A kind of preparation method and application of agarose-bonded silica gel hybrid monolithic column

A technology for bonding silica gel and agarose, applied in chemical instruments and methods, separation methods, other chemical processes, etc., can solve the problems of difficult preservation, easy to swell, etc., achieve good mechanical strength, improve hydrophilicity, and skeleton structure. stable effect

Inactive Publication Date: 2018-05-01
LUOYANG INST OF SCI & TECH
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  • Claims
  • Application Information

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Problems solved by technology

[0003] The purpose of the present invention is to provide a method for preparing an agarose-bonded silica gel hybrid monolithic column. The method has the advantages of simple operation and convenient surface modification of the prepared monolithic column, and solves the problem of immunophilic agarose gel as a carrier. And the column is not easy to store, prone to swelling and other defects, the prepared monolithic column has the advantages of good mechanical strength, hydrophilicity and good reproducibility, and has high selectivity and high enrichment factor for the enrichment of biomolecules

Method used

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  • A kind of preparation method and application of agarose-bonded silica gel hybrid monolithic column
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  • A kind of preparation method and application of agarose-bonded silica gel hybrid monolithic column

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[0023] A method for preparing an agarose-bonded silica gel hybrid monolithic column. The monolithic column uses a sol-gel method to mix tetramethoxysilane, agarose, polyethylene glycol, and urea in an ice bath to prepare a prepolymerized system, then put the pre-polymerization system into a quartz capillary to prepare an affinity monolithic column, and finally modify the affinity monolithic column in three steps to obtain an agarose-bonded silica gel hybrid monolithic column with amide groups on the surface.

[0024] The preparation method of the agarose-bonded silica gel hybrid monolithic column specifically comprises the following steps:

[0025] Step 1, oxidizing agarose, the oxidation process is: agarose is reacted for 6 hours in a nitric acid solution with a concentration of 3 mol / L, a reaction temperature of 70°C and light;

[0026] Step 2: Add urea and polyethylene glycol to the centrifuge tube, add oxidized agarose at the same time, stir evenly in an ice bath, then dro...

Embodiment 1

[0028] a) Capillary pretreatment

[0029] Cut off a certain length of capillary, rinse with water for 40 minutes, 1M NaOH for 80 minutes (forward flushing for 40 minutes and backflushing for 40 minutes to ensure that NaOH fills the entire capillary), plug both ends, and age at 40°C for 12 hours; then rinse with water for 40 minutes, 1M HCL for 80 minutes, and water Until the solution in the capillary becomes neutral; finally blow with nitrogen at 160°C for 6h.

[0030] 2) Oxidation treatment of agarose

[0031] Accurately weigh 2.00g of agarose and 13ml of 3mol / L nitric acid, stir and react for 7h, and then oxidize the agarose. The infrared spectrum of the oxidized agarose is as follows: figure 1 shown.

[0032] 3) Preparation of agarose-bonded organic-silica hybrid monolithic column

[0033] Accurately weigh 0.1706g of polyethylene glycol (PEG), 0.2504g of urea and mix with 3ml of nitric acid oxidized agarose solution, stir evenly in an ice bath, add about 1ml of 2 drops o...

Embodiment 2

[0039] Application of agarose-bonded silica hybrid monolithic column

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Abstract

The invention discloses a preparation method of an agarosebonded silica gelhybridized monolithic column. The preparation method comprises the steps that tetramethyl orthosilicate, agarose, polyethylene glycol and urea are mixed under an ice-bath condition to prepare a prepolymerization system by adopting a sol-gel method, then the prepolymerization system is put in a quartz capillary tube to prepare an affinity monolithic column, and finally the affinity monolithic column is subjected to three-step modification to obtain the agarosebonded silica gelhybridized monolithic column containing amide groups on the surface. The preparation method of the agarosebonded silica gelhybridized monolithic column has the advantages of being simple in operation and making the surface of the prepared monolithic column convenient to modify, the problem that an immunoaffinity column using agarose gel as a carrier is not easily stored and easily produces swelling is solved, and the prepared monolithic column has the advantages of being good in mechanical strength, stable in framework structure, good in permeability, uniform in texture, good in hydrophily and reproducibility and the like and has high selectivity and high concentration rate to biomolecule.

Description

technical field [0001] The invention belongs to the field of preparation of chromatographic columns, and in particular relates to a preparation method and application of an agarose-bonded silica gel hybrid monolithic column. Background technique [0002] The preparation of the monolithic column adopts the method of in-situ polymerization, which avoids the cumbersome steps of particle packing and firing the plug for the packed column, and has a higher column capacity, so it has received extensive attention in recent years. Due to its unique pore structure, the monolithic column has better permeability than conventional packed columns and can achieve rapid separation. According to the different matrix materials, the monolithic column can be divided into silica gel monolithic column, organic polymeric monolithic column, organic-silica gel Hybrid monolithic columns. The advantage of the silica gel monolithic column is good mechanical strength, high temperature resistance and or...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): B01D15/20B01D15/38B01J20/286B01J20/30G01N30/08
CPCB01D15/206B01D15/3804B01J20/286B01J2220/52B01J2220/84G01N30/08
Inventor 张少文彭传云吴春来王前进王万慧付巧鸽邓美萍万珊
Owner LUOYANG INST OF SCI & TECH
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