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Method for detecting AGEs (Advanced Glycation End Products) and application

A technology for advanced glycation and end-products, applied in the field of food safety testing and analytical chemistry, to achieve the effects of good stability, wide linear range and simple operation

Inactive Publication Date: 2018-09-07
NANJING NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] In order to overcome the deficiencies of the above-mentioned prior art, the present invention provides a method and application for detecting advanced glycation end products, which solves the problem that there is no high-efficiency, multi-residue detection method for AGEs at present, and has a higher High detection sensitivity and wide linear range

Method used

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  • Method for detecting AGEs (Advanced Glycation End Products) and application
  • Method for detecting AGEs (Advanced Glycation End Products) and application
  • Method for detecting AGEs (Advanced Glycation End Products) and application

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Experimental program
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Embodiment 1

[0031] The preparation of embodiment 1 electrochemical immunosensor

[0032] First, drop-coat 6 μL of graphene-chitosan solution on the surface of the polished and cleaned glassy carbon electrode, and after drying, drop-coat 10 μL of N(ε) carboxymethyllysine (CML) solution on the surface, and then dry it. That is, the electrochemical immunosensor is obtained.

Embodiment 2

[0033] Embodiment 2 is to the detection of N (ε) carboxymethyllysine (CML)

[0034] Prepare a series of ethanol solutions (volume ratio of ethanol to water: 2:3) of CML containing a series of different concentrations (including blank) as standard solutions.

[0035] Prepare electrochemical immunosensor with the method for embodiment 1, it is placed in the acetonitrile solution of 5mmol / L ferrocene and carries out differential pulse voltammetry (DPV) scanning, record response current, denoted as I 0 . Then immerse it in a phosphate buffer solution with a total volume of 50 μL containing 8 μL of pentosidine monoclonal antibody and a series of CML standard solutions of different concentrations, place it in an incubator at 37°C for 40 min, and rinse it with phosphate buffer solution After placing in the acetonitrile solution of 5mmol / L ferrocene, carry out differential pulse voltammetry (DPV) scanning, DPV curve figure is as follows figure 1 shown. The concentration of the curv...

Embodiment 3

[0036] Embodiment 3 is to the detection of N (ε) carboxyethyl lysine (CEL)

[0037] Prepare a series of ethanol solutions (volume ratio of ethanol to water: 2:3) of CEL containing a series of different concentrations (including blank) as standard solutions.

[0038] Prepare electrochemical immunosensor with the method for embodiment 1, it is placed in the acetonitrile solution of 5mmol / L ferrocene and carries out differential pulse voltammetry (DPV) scanning, record response current, denoted as I 0 . Then immerse it in a phosphate buffer solution with a total volume of 50 μL containing 8 μL of pentosidine monoclonal antibody and a series of CEL standard solutions of different concentrations, place it in an incubator at 37°C for 40 min, and rinse it with phosphate buffer solution After placing in the acetonitrile solution of 5mmol / L ferrocene, carry out differential pulse voltammetry (DPV) scanning, DPV curve figure is as follows image 3shown. The concentration of the curve...

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Abstract

The invention discloses a method for detecting AGEs (Advanced Glycation End Products) and application. The method comprises the following steps: firstly preparing a glassy carbon electrode modified bygraphene-chitosan / N(epsilon)carboxymethyl lysine, and then realizing multi-residue detection on five AGEs comprising the N(epsilon)carboxymethyl lysine, N(epsilon)carboxyethyl lysine, pentosidine, pyruvaldehyde imidazolinone and argpyrimidine by combining an indirect competition method on the basis of a principle that cross immunoreaction can be generated by a pentosidine monoclonal antibody andmultiple AGEs. According to the method for detecting the AGEs, disclosed by the invention, the problem that a current AGEs detecting method just can be used for detecting a single substance is solved,higher sensitivity (of which the detecting limit is 0.06 to 0.1 ng / mL) and wider linear range (0.1 to 1000 ng / mL) are obtained, the operation is simple, the cost is lower, the demands on clinical AGEs detection of a human body can be completely fitted, and important practical application value is obtained.

Description

technical field [0001] The invention belongs to the technical field of food safety detection and analytical chemistry, and in particular relates to a method and application for detecting advanced glycation end products. Background technique [0002] Advanced glycation end products (AGEs) are the final products of a series of irreversible and non-enzymatic reactions between carbohydrates and proteins, lipids, amino acids, etc. in the human body. AGEs are closely related to the development of a variety of chronic diseases that are widely concerned by the society, such as diabetes and its complications, uremia, Alzheimer's disease, cataract, primary osteoarthritis, atherosclerotic lesions, etc. Therefore, the clinical detection of AGEs is of great significance and value for the early diagnosis and prevention of the above diseases. [0003] The types of AGEs are extremely diverse, and only a few of them are currently known, such as N(ε)carboxymethyllysine (CML), N(ε)carboxyethy...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577
CPCG01N33/577
Inventor 赵波王雅娟邵科峰赵虎
Owner NANJING NORMAL UNIVERSITY
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