CN1260567C - Surface silanization regenerated cellulose microsphere filling and preparation method and use thereof - Google Patents

Abstract

The invention relates to a surface silanized regenerated cellulose microsphere filler, which is composed of 48-90wt% cellulose, 9-50wt% konjac glucomannan and 0.5-4wt% silane. Mix 4-6% cellulose NaOH/thiourea aqueous solution and 2-4% konjac glucomannan NaOH/thiourea aqueous solution to obtain a mixed solution, and then add Span80 liquid paraffin solution to the above mixed solution , stir, then add 1-10wt% CaCl ₂ aqueous solution and stir, finally add HCl aqueous solution, filter, wash to obtain microspheres, after the obtained microspheres are freeze-dried, use N,N-dimethylacetamide as the medium, add catalyst Pyridine, adding chlorosilane at 20-100°C and stirring for 3-5 hours, filtering and washing to obtain regenerated cellulose microsphere filler microspheres with a particle size of 30-100 μm. The microsphere filler prepared by the invention has small particle size, high alkali resistance, low adsorption to biomacromolecules, and high efficiency of the filled chromatographic column.

Description

Surface silanized regenerated cellulose microsphere filler and its preparation method and use

technical field

The invention relates to a surface silanized regenerated cellulose microsphere filler, a preparation method and application thereof. It belongs to the technical field of polymer chemistry and also belongs to the technical field of chromatography.

Background technique

Cellulose and konjac glucomannan are rich natural polymers and are environmentally friendly materials; moreover, because they contain rich hydroxyl groups, they can be derivatized to prepare many functional materials; therefore, their research and development are increasingly attractive Attention, especially the development of high value-added products has become a research hotspot in the past ten years. On the other hand, separation, fractionation or purification of biomacromolecules or synthetic polymers using preparative gel permeation chromatography columns is of great importance in both industry and research. However, since the commonly used chromatographic column fillers are cross-linked polystyrene, dextran, agarose, polyacrylamide and porous silica gel, their production process is complicated and the production conditions are strict, so they are expensive and have poor biocompatibility, making it difficult to Meet the requirements of biological products. The use of cheap cellulose and konjac glucomannan as raw materials to produce preparative gel permeation chromatography column fillers can not only reduce costs, but also facilitate the development and utilization of abundant renewable resources.

The new developments in the research on cellulose chromatographic column fillers mainly contain the use of cellulose/17.5% NaOH aqueous solution (Japanese Open Patent Gazette, A, flat 2-235944, 1990), 9.2% cellulose viscose (Japanese Open Patent Gazette, A, Zhao 63-92603, 1988), cellulose/calcium thiocyanate solution (J.Chromatogr.A, 720,151, 1996), cellulose cuproammonia solution (Chinese patent, publication number, 98113654.0, 1998) and others Additives are mixed to prepare porous cellulose particle fillers, and viscose is used to prepare microsphere regenerated cellulose ion exchangers (ion exchange and adsorption, 14 (1), 23, 1998), and there are also cellulose and konjac glucomannan in NaOH/ Preparation of porous filler in thiourea solvent system for fractionation of polysaccharides (Chinese Patent, Publication No. 1424139A, 2003). The disadvantages of these methods are that the cellulose solvent system is more expensive or pollutes the environment, or the scope of application is narrow, such as not being able to be used in strong alkaline conditions, or the filler is irregular spherical and the particle size is large so that the filled color The performance of the general column is reduced, or the filler adsorbs the graded polymer, which leads to abnormal chromatographic behavior, etc. In addition, cellulose derivative gel (J.Chromatogr.A, 919, 29, 2001), cellulose derivative microspheres (J.Chromatogr.A, 552, 389, 1991; Journal of Sichuan University (Engineering Science Edition), 32, 48, 2000), cellulose-coated inorganic fillers (J.Chromatogr.A, 904, 17, 2000; J.Chromatogr.Sci., 40, 315, 2002) are also used in gel permeation chromatography column fillers, However, its preparation process is complicated and unstable, which limits its application in separation, fractionation and purification of macromolecular substances.

Contents of the invention

The problem to be solved by the present invention is to provide a surface silanized regenerated cellulose microsphere filler and its preparation method and application. The adsorption of biological macromolecules is low, so that the efficiency of the packed chromatographic column is high.

The technical solution provided by the invention is: a surface silanized regenerated cellulose microsphere filler, which is composed of: 48%-90% cellulose, 9%-50% konjac glucomannan, 0.5%-4 % of silane, the above percentages are mass percentages.

The regenerated cellulose microsphere filler is interwoven with cellulose and konjac glucomannan to form microspheres with a mesh structure, the particle diameter is 30-100 μm, the average pore diameter is 570-1130 nm, and the pore surface of the microspheres is modified by silane.

