CN101869828A - Method for preparing temperature stimuli responsive intelligent chromatographic column - Google Patents
Method for preparing temperature stimuli responsive intelligent chromatographic column Download PDFInfo
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- CN101869828A CN101869828A CN 201010209655 CN201010209655A CN101869828A CN 101869828 A CN101869828 A CN 101869828A CN 201010209655 CN201010209655 CN 201010209655 CN 201010209655 A CN201010209655 A CN 201010209655A CN 101869828 A CN101869828 A CN 101869828A
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Abstract
The invention discloses a method for preparing a temperature stimuli responsive intelligent chromatographic column, which comprises the following steps of: A, uniformly mixing N-Isopropyl acrylamide, N,N'-methylene-bis-acrylamide, polyethylene glycol-20,000, 10 percent w/v ammonium persulfate and deionized water by using ultrasonic waves; B, deoxidizing reaction products in the step A, adding 10 percent v/v N,N,N',N'-tetramethyl ethylene diamine, and uniformly mixing fully; and C, pouring reaction products in the step B into a stainless steel chromatographic column tube, performing reaction at the temperature of between -18 and 60 DEG C for 10 to 100 hours, arranging chromatographic column heads at two ends of the column tube, connecting the column tube to a high pressure pump, and washing the chromatographic column body by using ultrapure water as a mobile phase. The method has the advantage of simple process; the prepared monolithic column has good pore structure, high mechanical strength and high column efficiency; during use, the water is taken as the mobile phase, so the waste and pollution caused by using a large quantity of organic solvent can be avoided; and in the separation process, the action force between reagents and the monolithic column can be controlled only by changing temperature, so the separation is simple and feasible, and the separation effect is good.
Description
Technical field
The present invention relates to the parting material preparation field, especially a kind of preparation method of temperature stimuli responsive intelligent chromatographic column.
Background technology
At present, chromatographic column is generally the organic polymer integral post, it is the bar-shaped integral body that is generated by in-situ preparing such as monomer, crosslinking agent, initator, pore-foaming agents, has that preparation is simple, a favorable reproducibility, advantage that porous is superior, can realize to object fast, efficiently separation.But it still exists pore size and pore-size distribution to be difficult to control, internal structure heterogeneity, separates shortcomings such as small-molecule substance post effect is low; Especially, with this type of chromatographic column separate drug, flowing is necessary for organic solvent mutually, and cost is very high and easily environment is polluted.
Stimulation-response type polymer be meant self to external world the slight change of environment make response and produce that corresponding physical arrangement and chemical property change or even a family macromolecule material of sudden change; Wherein, environmental stimuli can be temperature, pH, ionic strength, electric field, light etc.Under the prior art condition, though progress in Intelligent Hydrogel can be avoided the use of organic solvent in theory, have very application prospects, because it exists bad mechanical property, response speed to wait shortcoming slowly, practical application has been subjected to very big restriction again.
At these deficiencies, Ayano[Eri Ayano, Kyoko Nambu, ChikakoSakamoto, Hideko Kanazawa, Akihiko Kikuchi, Teruo Okano.Aqueous chromatography system using pH-and temperature-responsive stationary phase with ion-exchange groups.Journalof Chromatography A, 2006,1119:58-65] and Zhang Rongyue [Rongyue Zhang, Gengliang Yang, Peiyong Xin, Li Qi, Yi Chen.Preparation ofpoly (N-isopropylacrylamide)-grafted polymer monolith forhydrophobic interaction chromatography of proteins.Journal ofChromatography A, 2009,1216 (12): 2404-2411] etc. intelligent gel is bonded on silica gel or the polyalcohol integral pole as the fixing separation that is used for hormone medicine mutually of high performance liquid chromatography, its with water as mobile phase, the temperature of phase water just can be controlled the active force between medicine and the fixing phase as long as change is flowed, thereby realize separating, avoided waste and pollution because of using a large amount of organic solvents to cause.But, still there are a lot of shortcomings in this method: the bonding process complex operation, be bonded on the carrier again after need preparing intelligent gel earlier, and factor such as the wherein preparation of gel, used carrier and bonding conditions all can influence separating power, implement comparatively difficulty, separating effect is difficult to guarantee.
Summary of the invention
The technical problem to be solved in the present invention provides a kind of preparation method of temperature stimuli responsive intelligent chromatographic column, technology is simple, prepared integral body firmly has good pore structure, mechanical strength height, post effect height, be the phase that flows with water during use, can avoid waste and pollution because of using a large amount of organic solvent to cause, separation process is as long as the change temperature just can be controlled the active force between medicine and the integral post, simple good separating effect.
