Preparation method and application of micro-enzyme reactor based on magnetic functionalized graphene oxide

A technology of enzyme reactor and graphene, which is applied in the direction of enzyme production/bioreactor, biochemical equipment and method, microbial measurement/inspection, etc., can solve the problem of high-efficiency loading of unfavorable enzymes, and achieve high reuse rate, heavy weight Good performance, simple and fast effect

Inactive Publication Date: 2013-10-23
NANCHANG UNIV
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Problems solved by technology

Traditional enzyme immobilization methods mainly include physical adsorption, sol-gel embeddi...

Method used

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  • Preparation method and application of micro-enzyme reactor based on magnetic functionalized graphene oxide
  • Preparation method and application of micro-enzyme reactor based on magnetic functionalized graphene oxide
  • Preparation method and application of micro-enzyme reactor based on magnetic functionalized graphene oxide

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Embodiment 1

[0025] Fabrication of PDMS chip: Using the SU-8 positive mold (Boao Biological Co., Ltd.) as a template, make a typical cross-shaped PDMS microfluidic chip channel, such as figure 1 shown. The specific production process is as follows: Take a certain amount of PDMS monomer and curing agent according to 10:1 (mass ratio), mix evenly, degas, pour on the SU-8 template, and cure at 70 oC for 2 hours. After cooling, peel off the PDMS chip containing the cross-shaped channel from the template, cut it into the desired shape with a blade, and punch holes in the buffer pool, sample pool and sample waste pool with a puncher to form a diameter of 3 mm holes. At the same time, using the smooth glass plate as a template, follow the same steps to prepare a PDMS chip without microchannels as a cover slip. The PDMS chip containing the cross channel and the PDMS cover sheet without the channel were ultrasonically cleaned with secondary water, methanol, and secondary water for 10 min, respect...

Embodiment 2

[0027] (1) Graphene oxide was prepared by the Hummers method: 0.5 g graphite powder and 0.5 g NaNO 3 Added to 23 mL to a concentration of 18 mol·L -1 H 2 SO 4 , slowly add 3 g KMnO 4 , after fully stirring and mixing, react in a water bath at 35 °C for 1 h to form a beige paste. Add 40 mL of water, stir at room temperature for 30 min, then dilute with water to 140 mL. Add 3 mL of 30% H by weight dropwise 2 o 2 , the solution turned from dark brown to bright yellow. Filter the product while it is hot, and wash the product by centrifugation with ultrapure water until the supernatant is neutral. -1 Centrifuge at 8000 rpm for 2 min, disperse the centrifuged product in ultrapure water, and -1 Centrifuge at a rotational speed for 2 min, disperse the centrifuged product in ultrapure water, and sonicate for 2 h to obtain a GO solution.

[0028] (2) Fe 3 o 4 Preparation of NPs-functionalized GO nanomaterials: Dissolve 40 mg of GO in 20 mL of ultrapure water and sonicate fo...

Embodiment 3

[0036] Based on GO / Fe 3 o 4 Application of microenzyme reactor prepared by / AChE in pesticide detection:

[0037] (1) Effects of detection potential, separation voltage, and substrate ATCh concentration on the performance of the microenzyme reactor

[0038] Image 6 A is the effect of the detection potential on the performance of the microenzyme reactor. When the detection potential is lower than -0.3 V, the peak current of TCh is small; as the detection potential increases, the peak current increases. When the detection potential is higher than +0.1 V, the peak current increases slowly; if the detection potential continues to increase, the background current also increases. In addition, the carbon fiber working electrode is easy to soften when subjected to high voltage. In order to prolong the service life of the working electrode and comprehensively consider the stability, reproducibility and signal-to-noise ratio of the detection signal, +0.1 V was selected as the detec...

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Abstract

The invention discloses a preparation method of a micro-enzyme reactor based on magnetic functionalized graphene oxide, and application of the micro-enzyme reactor in pesticide detection, belonging to the technical field of micro-fluidic control chips. The micro-enzyme reactor which can be applied to pesticide detection is prepared by firstly, loading Fe3O4 magnetic nano particles on the surface of graphene oxide by using a one-step in-situ synthesis method, so as to prepare a graphene oxide/Fe3O4 nano composite material with both good magnetism and biocompatibility, secondly, fixing acetyl choline esterase on the surface of the graphene oxide/Fe3O4 through action of pi-pi, hydrogen bonds and water repelling and the like, and fixing a graphene oxide/Fe3O4/acetyl choline esterase compound inside a PDMS (Polydimethylsiloxane) micro chip channel through the action of an external magnetic field. Based on the inhibition principle of pesticide to acetyl choline esterase, the rapid quantitative detection on dimethoate is realized. The micro-enzyme reactor which is prepared based on a graphene oxide/Fe3O4/acetyl choline esterase composite material is rapid in action, simple to operate and good in reproducibility, and an effective means is provided for rapid and sensitive detection on pesticide.

Description

technical field [0001] The invention relates to a preparation method of a microenzyme reactor based on magnetically functionalized graphene oxide and its application in pesticide detection, belonging to the technical field of microfluidic chips. Background technique [0002] my country is a traditional agricultural country, and the amount of pesticides used has always ranked first in the world. The widespread use of pesticides not only causes serious pollution to the environment, but also causes harm to human health, which has aroused widespread concern in the society. At present, many analytical methods have been applied to the detection of pesticide residues. Gas chromatography and high performance liquid chromatography have high accuracy and sensitivity for the detection of pesticide residues, but there are disadvantages such as high cost and long time consumption. Therefore, it is imminent to seek a fast, sensitive and convenient method for the detection of pesticide re...

Claims

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Application Information

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IPC IPC(8): C12M1/40C12Q1/46
Inventor 梁汝萍王晓妮邱建丁
Owner NANCHANG UNIV
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