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Preparation method and application of agarose bonded silica gel hybridized monolithic column

A technology of bonding silica gel and monolithic column, applied in chemical instruments and methods, separation methods, other chemical processes, etc., can solve the problems of easy swelling, difficult to store, etc., and achieve improved hydrophilicity, good mechanical strength, and skeleton structure. stable effect

Inactive Publication Date: 2017-01-18
LUOYANG INST OF SCI & TECH
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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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  • Preparation method and application of agarose bonded silica gel hybridized monolithic column
  • Preparation method and application of agarose bonded silica gel hybridized monolithic column
  • Preparation method and application of agarose bonded silica gel hybridized monolithic column

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preparation example Construction

[0023] A method for preparing agarose-bonded silica gel hybrid monolithic column. The monolithic column adopts a sol-gel method to mix tetramethoxysilane, agarose, polyethylene glycol, and urea under ice bath conditions to prepare a prepolymerized column. System, then the pre-polymerization system is loaded into the quartz capillary to prepare the affinity monolithic column, and finally the affinity monolithic column is modified in three steps to prepare the agarose-bonded silica gel hybrid monolithic column containing amide groups on the surface.

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

[0025] Step 1. Oxidize the agarose, the oxidation process is: the agarose is reacted for 6h under the concentration of 3mol / L nitric acid solution, the reaction temperature is 70℃ and the light conditions;

[0026] Step 2. Take urea and polyethylene glycol into the centrifuge tube, add the oxidized agarose a...

Embodiment 1

[0028] A) Capillary pretreatment

[0029] Intercept a certain length of capillary, rinse with water for 40min, 1M NaOH for 80min (forward flush 40min, backflush 40min to ensure that the entire capillary is filled with NaOH), then plug both ends, age at 40℃ for 12h; then rinse with water for 40min, 1MHCL for 80min, water rinse Until the solution in the capillary becomes neutral; finally, nitrogen is blown at 160℃ for 6h.

[0030] 2) Oxidation treatment of agarose

[0031] Accurately weigh 2.00g agarose and 13ml 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 polyethylene glycol (PEG), 0.2504g urea and 3ml nitric acid oxidized agarose solution and mix, stir evenly under ice bath conditions, and then drop 2 drops of concentrated ammonia water about 1ml, stir and add 1ml tetra...

Embodiment 2

[0039] Application of agarose bonded silica gel 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 particularly 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 in-situ polymerization, which avoids the tedious steps of particle packing and firing the plunger of the packed column, and has a higher column capacity, so it has received extensive attention in recent years. Due to its unique pore structure, monolithic columns have better permeability than conventional packed columns and can realize rapid separation. According to the different matrix materials, monolithic columns can be divided into silica monoliths, organic polymerization monoliths, and organic-silica gels. Hybrid monolithic column. The advantages of silica gel monolithic columns are good mechanical strength, high temperature resistance and organic solvent resistance; disadvantag...

Claims

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Application Information

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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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