Enzyme-free pyruvic acid sensing detection method based on H2O2

A technology of pyruvate sensing and detection method, which is applied in the field of electrochemical sensing, can solve the problems of easy inactivation, difficult development of pyruvate biosensors, poor stability of natural enzymes, etc., and achieves a wide linear detection range, high efficiency and real-time Detection, long service life effect

Active Publication Date: 2021-03-19
NANJING UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The poor stability and easy inactivation of the natural enzyme pyruvate oxidase is one of the important reasons why the development of pyruvate biosensors is currently limited and difficult to be widely used.

Method used

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  • Enzyme-free pyruvic acid sensing detection method based on H2O2
  • Enzyme-free pyruvic acid sensing detection method based on H2O2

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032]Example 1 Draw a standard curve

[0033](1) 50 mM PBS phosphate tendon (pH 6.5) is prepared as an electrode buffer;

[0034](2) Constructing a three-electrode system, wherein the working electrode is a Pusher blue repair electrode, and the electrolyte solution is 50ml of electrode buffer, and the applied potential is -0.05V, performs timing current scan, activates 30 min, to the current baseline stable, record the corresponding current Value i0;

[0035](3) Add H in electrode buffer2O2, Make it a final concentration of 0.2 mm, and the current value I is recorded after the current value is stable.1;

[0036](4) Apxtone is added to the electrode buffer and the sodium pyruvate of different concentration gradients, so that the final concentration is 0, 1, 2, 4, 6, 8 and 10 mm, record current changes, and accurately recorded Current value I when pyruvate solution 25S, 50S, 100S, and 150S2. Such asfigure 1 As shown, after adding pyruvate sodium, the current value has slowly decreases, that is, ...

Embodiment 2

[0038]Example 2 actual application

[0039]Apyclhancetate is added to acetate in acetate, pneumoniae, and acetate, acetate, respectively, a pyreonate, respectively, an exogenous amount of acetate, and a preparation method provided in accordance with the detection method provided in accordance with the present invention. Perform test, at 25 ml of 0.2mm h2O21 ml of the sample to be tested is added to the electrode buffer, and the current change in real-time recording of the Prussian blue dressing electrode. Sodium pyruvate concentration in sample*= (25.27 × W-0.32) × 26.

[0040](1) Clostridium Acetobutylicum at a 40 ml fermentation system for anaerobic fermentation of anaerobic fermentation, the main product of the fermentation broth was detected by the high efficiency gas chromatograph, which was 12.1 g / L butanol, 6.3 g / L. Taric acid, 2.0 g / l ethanol, and 1.6 g / L acetone, sodium pyruvate having a final concentration of 11.0 g / L, for sample 1.

[0041](2) Klebsiella pneumoniae in 40...

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Abstract

The invention provides an enzyme-free pyruvic acid sensing detection method based on H2O2. The method comprises the following steps: adopting a three-electrode system, wherein an electrolyte solutionof the three-electrode system contains H2O2; adding sodium pyruvate with different concentrations into the electrolyte solution, and acquiring a standard curve of current response and sodium pyruvateconcentration by utilizing a chronoamperometry, wherein the working electrode of the three-electrode system is an H2O2 sensitive electrode; and comparing the standard curve according to the current response value generated by the to-be-detected sample solution containing H2O2 and pyruvic acid to obtain the concentration of sodium pyruvate. According to the method, the H2O2 consumption rate is monitored on line in real time by utilizing a Prussian blue modified electrode based on a carbonic acid removing reaction of pyruvic acid-H2O2, so that the concentration of pyruvic acid is quantitativelyobtained. The pyruvic acid concentration detection strategy adopting the method has a wide linear detection range, can effectively avoid the problems of poor repetitive effect and the like caused by poor stability of pyruvate oxidase, and can realize rapid and efficient real-time detection in a complex biological system, and the detection limit can reach 0.5 mM.

Description

Technical field[0001]The present invention belongs to the field of electrochemical sensing, and specific to acetone-H-H2O2Enzyme pyruvate sensitive electrode detection method of de-carbonate reaction.Background technique[0002]Pyruvic acid is a key intermediate product that is undergoing the biological metabolism. It has a pivotal position in the growth of strain metabolism and even the fermentation stage, its concentration detection has an important reference value of reaction microbial cell growth metabolism and fermentation. In addition, the pyruvate concentration in the blood can be used as one of the diagnostic indicators of vitamin B1 deficiency, and the diagnosis and treatment of diabetic ketoacidosis and the anionic litterity poisoning is also important. Therefore, the rapid real-time monitoring of pyruvate concentration has a very guiding role and economic benefits in the field of biological fermentation production, medical research and food safety.[0003]Currently, the metho...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/327
CPCG01N27/327
Inventor 周杰姜岷杨璐董维亮陈敏皎储震宇金万勤
Owner NANJING UNIV OF TECH
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