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Method for determination of metal ion adsorption stability parameters on fixed-metal affinity column

A technology of metal ions and stability constants, applied in the measurement of color/spectral characteristics, etc., can solve the problems of large error in measurement results, cumbersome detection process, poor reproducibility, etc., and achieve the effect of data stability

Inactive Publication Date: 2013-10-23
NORTHWEST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The commonality of the above methods is that none of them can directly measure K at the same time. ML and Λ 0 These two parameters, and both adopt the off-line detection method, which makes the detection process cumbersome, the error of the measurement result is large, and the reproducibility is poor.

Method used

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  • Method for determination of metal ion adsorption stability parameters on fixed-metal affinity column
  • Method for determination of metal ion adsorption stability parameters on fixed-metal affinity column
  • Method for determination of metal ion adsorption stability parameters on fixed-metal affinity column

Examples

Experimental program
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Effect test

preparation example Construction

[0032] ( ) Preparation of epoxy silica gel: Weigh 2 g of dry acidified silica gel, put it in a flask, add 40 mL of 0.1 mol / L NaAc-HAc buffer solution (pH 4.0), sonicate for 2 min, and add 2 mL of γ-GLDP dropwise , continued the reaction at 90 °C for 2 hr with stirring, cooled, filtered and washed with water until neutral, then dried and stored.

[0033] ( ) Preparation of aminocarboxylic acid chelating agent: take 30 mL 1 mol / L Na 2 CO 3 , add 1.5 g of ammonia carboxyl agent, adjust the pH to 8.5 after dissolving, and add 2 g of epoxy silica gel to it. Stir and react continuously at 60-65°C for 12 h, filter and wash with water, 10% HAc and water until neutral to obtain aminocarboxylic acid chelating agent.

[0034] ( ) Filling of aminocarboxylic acid chelating agent: Take 2 g of aminocarboxylic acid chelating agent synthesized above, mix it with 40 mL isopropanol as solvent, pour it into the packing column, and automatically fill it under 400 kg pressure for 20 min ( ...

Embodiment 1

[0053] Embodiment 1 Accuracy and precision test

[0054] Preparation of L-Glu affinity column:

[0055] ( ) Preparation of epoxy silica gel: Weigh 2 g of dry acidified silica gel, put it in a flask, add 40 mL of 0.1 mol / L NaAc-HAc buffer solution (pH 4.0), sonicate for 2 min, and add 2 mL of γ-GLDP dropwise , continued the reaction at 90 °C for 2 hrs with stirring, cooled, filtered and washed with water until neutral, then dried and stored;

[0056] ( ) Preparation of L-Glu chelating agent: Take 30 mL 1 mol / L Na 2 CO 3 , add 1.5 g L-Glu, adjust the pH to 8.5 after dissolving, and add 2 g epoxy silica gel to it. Stir continuously at 60-65°C for 12 h, filter and wash with water, 10% HAc and water until neutral to obtain L-Glu chelating agent;

[0057] ( ) Filling of L-Glu chelating agent: take 2 g of the synthesized Glu-silica gel filler, mix it with 40 mL of isopropanol as solvent, pour it into the packing column, and automatically fill it under 400 kg pressure for 20...

Embodiment 2

[0065] The method of the present invention is used for the determination of the bonding parameters of different metal ions on the L-glutamic acid (L-Glu) chromatographic column

[0066] Connect the L-Glu chromatographic column to the chromatographic system and use 5 mmol / L NaNO 2 -20 mmol / L NaAC-HAC (pH5.0), 5 mmol / L NaNO 2 -10 mmol / L Na-PB (pH5.0), 5 mmol / L NaNO 2 -15 mmol / L Tris-HCl (pH3.0) buffer solution was passed through the chromatographic column at a flow rate of 0.5 mL / min at a wavelength of 356 nm until the elution curve reached the maximum absorption plateau, and blank breakthrough curves under different buffer systems were obtained. Respectively at 800nm, 510nm, 396nm wavelength at a flow rate of 0.5 mL / min, in 20 mmol / L NaAC-HAC (pH5.0), 10 mmol / L Na-PB (pH5.0), 15 mmol / L Tris- 0.3 mmol / L, 0.35 mmol / L, 0.4 mmol / L, 0.45 mmol / L, 0.5 mmol / L CuSO 4 、NiSO 4 、CoCl 2 Breakthrough curve of solution on L-Glu column. After each breakthrough curve of a metal ion is mea...

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Abstract

The invention discloses an evaluation method for determination of metal ion bonding stability on a fixed-metal affinity chromatography column. The evaluation method comprises preparation of epoxy silica gel, preparation of an amino-carboxylic acid chelating agent and determination of complexation stability constant and saturation adsorption quantity. An amino-carboxylic acid chromatographic column is connected to a chromatogram system, and metal ions of different concentrations are bonded to the amino-carboxylic acid chelating agent through use of frontal chromatography by different buffer solution systems. Breakthrough curves of the metal ions of different concentrations are obtained when bonding balances are achieved, and breakthrough times of the metal ions of different concentrations are calculated through use of Peakfit software. Apparent molar adsorption quantities are calculated based on the breakthrough times. The metal ion concentrations and the apparent molar adsorption quantities are substituted into Langmuir adsorption model, and then the complexation stability constant and the saturation adsorption quantity of fixed metal ions on the amino-carboxylic acid chelating chromatographic column can be determined. The evaluation method is simple and fast, can determine simultaneously multiple adsorption parameters, can more really and accurately reflect the adsorption stability of fixed metal ions on the amino-carboxylic acid chelating agent.

Description

technical field [0001] The invention relates to a method for evaluating the bonding stability of metal ions on a fixed metal affinity chromatographic column, in particular to a method for determining the adsorption stability parameters of fixed metal ions on aminocarboxylic acid chelating agents. Background technique [0002] There are fatal flaws in the actual use of fixed metal affinity chromatographic columns. During the elution of the sample, the metal affinity properties of the column showed instability. The adsorbed sample can be released from the metal affinity column by changing the pH and ionic strength of the solution or adding a competitor, but it is also accompanied by the disadvantage of metal ion leakage. This results in short column life, poor reproducibility and easy contamination of the detection chamber. Especially in the large-scale production of food, medicine and other products, the leakage of metal ions will not only bring environmental pollution, but...

Claims

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Application Information

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IPC IPC(8): G01N21/31
Inventor 李蓉陈斌陈晓丽马晓迅陈国亮
Owner NORTHWEST UNIV
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