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

A biological matrix and silver ion technology, applied in the field of biochemical detection, can solve problems such as residues and non-specific adsorption 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—"protein replacement method", which can improve the enrichment efficiency of silver ions in biological matrices by "protein replacement method", and is used for the decomposition of silver ions in biological matrix samples. Polymerization and desorption to solve the problem of non-specific adsorption and serious residue of silver ions in the matrix

Method used

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

Examples

Experimental program
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Effect test

Embodiment 1

[0067] Silver ion enrichment method in embodiment 1 urine sample

[0068] The pretreatment method of urine samples (protein precipitation method) includes the following specific steps: take 200 μL of blank urine, add 100 μL of 0.7% nitric acid, then add 100 μL of fetal bovine serum, mix well, then add 1600 μL of 1% nitric acid, and shake for 3 minutes , pipette 100 μL into a high-density polypropylene EP tube, and centrifuge at 18,000 g for 5 min. The present invention enriches silver ions in urine through the "protein replacement method", and utilizes that the polymerization ability of the protein-silver ion chelate is stronger than the non-specific adsorption capacity of silver ions and the tube wall, and at the same time, the above chelate can pass through Changes in pH effect polymerization and depolymerization. By optimizing the nitric acid addition ratio before protein replacement to adjust the pH, the protein ratio in protein replacement, and the pH change after protei...

Embodiment 2I

[0071] Embodiment 2ICP-MS quantitative kit and its application method

[0072] 1. Kit composition: silver standard, silver control, indium internal standard, high-density polypropylene EP tube, nitric acid solution I (0.7%), nitric acid solution II (1%), fetal bovine serum.

[0073] The use of the kit and the concentration of the working solution are as follows:

[0074] Silver standard stock solution (CSS): 1000 μg mL -1 ,

[0075] 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).

[0076] Silver standard urine 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 clinically collected urine from healthy subjects.

[0077] Silver quality control working solution:...

Embodiment 3

[0098] Embodiment 3 kit related index verification test

[0099] 1. Standard curve drawing

[0100] Prepare standard curve urine series concentration samples and quality control samples according to the method 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, and take the serial standard urine sample concentration (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 between the analyte and the internal standard in the series of standard urine samples into the linear regression equation for calculation, and compare the concentration values ​​obtained from the stan...

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Abstract

The invention discloses a method for enriching silver ions in a biological matrix, a quantitative kit and a detection method, and the method for enriching the silver ions comprises the following steps: S1, adding a nitric acid solution I into a biological matrix sample to enable the pH value of the system to be 6.5-8.0; S2, adding a protein solution; and S3, adding a nitric acid solution II to enable the pH value of the system to be 1.0-3.0. 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%; and the protein solution is a fetal calf serum protein solution. According to the silver ion enrichment method provided by the invention, efficient enrichment of the silver ions in the biological matrix sample is realized through a protein replacement method, and the accuracy, the operation precision and the parallelism of ICP-MS quantitative detection of the silver ions can be effectively improved.

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 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 cells to repeat the ba...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/62G01N1/34G01N1/38
CPCG01N27/62G01N1/34G01N1/38
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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