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An in situ synthesis of gold nanoparticles for colorimetric detection of lipase enzyme activity

A nano-gold colorimetric method and lipase enzyme activity technology, which is applied in the field of analytical chemistry, can solve the problems of the undisclosed nano-gold colorimetric method for detecting lipase enzyme activity, and achieve suitable popularization and application, simplification of operation steps, and dissolution good performance

Active Publication Date: 2019-02-12
NANJING TECH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The prior art does not disclose a technical solution for the detection of lipase activity based on in situ synthesis of gold nanoparticles colorimetric method

Method used

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  • An in situ synthesis of gold nanoparticles for colorimetric detection of lipase enzyme activity
  • An in situ synthesis of gold nanoparticles for colorimetric detection of lipase enzyme activity
  • An in situ synthesis of gold nanoparticles for colorimetric detection of lipase enzyme activity

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] (1) Add the final concentration of Tween solution to 2mL A and B centrifuge tubes respectively to be 1% (V / V);

[0027] (2) Add buffer solution to the A and B centrifuge tube solutions obtained in step (1) respectively to adjust the pH, and adjust the pH value to 8.5; add the lipase solution to be tested to the B centrifuge tube, and the A centrifuge tube does not need Add the lipase solution to be tested; mix well, and adjust the reaction temperature to 45°C; the UV-visible spectrum of the solution after a certain period of time can be obtained by adjusting the reaction time of the lipase solution to be tested (see figure 2 ), with ΔA 530 Be the ordinate, the time is the abscissa, draw the kinetic curve of lipase activity (see image 3 );

[0028] (3) to the A, B centrifuge tube solution that obtains in step (2), the final concentration that adds chloroauric acid solution is 1.5mmol / L, mixes, and regulating reaction temperature is 25 ℃ and the time is 210min; In A c...

Embodiment 2

[0033] Compared with embodiment 1, the difference is only that the operating parameters of steps 1-3 are different, and the present embodiment is specifically:

[0034] (1) Add the final concentration of Tween solution to 2mL A and B centrifuge tubes respectively to be 0.1% (V / V);

[0035] (2) Add buffer solution to the A and B centrifuge tube solutions obtained in step (1) respectively to adjust the pH, and adjust the pH value to 6; add the lipase solution to be tested to the B centrifuge tube, and the A centrifuge tube does not need Add the lipase solution to be tested; mix well, and adjust the reaction temperature to 30°C; by adjusting the reaction time of the lipase solution to be tested, the ultraviolet-visible spectrum of the test solution after a certain period of time can be obtained, expressed as ΔA 530 is the ordinate, the time is the abscissa, and draws the kinetic curve of the lipase enzyme activity;

[0036] (3) Adding the final concentration of chloroauric acid ...

Embodiment 3

[0038] Compared with embodiment 1, the difference is only that the operating parameters of steps 1-3 are different, and the present embodiment is specifically:

[0039] (1) Add the final concentration of Tween solution to 2mL A and B centrifuge tubes respectively to be 3% (V / V);

[0040] (2) Add buffer solution to the A and B centrifuge tube solutions obtained in step (1) respectively to adjust the pH, and adjust the pH value to 9; add the lipase solution to be tested to the B centrifuge tube, and the A centrifuge tube does not need Add the lipase solution to be tested; mix well, and adjust the reaction temperature to 60°C; by adjusting the reaction time of the lipase solution to be tested, the ultraviolet-visible spectrum of the test solution after a certain period of time can be obtained, expressed as ΔA 530 is the ordinate, the time is the abscissa, and draws the kinetic curve of the lipase enzyme activity;

[0041] (3) to the A, B centrifuge tube solution that obtains in ...

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Abstract

The present invention relates to an in-situ synthesized gold nanoparticle colorimetric method for detecting lipase enzyme activity. According to the present invention, the characteristic that the tween self-oxidation produces hydrogen peroxide is utilized to reduce chloroauric acid into gold nanoparticles so as to be adopted as the coloring signal element, and the tween is concurrently adopted as the lipase substrate and the protection agent of the in-situ synthesized gold nanoparticles; the colorimetric analysis method with characteristics of low cost, simple operation, convenience and rapidness are developed; and the adopted tween is amphiphilic, the emulsification process is not required, the stable in-situ synthesized gold nanoparticles have the strong high-concentration salt ion resistance, such that the in-situ synthesized gold nanoparticle colorimetric method is suitable for promotion and application.

Description

technical field [0001] The invention belongs to the field of analytical chemistry, and specifically relates to a chemical ratio for detecting lipase enzyme activity in which in-situ synthesized nano gold is used as a signal element, and Tween is used as a lipase substrate and as a protective agent for in situ synthesized nano gold. color method. Background technique [0002] Lipase (Lipase, EC 3.1.1.3) is a long-chain triacylglyceride hydrolase that can only act in heterogeneous or organic phases. It mainly catalyzes reactions such as esterification, hydrolysis, aminolysis, and transesterification. It is widely used in many industrial fields such as food, detergent, leather, textiles, cosmetics, papermaking and pharmaceuticals. As the application fields of lipase continue to expand, it is particularly important to establish a method for detecting lipase activity with low cost, simple operation, and convenience. [0003] As we all know, there are many methods for detecting ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/78
Inventor 田丹碧汤燕黄和江凌张卫丁亚琦
Owner NANJING TECH UNIV