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Urease inhibitor determination method based on fluorescence gold nano cluster

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

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

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0030] 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

[0032]Add 0.05 mL of urease solution (pH=6.0) with a concentration of 1.5 U / mL to 0.2 mL of urease solution containing mercaptoethylamine (8 μmol / L) or p-benzoquinone (50 μmol / L) at a concentration of 1 mol / L In urea solution (pH=6.0), after mixing evenly, react in a constant temperature water bath at 25°C for 40 min. (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 min. 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 1 A in A) or 50 μmol / 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 0.05 mL of urease solution (pH=6.0) with a concentration of 1.5 U / mL into 0.2 mL of urea solution (pH=6.0) with a concentration of 1 mol / L containing different concentrations of mercaptoethylamine. C for 40 min in a constant temperature water bath. Add 0.2 mL of the solution prepared in Example 1 to the above reaction solution, react in a constant temperature water bath at 25°C for 3 min, and measure 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 a fluorescence gold nano cluster. The urease inhibitor determination method is characterized in that urea is catalyzed to generate ammonium and carbon dioxide by utilizing the specificity of urease, the pH value of the system can be increased through the newly generated ammonium, the fluorescence of the gold nano cluster protected by N-acetyl-L-cysteine is extinguished, the urease inhibitor can prevent the process that the urea is decomposed by the catalyzing of the urease, the extinguishment of the fluorescence is inhibited, and the urease inhibitor determination method can be used for detecting the urease inhibitor. An F650 value is determined, the inhabitation rate is calculated, and the contents of IC50 of cysteamine and p-benzoquinone, respectively 2.8mu mol / L and 11.9mu mol / L, can be obtained through the fitting by software. The urease inhibitor determination method can be used for the high-throughput screening of the urease inhibitor.

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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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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