填充型纳流液相色谱柱技术的研究进展

纳流液相色谱是高效液相色谱微型化的重要形式,由于其具有溶剂消耗少、样品需求量小、检测灵敏度高以及与质谱联用兼容性好等特点,在生命科学、生物医药以及微纳分析等领域得到越来越广泛的应用。填充柱是最常用的纳流色谱柱类型,通常由球形色谱填料颗粒填充入毛细管内制成。样品组分是在色谱柱上实现分离,色谱柱的性能对色谱分析的选择性和分辨率至关重要,因此制备高质量的纳流色谱填充柱对获得高效的纳流液相色谱分离尤为关键。纳流色谱柱的制备包含两个重要技术环节：柱塞技术和填充工艺。近年来,随着柱塞与填充技术的进步,填充型纳流液相色谱柱的稳定性和分离性能得到了显著提升。本文对近十年来纳流液相色谱柱技术的研究进展,主要是柱塞技术和填充技术的研究和应用进展进行了概述,介绍了其在组学研究和生物制药领域中的应用,并对其发展前景进行了展望。     

串联微柱在超临界流体色谱中的应用

得力于气相和液相色谱的进步,一种新的色谱技术,超临界流体色谱(SFC)异军突起,而各种柱技术又推动了这种技术的发展速度。在SFC中开管柱和填充柱都在使用,且各具特色。但除了单位柱长具有较大的压降、柱子使用长度受到限制外,填充柱显示出更多的优点。 微柱,即填充石英毛细管柱,内径小于1mm。它以弹性石英毛细管作为柱管,管内填充色谱填料。微柱的细内径和固有的高渗透性使其兼有填充柱和     

高效制备液相色谱柱技术的研究进展

本文概述了高效制备液相色谱柱的柱型结构、填料以及柱填充方法等研究的最新进展,讨论了制备柱与分析柱的不同特征,对目前普遍使用的压缩型制备柱的类型、结构及填充方法作了较为全面的评述,总结比较了工业化制备色谱填料不同于分析色谱填料的特点,探讨了高效制备液相色谱柱技术的应用和发展前景.     

硅胶整体柱的研究进展

自液相色谱技术创立以来,色谱填充材料经历了4个发展阶段[1],包括从形状不规则的填充颗粒到球形颗粒填料、从大的颗粒到小的填充颗粒、从不纯的填充材料到高纯的球形硅胶颗粒以及目前整体柱的产生,一直在向着提高柱效和实现快速分离的方向发展。与整体柱相比,传统的液相色谱柱存在种种弊端和限制[2,3],人们为了提高颗粒型填充柱的柱效而使用越来越小的填料,但柱压降随填料粒度的减小却急剧上升。因此,综合柱效和分离速度两方面因素,较为折中的5μm填料被广泛使用。换句话讲,传统的高效液相色谱(HPLC)填充柱对分离对象很难同时实现既高…     

液相色谱动力学过程的研究—液相色谱系统质量平衡模型的建立和矩的推导

]本文在前人建立的填充柱液相色谱质量平衡模型的基础上,考虑到柱外效应（包括进样系统,进样器与填充柱及检测器与填充柱间的连接管和检测池）的影响,建立了整个色谱系统的质量平衡模型。在此基础上,通过Laplace变换的方法,导出了色谱流出曲线的一级矩和二、三级中心矩的表达式,并对结果作了讨论。     

毛细管气相色谱用固定液的进展

一、前言 自从1979年弹性石英毛细管柱问世之后.毛细管气相色谱得到了迅速的发展。以毛细管柱代替填充柱的趋势日益明显.特别是1983年大内径厚液膜毛细管柱的发展和应用使这种趋势更加肯定了。当然目前在国内填充柱气相色谱仍在例行分析中占主导地位,但是随着毛细管色谱技术在国内的进一步发展,填充柱会逐步地被毛细管柱所取代。     

硅胶整体柱芯的制备方法研究

目前,新型色谱填充材料发展很快,但传统的高效液相色谱(HPLC)颗粒型填充柱存在缺陷,难以实现既高效又快速的分离,Minakuchi等制备了连续的硅胶整体柱,该柱同时具有通孔和中孔双孔结构,孔容量大大高于填充柱,因此,该柱具有高速、低压、高效、适于溶剂及流速梯度洗脱等优点,适合高通量分析和快速分离,     