For solving the problems of the technologies described above, the technical solution used in the present invention is:
A kind of preparation method of temperature stimuli responsive intelligent chromatographic column the steps include:
A, with N-N-isopropylacrylamide, N, N '-methylene-bisacrylamide, polyethylene glycol-20000,10% w/v ammonium persulfate, the ultrasonic mixing of deionized water;
B, on go on foot and add 10% v/v N after the product deoxygenation, N, N ', N '-tetramethylethylenediamine, fully mixing;
C, on go on foot in the product impouring stainless steel chromatogram column jecket, in-18~60 ℃, react 10~100h, the chromatographic column column cap is installed at the column jecket two ends, is connected in then on the high pressure pump, washes the chromatogram cylinder with ultra-pure water mutually for flowing, promptly;
Wherein the usage ratio of each component is: N-N-isopropylacrylamide 0.1~0.3g, N, N '-methylene-bisacrylamide 0.1~0.2g, polyethylene glycol-20,000 0.01~0.08g, 10% w/v ammonium persulfate, 1.4~100 μ L, deionized water 1.4mL, 10% v/v N, N, N ', N '-tetramethylethylenediamine 1.4~14 μ L.
Among the present invention, flow rate of mobile phase is 1mL/min among the step C, and chromatogram cylinder back pressure is 18bar.
The usage ratio of each component can be N-N-isopropylacrylamide 0.3g, N, N '-methylene-bisacrylamide 0.2g, polyethylene glycol-20000 0.08g among the present invention, 10% w/v ammonium persulfate, 10 μ L, deionized water 1.4mL, 10% v/v N, N, N ', N '-tetramethylethylenediamine 10 μ L; Reaction temperature among the step C is-18 ℃, and the reaction time is 24h.
The usage ratio of each component can also be N-N-isopropylacrylamide 0.3g, N, N '-methylene-bisacrylamide 0.2g, polyethylene glycol-20000 0.08g among the present invention, 10% w/v ammonium persulfate, 10 μ L, deionized water 1.4mL, 10% v/v N, N, N ', N '-tetramethylethylenediamine 10 μ L; Reaction temperature among the step C is 25 ℃, and the reaction time is 24h.
The usage ratio of each component can also be N-N-isopropylacrylamide 0.3g, N, N '-methyl bisacrylamide 0.2g, polyethylene glycol-20000 0.01g among the present invention, 10% w/v ammonium persulfate, 100 μ L, deionized water 1.4mL, 10% v/v N, N, N ', N '-tetramethylethylenediamine 14 μ L; Reaction temperature among the step C is 60 ℃, and the reaction time is 100h.
The usage ratio of each component can also be N-N-isopropylacrylamide 0.2g among the present invention, N, N '-methylene-bisacrylamide 0.15g, polyethylene glycol-20000 0.045g, 10%w/v ammonium persulfate 50 μ L, deionized water 1.4mL, 10% v/v N, N, N ', N '-tetramethylethylenediamine 7.9 μ L; Reaction temperature among the step C is 39 ℃, and the reaction time is 55h.
The usage ratio of each component can also be N-N-isopropylacrylamide 0.1g among the present invention, N, N '-methylene-bisacrylamide 0.1g, polyethylene glycol-20000 0.01g, 10% w/v ammonium persulfate, 1.4 μ L, deionized water 1.4mL, 10% v/v N, N, N ', N '-tetramethylethylenediamine 1.4 μ L; Reaction temperature among the step C is-18 ℃, and the reaction time is 10h.
Adopt the beneficial effect that technique scheme produced to be: technology of the present invention is simple, easy operating, and favorable reproducibility, mild condition, cost of material is cheap, and contaminative is little, and is environmentally friendly; The prepared chromatographic column of the present invention has micron-sized through hole and three-dimensional skeleton continuously, compares advantage such as have the structure homogeneous, the aperture is little with the chromatographic column of conventional method preparation; Referring to accompanying drawing, the prepared monolithic chromatogram post of the present invention is the phase that flows with water, by to temperature controlling, realized the efficient separation that has different hydrophobic steroid medicines and three kinds of benzene series things to six kinds, the post that wherein separates small-molecule substance is imitated apparently higher than traditional organic polymer integral post, among Fig. 7,8, be followed successively by: 1: hydrocortisone according to peak sequence; 2: cortisone acetate; 3: hydroprednisone acetate; 4: fluocinolone acetonide; 5: betamethasone acetate; 6: beclomeasone propionate among Fig. 9,10, is followed successively by according to peak sequence: 1: acetophenone; 2: propiophenone; 3: phenyl propyl ketone; Chromatographic condition is: flowing is water mutually, and flow velocity is 1mL/min, and the detection wavelength is 254nm.
