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Intracellular nano-silver and silver ion quantitative determination method

A determination method and technology for silver ions are applied in the field of quantitative determination of nano-silver and silver ions, which can solve the problems of inability to quantitatively measure nano-silver and inability to distinguish nano-silver.

Active Publication Date: 2020-05-15
HKUST SHENZHEN RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] The purpose of the present invention is to provide a quantitative assay method for intracellular nano-silver and silver ions, aiming to solve the problem of inability to effectively combine the signal of nano-silver and the signal of silver ion in the existing assay method without destroying the cells. Distinguished, unable to quantitatively measure both nano-silver and silver ions in the cell at the same time

Method used

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  • Intracellular nano-silver and silver ion quantitative determination method
  • Intracellular nano-silver and silver ion quantitative determination method
  • Intracellular nano-silver and silver ion quantitative determination method

Examples

Experimental program
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Effect test

Embodiment 1

[0063] cell culture

[0064] The zebrafish larval cells derived from one day old were used as the cells to be tested. Under the condition of 28°C, the DMEM medium supplemented with 10% fetal bovine serum was used as the culture medium to culture the cells to be tested, and subcultured to logarithmic Cells in the growth phase were used for subsequent experiments.

Embodiment 2

[0066] Determination of AIE Fluorescent Probe Concentration

[0067] Using Thiazolium Blue Colorimetric Method (MTT Method) to Investigate the Toxicity of AIE Fluorescent Probe

[0068] The cells to be tested in the logarithmic growth phase were collected, transferred to a 96-well plate at a density of 5000 cells / well, and cultured overnight at 28°C; the old medium was removed, and the cell pellet was collected. In the cell pellets, new culture medium containing different concentrations of AIE fluorescent probes were added, and cultured at 28°C for 6 hours; then 10 μL of MTT solution with a concentration of 5 mg / mL was added to each well, and the Incubate at 37°C for 4 hours; add 100 μL of a mixed solution consisting of 10% sodium lauryl sulfate and 0.01mol / L hydrochloric acid to each well, and incubate at 37°C for 6 hours to determine its Absorbance at 595nm, wherein, the zeroing sample is a mixture of culture medium, MTT, and dimethyl sulfoxide; the control sample is a mixt...

Embodiment 3

[0072] Determination of the concentration of silver ion fluorescent probe TEZ-TPE-1

[0073] Toxicity of Silver Ion Fluorescent Probe TEZ-TPE-1 Using Thiazolium Blue Colorimetric Method (MTT Method)

[0074] The cells to be tested in the logarithmic growth phase were collected, transferred to a 96-well plate at a density of 5000 cells / well, and cultured overnight at 28°C; the old medium was removed, and the cell pellet was collected. In the cell pellet, silver ion fluorescent probes containing different concentrations (0 μmol / L, 5 μmol / L, 10 μmol / L, 20 μmol / L, 30 μmol / L, 50 μmol / L, 70 μmol / L, 100 μmol / L) were added respectively. Incubate the new medium for TEZ-TPE-1 at 28°C for 6 hours; add 10 μL of MTT solution with a concentration of 5 mg / mL to each well, and incubate at 37°C for 4 hours; Add 100 μL of a mixed solution consisting of 10% sodium lauryl sulfate and 0.01 mol / L hydrochloric acid to each well, incubate at 37°C for 6 hours, and measure its absorbance at 595 nm, wh...

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Abstract

The invention provides an intracellular nano-silver and silver ion quantitative determination method which is simple and convenient. The method comprises: establishing a regression equation of the mass of silver ions and the fluorescence intensity of the silver ions, further detecting the fluorescence intensity of the nano-silver and the fluorescence intensity of the silver ions, and calculating the concentration of the silver ions by combining the regression equation; determining the total concentration of the nano-silver and the silver ions, and calculating to obtain the concentration of thenano-silver. The method has the advantages that under the condition that the cells are not damaged, different fluorescent probes are adopted to effectively distinguish signals of nano-silver from signals of silver ions. Meanwhile, by establishing a regression equation of the silver ions, the nano-silver and the silver ions in the cells are quantitatively determined, and the method is high in detection speed, high in detection sensitivity, high in selectivity, free of damage to cell thalli and capable of achieving nondestructive analysis on the cells.

Description

technical field [0001] The invention relates to the field of quantitative determination of nano silver and silver ions, in particular to a quantitative determination method of intracellular nano silver and silver ions. Background technique [0002] With the rapid development of nanotechnology, nano-products are increasing in daily life. Among them, silver nanoparticle (AgNPs) has become one of the most widely used nanomaterials due to its superior antibacterial and bactericidal properties, and is widely used in medical products. Various fields such as textile industry and water purification. However, when nanomaterials are widely used, they are inevitably released into environmental water bodies and ingested by aquatic organisms, thereby causing toxicity to aquatic organisms. [0003] Due to their nanoscale size effects, nanomaterials are likely to be the natural defense system of organisms, entering cells and disrupting cell functions. As the most basic structural and fun...

Claims

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Application Information

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IPC IPC(8): G01N21/64
CPCG01N21/6428G01N21/6486
Inventor 王文雄颜能
Owner HKUST SHENZHEN RES INST
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