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A kind of preparation method of metal chelate magnetic beads connected with tricarboxymethylethylenediamine

A tricarboxymethylethylenediamine, metal chelating technology, applied in the directions of alkali metal compounds, chemical instruments and methods, alkali metal oxides/hydroxides, etc., can solve problems such as inability to remove metal ions, and achieve easy The effect of magnification, simple preparation process and convenient use

Inactive Publication Date: 2018-07-13
LUOYANG INST OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Chinese patent application "a metal chelated nano-magnetic bead, its preparation method and its application" (application number CN200610002030.1) discloses a nano-magnetic bead connected with IDA. After the magnetic bead is chelated with metal ions, the metal ions (such as six-coordinated metals Ni, Cu, Co, and Ca, etc.) there are three empty orbitals that can bind to proteins with metal binding sites, so nano-magnetic beads made of IDA as a chelating group have a strong ability to adsorb proteins , can be used well for the separation and identification of metal-binding proteins in solution, but cannot effectively remove metal ions from macromolecules

Method used

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Examples

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preparation example Construction

[0017] A method for preparing metal chelate magnetic beads connected with tricarboxymethylethylenediamine, comprising the following steps:

[0018] Step 1. Activation of hydroxyl magnetic beads: adding lye to the hydroxy magnetic beads for activation to obtain a mixture I containing activated hydroxy magnetic beads and lye for later use; the amount of lye added is such that the pH of the mixture I reach 12-14;

[0019] Step 2, epoxidation: According to the volume ratio of mixture 1, dimethyl sulfoxide, epichlorohydrin and sodium hydroxide solution, the ratio of 1:2:1:4 is added to the mixture 1 obtained in the above step 1. sulfoxide, epichlorohydrin and sodium hydroxide with a concentration of 1 mol / L, stirred at 60°C for 12-14 hours, removed the solution, and washed the obtained magnetic beads until neutral to obtain epoxy-activated magnetic beads. spare;

[0020] Step 3: Add 3ml of sodium bicarbonate solution and 2ml of ethylenediamine to every 10mg of the epoxy-activated...

Embodiment 1

[0029] Add 6 mol / L sodium hydroxide solution to 1ml of 10mg / ml hydroxyl magnetic beads and fully activate it to obtain a mixture I containing activated hydroxyl magnetic beads and lye; the amount of 6 mol / L sodium hydroxide solution is enough to make the mixture The pH value of I reaches 12; add 2ml dimethyl sulfoxide, 1 ml epichlorohydrin and 4ml 1 mol / L sodium hydroxide solution to the mixture plus I (1 ml epichlorohydrin and 4ml sodium hydroxide solution slowly Under stirring, add in 5 times within 14 hours, with an interval of 2.8 hours each time), stir slowly at 60°C overnight and then overnight, then wash with water, wash with dilute acetic acid, and wash with water to obtain epoxy-activated magnetic beads. Then add 3ml of 0.2mol / L sodium bicarbonate solution and 2ml of ethylenediamine to the epoxy-activated magnetic beads, shake slowly at 50°C for 20 hours, remove the solution, and wash the obtained magnetic beads with water, dilute acetic acid and water successively, an...

Embodiment 2

[0031]Add 5 mol / L sodium hydroxide solution to 1ml 10mg / ml hydroxyl magnetic beads and fully activate to obtain mixture I containing activated hydroxyl magnetic beads and lye; the amount of 5mol / L sodium hydroxide solution is enough to make mixture I The pH value reaches 13; add 2ml dimethyl sulfoxide, 1 ml epichlorohydrin and 4ml 1 mol / L sodium hydroxide solution to the mixture plus I (1 ml epichlorohydrin and 4ml sodium hydroxide are slowly stirred , added in 5 times within 13 hours, with an interval of 2.6 hours each time), stirred slowly at 60°C overnight and then removed the solution. The obtained magnetic beads were first washed with water, then washed with dilute acetic acid, and finally washed with water until neutral, and the obtained epoxy Activate the magnetic beads. Then add 3ml 0.1mol / L sodium bicarbonate solution and 2ml ethylenediamine to the epoxy-activated magnetic beads, shake slowly at 50°C for 20 hours, then wash with water, dilute acetic acid, and water, a...

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Abstract

The invention relates to a preparation method of metal-chelated magnetic beads connecting tricarboxymethyl ethylenediamine. The preparation method comprises the steps: adding an alkali liquid to hydroxyl magnetic beads, and activating to obtain a mixture I containing the activated hydroxyl magnetic beads and the alkali liquid; adding dimethyl sulfoxide, epoxy chloropropane and sodium hydroxide with the concentration of 1 mol / L to the obtained mixture I, stirring for 12-14 hours at the temperature of 60 DEG C, and thus obtaining epoxy activated magnetic beads; adding a sodium bicarbonate solution and ethanediamine to the obtained epoxy activated magnetic beads, and carrying out slow oscillation for 20 hours at the temperature of 50 DEG C, to obtain processed epoxy activated magnetic beads; and adding a sodium hydroxide solution and a sodium bicarbonate solution to the obtained processed epoxy activated magnetic beads, mixing evenly, then adding bromoacetic acid, and stirring at room temperature, to prepare the metal-chelated magnetic beads connecting tricarboxymethyl ethylenediamine. The method is simple, and the prepared metal-chelated magnetic beads have strong metal binding ability and large combination capacity, and can be used for removing metal ions in solutions and biological macromolecules.

Description

technical field [0001] The invention relates to a metal chelating magnetic bead, in particular to a method for preparing a metal chelating magnetic bead connected with tricarboxymethylethylenediamine. Background technique [0002] In solution, metal ions are often combined with biomacromolecules such as proteins, nucleic acids, sugars, etc., affecting our analysis of the structure and function of these biomacromolecules. Existing methods for demetallization are by adding high concentrations of low molecular weight chelating agents, such as EDTA and EGTA, or by metal chelating agarose gel. The former removes the metal complexes of the chelating agent and at the same time the metal ions in the buffer will recombine with the macromolecules, affecting the demetallization effect. Currently, agarose gels are widely used to demetallize biomacromolecules. The particle size of agarose is uniform, and there are reactive active groups with appropriate density on the surface, which is...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): B01J20/22B01J20/28B01J20/30
Inventor 袁东张红晓张羽
Owner LUOYANG INST OF SCI & TECH
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