Description of drawings
The present invention is further detailed explanation below in conjunction with the drawings and specific embodiments.
Fig. 1 is the inner pattern sem photograph of the prepared chromatographic column of embodiment 1 I;
Fig. 2 is the inner pattern sem photograph of the prepared chromatographic column of embodiment 1 II;
Fig. 3 is the inner pattern sem photograph of the prepared chromatographic column of embodiment 2;
Fig. 4 is the inner pattern sem photograph of the prepared chromatographic column of embodiment 3;
Fig. 5 is the inner pattern sem photograph of the prepared chromatographic column of embodiment 4;
Fig. 6 is the inner pattern sem photograph of the prepared chromatographic column of embodiment 5;
Fig. 7 utilizes the prepared chromatographic column of the present invention to separate the chromatogram of steroid medicine under hydrophobic pattern, and wherein column temperature is 25 ℃;
Fig. 8 utilizes the prepared chromatographic column of the present invention to separate the chromatogram of steroid medicine under hydrophobic pattern, and wherein column temperature is 55 ℃;
Fig. 9 utilizes the prepared chromatographic column of the present invention to separate the chromatogram of benzene series thing under hydrophobic pattern, and wherein column temperature is: 25 ℃;
Figure 10 utilizes the prepared chromatographic column of the present invention to separate the chromatogram of benzene series thing under hydrophobic pattern, and wherein column temperature is 55 ℃.
The specific embodiment
The explanation that following examples are detailed the present invention.
A kind of preparation method of temperature stimuli responsive intelligent chromatographic column, its step is:
A, with 0.3g N-N-isopropylacrylamide, 0.20g N, N '-methylene-bisacrylamide, 0.08g polyethylene glycol-20000,10 μ L, 10% w/v ammonium persulfate, the ultrasonic mixing of 1.4mL deionized water;
Wherein, the N-N-isopropylacrylamide is as polymerization single polymerization monomer, N, N '-methylene-bisacrylamide as polymeric cross-linker, polyethylene glycol-20000 as pore-foaming agent, 10% w/v ammonium persulfate as initator;
B, on go on foot and add 10 μ L, 10% v/v N after the product deoxygenation, N, N ', N '-tetramethylethylenediamine, fully mixing;
Wherein, 10% v/v N, N, N ', N '-tetramethylethylenediamine is as catalyst;
C, on go on foot in the product impouring stainless steel chromatogram column jecket, in-18 ℃, react 24h, the chromatographic column column cap is installed at the column jecket two ends, is connected in then on the high pressure pump, washes the chromatogram cylinder with the 200mL ultra-pure water mutually for flowing, promptly; Wherein flow rate of mobile phase is 1mL/min, and chromatogram cylinder back pressure is 18bar.
Fig. 1,2 is the inner pattern sem photograph of the chromatographic column of present embodiment preparation, as can be seen, the prepared chromatographic column of present embodiment has micron-sized through hole and three-dimensional skeleton continuously, compares advantage such as have the structure homogeneous, the aperture is little with the chromatographic column of conventional method preparation.
Referring to accompanying drawing 7,8,9,10, the prepared monolithic chromatogram post of present embodiment is the phase that flows with water, by to temperature controlling, realized the efficient separation that has different hydrophobic steroid medicines and three kinds of benzene series things to six kinds, the post that wherein separates small-molecule substance is imitated apparently higher than traditional organic polymer integral post; Among Fig. 7,8, be followed successively by: 1: hydrocortisone according to peak sequence; 2: cortisone acetate; 3: hydroprednisone acetate; 4: fluocinolone acetonide; 5: betamethasone acetate; 6: beclomeasone propionate; Among Fig. 9,10, be followed successively by: 1: acetophenone according to peak sequence; 2: propiophenone; 3: phenyl propyl ketone; Chromatographic condition is: flowing is water mutually, and flow velocity is 1mL/min, and the detection wavelength is 254nm.
The difference of the second embodiment of the invention and first embodiment is: wherein the consumption of each component is: N-N-isopropylacrylamide 0.3g, N, N '-methylene-bisacrylamide 0.2g, polyethylene glycol-20000 0.08g, 10% w/v ammonium persulfate, 10 μ L, deionized water 1.4mL, 10% v/vN, N, N ', N '-tetramethylethylenediamine 10 μ L; Reaction temperature among the step C is 25 ℃, and the reaction time is 24h.