The present invention also provides a preparation method for the above-mentioned regenerated cellulose microsphere filler. According to the mass ratio of cellulose:konjac glucomannan of 10:1 to 48:50, 4-6% cellulose NaOH/thiourea aqueous solution Mix with 2-4% konjac glucomannan NaOH/thiourea aqueous solution and defoam to obtain a mixed solution. The NaOH/thiourea aqueous solution used is 4-8% NaOH/4-6% thiourea aqueous solution; ～10wt% Span80 liquid paraffin solution is added in the above mixed solution, the amount of Span80 liquid paraffin solution used is 50～200% of the volume of the above mixed solution, stir for 0.5～6 hours, then add 1～10wt% CaCl ₂ aqueous solution and stir 10-120 minutes, finally add 1-5wt% HCl aqueous solution, filter, wash with acetone, absolute ethanol and water successively to obtain microspheres, after freeze-drying the obtained microspheres, use N,N-dimethylacetamide as Medium, add catalyst pyridine, add 100-300wt% microsphere mass chlorosilane at 20-100°C and stir for 3-5 hours, filter, wash with toluene, acetone and absolute ethanol to obtain regenerated fibers with a particle size of 30-100 μm Plain microsphere filler microspheres.

The application of the regenerated cellulose microsphere filler of the present invention in the separation, classification or purification of high molecular substances in water, alkaline water or organic solvent systems.

The pore size of the microsphere filler is controlled by the content of konjac glucomannan, but it does not dissolve out as a pore-forming agent during the solidification and regeneration process, but interweaves with cellulose to form a network structure. Porous fillers with different pore sizes, especially large pore sizes and wide pore size distributions, can be obtained by changing the ingredient ratio of cellulose and konjac glucomannan. Microspheres with different particle sizes can be obtained by changing the amount of emulsifier Span80 liquid paraffin solution. By changing the amount of trimethylchlorosilane, microsphere fillers with different degrees of microsphere surface modification can be obtained. Since the surface and internal pore surface of the cellulose/konjac glucomannan blend microspheres are modified by trimethylchlorosilane, not only the filler maintains the biocompatibility of the cellulose/konjac glucomannan blend microspheres and other properties, and greatly reduce and improve the adsorption performance of the filler to biomacromolecules, and at the same time enhance the alkali resistance of the filler, so the chromatographic column filled with this filler can be used at -40-100°C in water, It is normally used in alkaline water and organic solvent system, especially for separation, fractionation and purification of high molecular weight substances with molecular weight below 50×10 ⁴ . For a 500×20mm preparative chromatographic column, the daily preparation amount is 6-15 grams of high molecular substances.

In the present invention, for the first time, under the new NaOH/thiourea solvent system, cotton linters and konjac glucomannan are used as raw materials to prepare regenerated cellulose microspheres by the emulsion method, and the surface of the microspheres is modified with chlorosilane to obtain low adsorption and Gel permeation chromatography column packing with strong alkali resistance. The present invention is substantially different and represents a significant improvement over the known art. The process of the present invention is simple, rapid, easy to operate, low in cost and wide in application range, and the prepared microsphere filler has small particle size (30-100 μm), high alkali resistance, and low adsorption to biological macromolecules. The column has high performance and is suitable for preparative gel permeation chromatography columns produced in large quantities. It is easy for industrial production and preparation.

Description of drawings

Fig. 1 is the electron micrograph of the surface silanized regenerated cellulose microsphere filler that the embodiment of the present invention 1 makes;

Fig. 2 is an electron micrograph of the surface silanized regenerated cellulose microsphere filler prepared in Example 2 of the present invention.

Detailed ways

The technical scheme of the present invention is further described below in conjunction with specific embodiment:

Example 1

Mix 3% cellulose and 2% konjac glucomannan NaOH/thiourea aqueous solution according to the ratio of cellulose: konjac glucomannan is 6:4 and defoam, take 40 grams and put it into a 250mL three-port bottle, add 40mL of 2% Span80 liquid paraffin solution into the system, stir at 200 rpm for 2 hours, then add 50mL of 5% _CaCl2 aqueous solution and stir for 1 hour, then add 100mL of 2% aqueous HCl and stirred for half an hour. Stirring was stopped, filtered, and the obtained solid was washed with acetone, absolute ethanol and distilled water, and the product was lyophilized. Take 4 grams of the dried above product and put it into a 100 mL three-necked flask, add 30 mL of N,N dimethylacetamide and 10 mL of pyridine, blow nitrogen at 20°C and stir at a speed of 100 rpm for 1 hour, then add 4 grams of Chlorotrimethylsilane, stirred for 2 hours. The obtained product was filtered and washed with toluene, acetone and absolute ethanol to obtain the desired filler, with a particle size of 50 to 80 μm and a silane content of 2.1%. The scanning electron microscope picture is shown in Figure 1.

