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Feed-limited chromatographic packing material, preparation method thereof, stationary phase containing same and application thereof

A technology of chromatographic packing and stationary phase, which is applied in the field of restricted chromatographic packing and its preparation, can solve the problems of narrow application range, limited application range and narrow application range of chromatographic packing, and achieve strong retention and separation ability, repeatable High reliability and low cost effect

Active Publication Date: 2021-05-14
纳谱分析技术(苏州)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In recent years, restricted fillers have been reported in the literature (Anal.Bioanal.Chem., 2006, 384: 1462-1469; J.Chromatogr.B, 2004, 801: 141-156), but the methods provided in the literature are not in the synthesis It is necessary to introduce hydrophilic groups on the outer surface of silica gel by enzymatic hydrolysis, resulting in high cost
CN102335597A provides a kind of entry-limited poly(styrene-co-divinylbenzene) coated silica gel chromatographic filler and its preparation method, which coats poly(styrene-co-divinylbenzene) on the surface of silica gel, and then coats The surface of the layer is grafted with polyglycidyl methacrylate, and after hydrolysis to obtain a hydrophilic surface, the obtained chromatographic packing has a narrow application range and can effectively separate hydrophobic compounds in complex biological samples.
CN103626934A discloses a limited entry type poly(methacrylic acid-3-sulfonic acid propyl ester) modified silica gel chromatographic filler, which grafts double-layer polymers on the surface of silica gel, and the inner layer is poly(methacrylic acid-3-sulfonic acid) Propyl ester-co-ethylene glycol dimethacrylate), the outer layer is poly(monoglycerol methacrylate), and its chromatographic packing can be used as a chromatographic stationary phase for biological samples and environmental sample molecules, and is used for the separation of cationic compounds Direct sample extraction, narrow application range
When preparing limited-access medium-type chromatographic packing, physical shielding is often used to exclude biological macromolecules by using the pore structure size in the medium. This type of limited-access chromatographic packing generally requires a pore size of less than 8nm. Using this The way of physical shielding greatly limits its scope of application

Method used

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  • Feed-limited chromatographic packing material, preparation method thereof, stationary phase containing same and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0060] The preparation method of a kind of limited advance type chromatographic filler is as follows:

[0061] (1) Disperse 10 g of silica gel microspheres in 100 mL of phosphate buffer (10 mM, pH=10), and stir for 1 hour to prepare a suspension of silica gel microspheres for use, wherein the silica gel microspheres have a particle size of 5 μm and a pore size of 10nm;

[0062] (2) 32g 4-vinylbenzyl glycidyl ether and 2.5g 80% divinylbenzene are mixed, add to 200g and dissolve 2g 70% Triton X-100 (a kind of nonionic surfactant ), 2.1g of potassium persulfate and 15g of 50% potassium pyrosulfite in aqueous solution, the above-mentioned mixed solution was degassed under nitrogen for 5min, after airtight, it was stirred under a water bath at 40°C for 10 hours to complete the polymerization reaction to obtain a white opaque product, which was obtained by Filter to obtain nano-scale polymer latex particles, then mix and stir the above-mentioned nano-polymer latex particles with 10...

Embodiment 2

[0069] The only difference from Example 1 is that in this example, octadecyltrimethoxysilane is replaced by octadecyltrimethoxysilane in step (4) to obtain silica gel microspheres with octaalkyl groups.

Embodiment 3

[0071] The only difference with Example 1 is that in this example, the functional group in step (4) changes from octadecyl to amino, and the steps are as follows:

[0072] Disperse 3 g of silica gel microspheres with polystyrene microspheres adsorbed on the surface obtained in step (3) into 50 mL of anhydrous toluene, then add 5.2 g of 3-aminopropyltriethoxysilane, and react at 110 ° C for 24 h, Wash with toluene and acetonitrile three times in turn, and then dry to obtain silica gel microspheres with amino groups.

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Abstract

The invention provides a kind of feed-limited chromatographic filler and its preparation method and its stationary phase and its application. The preparation method comprises the following steps: (1) making a negatively charged porous material and a positively charged polymer The microspheres react to obtain a porous material with polymer microspheres adsorbed on the surface; (2) introducing functional groups into the internal pores of the porous material obtained in step (1) to obtain a porous material with functional groups; (3) Under the premise of not destroying the functional groups in the pores of the porous material, remove the polymer microspheres adsorbed on the surface of the porous material; (4) chemically modify the outer surface of the porous material obtained in step (3) to make the outer surface hydrophilic , to obtain the restricted-advance chromatographic filler. The feed-limited chromatographic filler provided by the invention has a high exclusion rate for proteins, and shows strong retention and separation capabilities for small molecular substances.

Description

technical field [0001] The invention belongs to the technical field of chromatographic separation materials, and relates to a limited-advance chromatographic filler, a preparation method thereof, a stationary phase containing the same and an application thereof. Background technique [0002] The complex matrix of biological and environmental samples and the low concentration of analytes determine that they must be properly pretreated before determination. Solid-phase extraction has become the most commonly used method for biological fluid and Sample preparation methods for environmental water samples. When solid-phase extraction is used to separate and enrich small molecule analytes in biological tissue samples, a problem often encountered is that biological macromolecules in the sample, such as proteins and nucleic acids, occur when encountering hydrophobic reversed-phase solid-phase extraction packing materials. The denaturation of biological macromolecules, the denature...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): B01J20/282B01J20/30B01D15/10
CPCB01D15/10B01J20/282
Inventor 刘晓东刘晓艳
Owner 纳谱分析技术(苏州)有限公司
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