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Electrochemical analysis method constructed based on bovine serum albumin-ruthenium dioxide nanoparticles and used for detecting intracellular hydrogen peroxide

A technology for bovine serum albumin and electrochemical analysis, applied in the field of electrochemistry, can solve problems such as being easily affected by interferents, difficult to detect with high sensitivity, and time-consuming, and achieve fast detection speed, good reproducibility, and high sensitivity Effect

Pending Publication Date: 2021-08-13
FUJIAN MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] Currently most H 2 o 2 Detection methods, such as chromatography, ultraviolet spectrophotometry, fluorescence, etc., are generally time-consuming, susceptible to interference and difficult to detect in real time and highly sensitive

Method used

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  • Electrochemical analysis method constructed based on bovine serum albumin-ruthenium dioxide nanoparticles and used for detecting intracellular hydrogen peroxide
  • Electrochemical analysis method constructed based on bovine serum albumin-ruthenium dioxide nanoparticles and used for detecting intracellular hydrogen peroxide
  • Electrochemical analysis method constructed based on bovine serum albumin-ruthenium dioxide nanoparticles and used for detecting intracellular hydrogen peroxide

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0034] BSA-RuO 2 The preparation method of NPs was as follows: 1.6 mL of bovine serum albumin with a concentration of 18.75 mg / mL was mixed with 0.4 mL of ruthenium trichloride with a concentration of 75 mmol / L, stirred at 37 °C for 90 minutes to ensure uniform mixing, and then The pH of the above system was adjusted to 11 with sodium hydroxide, and the reaction was fully stirred at 70° C. for 2.5 hours. After the reaction, the solution was black. After the reaction, the solution was put into an ultrafiltration tube with a molecular weight cut-off of 3k, subjected to centrifugal ultrafiltration at 6000 r / min, and washed with water three times. After the volume was fixed to 2 mL, it was bovine serum albumin-ruthenium dioxide nanoparticles (2 mg / mL) required for the experiment, and stored at 4°C. figure 1 for BSA-RuO 2 Preparation process of NPs and detection of intracellular H by electrochemical method 2 o 2 schematic diagram.

example 2

[0036] Free radical scavengers can successfully detect superoxide radicals and hydroxyl radicals generated in vivo or in vitro. 5,5-Dimethyl-1-pyrroline N-oxide is a commonly used hydroxyl radical scavenger, such as figure 2 As shown in the electron spin resonance spectrum, the BSA-RuO prepared in Example 1 2 NPs can catalyze H 2 o 2 Generate hydroxyl radicals to interact with 5, 5-dimethyl-1-pyrroline N-oxide, thus proving that BSA-RuO 2 NPs can efficiently catalyze H 2 o 2 .

example 3

[0038] Utilize cyclic voltammetry to examine the BSA-RuO that example 1 makes 2 NPs / GCE versus H 2 o 2 electrochemical catalysis, such as image 3 As shown, from a to d are respectively unmodified GCE, H 2 o 2 + GCE, BSA-RuO 2 NPs / GCE and H 2 o 2 + BSA-RuO 2 Cyclic voltammetry curves of NPs / GCE at a scan rate of 50 mV / s and pH=7.0 (0.05 mol / L PBS). In the absence of H 2 o 2 case of GCE(a) and BSA-RuO 2 NPs / GCE (c) did not detect obvious redox peaks. Under bare GCE (b) condition for H 2 o 2 The response is negligible, indicating that unmodified BSA-RuO 2 GCE of NPs applied to electrochemical H 2 o 2 Detection performance is poor. As shown in (d), BSA-RuO 2 NPs / GCE versus H 2 o 2 There is an obvious catalytic effect, and an obvious reduction peak appears at about -0.5 V, with a peak current of 50.54 µA.

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Abstract

The invention discloses an electrochemical analysis method constructed based on bovine serum albumin-ruthenium dioxide nanoparticles and used for detecting intracellular hydrogen peroxide. An electron spin resonance spectrum is utilized to prove the catalytic process that BSA-RuO2NPs catalyzes H2O2 to generate hydroxyl radicals, a BSA-RuO2NPs modified glassy carbon electrode has an obvious catalytic effect on H2O2, the reduction peak is about -0.5 V, and the peak current has a linear relationship (R2=0.9965) in a range of 20-200 mV / s along with a scanning speed. When the differential pulse voltammetry is applied to the catalytic process of the BSA-RuO2NPs modified rotating disk electrode on H2O2, the peak current and the H2O2 concentration have a linear relationship (R2=0.9967) at 0.40-3850 [mu] mol / L, and the lowest detection limit is 0.18 [mu] mol / L. The electrochemical analysis method is successfully applied to detection of H2O2 in cells.

Description

technical field [0001] The invention relates to an electrochemical method constructed based on bovine serum albumin-ruthenium dioxide nanoparticles, which is used for detecting the hydrogen peroxide content in cells, and belongs to the field of analytical chemistry and nanotechnology. Background technique [0002] Reactive oxygen species (ROS) include some oxygen free radicals, such as superoxide anion, hydroxyl radicals, etc., as well as some molecules with high activity such as hydrogen peroxide (H 2 o 2 ) and singlet oxygen. Compared with other reactive oxygen species, hydrogen peroxide is relatively stable and easily diffuses between cells and their surrounding environment. As a messenger molecule, hydrogen peroxide is an intermediate product of cellular oxygen metabolism commonly found in aerobic organisms. Current research shows that H in cells 2 o 2 The increase of the content can directly or indirectly induce the malignant transformation of cells, and its concent...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/327
CPCG01N27/3275G01N27/3277G01N27/3278
Inventor 陈伟何少斌邓豪华彭花萍杨柳
Owner FUJIAN MEDICAL UNIV
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