HSAB theory based sulfhydryl cotton post-modification material as well as preparation and application thereof

A modified material, mercapto cotton technology, applied in the field of analytical chemistry, to achieve good specificity, stable performance, and low back pressure

Inactive Publication Date: 2015-05-06
WUHAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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  • HSAB theory based sulfhydryl cotton post-modification material as well as preparation and application thereof
  • HSAB theory based sulfhydryl cotton post-modification material as well as preparation and application thereof
  • HSAB theory based sulfhydryl cotton post-modification material as well as preparation and application thereof

Examples

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Embodiment 1

[0022] Embodiment 1 mercapto cotton post-modification material SCF-Ni 2+ 、SCF-Cu 2+ , SCF-Pd 2+ , Preparation of SCF-Au

[0023] SCF-Ni 2+ The preparation (schematic diagram see figure 1): Treat mercapto cotton with tris(2-carboxyethyl)phosphine (TCEP), open possible disulfide bonds on mercapto cotton to expose more mercapto groups, and obtain pre-reduced mercapto cotton. In the centrifuge tube, add 484.5mg trishydroxymethylaminomethane (Tris), 234.0mg sodium chloride (NaCl), 950.8mg nickel chloride hexahydrate (NiCl 2 ·6H 2 (0) and 40mL deionized water to obtain a nickel solution (Tris / NaCl / NiCl) with a suitable pH of 7.2 2+ =1:1:1, the amount ratio of substances); then add 400mg pre-reduced mercapto cotton to the mixed solution, fully infiltrate, seal the system, and react in a shaker at 45°C for 10h; after the product is washed with deionized water (full Wash off residual nickel ions), dry in a vacuum oven at 45°C for 4h to obtain SCF-Ni 2+ .

[0024] SCF-Cu 2+ Pr...

Embodiment 2

[0027] Example 2SCF-Ni 2+ Fiber-packed SPE for selective extraction of histidine-tagged green fluorescent protein (His-tagged GFP)

[0028] Accurately weigh the SCF-Ni prepared in 5mg embodiment 1 2+ In a 1mL commercial pipette tip, use stainless steel wire to compact slightly (the length of the material filling is about 6mm), and the tightness of the filling must ensure good permeability and small dead volume ( figure 1 ). First, take 500 μL of the sample solution (20mM Tris, 100mM NaCl, pH 8.0) in a centrifuge tube, and use SCF-Ni 2+ The pipette tip of the material repeatedly pushes and pulls the sample solution 5 times to fully balance the extracted material. Take 500 μL of the sample solution in a centrifuge tube, add 1 μg of histidine-tagged green fluorescent protein (His-tagged GFP) and 1 μg of non-histidine-tagged red fluorescent protein (normal mouse IgG labeled by red-emitting Cy5), and use Balanced SCF-Ni 2+ The mixture was repeatedly extracted 40 times. After ...

Embodiment 3

[0030] Example 3SCF-Ni 2+ Application of fiber-packed SPE to the separation and purification of histidine protein in Escherichia coli cell lysate

[0031] The cultured Escherichia coli cells were collected, resuspended with binding buffer (20 mM Tris, 100 mM NaCl, pH 8.0), placed in an ice bath and subjected to sonication until no obvious cells were found. Centrifuge at 12000×g for 20 min at 4°C, collect the supernatant (lysate sample), and store in a -20°C refrigerator.

[0032] Accurately weigh the SCF-Ni prepared in 5mg embodiment 1 2+ In a 1mL commercial pipette tip, use stainless steel wire to compact slightly (the length of the material filling is about 6mm), and the tightness of the filling must ensure good permeability and small dead volume. First, take 500 μL of the sample solution (20mM Tris, 100mM NaCl, pH 8.0) in a centrifuge tube, and use SCF-Ni 2+ The pipette tip of the material repeatedly pushes and pulls the sample solution 5 times to fully balance the extra...

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Abstract

The invention discloses an HSAB theory based sulfhydryl cotton post-modification material as well as preparation and application thereof, and belongs to the technical field of analytical chemistry. The HSAB theory based sulfhydryl cotton post-modification material is sulfhydryl cotton for modifying Ni<2+>, Cu<2+>, Pd<2+> or Au on the sulfhydryl cotton, wherein the preparation of the sulfhydryl cotton for modifying Ni<2+>, Cu<2+> or Pd<2+> comprises the following steps: dissolving tris(hydroxymethyl) aminomethane, sodium chloride, Ni<2+> salt, Cu<2+> salt or Pd<2+> salt in water; adding TCEP-treated sulfhydryl cotton, and sealing for reaction after full infiltration; cleaning and drying the products with deionized water. The preparation of the sulfhydryl cotton for modifying Au comprises the following steps: adding TCEP-treated sulfhydryl cotton to nano-gold solution, and sealing for reaction after full infiltration; drying the products after cleaning with deionized water. The HSAB theory based sulfhydryl cotton post-modification material has the advantages of simple operation, less solvent consumption and fast speed, and is especially suitable for the analysis of biological samples.

Description

technical field [0001] The invention belongs to the technical field of analytical chemistry, and in particular relates to a post-modification material of mercapto cotton based on the theory of soft and hard acids and bases and its preparation and application. Background technique [0002] Cotton is a natural fiber material with good biocompatibility, permeability, hydrophilicity, mechanical properties and chemical stability that can exist stably in aqueous solution or organic solvent. In recent years, it has been widely used as a solid phase extraction (solid phase extraction, SPE) adsorbent in sample pretreatment. But the single functional group on the surface of cotton limits its application. [0003] Sulfhydryl cotton (SCF) is a fiber material prepared by esterification of cotton, which is usually used for the enrichment of trace metals in water samples. At present, mercapto cotton has been commercialized, and the method for preparing mercapto cotton is simple and cheap...

Claims

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Application Information

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IPC IPC(8): D06M11/00D06M11/83D06M13/282D06M13/368D06M11/13G01N1/34B01D15/08D06M101/06
Inventor 冯钰锜何小梅余琼卫
Owner WUHAN UNIV
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