Fig. 3 is the inner pattern sem photograph of the chromatographic column of present embodiment preparation, and as can be seen, the prepared chromatographic column of present embodiment has micron-sized through hole and three-dimensional skeleton continuously equally.
With water is the phase that flows, and by to temperature controlling, utilizes the prepared monolithic chromatogram post of present embodiment can realize efficient separation to six kinds of steroids medicines and three kinds of benzene series things among the embodiment 1 equally, separation condition also with.
The difference of the third embodiment of the invention and first embodiment is: wherein the consumption of each component is: N-N-isopropylacrylamide 0.3g, N, N '-methylene-bisacrylamide 0.2g, polyethylene glycol-20000 0.01g, 10% w/v ammonium persulfate, 100 μ L, deionized water 1.4mL, 10% v/vN, N, N ', N '-tetramethylethylenediamine 14 μ L; Reaction temperature among the step C is 60 ℃, and the reaction time is 100h.
Fig. 4 is the inner pattern sem photograph of the chromatographic column of present embodiment preparation, and as can be seen, the prepared chromatographic column of present embodiment has micron-sized through hole and three-dimensional skeleton continuously equally.
With water is the phase that flows, and by to temperature controlling, utilizes the prepared monolithic chromatogram post of present embodiment can realize efficient separation to six kinds of steroids medicines and three kinds of benzene series things among the embodiment 1 equally, separation condition also with.
The difference of the fourth embodiment of the invention and first embodiment is: wherein the consumption of each component is: N-N-isopropylacrylamide 0.2g, N, N '-methylene-bisacrylamide 0.15g, polyethylene glycol-20000 0.045g, 10% w/v ammonium persulfate, 50 μ L, deionized water 1.4mL, 10% v/vN, N, N ', N '-tetramethylethylenediamine 7.9 μ L; Reaction temperature among the step C is 39 ℃, and the reaction time is 55h.
Fig. 5 is the inner pattern sem photograph of the chromatographic column of present embodiment preparation, and as can be seen, the prepared chromatographic column of present embodiment has micron-sized through hole and three-dimensional skeleton continuously equally.
With water is the phase that flows, and by to temperature controlling, utilizes the prepared monolithic chromatogram post of present embodiment can realize efficient separation to six kinds of steroids medicines and three kinds of benzene series things among the embodiment 1 equally, separation condition also with.
The difference of the fifth embodiment of the invention and first embodiment is: wherein the consumption of each component is: N-N-isopropylacrylamide 0.1g, N, N '-methylene-bisacrylamide 0.1g, polyethylene glycol-20000 0.01g, 10% w/v ammonium persulfate, 1.4 μ L, deionized water 1.4mL, 10% v/vN, N, N ', N '-tetramethylethylenediamine 1.4 μ L; Reaction temperature among the step C is-18 ℃, and the reaction time is 10h.
Fig. 6 is the inner pattern sem photograph of the chromatographic column of present embodiment preparation, and as can be seen, the prepared chromatographic column of present embodiment has micron-sized through hole and three-dimensional skeleton continuously equally.
With water is the phase that flows, and by to temperature controlling, utilizes the prepared monolithic chromatogram post of present embodiment can realize efficient separation to six kinds of steroids medicines and three kinds of benzene series things among the embodiment 1 equally, separation condition also with.
Foregoing description only proposes as the enforceable technical scheme of the present invention, not as the single restrictive condition to its technical scheme itself.
Claims (7)
1. the preparation method of a temperature stimuli responsive intelligent chromatographic column, its characterization step is:
A, with N-N-isopropylacrylamide, N, N '-methylene-bisacrylamide, polyethylene glycol-20000,10% w/v ammonium persulfate, the ultrasonic mixing of deionized water;
B, on go on foot and add 10% v/v N after the product deoxygenation, N, N ', N '-tetramethylethylenediamine, fully mixing;
C, on go on foot in the product impouring stainless steel chromatogram column jecket, in-18~60 ℃, react 10~100h, the chromatographic column column cap is installed at the column jecket two ends, is connected in then on the high pressure pump, washes the chromatogram cylinder with ultra-pure water mutually for flowing, promptly;
Wherein the usage ratio of each component is: N-N-isopropylacrylamide 0.1~0.3g, N, N '-methylene-bisacrylamide 0.1~0.2g, polyethylene glycol-20,000 0.01~0.08g, 10% w/v ammonium persulfate, 1.4~100 μ L, deionized water 1.4mL, 10% v/v N, N, N ', N '-tetramethylethylenediamine 1.4~14 μ L.
