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Method for detecting tripolycyanamide based on nanogold mimetic peroxidase

A technology of peroxidase and melamine, which is applied in the field of analytical chemistry and nanometers, can solve the problems of time-consuming, poor sensitivity, and limited application, and achieve the effects of low processing requirements, simple and fast preparation process, and high detection sensitivity

Inactive Publication Date: 2012-11-07
FUJIAN MEDICAL UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Most of these methods show quite high sensitivity, but the complex modification process of gold nanoparticles limits their application to some extent.
The technique of colorimetric detection of melamine during the formation of gold nanoparticles has also been reported, but it is very time-consuming
Simpler methods for the detection of melamine have been established using unmodified gold nanoparticles (actually citrate-coated gold nanoparticles) as chromogenic probes, but all were less sensitive

Method used

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  • Method for detecting tripolycyanamide based on nanogold mimetic peroxidase
  • Method for detecting tripolycyanamide based on nanogold mimetic peroxidase
  • Method for detecting tripolycyanamide based on nanogold mimetic peroxidase

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Effect test

Embodiment 1

[0036] 500 μL of 0.1 g / L chloroauric acid aqueous solution was diluted with 39.5 ml of water, and 0.8 ml of 0.1 g / L sodium borohydride aqueous solution was added under vigorous stirring (the addition time was controlled within 5 minutes), and the color of the reaction solution From light yellow to wine red, continue to stir rapidly in the dark for 1 hour. The average particle size of the obtained nano-gold is 8.1 ± 1.1 nm (see figure 1 ), with a maximum absorption wavelength of 518 nm (see figure 2 ), stored at 4 ℃, the nano-gold can be kept for at least two months and is relatively stable. All glassware used in the above process was soaked in aqua regia, washed thoroughly with double distilled water, and dried.

Embodiment 2

[0038] Take 0.05 ml of the nano-gold solution prepared in Example 1, add 0.25 ml of hydrogen peroxide with a concentration of 8 mol / L, and 0.05 ml of 3,3',5,5'-tetramethylbenzidine with a concentration of 16 mmol / L Hydrochloride, 0.65 ml of phosphate buffer solution (pH=4) with a concentration of 10 mmol / L, after mixing, incubate at 37 °C for 10 minutes, the mixed solution turns blue, and the maximum absorption wavelength is 652 nm (see image 3 ).

Embodiment 3

[0040] Add 0.1 ml of the nano-gold solution prepared in Example 1 to 0.3 ml of phosphate buffer solution containing 0.02 mg / L melamine (the concentration of phosphate buffer solution is 0.01 mol / L, pH=3), mix and incubate at 40 °C 5 minutes, centrifuge the mixed solution for 5 minutes (0-15000 rpm), take 0.2 ml of supernatant, add 0.25 ml of hydrogen peroxide with a concentration of 8 mol / L, 0.05 ml of 3,3',5 , 5'-Tetramethylbenzidine hydrochloride, 0.5 ml of 10 mmol / L phosphate buffer solution (pH=4), after mixing, incubate at 37°C for 10 minutes, and measure the absorbance at 652 nm wavelength. In the blank control, 0.3 ml of phosphate buffer solution (the concentration of phosphate buffer solution is 0.01 mol / L, pH=3) was used instead of melamine solution. Such as Figure 4 As shown, with the increase of centrifugal speed, ΔA 652 (Blank control experiment A 652 Subtract Chromogenic Experiment A 652 ) increases, and when the speed reaches 10000 rpm, ΔA 652 Reaches the m...

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Abstract

The invention discloses a method for detecting tripolycyanamide based on nanogold mimetic peroxidase. Nanogold stimulates the change of the activity of the peroxidase after being interacted with tripolycyanamide, hydrogen peroxide is catalyzed by nanogold to oxidize 3,3',5,5'-tetramethylbenzidine dihydrochloride so as to be conducted for color development, therefore, the change of the color of a solution and the characteristics of an ultraviolet absorption spectrum are shown. The detection limit of the visual observation is 0.02mg / L. The linear range detected by the optical density is 5-80 mug / L, and the detection limit is 1.2 mug / L. The content of tripolycyanamide in a milk powder sample can be detected well by the method after being simply preprocessed.

Description

technical field [0001] The invention relates to a rapid content determination method for melamine using nano gold as a simulated peroxidase, belonging to the fields of analytical chemistry and nanotechnology. Background technique [0002] It can provide colorimetric detection of naked eye identification signal, which has the advantages of simple, fast, suitable for real-time and on-site detection. Colorimetric sensors based on gold nanoparticles have received extensive attention in recent years, most of which are based on the color change produced by plasmonic coupling during the aggregation or aggregation-redispersion of gold nanoparticles. Gold nanoparticles have been increasingly used in colorimetric detection of cells, proteins, DNA, metal ions and small molecules. [0003] Melamine (1,3,5-triazine-2,4,6-triamine, C 3 h 6 N 6 ) is a common chemical substance, mainly used in the production of melamine formaldehyde resin. Because of its high nitrogen content (66.6%) a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/78G01N21/31
Inventor 陈伟邓豪华林新华李光文洪磊
Owner FUJIAN MEDICAL UNIV
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