.0 g of polyethylene glycol was placed in ethylene glycol solution, stirred vigorously for 30 minutes, then transferred to an autoclave, reacted in an oven at 200°C for 8 hours, and cooled to room temperature. Finally, after the synthesized product was washed with ethanol, it was dried in a vacuum oven for 6 hours;

[0065] (2) Polydopamine (PDA) wrapped Fe 3 o 4 Preparation: Fe 3 o 4 Dissolve magnetic nanoparticles and dopamine (DA) in water to obtain a mixed solution. The concentration of dopamine in the mixed solution is 0.5 mg / mL, and the percentage is the volume ratio of the mass of dopamine to water; in 100 mL of H 2 Add 5 mmol / L tris-hydroxymethylaminomethane solution in O to the mixed solution, then adjust the pH value of the mixed solution to 7.0 with 0.1 mol / L HCl solution; stir for 8 hours, and then use 10 mL of water Wash and dry in a vacuum oven for ...

Embodiment 3

[0068] (1) Fe 3 o 4 Preparation of magnetic nanoparticles: take FeCl 3 ·6H 2 O, anhydrous sodium acetate, 1.0 g of polyethylene glycol was placed in ethylene glycol solution, stirred vigorously for 30 minutes, then transferred to an autoclave, reacted in an oven at 200°C for 8 hours, and cooled to room temperature. Finally, after the synthesized product was washed with ethanol, it was dried in a vacuum oven for 6 hours;

[0069] (2) Polydopamine (PDA) wrapped Fe 3 o 4 Preparation: Fe 3 o 4 Dissolve magnetic nanoparticles and dopamine (DA) in water to obtain a mixed solution. The concentration of dopamine in the mixed solution is 1.0 mg / mL, and the percentage is the volume ratio of the mass of dopamine to water; in 100 mL of H 2 Obtain 5mmol / L tris-hydroxymethylaminomethane solution in O, add it to the mixed solution, and then adjust the pH value of the mixed solution to 8.0 with 0.1mol / L HCl solution; stir for 24 hours, and use 10-30mL washed with ethanol, and dried i...

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Abstract

The invention discloses enzyme-based magnetic nanoparticles, a preparation method, an application and a detection method for glucose. The preparation method comprises the following steps of 1) mixing Fe3O4 magnetic nanoparticles and dopamine with water according to a mass ratio of Fe3O4 magnetic nanoparticles to dopamine being (1:1)-(1:4) uniformly, adding the above mixture in an alkali buffering system, and stirring for 8-48 hours to obtain Fe3O4 coated by polydopamine; 2) mixing a horseradish peroxidase solution, a glucose oxidase solution and Fe3O4 coated by polydopamine with dopamine uniformly to obtain a solution B, and reacting for 5-25 hours. In the solution B, the concentrations of horseradish peroxidase, glucose oxidase and Fe3O4 coated by polydopamine and dopamine are 0.025-0.2 mg/mL, 0.025-0.2 mg/mL, 0.5-25 mg/mL and 1-5 mg/mL respectively. The enzyme-based magnetic nanoparticles can detect glucose and can be used repeatedly, and waste can be treated by combustion or high temperature, so that zero-pollution can be achieved.

Description

technical field [0001] The invention relates to an enzyme-based magnetic nanoparticle, a preparation method, an application and a glucose detection method. Background technique [0002] Diabetes is a social health problem caused by a metabolic disorder. The content of glucose in normal human blood ranges from 4.4 to 6.6mmol / L (80 to 120mg / dL). Excessive blood sugar concentration will lead to hyperglycemia or insufficient insulin, which will lead to diabetes. In recent years, in the prior art, the method of immobilizing enzymes is used to detect glucose. [0003] Glucose enzymes can be divided into glucose oxidase (GOx) and glucose dehydrogenase (GDH). Using glucose oxidase to measure glucose concentration has become a widespread and effective method. At present, the most commonly used glucose detection method in clinic is the glucose oxidase analysis method, which has the characteristics of good specificity, fast reaction rate and anti-uric acid and ascorbic acid interfere...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/14C12Q1/28C12Q1/26
Inventor 王银玲宋倩李茂国周运友
Owner ANHUI NORMAL UNIV
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