Electric potential type biosensor and detection method

A biosensor, potentiometric technology, applied in biochemical equipment and methods, instruments, measuring devices, etc., can solve the problems of high cost of selective ionophore, complicated calculation method of urea concentration, and complicated structure of indicating electrode, etc. The effect of large-scale manufacturing, simple structure, and reduced inspection costs

Pending Publication Date: 2019-06-28
LEADWAY HK
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Problems solved by technology

However, this potentiometric measurement method in these two US patents also has obvious disadvantages: NH 4 + Selective ionophores are expensive; according to the Nernst equation, under certain conditions, the potential difference between the indicator electrode and the reference electrode has a linear relationship with the logarithm of the urea concentration in the blood, and the urea in the blood is calculated based on this Concentration, which will lead to more complicated calculation method of urea concentration in blood; the structure of the indicator electrode is more complicated, and it is more difficult to manufacture
This calculation method is more complicated
Furthermore, since it involves calculating the concentration of a given ion in solution using a urease electrode, the detection result is susceptible to interference from other ions in solution

Method used

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  • Electric potential type biosensor and detection method
  • Electric potential type biosensor and detection method
  • Electric potential type biosensor and detection method

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0076] Embodiment 1 reaction reagent preparation

[0077] Prepare the second reaction reagent on the indicating electrode of the present invention and the first reaction reagent on the reference electrode, as follows:

[0078] The reaction reagent of the present invention: the second reaction reagent on the indicating electrode contains 0.05~0.5M PBS (pH6.0~7.5), trehalose 0.25%~1% (w / w), Triton X-100 0.1%~0.6% ( v / v), urease (2280~8712U / mL); the first reaction reagent on the reference electrode contains 0.05~0.5M PBS (pH6.0~7.5), Triton X-100 0.1%~0.6% (v / v ), trehalose 0.25% ~ 1% (w / w), ruthenium compound, potassium ferricyanide K 3 [Fe(CN) 6 ] or potassium ferrocyanide K 4 [Fe(CN) 6 ] 1.0 to 4.5% (w / w).

[0079] Reaction reagent 1 of the present invention: the second reaction reagent on the indicator electrode contains 0.1M PBS (pH 7.2), trehalose 1%, Triton X-100 1%, urease (4491.2U / ml); the first reaction reagent on the reference electrode The reaction reagent conta...

Embodiment 2

[0086] Embodiment 2: Detection of potential difference

[0087] use as figure 1 or figure 2 The potential type biosensor (here referred to as figure 2 The potential-type biosensor in the sample is used as an example) to detect blood samples, which can be selected from plasma, serum or whole blood. Here, whole blood samples are used as an example to illustrate, and the detailed steps are as follows:

[0088] (1) Insert the electrical connection end of the potential type biosensor into a detection instrument through the first contact and the second contact of the potential type biosensor (there are many types of the detection instrument, here for electrochemical In the electrical connector of the workstation Biologic VSP EC-Lab as an example);

[0089] (2) Select a variety of whole blood samples with different urea concentrations (here four different concentrations of 4.5mM, 14.5mM, 24.5mM and 41.5mM are used for illustration), without applying external voltage to the indic...

Embodiment 3

[0101] Example 3: Selecting an Electron Transporter

[0102] Let the electron carrier in the reaction reagent 1 of the present invention be replaced by potassium ferricyanide or potassium ferrocyanide respectively, when changing to potassium ferricyanide, the second reaction reagent on the indicator electrode and the first reaction reagent on the reference electrode Reaction reagent is as shown in the reaction reagent 2 of the present invention in embodiment 1; When changing to potassium ferrocyanide, the second reaction reagent on the indicator electrode and the first reaction reagent on the reference electrode are as in embodiment 1 Reagent 3 of the present invention is shown.

[0103] Select four kinds of whole blood samples with different urea concentrations (5.4mM, 17.5mM, 25.4mM and 31.8mM), and detect according to the potential difference detection method in Example 2, and each kind of whole blood samples with different urea concentrations is detected multiple times in ...

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Abstract

The invention provides an electric potential type biosensor and method for detecting the concentration of urea in a sample. The electric potential type biosensor comprises an indication electrode anda reference electrode, a second reaction reagent in the indication electrode includes urease but not an electron transfer body, and a first reaction reagent in the reference electrode includes the electron transfer body but not the urease. Under the condition that an external voltage is not applied, urea in the urease catalysis sample makes reaction after that blood and the like are added, an electric potential difference is thus generated between the indication electrode and the reference electrode, and is in linear relation with the concentration of urea in the sample, and the concentrationof urea in the sample can be calculated according to the linear relation.

Description

technical field [0001] The invention relates to a potential biosensor and a method for detecting urea concentration in a sample, and belongs to the technical field of electroanalysis and chemical detection. Background technique [0002] Urea (urea) is the final product of the catabolism of protein and its constituent amino acids, which is not combined with plasma protein. In protein catabolism, protein is decomposed into amino acids, and ammonia formed by deamination of amino acids enters the urea cycle in the liver and produces urea with carbon dioxide. The urea produced in the liver is mainly excreted by the kidneys, and a small part is excreted by sweat. In the kidneys, urea is filtered by the glomeruli and excreted in the urine. If kidney function is impaired, the glomerular filtration rate decreases and the concentration of urea in the blood increases. Therefore, the urea content in the blood is one of the most important physiological indicators for evaluating renal ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/26G01N27/30G01N27/327
CPCG01N27/327C12N9/80C12Y305/01005G01N27/3272
Inventor 孙玉龙项新亮程艳张莉
Owner LEADWAY HK
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