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ECL (electrochemiluminescence) determining method for ATP (adenosine triphosphate)

A technology of adenosine triphosphate and adenosine triphosphate, which is applied in the field of adenosine triphosphate electrochemiluminescence determination, can solve the problems of expensive instruments, high sensitivity, and complicated operations.

Inactive Publication Date: 2018-10-16
FUJIAN MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the detection methods of adenosine triphosphate include high performance liquid chromatography, electrophoresis, mass spectrometry and luciferase method, etc., but the above methods have problems such as complicated operation, expensive equipment, and high sensitivity.

Method used

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  • ECL (electrochemiluminescence) determining method for ATP (adenosine triphosphate)
  • ECL (electrochemiluminescence) determining method for ATP (adenosine triphosphate)
  • ECL (electrochemiluminescence) determining method for ATP (adenosine triphosphate)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Add 0.6 mL of 0.5 mol / L sodium hydroxide and 0.4 mL of 20 mg / mL chloroauric acid solution to 4 mL of 0.08 mol / L N-acetyl-L-cysteine ​​solution, mix After uniformity, they were incubated in a constant temperature water tank at 37 °C for 3 h. After the reaction, the reaction solution was purified by dialysis with a dialysis bag with a molecular cut-off of 3500 to obtain an aqueous solution of N-acetyl-L-cysteine-gold nanoclusters, which was freeze-dried to obtain N-acetyl-L-cysteine Acid-gold nanocluster powder.

Embodiment 2

[0051] Glassy carbon electrodes with a diameter of 3 mm were coated with 1.0 μm, 0.3 μm and 0.05 μm Al 2 o 3 The powder is polished and polished in turn until it reaches a smooth mirror surface, and then put in HNO in turn 3 solution, absolute ethanol, ultrasonic cleaning in deionized water for 3 minutes, N 2 blow dry. Take 5 μL of the N-acetyl-L-cysteine-gold nanocluster solution prepared in Example 1 and drop it on the surface of the treated glassy carbon electrode, and dry it at room temperature to obtain N-acetyl-L-cysteine-gold nanocluster solution Gold nanoclusters modified glassy carbon electrodes. The N-acetyl-L-cysteine-gold nanocluster modified glassy carbon electrode was soaked in 0.1 mol / L sodium borohydride solution for 5 min to obtain the gold nanocluster probe modified electrode. The above-mentioned gold nanocluster probe modified electrode was used as the working electrode, the platinum wire was used as the counter electrode, and the Ag / AgCl electrode was u...

Embodiment 3

[0053] Glassy carbon electrodes with a diameter of 3 mm were coated with 1.0 μm, 0.3 μm and 0.05 μm Al 2 o 3 The powder is polished and polished in turn until it reaches a smooth mirror surface, and then put in HNO in turn 3 solution, absolute ethanol, ultrasonic cleaning in deionized water for 3 minutes, N 2blow dry. Take 5 μL of the N-acetyl-L-cysteine-protected gold nanocluster solution prepared in Example 1 and drop it on the surface of the treated glassy carbon electrode, dry it at room temperature, and further soak the electrode in 0.1 mol / L react in sodium borohydride solution for 5 minutes to obtain a gold nanocluster probe modified glassy carbon electrode. A three-electrode system was adopted, with gold nanocluster probe-modified glassy carbon electrode as the working electrode, platinum wire electrode as the counter electrode, and Ag / AgCl as the reference electrode. 4 -H 2 SO 4 (1:1 V / V) solution, manganese dioxide was electrodeposited, the deposition potential...

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Abstract

The invention discloses an ECL (electrochemiluminescence) determining method for ATP (adenosine triphosphate). According to the ECL determining method for ATP, gold nanoclusters with high quantum yield are taken as an ECL probe, a manganese dioxide nanomaterial is taken as a quenching agent, when ATP exists, an aptamer can be specifically bound with ATP and DNAzyme cannot be formed, glutathione and manganese dioxide on an electrode surface are subjected to a redox reaction, ECL signals of the gold nanocluster probe are restored gradually with increase of ATP concentration, and content of ATP is determined. Logarithm of ATP concentration in the concentration range of 3.6*10<-12>-3.6*10<-3> mol / L has a linear relation with the change value of ECL intensity, and limit of detection is 1.4*10<-12> mol / L. The method is simple to operate, low in cost, high in sensitivity and good in reproducibility and can be used for determining ATP in an actual sample, thereby having better clinical application prospect.

Description

technical field [0001] The invention relates to an electrochemiluminescence assay method for adenosine triphosphate, which belongs to the field of analytical chemistry and nanotechnology. Background technique [0002] Adenosine triphosphate (ATP) is the provider of energy required by various life activities of organisms and an important hub of energy metabolism and transformation in organisms. The main function of adenosine triphosphate is to provide energy for the body, participate in the metabolism of fat, sugar, protein and nucleic acid in the body, and play an irreplaceable role in maintaining the normal function of the organism. Therefore, in clinical testing, ATP is used as a biomarker for many diseases. At present, the detection methods of ATP include high performance liquid chromatography, electrophoresis, mass spectrometry and luciferase method, etc., but the above methods have problems such as complicated operation, expensive equipment, and high sensitivity. Ther...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/76G01N27/327
CPCG01N21/76G01N27/3277G01N27/3278
Inventor 彭花萍黄种南陈伟邓豪华吴伟华
Owner FUJIAN MEDICAL UNIV
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