共查询到20条相似文献,搜索用时 93 毫秒
1.
2.
高效液相色谱法分离蛋白的大孔硅质固定相的制备及分离机理 总被引:5,自引:4,他引:5
介绍了不同孔径的大孔硅胶基质的制备,二醇基和氨基固定相的合成,及其蛋白的分离特性。探讨流动相中的盐浓度和PH对蛋白保留时间的影响,以及在此两种固定相上蛋白质分离机理的比较。由于大孔填料具有小的比表面积、性容量也低,从而有利于作蛋白质等生物大分子的高效快速分离分析。 相似文献
3.
介绍了不同孔径的大孔硅胶基质的制备、二醇固定相的合成及其对蛋白质的色谱分离;考察了焙烧温度对硅胶孔隙度的影响;探讨了流动相中盐浓度和pH对蛋白质保留时间、柱效、分离度的影响以及硅胶孔径对键合密度和分离性能的影响。该种大孔固定相相对于常规多孔固定相具有小的比表面积、柱容量低、从而有利于生物大分子的微量分离分析。 相似文献
4.
5.
将粒径为3.0μm无孔单分散亲水性交联聚甲基丙烯酸环氧丙酯树脂(PGMA/EDMA)的表面经不同的化学方法改性,制备了两种不同配基的弱阴离子交换(WAX-Ⅰ和WAX-Ⅱ)色谱填料。在同等色谱条件下,比较了两种填料对蛋白质的分离性能,发现WAX-Ⅰ型填料分离性能优于WAX-Ⅱ。详细考察了WAX-Ⅰ型弱阴离子交换色谱填料流动相pH值、流速及有机溶剂等对蛋白质保留的影响。实验结果表明,当流动相流速为3mL/min时,4种标准蛋白可在2min内基线快速分离,蛋白质的保留符合阴离子交换色谱规律。该填料可以用于生物工程产品的快速分离和纯化。 相似文献
6.
快速弱阴离子交换无孔聚合物固定相的制及用于蛋白质的高效液相色谱分离 总被引:1,自引:0,他引:1
将粒径为3.0 μm无孔单分散亲水性交联聚甲基丙烯酸环氧丙酯树脂(PGMA/EDMA)的表面经不同的化学方法改性,制备了两种不同配基的弱阴离子交换(WAX-Ⅰ和WAX-Ⅱ)色谱填料.在同等色谱条件下,比较了两种填料对蛋白质的分离性能,发现WAX-Ⅰ型填料分离性能优于WAX-Ⅱ.详细考察了WAX-Ⅰ型弱阴离子交换色谱填料流动相pH值、流速及有机溶剂等对蛋白质保留的影响.实验结果表明,当流动相流速为3mL/min时,4种标准蛋白可在2 min内基线快速分离,蛋白质的保留符合阴离子交换色谱规律.该填料可以用于生物工程产品的快速分离和纯化. 相似文献
7.
8.
以具有双孔结构的聚甲基丙烯酸环氧丙酯(PGMA)微球为基质,以葡萄糖进行表面亲水改性,制备了强阳离子交换色谱填料,并将其用于复杂生命体系中生物大分子的快速而高效的分离、分析与纯化。葡萄糖亲水改性增进了填料的生物相容性,提高了蛋白质样品的回收率;双孔结构及较高的比表面积赋予填料良好的柱渗透性和样品负载量。以标准蛋白质为样品,考察了该填料对生物样品的分离性能。以100 mm×4.6 mm的色谱柱分离4种蛋白质,在6 min内实现了基线分离;以溶菌酶为样品,填料的吸附容量为39.5 g/L,在蛋白质快速分离纯化分析中显示了良好的应用前景。 相似文献
9.
随着科学技术的发展,人们需要分离分析的样品越来越复杂,尤其是多肽、蛋白质类生物样品的复杂性使得单一模式色谱难以满足分离分析的要求。混合模式色谱因其独特的分离性能,可以在一次分离中获得与多维色谱相当的分离效果,而且可以避免多维色谱系统结构复杂、流动相兼容性差、分析时间长等问题,成为近年来的研究热点之一。混合模式色谱的研究重点是色谱固定相的设计与开发。混合模式色谱固定相包括反相/离子交换混合固定相、反相/亲水混合固定相、亲水/离子交换混合固定相、两性离子交换混合固定相及三相混合固定相。本文综述了近年来混合模式色谱填料的研究及应用进展,并对混合模式色谱及固定相的发展前景进行了展望。 相似文献
10.
二乙烯苯与乙烯基吡咯烷酮共聚硅质填料的制备及对蛋白质的分离 总被引:1,自引:0,他引:1
大孔硅胶基质与二乙氧基甲基乙烯基硅烷反应后,再与二乙烯,乙烯基吡咯烷酮共聚,制得新型反相色谱填料。实验结果表明,该填料对蛋白质的分离性能好,柱效高,速度快,惰性好。为生物大分子的高效液相色谱分离提供了一种新型填料。 相似文献
11.
12.