Example 2

4% cellulose and 2% konjac glucomannan NaOH/thiourea aqueous solution are mixed by cellulose: konjac glucomannan is a ratio of 7:3 and defoamed, and 40 grams are put into 250mL three mouthfuls bottle, add 50 mL of 4% Span80 liquid paraffin solution into the system, stir at 200 rpm for 1 hour, then add 80 mL of 5% CaCl ₂ aqueous solution and stir for 1 hour, then add 80 mL of 5% aqueous HCl and stirred for half an hour. Stirring was stopped, filtered, and the resulting solid was washed with acetone, absolute ethanol and distilled water, and the product was freeze-dried. Take 4 grams of dried product and put it into a 100mL three-necked flask, add 30mL N, N dimethylacetamide and 10mL pyridine, blow nitrogen at 40C and stir at a speed of 100 rpm for 1 hour, then add 8 grams of triethyl Chlorosilane, stirred for 2 hours. The obtained product is filtered and washed with toluene, acetone and absolute ethanol, which is the desired filler, with a particle size of 30 to 50 μm and a silane content of 2.8%. The scanning electron micrograph is shown in Figure 2.

Example 3: The surface silanized regenerated cellulose microsphere filler prepared in Example 1 was packed into a glass tube of 550×20 mm to form a gel bed of 500×20 mm, thereby preparing a preparative gel permeation chromatography column. The theoretical pedal number of this preparative gel permeation chromatography column is 2730. The column is used for fractionating tetrahydrofuran solution of polycaprolactone with a weight average molecular weight _Mw of 8.31×10 ⁴ and a dispersion index d of 1.55. The mobile phase is tetrahydrofuran and the flow rate is 1.2 mL min-1. ^The polymer was ^divided into 6 ^fractions to ^obtain fractional products with different molecular weights ^{. 4} , the dispersion index is 1.2. The weight of each fraction accounts for 10-15% of the injection volume, and the total yield reaches 96.2%. It is indicated that the surface silanized regenerated cellulose microsphere filler can be used for fractionating polymers in organic solvent systems.

Example 4: The surface silanized regenerated cellulose microsphere filler prepared in Example 2 was packed into a glass tube of 550×20 mm to form a gel bed of 500×20 mm, thereby preparing a preparative gel permeation chromatography column. The column is used to fractionate NaOH aqueous solution of dextran with a weight average molecular weight _Mw of 12.1×10 ⁴ and a dispersion index d of 1.70. The mobile phase is 0.05M NaOH aqueous solution and the flow rate is 1.2mL min-1. ^{The polymer} was ^divided into 6 ^fractions to obtain fractional products with different molecular weights ^{. 4} , the dispersion index is 1.3, and the total yield is 95.1%. It shows that the surface silanized regenerated cellulose microsphere filler has high alkali resistance, low adsorption to biomacromolecules, and high performance of the filled chromatographic column.

Claims (3)

1. surface silicon alkanisation regenerated cellulose microballoon filler, it consists of: 48%～90% cellulose, 9%～50% konjaku glucomannan, 0.5%～4% silane, above number percent are mass percent; Being interweaved with forming by cellulose and konjaku glucomannan has mesh-structured microballoon, and its particle diameter is that 30～100 μ m, average pore size are 570～1130nm, hydride modified microballoon hole surface.

2. the preparation method of the described regenerated cellulose microballoon of claim 1 filler, it is characterized in that: by cellulose: konjaku glucomannan is 10: 1 to 48: 50 a mass ratio, cellulose NaOH/ thiourea solution with 4～6% and 2～4% konjaku glucomannan NaOH/ thiourea solution mix and deaeration obtains mixed solution, and used NaOH/ thiourea solution is 4～8%NaOH/4～6% thiourea solution; The Span80 whiteruss solution that will contain 1～10wt% then joins in the above-mentioned mixed solution, and the amount of used Span80 whiteruss solution is 50～200% of an above-mentioned mixed liquor volume, stirs 0.5～6 hour, adds the CaCl of 1～10wt% again ₂Aqueous solution stirred 10～120 minutes, the HCl aqueous solution that adds 1～5wt% at last, filter, obtain microballoon with acetone, absolute ethyl alcohol and water washing successively, thus obtained microsphere is after freeze drying, with N, the N-dimethyl acetamide is a medium, adds the catalyzer pyridine, adds the chlorosilane of 100～300wt% microspheres quality and stirs 3～5 hours at 20-100 ℃, filter, obtain the regenerated cellulose microballoon filler microballoon that particle diameter is 30～100 μ m with toluene, acetone and absolute ethanol washing.

3. the application in the described regenerated cellulose microballoon of claim 1 filler separation in water, buck or organic solvent system, classification or the purified high-molecular material.