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Polymer liquid film potential sensor for detecting enzymatic activity and detection method thereof

A potential sensor and detection method technology, applied in the field of sensors, can solve problems such as difficulty in realizing the joint detection of multiple product ions, limited enzyme species, inability to capture potential signals, etc., to avoid changes in test performance, low cost, and anti-interference ability. strong effect

Active Publication Date: 2011-11-09
YANTAI INST OF COASTAL ZONE RES CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

It should be pointed out that in this type of method, the detection is carried out separately after the reaction is terminated, so the potential signal of the transient intermediate product in the reaction process cannot be captured; the sensitive membrane is activated by a certain final product ion before the measurement, so it is difficult to realize Co-detection of multiple product ions
Due to this limitation, the enzyme species successfully detected by polymer liquid membrane ion selective electrodes are very limited, especially the highly sensitive detection of important immune-related enzymes such as peroxidase needs to be realized urgently

Method used

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  • Polymer liquid film potential sensor for detecting enzymatic activity and detection method thereof
  • Polymer liquid film potential sensor for detecting enzymatic activity and detection method thereof
  • Polymer liquid film potential sensor for detecting enzymatic activity and detection method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0029] The potential sensor includes a working electrode 1, an external reference electrode 2, an internal reference electrode 3 and a potentiometer 4. The potentiometer 4 is respectively connected to the internal reference electrode 3 and the external reference electrode 4 through wires, and the internal reference electrode 3 is inserted into the In the working electrode 1 filled with liquid 8, the working electrode 1 and the external reference electrode 2 are inserted into the detection cell 5 filled with the detection liquid 6. The working electrode 1 is an electrode carrier with a polymer sensitive film 7 fixed on it. A magnetic stirrer 9 with a stirring effect is provided, and a magnetic stirrer is provided at the bottom of the detection pool to rotate the magnetic stirrer. The electrode carrier on which the polymer sensitive membrane 7 is fixed is a polyvinyl chloride tube.

[0030] Detection method:

[0031] With aniline as the substrate, the enzyme to be tested is hor...

Embodiment 2

[0040] The difference from Example 1 is:

[0041] With phenol as the substrate, the enzyme to be tested is horseradish peroxidase.

[0042] a. The anion exchanger tri-dodecylmethyl ammonium chloride doped polymer film is used as the working electrode sensitive film, the silver-silver chloride electrode is the internal and external reference electrode, and the internal reference electrode is connected with the electrochemical workstation CHI760D The working electrode joint is connected, the external reference electrode is connected to the reference electrode joint, and the "open circuit potential-time" technique is selected to measure the potential value.

[0043] The preparation of the working electrode is: according to the mass percentage 1.5%: 66%: 32.5%, a total of 345 mg of tri-dodecylmethyl ammonium chloride, polyvinyl chloride, and dioctyl sebacate are dissolved in 3.5 mL of tetrahydrofuran solution , placed in a flat-bottomed glass ring (with an inner diameter of 3.6 c...

Embodiment 3

[0048] The difference from Example 1 is:

[0049] With 3′,5,5′-tetramethylbenzidine as substrate, the enzyme to be tested is horseradish peroxidase.

[0050] In the sensor, the polyethylene tube fixed with the polymer sensitive film is sleeved on the electrode head of the rotating disk electrode, and the internal reference electrode is connected to the reference electrode interface of the rotating disk electrode.

[0051] After the rotational speed of the rotating disc electrode was adjusted to 5000rpm, add 20 μL of 0.1M 3,3′,5,5′-tetramethylbenzidine (final concentration 2mM) to 1mL of the detection solution, and after the isoelectric value was stable, add 1μL of 1M H 2 o 2 (final concentration 1mM), then add different concentrations of horseradish peroxidase solutions, and record the potential increase 300s after the oxidation reaction is initiated. Use six identical electrodes to measure the final concentration of 10 under the same conditions -6 U / mL, 10 -5 U / mL, 10 -4...

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Abstract

The invention relates to a sensor, in particular to a polymer liquid film potential sensor for detecting enzymatic activity and a detection method thereof. A potential detector is respectively connected with an internal reference electrode and an external reference electrode through wires, the internal reference electrode is inserted in a working electrode filled with internal liquid, the working electrode and the external reference electrode are inserted in a detection pool with detection liquid, the working electrode is an electrode carrier on which a polymer sensitive film is fixed, and a power device with a solution disturbance function is arranged in the detection pool. According the detection method, the potential detector is used for detecting the change of the potential of the polymer sensitive film at the bottom of the working electrode before and after enzymatic reaction, and the enzymatic activity concentration can be detected on the basis of a standard working curve according to a potential change signal. The potential sensor has the advantages of high generality, good selectivity, high sensitivity and low cost and is suitable for detection under a complicated matrix condition.

Description

technical field [0001] The invention relates to a sensor, in particular to a polymer liquid membrane potential sensor for measuring enzyme activity and a detection method thereof. Background technique [0002] As high-efficiency biocatalysts, enzymes are widely distributed in nature and play important physiological roles. In addition to their applications in immunoassays and other fields, the determination of enzyme activity is an important topic in the field of analytical science. Existing enzyme detection techniques mainly include amperometric analysis, photometric analysis, fluorescence analysis, chemiluminescence analysis, mass analysis, surface plasmon resonance analysis, etc., but these methods are either costly or have strict requirements on sample color and turbidity. Or it is sensitive to the measurement environment such as vibration. In order to overcome the above problems, it is necessary to develop a more efficient method for the determination of enzyme activity....

Claims

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Application Information

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IPC IPC(8): G01N27/413C12Q1/26C12Q1/48C12Q1/34C12Q1/25
Inventor 秦伟王学伟丁家旺
Owner YANTAI INST OF COASTAL ZONE RES CHINESE ACAD OF SCI
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