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A method for determining the adsorption stability parameters of metal ions on a 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: 2016-04-13
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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  • A method for determining the adsorption stability parameters of metal ions on a fixed metal affinity column
  • A method for determining the adsorption stability parameters of metal ions on a fixed metal affinity column
  • A method for determining the adsorption stability parameters of metal ions on a fixed metal affinity column

Examples

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Effect test

preparation example Construction

[0032] ( ) Preparation of epoxy silica gel: Weigh 2g of dry acidified silica gel, put it in a flask, add 40mL0.1mol / LNaAc-HAc buffer solution (pH4.0), sonicate for 2min, add 2mLγ-GLDP dropwise, and stir at 90°C The reaction was continued for 2 hours, cooled, filtered and washed with water until neutral, then dried and stored.

[0033] ( ) Preparation of aminocarboxylic acid chelating agent: take 30mL1mol / LNa 2 CO 3 , add 1.5g of ammonia carboxyl agent, adjust the pH to 8.5 after dissolving, and add 2g of epoxy silica gel to it. Stir and react continuously at 60-65°C for 12 hours, 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 2g of aminocarboxylic acid chelating agent synthesized above, mix it with 40mL isopropanol as solvent, pour it into the packing column, and automatically fill it under 400kg pressure for 20min (10min isopropanol , 10min ...

Embodiment 1

[0053] Embodiment 1 Accuracy and precision test

[0054] Preparation of L-Glu affinity column:

[0055] ( ) Preparation of epoxy silica gel: Weigh 2g of dry acidified silica gel, put it in a flask, add 40mL0.1mol / LNaAc-HAc buffer solution (pH4.0), sonicate for 2min, add 2mLγ-GLDP dropwise, and stir at 90°C The reaction was continued for 2 hr, cooled, filtered and washed with water until neutral and then dried and stored;

[0056] ( ) Preparation of L-Glu chelating agent: take 30mL1mol / LNa 2 CO 3 , add 1.5g L-Glu, adjust the pH to 8.5 after dissolving, and add 2g epoxy silica gel to it. Stir and react continuously at 60-65°C for 12 hours, 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 2g of Glu-silica gel filler synthesized above, mix it with 40mL isopropanol as solvent, pour it into the packing column, and automatically fill it under 400kg pressure for 20min (10min isopr...

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 5mmol / L NaNO 2 -20mmol / L NaAC-HAC (pH5.0), 5mmol / L NaNO 2 -10mmol / L Na-PB (pH5.0), 5mmol / L NaNO 2 -15mmol / LTris-HCl (pH3.0) buffer solution, passed through the chromatographic column at a flow rate of 0.5mL / min at a wavelength of 356nm until the outflow 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 0.5mL / min, in 20mmol / LNaAC-HAC (pH5.0), 10mmol / LNa-PB (pH5.0), 15mmol / LTris-HCl (pH3.0) 0.3mmol / L, 0.35mmol / L, 0.4mmol / L, 0.45mmol / L, 0.5mmol / LCuSO 4 、NiSO 4 、CoCl 2 Breakthrough curve of solution on L-Glu column. After each breakthrough curve of a metal ion is measured, the me...

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Abstract

The invention discloses a method for evaluating the stability of metal ion bonding on a fixed metal affinity chromatographic column, which includes the preparation of epoxy silica gel, the preparation of aminocarboxylic chelating agents, and the measurement of complex stability constant and saturated adsorption capacity. Connect the aminocarboxylic chromatography column to the chromatography system, and use cutting-edge chromatography to bond different concentrations of metal ions to the aminocarboxylic chelating agent in different buffer systems. When the bonding reaches equilibrium, respective breakthrough curves are obtained. Use Peakfit software calculates their respective breakthrough times; calculates the apparent molar adsorption capacity based on the breakthrough time; substitute the metal ion concentration and apparent molar adsorption capacity into the Langmuir adsorption model to measure the fixed metal ions on the aminocarboxyl chelating column The complexation stability constant and saturation adsorption capacity. The method is fast and simple, can measure multiple adsorption parameters at the same time, and more truly and accurately reflects the adsorption stability of fixed metal ions on the aminocarboxyl 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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Patent Type & Authority Patents(China)
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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