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Modified-electrode-based tyrosinase biosensor as well as preparation method and application thereof

A biosensor and tyrosinase technology, applied in the field of tyrosinase biosensor and its preparation, can solve the problems of affecting enzyme activity, unstable fixation, etc., and achieve the effects of improving activity, rapid production and long service life

Inactive Publication Date: 2013-01-02
HUNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Commonly used enzyme immobilization methods include: adsorption immobilization method, embedding immobilization method, covalent immobilization method and cross-linking immobilization method. These methods have the disadvantages that the immobilization is not firm, the cross-linking agent used in the immobilization affects the activity of the enzyme, and it is easy to use. Defects such as environmentally harmful substances

Method used

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  • Modified-electrode-based tyrosinase biosensor as well as preparation method and application thereof
  • Modified-electrode-based tyrosinase biosensor as well as preparation method and application thereof
  • Modified-electrode-based tyrosinase biosensor as well as preparation method and application thereof

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Embodiment 1

[0052] a kind of like figure 1 The present invention shown is based on the modified electrode tyrosinase biosensor, the biosensor includes a glassy carbon electrode 1, the detection end surface of the glassy carbon electrode 1 is deposited with nano-gold particles 2, and the outer surface of the nano-gold particles 2 is adsorbed with a layer The L-lysine film 3 is loaded with a layer of ordered mesoporous carbon-supported gold nanoparticles 4 on the outside of the L-lysine film 3 , and tyrosinase 5 is physically adsorbed on the ordered mesoporous carbon-supported gold nanoparticles 4 .

[0053] The preparation method of the tyrosinase biosensor based on the modified electrode of the present embodiment comprises the following steps:

[0054] 1. Preparation of ordered mesoporous carbon.

[0055] 1.1 Synthesis of mesoporous silicon template SBA-15: 8.0g block copolymer Pluronic P123 was placed in hydrochloric acid (270g H 2 O and 320mL, 1.54M hydrochloric acid), stirred in a wa...

Embodiment 2

[0070] Embodiment 2: detection application of tyrosinase biosensor

[0071] The modified electrode-based tyrosinase biosensor of Example 1 of the present invention is used as a working electrode, a saturated calomel electrode is used as a reference electrode, and a platinum electrode is used as a counter electrode to establish a three-electrode system, and connect the three-electrode system to an electrochemical workstation , to detect the concentration of hydroquinone and catechol in the solution to be tested.

[0072] Working conditions: the electrolyte used is phosphate buffered saline solution (PBS) of 1 / 15mol / L, and the current response value of the tyrosinase biosensor to the same concentration of catechol is measured in the range of pH=4.5~9.2, and optimized The result is as Figure 9 and Figure 10 As shown, pH=6.98 is selected as the optimum pH value, because at this pH value, its response current is large and stable.

[0073] The detection principle of the tyrosin...

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Abstract

The invention discloses a modified-electrode-based tyrosinase biosensor comprising a glassy carbon electrode, wherein nanogold particles are deposited on the surface of a detection end of the glassy carbon electrode; an L-lysine film is adsorbed on the outer surfaces of nanogold particles; ordered mesoporous carbon supported nanogold is loaded outside the L-lysine film; and tyrosinase is adsorbed on the ordered mesoporous carbon supported nanogold. The preparation method comprises the following steps of: firstly preparing suspension liquid of the ordered mesoporous carbon supported nanogold, depositing the nanogold particles on the glassy carbon electrode by using an electrochemical method, immersing the detection end of the nanogold modified electrode in an L-lysine solution, forming a layer of L-lysine film on the outer surface of each nanogold particle by using the electrochemical method, dropwise adding the suspension liquid of the ordered mesoporous carbon supported nanogold to the surface of the detection end, drying, and dropwise adding a tyrosinase solution in air and so as to obtain the tyrosinase biosensor. The tyrosinase biosensor disclosed by the invention has the advantages of low cost, simpleness in manufacturing, long service life, high enzyme activity, high detection precision and efficiency and the like.

Description

technical field [0001] The invention relates to a biosensor and its preparation method and application, in particular to a tyrosinase biosensor and its preparation method and application. Background technique [0002] At present, the main methods for the determination of organic pollutants such as hydroquinone and catechol include chromatography, ultraviolet spectroscopy, synchronous fluorescence spectroscopy, spectrophotometry, derivative photometry, flow injection analysis, etc. However, these methods All have defects such as complicated pretreatment, long time-consuming, large sample matrix effect, long analysis cycle, etc., require high operating level of staff and expensive instruments, so it is difficult to popularize and apply in small and medium-sized enterprises. For example, when spectrophotometry is used for detection, due to the requirements for substrate turbidity and the influence of light-interfering substances, its accuracy and scope of use are limited; while...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/327
Inventor 汤琳周耀渝曾光明龚继来李贞刘媛媛章毅杨贵德严洲雷晓霞
Owner HUNAN UNIV
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