A method for quantitatively detecting 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, good specific effect

Inactive Publication Date: 2018-06-08
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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  • A method for quantitatively detecting l-cysteine
  • A method for quantitatively detecting l-cysteine
  • A method for quantitatively detecting l-cysteine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Construction of Glucose / Oxygen Biofuel Cell

[0019] 1. Preparation of Bioanode

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

[0021] 2. Preparation of Biocathode

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

[0023] Insert bioanode and biocathode into 5mL acetic acid-sodium acetate buffer solution (pH 5.5) to assemble biofuel cell, 30mM glucose as fuel, and add 2mM ABTS as electron mediator, use CHI660E electrochemical workstation, adopt linear sweep voltamm...

Embodiment 2

[0025] Utilizing the constructed glucose / oxygen biofuel cell, in Cu 2+ Highly sensitive detection of L-cysteine ​​is achieved in the presence of:

[0026] Add 4μM of 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, take the concentration of L-cysteine ​​as the abscissa, and the variation of the open circuit potential as the ordinate, draw a standard curve, a straight line can be obtained, and the slope can be obtained, which is the slope of the working curve of L-cysteine. The linear relationship between the change of potential and the concentration of L-cysteine ​​(see image 3 ).

[0027] The working curve of L-cysteine ​​is as follows figure 2 As shown, it can be seen from the figure that the R of the working curve 2 The value reaches 0.997, and the linear fit is very good.

[...

Embodiment 3

[0030] Determination of L-cysteine ​​content in a local brand L-cysteine ​​compound tablet:

[0031] 1. Sample treatment: Take an appropriate amount of L-cysteine ​​compound tablets of a certain local brand, grind and dissolve in ultrapure water, filter with a 0.22 μm disposable filter membrane, and use 100mM acetic acid-sodium acetate buffer ( pH 5.5) was diluted to volume.

[0032] 2. The detection process of L-cysteine: Assemble the fuel cell with 5mL acetic acid-sodium acetate buffer solution (pH 5.5) containing 30mM glucose and 2mM ABTS as the base solution, and add 4mM Cu to the base solution 2+ The measured open circuit potential OCP of the system 1 , and then add different volumes of L-cysteine ​​samples to the bottom solution successively, and measure the open circuit potential OCP 2 .

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

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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 measuring L-cysteine ​​by using a glucose / oxygen biofuel cell. Background technique [0002] L-cysteine ​​is an important amino acid found in many proteins, glutathione. It has very important functions in many biological processes, and abnormal levels of it are associated with 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 for bakery products, as an essential ingredient of dough improver. Therefore, the quantitative detection of L-cysteine ​​is of great significance not only for physiological processes and clinical diagnosis, but also for the food processing industry. [0003] At present, the commonly used methods for the quantitative detection of L-cysteine ​​include spectrophotometry, high per...

Claims

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Application Information

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Patent Type & Authority Patents(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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