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Urease Inhibitor Determination Method Based on Fluorescent Gold Nanoclusters

A technology of urease inhibitor and fluorescent gold nanometer, which is applied in analytical chemistry and nanometer fields, can solve problems such as environmental pollution and economic loss, and achieve the effect of simple and fast preparation process and suppression of fluorescence quenching

Inactive Publication Date: 2016-06-29
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 Inhibitor Determination Method Based on Fluorescent Gold Nanoclusters
  • Urease Inhibitor Determination Method Based on Fluorescent Gold Nanoclusters
  • Urease Inhibitor Determination Method Based on Fluorescent Gold Nanoclusters

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0030] Add 0.6mL of 0.5mol / L sodium hydroxide solution and 0.4mL of 0.02g / L chloroauric acid solution into 4mL of 0.08mol / L N-acetyl-L-cysteine ​​solution , mixed, placed in a 37 ° C constant temperature water bath to react for 2.5h. 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

[0032] Add 0.05 mL of urease solution (pH=6.0) with a concentration of 1.5 U / mL to 0.2 mL of urea solution with a concentration of 1 mol / L containing mercaptoethylamine (8 μmol / L) or p-benzoquinone (50 μmol / L) respectively ( pH=6.0), mix well and react in a constant temperature water bath at 25°C for 40min. (2) Add 0.2 mL of the solution prepared in Example 1 to the above reaction solution, and react in a constant temperature water bath at 25°C for 3 minutes. Set up a group of blank control groups without urease inhibitors. After the reaction was completed, observe under a UV lamp, and add 8 μmol / L mercaptoethylamine ( figure 1A in A) or 50μmol / L p-benzoquinone ( figure 1 After B), the gold nanoclusters have red fluorescence, while the red fluorescence of the control group is quenched ( figure 1 in C). figure 2 is the corresponding fluorescence emission spectrum.

example 3

[0034] Add 0.05mL of urease solution (pH=6.0) with a concentration of 1.5U / mL into 0.2mL of urea solution (pH=6.0) with a concentration of 1mol / L containing different concentrations of mercaptoethylamine, mix well and incubate at 25°C React in a constant temperature water bath for 40 min. The solution prepared by 0.2mL of Example 1 was added to the above-mentioned reaction solution, reacted in a constant temperature water bath at 25°C for 3min, and measured the emitted light intensity value F 650 value to calculate the inhibition rate. The result is as image 3 As shown, the IC of mercaptoethylamine was obtained by software fitting 50 It is 2.8 μmol / L.

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Abstract

The invention discloses a Urease Inhibitor Determination Method Based on Fluorescent Gold Nanoclusters , which is characterized in that urease is used to specifically catalyze urea to generate ammonia and carbon dioxide, the newly generated ammonia can increase the pH value of the system, and quench the fluorescence of gold nanoclusters protected by N-acetyl-L-cysteine, while Urease inhibitors can prevent urease from catalyzing the decomposition of urea and inhibit fluorescence quenching, so they can be used in the detection of urease inhibitors. The F650 value was measured, the inhibition rate was calculated, and the IC50 of mercaptoethylamine and p-benzoquinone were obtained by software fitting as 2.8 μmol / L and 11.9 μmol / L, respectively. The invention can be applied to high-throughput screening of urease inhibitors.

Description

technical field [0001] The invention relates to a method for measuring a urease inhibitor 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. Urease inhibitors are drugs that dissolve urinary stones and stop new crystals from forming in the urine. 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 order to improve the utilization r...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/573G01N33/533
CPCG01N33/533G01N33/573
Inventor 陈伟邓豪华林小青王艳红沈奕珉
Owner FUJIAN MEDICAL UNIV
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