Native and modified alumina,titania and zirconia in normal and reversed-phase high-performance liquid chromatography 总被引:2,自引:0,他引:2
Summary Chromatographic properties of silica, alumina, titania and zirconia have been investigated in normal phase mode in the separation of test mixtures of basic, neutral and acidic compounds. In contrast to silica the chromatographic behaviour revealed the basic properties of the alumina, titania and zirconia surfaces. Therefore, separation of basic compounds on these packings seems very promising. Lypophilic packings have been synthesized by modification of titania, zirconia and alumina with organosilanes and polymers and tested for the separation of basic compounds and proteins. High hydrolytic stability of the modified packings was observed during separations with strong alkali and acidic eluents. 相似文献
13.
Very fast reversed-phase separations of biomacromolecules are performed using columns made with superficially porous silica microsphere column packings ("Poroshell"). These column packings consist of ultra-pure "biofriendly' silica microspheres composed of solid cores and thin outer shells with uniform pores. The excellent kinetic properties of these new column packings allow stable, high-resolution gradient chromatography of polypeptides, proteins, nucleic acids, DNA fragments, etc. in a fraction of the time required for conventional separations. Contrasted with <2-microm non-porous particles, Poroshell packings can be used optimally with existing equipment and greater sample loading capacities, while retaining kinetic (and separation speed) advantages over conventional totally porous particles. 相似文献
14.
High-performance liquid chromatography on silica using eluents containing cetyltrimethylammonium (CTMA) bromide was investigated, and adsorption isotherms were determined for two silica packings of different pore diameter and surface area. It was found that about one CTMA ion was adsorbed per square nanometer of the silica surface at pH 7.5 and at a concentration of 6 mM CTMA bromide in 50% of methanol.Fourteen different silica packings were compared using a test mixture, and thirteen were found to exhibit the same selectivity towards the test mixture, which included acids, bases and non-ionic compounds, thus providing a chromatographic system that is largely independent of the origin of the column material.The retention mechanisms for the five test compounds are discussed. 相似文献
15.
16.
《液相色谱法及相关技术杂志》2012,35(5):839-851
Abstract The relationship between the textural properties (pore size, pore volume and surface area) of reversed-phase silica gel packings for HPLC and the dynamic loading capacity of large biomolecules was studied by using silica gels manufactured by similar processes. Several silica gels whose unbonded pore diameters range from 100 to 250 A and whose pore volumes range from 1.0 to 1.4 ml/g have been prepared and characterized. The bonded phase is monomeric C18. The textural properties of the bonded silica gels are also presented and related to the properties of the unbonded silica gels. Chromatographic evaluation with typical proteins in an underload-to-overload condition was performed in order to relate the influence of textural properties of silica gel to loading capacity and resolution. The packings with larger pore size and pore volume produced better column performance and higher loading of proteins. 相似文献
17.
分离蛋白质的高效强阳离子交换色谱填料的合成及性能研究 总被引:3,自引:0,他引:3
带端羟基的大孔硅胶与氯磺酸反应制得分离蛋白质的强阳离子交换色谱填料。填料分离度高,蛋白质活性回收率大于76%,可用于蛋白质及生物工程产品的分离。 相似文献
18.
Three chiral stationary phases, obtained by grafting silica gel with (-)-trans-1,2-cyclohexanediamine, were studied for the resolution of α-amino acids by ligand-exchange chromatography. The packings were prepared by bonding the chiral ligand to silica gel via different hydrocarbon spacers. Separation of the optical isomers was accomplished by eluents containing a constant concentration of copper(II) acetate (0.05mM). The elution sequence of amino acids was found to be dependent on the grafting reaction selected to prepare the chiral packings. 相似文献
19.
《液相色谱法及相关技术杂志》2012,35(10):1465-1480
Abstract The retention characteristics of two sets of chemically bonded non-polar silica packings with a high surface concentration of functional groups have been compared in the reversed-phase mode of liquid chromatography: (i) the conventional packings prepared by chemical modification with trimethylsilyl, heptyl, dodecyl and octadecyl groups, and (ii) mixed-phase materials where differences in the amount of organic bonded phase were achieved by bonding octadecyl and trimethylsilyl groups in different proportions. The retention data of two homologous series of solutes show that these two classes of packings are distinctly different; in particular, the capacity factors and selectivities are always higher on the mixed-phase bonded silica packings at the same percentage of carbon. 相似文献
20.
The pore character of packings for liquid chromatography, especially reversed-phase (RPLC) packings, has been studied by means of the nitrogen adsorption method (BET method). Micro-spherical silica gels with 9, 12, 30, 40 and 50 nm average pore diameter have been used as carriers. These silica gels have been modified with monochlorodimethyloctadecylsilane and several polymers (polyoctadecylmethacrylate-methylmethacrylate co-polymer, polyacrylamide gel, polyvinyl alcohol, poly-2-hydroxyethyl-methacrylate). A larger decrease in the specific surface area values was observed in the case of the polymer coating with the polyoctadecylmethacrylate-methylmethacrylate co-polymer compared with the derivatization by silanes or the modification with polymers without C18-groups. A new approach has been suggested to explain some questions concerning the interpretation of the data obtained during the measurements of the pore characteristics of the derivatized packings. An attempt has been made to reveal peculiarities connecting values of the measured surface of RP-packings with the specific surface area values of the initial silica, as well as with the chromatographically accessible surface. 相似文献