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Fluorescence biosensor for detecting high-sensitivity copper ionss and detection method thereof

A copper ion detection and biosensor technology, applied in the field of fluorescent biosensors, can solve problems such as low sensitivity and poor selectivity, achieve good reproducibility, reduce quenching degree, and improve selectivity

Inactive Publication Date: 2012-07-25
HARBIN INST OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to use the high sensitivity of quantum dot fluorescence detection combined with the high efficiency and specificity of enzyme-catalyzed reaction to solve the problems of low sensitivity and poor selectivity in current copper detection, and provide a high-sensitivity fluorescent biosensor for copper ion detection and its detection method

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  • Fluorescence biosensor for detecting high-sensitivity copper ionss and detection method thereof
  • Fluorescence biosensor for detecting high-sensitivity copper ionss and detection method thereof
  • Fluorescence biosensor for detecting high-sensitivity copper ionss and detection method thereof

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specific Embodiment approach 1

[0018] Embodiment 1: The fluorescent biosensor for copper ion detection in this embodiment is made of quantum dots, enzymes and enzyme substrates, wherein the concentration of quantum dots is 10 -7 ~10 -3 mol / L, the enzyme concentration is 0.01~10U / mL, and the enzyme substrate concentration is 10 -4 ~1 mol / L.

[0019] In this embodiment, the quantum dot is one or a mixture of CdSe, CdTe, CdS, PbS, CdSeZnS, CdSeCdS, and CdSeS, wherein CdSeZnS represents a quantum dot with a core-shell structure, CdSe is the core, and the outer shell Quantum dots coated with a layer of ZnS.

[0020] In this embodiment, the enzyme is glycerol phosphate oxidase, cytochrome C oxidase, alkaline phosphatase, glucose oxidase, alcohol oxidase, sarcosine oxidase, choline oxidase, ascorbate oxidase, xanthine oxidase One of enzymes, glutathione oxidase, D-amino acid oxidase.

[0021] In this embodiment, the enzyme substrate is one of glycerophosphate, cytochrome C, glucose, methanol, sarcosine, chol...

specific Embodiment approach 2

[0022] Specific embodiment 2: In this embodiment, copper ions in water are detected according to the following steps: 1. Enzyme is added to the quantum dot solution, and then the enzyme substrate is added to measure the change of the fluorescence intensity of the quantum dots. 2. The enzyme and copper ions of different concentrations are mixed first and then added to the quantum dot solution, and then the enzyme substrate is added to measure the change of the fluorescence intensity of the quantum dot. The detection limit of copper ions was calculated by plotting the relative fluorescence intensity-copper ion concentration. where the concentration of quantum dots is 10 -7 ~10 -3 mol / L, the enzyme concentration is 0.01~10U / mL, and the enzyme substrate concentration is 10 -4 ~1 mol / L.

[0023] The two steps of this embodiment are enzymatic reaction (and the inhibition of enzyme activity) and quantum dot fluorescence quenching respectively. The first step is the reaction betw...

specific Embodiment approach 3

[0027] Specific embodiment three: the concentration of quantum dots in the copper ion detection fluorescent biosensor of the present embodiment is 3.9 × 10 -5 mol / L, the enzyme concentration is 1U / mL, and the enzyme substrate concentration is 4.92×10 -3 mol / L.

[0028] In this embodiment, when the enzyme and the substrate are added to the quantum dot solution, the fluorescence of the quantum dot is significantly reduced, such as figure 1 shown. However, the enzyme and substrate itself have no significant effect on the fluorescence intensity of quantum dots, such as figure 1 As shown in (b), the fluorescence quenching of quantum dots should be caused by the hydrogen peroxide produced by the enzyme-catalyzed substrate.

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Abstract

The invention relates to a fluorescence biosensor for detecting high-sensitivity copper ions and a detection method thereof, which relate to the biosensor and a method for detecting the copper ions in water by utilizing the biosensor. In order to solve the problems of low sensitivity, poor selectivity and the like in existing copper detection, the fluorescence biosensor for detecting the copper ions is made of a quantum dot, enzyme and an enzyme substrate. The detection method comprises the following steps of: adding the enzyme into quantum dot solution, adding the enzyme substrate, and measuring the change of fluorescence intensity of the quantum dot; and mixing the enzyme and the copper ions with different concentration, adding the mixture into the quantum dot solution, adding the enzyme substrate, measuring the change of fluorescence intensity of the quantum dot, drawing according to relative fluorescence intensity-copper ions, and calculating a detection limit of the copper ions. The process is simple, the price is low, the reaction condition is moderate, the operation is easy, and the reproducibility is good. The method is a very promising detection technology, and is suitable for trace detection of the copper ions in environment and even bio-system.

Description

technical field [0001] The invention relates to a fluorescent biosensor and a method for detecting copper ions in water using it. Background technique [0002] Due to the sharp increase of industrial and agricultural waste and municipal solid waste, as well as the extensive use of pesticides and chemical fertilizers, the soil, water and other environments that humans rely on for survival have suffered serious heavy metal pollution, and the trend is increasing. Among all heavy metal ions, copper ion is an important trace element that plays an important role in various physiological environments. However, excessive intake of copper ions may cause eczema in the human body and harm the human kidney and central nervous system. Therefore, the analysis and detection of copper ions in environmental and biological samples is becoming more and more important. People use a variety of means to detect the concentration of trace copper ions, such as atomic absorption spectrometry, induc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
Inventor 郭彩欣戴志飞王金梁成晶
Owner HARBIN INST OF TECH
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