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Catalase fluorescence determination method based on gold nano-cluster probe

A technology of gold nanoclusters and catalase, which is applied in analytical chemistry and nanometer fields, can solve the problems of complex operation, insufficient precision, and poor repeatability, and achieve the effects of rapid preparation process, high sensitivity, and good specificity

Inactive Publication Date: 2015-02-04
FUJIAN MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For these methods, some require expensive instruments, some require special reagents, some are complicated to operate, and some methods themselves have poor repeatability and insufficient precision.

Method used

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  • Catalase fluorescence determination method based on gold nano-cluster probe
  • Catalase fluorescence determination method based on gold nano-cluster probe
  • Catalase fluorescence determination method based on gold nano-cluster probe

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] 0.025 ml of catalase solution with a concentration of 0.5 U / mL (containing HEPES——N-(2-hydroxyethyl)piperazine-N'-2-ethanesulfonic acid, concentration 20 mmol / L, pH=7.4 ) into 0.175 ml of a hydrogen peroxide solution with a concentration of 10 μmol / L (containing HEPES, a concentration of 20 mmol / L, pH=7.4), and reacted at 25°C for 90 minutes. 0.05 ml of Fe with a concentration of 0.9 mmol / L 2+ The solution (containing 40 mmol / L sulfuric acid) and 0.2 ml of the gold nanocluster solution prepared in Example 1 were sequentially added to the above reaction solution, shaken well and placed in a 25°C water bath for 10 minutes to react. Observed under ultraviolet light, the red fluorescence of the gold nanocluster solution without catalase was quenched after the Fenton reaction ( figure 1 A) in A), while the gold nanocluster solution recovered red fluorescence after adding catalase reaction ( figure 1 in B). figure 2 A gold nanocluster solution without adding catalase ( ...

example 3

[0037] Add 0.025 ml of 0.5 U / mL catalase solution (HEPES, 20 mmol / L pH=7.4) to 0.175 ml of 10 μmol / L hydrogen peroxide solution (HEPES, 20 mmol / L pH=7.4) in 25°C for 15-130 minutes. 0.05 ml of Fe with a concentration of 0.9 mmol / L 2+ The solution (containing 40 mmol / L sulfuric acid) and 0.2 ml of the gold nanocluster solution prepared in Example 1 were sequentially added to the above reaction solution, shaken well and placed in a 25°C water bath for 10 minutes to react. Such as image 3 As shown, after catalase catalyzed reaction for 90 minutes, the fluorescence intensity value F 650 The change is basically stable, so the catalase catalyzed reaction time is selected as 90 minutes.

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Abstract

The invention discloses a catalase fluorescence determination method based on a gold nano-cluster probe and relates to a catalase fluorescence determination method in which a gold nano-cluster protected by N-acetyl-L-cysteine serves as a fluorescence probe. The catalase fluorescence determination method is characterized in that H2O2 is catalyzed by Fe<2+> to generate hydroxyl radicals, so that the fluorescence of the gold nano-cluster is quenched; H2O2 can be catalyzed by the catalase and is decomposed into H2O and O2 to restrain quenching of the fluorescence of the gold nano-cluster, so that the change of fluorescence emission spectrum characteristics is represented, and the catalase can be detected. A linear relation is formed between a fluorescence intensity change value delta F650 and the concentration of catalase within a range of 0.01-0.3U / mL, and the detection limit is 0.002U / mL. The method is high in sensitivity and high in reproducibility, and can be used for determining the catalase in food, industry, environment and life systems.

Description

Technical field [0001] The present invention involves the measurement method of the hydrotide enzymes that use the gold nano-cluster of N-acetyl-L-cysteine as a fluorescent probe, which belongs to the field of analytical chemistry and nano-technology. Background technique [0002] Penoidase (also known as tentalase) is an oxidation enzyme widely existed in biological tissue.Penoidase is a type of tetramotinase. It consists of four sub -measuring sub -samples. Each subunit is 60,000 g / mol.Regiment, molecular weight is around 240000.The accumulation of active oxygen and free radicals in the organism can cause membrane fat peroxidation, which causes the organic disorders of the organism itself.Both polymers and non -enzymatic reactions can produce hydrogen peroxide (H 2 O 2 ), H 2 O 2 It is the preparation of active oxygen with toxicity.The main function of peroxide is catalytic H 2 O 2 Decomposition into water and oxygen, remove hydrogen peroxide in the body, so that the cells will...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
Inventor 陈伟邓豪华彭花萍兰青刘爱林
Owner FUJIAN MEDICAL UNIV
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