2. preparation method according to claim 1 is characterized in that: flow rate of mobile phase is 1mL/min among the step C, and chromatogram cylinder back pressure is 18bar.
3. preparation method according to claim 1, it is characterized in that: wherein the usage ratio of each component is: N-N-isopropylacrylamide 0.3g, N, N '-methylene-bisacrylamide 0.2g, polyethylene glycol-20000 0.08g, 10% w/v ammonium persulfate, 10 μ L, deionized water 1.4mL, 10% v/v N, N, N ', N '-tetramethylethylenediamine 10 μ L; Reaction temperature among the step C is-18 ℃, and the reaction time is 24h.
4. preparation method according to claim 1, it is characterized in that: wherein the usage ratio of each component is: N-N-isopropylacrylamide 0.3g, N, N '-methylene-bisacrylamide 0.2g, polyethylene glycol-20000 0.08g, 10% w/v ammonium persulfate, 10 μ L, deionized water 1.4mL, 10% v/v N, N, N ', N '-tetramethylethylenediamine 10 μ L; Reaction temperature among the step C is 25 ℃, and the reaction time is 24h.
5. preparation method according to claim 1, it is characterized in that: wherein the usage ratio of each component is: N-N-isopropylacrylamide 0.3g, N, N '-methylene-bisacrylamide 0.2g, polyethylene glycol-20000 0.01g, 10% w/v ammonium persulfate, 100 μ L, deionized water 1.4mL, 10% v/v N, N, N ', N '-tetramethylethylenediamine 14 μ L; Reaction temperature among the step C is 60 ℃, and the reaction time is 100h.
6. preparation method according to claim 1, it is characterized in that: wherein the usage ratio of each component is: N-N-isopropylacrylamide 0.2g, N, N '-methylene-bisacrylamide 0.15g, polyethylene glycol-20000 0.045g, 10% w/v ammonium persulfate, 50 μ L, deionized water 1.4mL, 10% v/v N, N, N ', N '-tetramethylethylenediamine 7.9 μ L; Reaction temperature among the step C is 39 ℃, and the reaction time is 55h.
7. preparation method according to claim 1, it is characterized in that: wherein the usage ratio of each component is: N-N-isopropylacrylamide 0.1g, N, N '-methylene-bisacrylamide 0.1g, polyethylene glycol-20000 0.01g, 10% w/v ammonium persulfate, 1.4 μ L, deionized water 1.4mL, 10% v/v N, N, N ', N '-tetramethylethylenediamine 1.4 μ L; Reaction temperature among the step C is-18 ℃, and the reaction time is 10h.
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Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
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CN103111258A (en) * | 2013-02-22 | 2013-05-22 | 山东大学 | Reaction column filled with river-sand loaded graphene oxide/titanium dioxide (GO/TiO2) filler |
JP2021525880A (en) * | 2018-06-06 | 2021-09-27 | ブリンク アーゲー | Device for fractionating suspension samples |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
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EP1589045A1 (en) * | 2004-04-20 | 2005-10-26 | Rohm And Haas Company | Polymeric adsorbent, and method of preparation and use |
CN101519474A (en) * | 2008-12-11 | 2009-09-02 | 淮阴师范学院 | Double-response water gel with high swelling property and synthetic method |
CN101745968A (en) * | 2008-11-27 | 2010-06-23 | 郑州东方三力耐火材料有限公司 | Gel injection molding and forming technology of specific zircon brick |
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Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
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EP1589045A1 (en) * | 2004-04-20 | 2005-10-26 | Rohm And Haas Company | Polymeric adsorbent, and method of preparation and use |
CN101745968A (en) * | 2008-11-27 | 2010-06-23 | 郑州东方三力耐火材料有限公司 | Gel injection molding and forming technology of specific zircon brick |
CN101519474A (en) * | 2008-12-11 | 2009-09-02 | 淮阴师范学院 | Double-response water gel with high swelling property and synthetic method |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
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CN103111258A (en) * | 2013-02-22 | 2013-05-22 | 山东大学 | Reaction column filled with river-sand loaded graphene oxide/titanium dioxide (GO/TiO2) filler |
JP2021525880A (en) * | 2018-06-06 | 2021-09-27 | ブリンク アーゲー | Device for fractionating suspension samples |
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