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Urease activity fluorescence determination method based on gold nano cluster

A gold nanocluster and urease activity technology, applied in analytical chemistry and nanometer fields, can solve problems such as environmental pollution and economic loss, and achieve the effects of high detection sensitivity and simple and fast preparation process.

Inactive Publication Date: 2014-12-17
FUJIAN MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In agriculture, when the activity of urease in the soil is too high, the urea in the fertilizer is rapidly decomposed into ammonia, which is discharged into the atmosphere, causing economic losses and environmental pollution

Method used

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  • Urease activity fluorescence determination method based on gold nano cluster
  • Urease activity fluorescence determination method based on gold nano cluster
  • Urease activity fluorescence determination method based on gold nano cluster

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0033] Add 0.6 mL of 0.5 mol / L sodium hydroxide solution and 0.4 mL of 0.02 g / L chloroauric acid solution to 4 mL of 0.08 mol / L N-acetyl-L-cysteine ​​solution , mixed well, and placed in a 37°C constant temperature water bath to react for 2.5 h. After the reaction, the reaction solution was purified with a dialysis bag with a molecular weight cut off of 3500. The obtained gold nanocluster solution is colorless under visible light, and produces strong red fluorescence under ultraviolet light irradiation. Stored in the dark at 4°C, it can remain relatively stable for at least one month.

example 2

[0035] Add 0.05 mL of urease solution (pH=6.0) with a concentration of 22 U / L to 0.2 mL of urea solution (pH=6.0) with a concentration of 1 mol / L, mix well and react in a constant temperature water bath at 25°C 40 min. 0.2 mL of the gold nanocluster solution prepared in Example 1 was added to the above solution, and reacted in a constant temperature water bath at 25°C for 3 min. Set up a group of blank control groups without urease. After the reaction, observed under ultraviolet light, the blank control group showed red fluorescence ( figure 1In A), while adding 22 U / L urease, the red fluorescence of gold nanoclusters was quenched ( figure 1 in B). figure 2 It is the fluorescence emission spectrum of the gold nanocluster solution of the blank control group and the urease group.

example 3

[0037] Add 0.05 mL of urease solution (pH=6.0) with a concentration of 22 U / L to 0.2 mL of urea solution (pH=6.0) of different concentrations, mix well and react in a constant temperature water bath at 25°C for 40 min. Add 0.2 mL of the gold nanocluster solution prepared in Example 1 to the above solution, react in a constant temperature water bath at 25°C for 3 min, and measure the emitted light intensity value F 650 . Depend on image 3 It can be seen that when the urea concentration is 1mol / L, the fluorescence quenching value ΔF 650 to reach maximum.

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Abstract

The invention discloses a urease activity fluorescence determination method based on a gold nano cluster. The method is characterized by comprising the steps of catalyzing urea to generate ammonium and carbon dioxide by utilizing the specificity of urease, wherein the newly generated ammonium can increase the pH value of a system, so that the fluorescence of the gold nano cluster protected by N-acetyl-L-cysteine is extinguished, the variation of fluorescence emission spectrum characteristics can be reflected, and the method can be used for determining the activity of the urease. F650 is in a linear relation with the concentration value of the urease in the range of 2.2 to 44U / L, and the detection limit is 0.55U / L. The urease activity fluorescence determination method is high in sensitivity, good in reproducibility and applicable to the determination of helicobacter pylori.

Description

technical field [0001] The invention relates to a method for measuring urease activity using gold nanoclusters protected by N-acetyl-L-cysteine ​​as fluorescent probes, and belongs to the fields of analytical chemistry and nanotechnology. Background technique [0002] Urease is a nickel-containing oligomerase that can efficiently and specifically catalyze the hydrolysis of urea to generate carbon dioxide and ammonia. In medicine, bacterial urease is a pathogenic factor that cannot be ignored. It can induce many diseases, such as pyelonephritis, hepatic coma, peptic ulcer and infectious urinary tract stones. In agriculture, when the activity of urease in the soil is too high, the urea in the fertilizer is rapidly decomposed into ammonia, which is discharged into the atmosphere, causing economic losses and environmental pollution. In addition, soy products such as soy milk, milk substitute powder, etc., if they contain urease, are harmful to the human body and will cause symp...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
Inventor 陈伟邓豪华吴钢伟刘银环彭花萍
Owner FUJIAN MEDICAL UNIV
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