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Method for quantitative detection of L-cysteine

A technology for quantitative detection of cysteine, applied in measuring devices, material analysis through electromagnetic means, instruments, etc., can solve the problems of miniaturization and high technical requirements of electrochemical workstations, achieve low cost, improve sensitivity, The effect of high sensitivity

Inactive Publication Date: 2016-06-22
QINGDAO INST OF BIOENERGY & BIOPROCESS TECH CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although this electrochemical sensor uses a relatively cheap instrument, its technically demanding defects are inevitable.
Moreover, it is difficult to miniaturize the electrochemical workstation

Method used

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  • Method for quantitative detection of L-cysteine
  • Method for quantitative detection of L-cysteine
  • Method for quantitative detection of L-cysteine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Construction of glucose / oxygen biofuel cell

[0019] 1. Preparation of bioanode

[0020] Add 4μL of multi-walled carbon nanotubes (2mg / mL), 5μL of vitamin K 3 And 10μLFAD-GDH solution (5mg / mL) were successively dripped on the glassy carbon electrode, and after drying, 5μL of Nafion solution (0.05wt%) was dripped. Dry at room temperature to prepare a bioanode based on FAD-GDH.

[0021] 2. Preparation of biocathode

[0022] 4μL of multi-walled carbon nanotubes (2mg / mL) and 10μL of laccase solution (5mg / mL) were successively drip-coated on the glassy carbon electrode and dried. Then, 2 μL of glutaraldehyde solution (2wt%) was added dropwise and dried at 4°C to prepare a biocathode based on laccase.

[0023] Insert the bioanode and biocathode into 5mL acetic acid-sodium acetate buffer (pH5.5) to assemble the biofuel cell, use 30mM glucose as fuel, and add 2mMABTS as electron mediator, using CHI660E electrochemical workstation, using linear sweep voltammetry Method (LSV), sweep fro...

Embodiment 2

[0025] Utilizing the constructed glucose / oxygen biofuel cell, in Cu 2+ In the presence of, high sensitivity detection of L-cysteine ​​is realized:

[0026] Add 4μM Cu to the battery of the above example 2+ , And then use a multimeter to detect the open circuit potential of the battery. Then add different concentrations of L-cysteine ​​and record the corresponding open circuit potential value. Finally, draw the standard curve with the concentration of L-cysteine ​​as the abscissa and the change of the open-circuit potential as the ordinate. A straight line can be obtained and its slope can be obtained, which is the slope of the working curve of L-cysteine. The linear relationship between the change in potential and the concentration of L-cysteine ​​(see image 3 ).

[0027] The working curve of L-cysteine ​​is as figure 2 As shown in the figure, the R of the working curve 2 The value reaches 0.997, and the linear fit is very good.

[0028] That is, the detection range is 20nM-3μM,...

Embodiment 3

[0030] Determination of L-cysteine ​​content in a local brand of L-cysteine ​​composite tablets:

[0031] 1. Sample processing: Take an appropriate amount of a local brand of L-cysteine ​​composite tablet, grind and dissolve it in ultrapure water, filter it with a 0.22μm disposable filter membrane, and use 100mM acetic acid-sodium acetate buffer ( pH5.5) Carry out dilution and constant volume.

[0032] 2. L-cysteine ​​detection process: use 5mL acetic acid-sodium acetate buffer (pH5.5) containing 30mM glucose and 2mM ABTS as the base solution to assemble the fuel cell, and add 4mMCu to the base solution 2+ Measure the open circuit potential OCP of the system 1 , And then successively add different volumes of L-cysteine ​​samples to the base solution, and measure the open circuit potential OCP respectively 2 .

[0033] 3. Calculation of L-cysteine ​​concentration: take the change of open circuit potential (OCP 2 –OCP 1 ) Calculate the dilution ratio within the range of the standard cu...

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Abstract

The invention relates to a method for detection of a physiological matter, and in particular, relates to a method for quantitative detection of L-cysteine by using a glucose / oxygen biofuel cell. A self powder supply property of the glucose / oxygen biofuel cell is utilized, at the same time, an open circuit electric potential of the cell is inhibited in the presence of Cu<2+>, Cu-S bonds are formed by the L-cysteine and Cu<2+>, and according to the change of the open circuit electric potential along with the concentration of the L-cysteine, the L-cysteine in a to-be-measured sample is quantitatively detected. With utilization of the principle, a linear relationship between the change quantity of the open circuit electric potential and the concentration of the L-cysteine is established, and high-sensitivity detection of the L-cysteine is realized. The method has the advantages of simple operation, high sensitivity and good specificity, and is applicable in detection of the L-cysteine in the fields of food industry, physiological processes and clinical medicine.

Description

Technical field [0001] The invention relates to a method for detecting physiological substances, in particular to a method for quantitatively determining L-cysteine ​​by using a glucose / oxygen biofuel cell. Background technique [0002] L-cysteine ​​is an important amino acid, present in many proteins and glutathione. It has very important functions in many biological processes, and its abnormal content level is related to many important diseases such as liver damage, skin damage, Alzheimer's disease and cardiovascular disease. In addition, L-cysteine ​​is widely used in the food processing industry, mainly used in bakery products, as an essential ingredient of dough improvers. Therefore, the quantitative detection of L-cysteine ​​is of great significance for both physiological processes and clinical diagnosis, as well as for the food processing industry. [0003] At present, the commonly used methods for quantitative detection of L-cysteine ​​include spectrophotometry, high perf...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/413
Inventor 刘爱骅侯传涛
Owner QINGDAO INST OF BIOENERGY & BIOPROCESS TECH CHINESE ACADEMY OF SCI
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