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Method for enriching silver ions in protein-containing biological matrix, quantitative kit and detection method

A biological matrix and silver ion technology, applied in the field of biochemical detection, can solve the problem of low enrichment efficiency of silver ions

Active Publication Date: 2020-10-27
CHANGSHA DUXACT BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

For this reason, the present invention proposes a method for enriching silver ions in biological matrices——"stepwise dilution method", which can improve the enrichment efficiency of silver ions in biological matrices through "stepwise dilution method", and is used for silver ion enrichment in biological matrix samples. Ion depolymerization and desorption to solve the problem of low enrichment efficiency of silver ions in the matrix

Method used

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  • Method for enriching silver ions in protein-containing biological matrix, quantitative kit and detection method
  • Method for enriching silver ions in protein-containing biological matrix, quantitative kit and detection method
  • Method for enriching silver ions in protein-containing biological matrix, quantitative kit and detection method

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Experimental program
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Effect test

Embodiment 1

[0062] Silver ion enrichment method in embodiment 1 blood sample

[0063] Blood sample pretreatment method ("step dilution method"), including the following specific steps:

[0064] Take 200 μL blank serum, add 40 μL 0.7% nitric acid (containing 1000ng·mL -1 Internal standard substance indium), after mixing evenly, add 2760 μL of 1% nitric acid, shake for 3 minutes, draw 100 μL into a high-density polypropylene EP tube, and centrifuge at 18000 g for 5 minutes.

[0065] The present invention enriches the silver ions in the serum through the "stepwise dilution method". Firstly, by adding a small amount of dilute nitric acid not exceeding the initial volume of the serum sample to be treated, the silver ions non-specifically adsorbed on the vessel wall are desorbed, and then a large amount of dilute nitric acid is added. Nitric acid (volume ratio of more than 10 times) changes the pH, thereby realizing sufficient depolymerization of the protein-silver ion chelate. Optimize the u...

Embodiment 2I

[0068] Embodiment 2ICP-MS quantitative kit and its application method

[0069] 1. Kit composition: silver standard, silver quality control, indium internal standard, high-density polypropylene EP tube, nitric acid solution I (0.7%), nitric acid solution II (1%).

[0070] The use of the kit and the concentration of the working solution are as follows: silver standard stock solution (CSS): 1000 μg mL -1 ,

[0071] Silver standard curve working solution: 300ng mL -1 (STD1), 600ng·mL -1 (STD2), 1200ng·mL -1 (STD3), 2400ng·mL -1 (STD4), 4800ng·mL -1 (STD5), 7200ng·mL -1 (STD6), 10000ng·mL -1 (STD7), the solvent is 0.7% nitric acid (with internal standard).

[0072] Silver standard serum sample: 0ng·mL -1 (S0), 15ng·mL -1 (S1), 30ng·mL -1 (S2), 60ng·mL -1 (S3), 120ng·mL -1 (S4), 240ng·mL -1 (S5), 360ng·mL -1 (S6), 500ng·mL -1 (S7), the solvent is the serum of healthy subjects collected clinically.

[0073] Silver quality control working solution: 300ng mL -1 (SLLOQ...

Embodiment 3

[0094] Embodiment 3 kit related index verification test

[0095] 1. Standard curve drawing

[0096] Configure standard curve serum concentration samples and quality control samples according to the method of the kit in Example 2. Inject the sample into the mass spectrometer system for analysis, record the response of the analyte and the internal standard, and calculate the response ratio of the analyte and the internal standard. Take the response ratio (Y) of silver and the internal standard as the ordinate, take the concentration of the serial standard serum samples (X) as the abscissa, and use the weight factor (1 / X 2 ), linear regression was performed by the least square method, and the linear regression equation and correlation coefficient were obtained. Substitute the response ratio of the analyte to the internal standard in the series of standard serum samples into the linear regression equation for calculation, and compare the concentration values ​​obtained from the ...

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Abstract

The invention discloses a method for enriching silver ions in a protein-containing biological matrix, a quantitative kit and a detection method. The method for enriching the silver ions comprises thefollowing steps: S1, adding a nitric acid solution I into a biological matrix sample to enable the pH value of the system to be 4.0-6.5; S2, adding a nitric acid solution II to enable the pH value ofthe system to be 1.0-3.0, wherein the concentrations of the nitric acid solution I and the nitric acid solution II are respectively 0.5%-2.0%; the volume of the biological matrix sample is greater than that of the nitric acid solution I; and the volume of the biological matrix sample is smaller than that of the nitric acid solution II. The silver ion quantitative kit comprises the nitric acid solution I, the nitric acid solution II, a protein solution, a silver standard substance, a silver quality control substance and an indium internal standard substance, wherein the concentrations of the nitric acid solution I and the nitric acid solution II are respectively 0.5%-2.0%.

Description

technical field [0001] The invention relates to the technical field of biochemical detection, in particular to a method for enriching silver ions in a protein-containing biological matrix, a quantitative kit and a detection method. Background technique [0002] Silver ions have a bactericidal effect. Drugs with silver ions as therapeutic ingredients mainly include dressings, liniments, gels, etc., which are mostly used for surface sterilization, physical neutralization, and burn treatment. The bactericidal mechanism of silver ions is mainly through the free Ag + Combining with the free sulfhydryl group (-SH) of proteins in microorganisms such as cells, changes the surface charge of proteins, denatures and coagulates proteins, destroys the activity of cell synthetases, and cells die due to loss of division and proliferation. The combination of silver ions and protein is reversible. When the bacterial cells lose their activity, the silver ions are freed from the bacterial cel...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/62G01N1/34G01N1/38G01N33/68
CPCG01N27/62G01N1/34G01N1/38G01N33/6848
Inventor 侯利平戴鹏唐智谢湘周玲欧阳忠华李芳芳谢秀芬李超鹏陈露露
Owner CHANGSHA DUXACT BIOTECH CO LTD
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