毛细管电色谱柱制备技术的进展

介绍了毛细管电色谱开管柱、填充柱和整体柱的各种制备技术及其优势与不足,特别是对于近期发展的毛细管电色谱整体柱的制备方法及其应用进行了系统综述。引用文献100篇。     

毛细管气相色谱法测定食品中环己基氨基磺酸钠

<正>毛细管气相色谱技术具有分离度高、重现性好和准确度高等优点,已广泛应用于各行各业的气相色谱的检测和分析中,正逐步替代填充柱色谱。但其在检测食品中环己基氨基磺酸钠时出现了标准曲线的相关性、精密度和准确度均较填充柱差的现象[1],虽然也有许多改进的方法[2-5],但无法提高测定方法的精密度和准确度,以致目前国家标准中仍以填充柱气相色谱法作为首选[6]。在用毛细管色谱     

毛细管离子色谱进展

从毛细管离子色谱柱制备和毛细管离子色谱仪器研制两方面评述了毛细管离子色谱目前的发展状况.毛细管离子色谱柱包括开管离子色谱柱,毛细管颗粒填充离子色谱柱以及最近几年发展起来的整体毛细管离子色谱柱.对毛细管离子色谱仪的总结包括微流量泵、小体积进样器、适合毛细管离子色谱系统的小体积抑制器、电导和光学检测器等.     

Preparation and use of packed capillary columns in chromatographic and related techniques

In this paper general considerations related to the various approaches and parameters influencing the preparation of highly efficient and stable capillary columns for use in liquid chromatography and related techniques are presented and the column production process is discussed in some detail. The different packing methods available for delivering a packing material into a capillary column are discussed from a practical viewpoint. Packing with a gas (dry packing), packing with a liquid solvent or a mixture of solvents (slurry packing), packing with supercritical carbon dioxide, electrokinetic packing, and sol-gel packing technologies are introduced and discussed throughout the paper. Practical recommendations for obtaining highly efficient (high plate numbers) and stable capillary packed columns are also addressed and discussed.     

不锈钢宽口径填充毛细管液相色谱柱

 设计了一种零死体积的二通式柱尾结构和一种使匀浆填料均匀进入色谱柱管内的储料池 ,研究了制备内径在 0 5mm～ 1 0mm的不锈钢宽口径填充毛细管液相色谱柱的方法。详述了以不同牌号、规格的反相ODS类固定相制备的不同柱长的色谱柱的性能。通过折合板高 /折合流速关系和不对称因子对柱性能进行了评价 ,结果表明 ,该方法制备的色谱柱柱效达到理论值的 75 %以上、RSD为 6% ,稳定性也很好。将其应用于抗癫痫药物和氯苯类化合物的分析 ,结果令人满意。     

Packing columns for capillary electrochromatography

Considering the current interest in capillary electrochromatography (CEC), performed in packed columns, we present the different methods used to pack capillary columns for use in CEC. General considerations on column packing are given and the column fabrication process is discussed in sufficient detail to allow instruction to those who are not experienced in the field. Five different packing methods are discussed to deliver packing material into the capillary column from a practical view point: slurry pressure packing, packing with supercritical CO2, electrokinetic packing, using centripetal forces, and packing by gravity. Entrapment of particulate material by sintering and sol-gel technology is also mentioned. Although slurry pressure packing procedures are most common, higher separation efficiencies are obtained using other packing approaches. Electrokinetic packing seems to be the simplest technique to deliver the packing material into the capillary columns. Nevertheless, as with the other packing techniques, skill and experience are required to complete all the steps involved in the fabrication of packed columns for CEC.     

Study of the influence of the aspect ratio on efficiency, flow resistance and retention factors of packed capillary columns in pressure- and electrically-driven liquid chromatography

The influence of the aspect ratio, rho (rho = column diameter/particle diameter), on column parameters such as efficiency, retention factors and flow resistance was studied in both high-performance liquid chromatography and capillary electrochromatography with packed capillary columns. In order to compare the true efficiencies of different columns, a procedure to account for external band broadening was applied. High efficiencies (reduced plate height h approximately 2) were obtained with capillary columns with internal diameters of 150-, 100-, and 75-microm, packed with 10-microm particles. In contrast to previous reports in the literature, no significant improvements in efficiency or flow resistance were observed when the aspect ratio of such columns was decreased. Our observations suggest that the wall effect in these types of columns is not significant. When the aspect ratio was decreased by increasing the particle size, a decrease in reduced plate height was observed. However, the results of flow resistance measurements showed that the latter effect should be attributed to differences in packing and particle batch quality rather than to differences in the aspect ratio.     

Ultrahigh-pressure liquid chromatography using a 1-mm id column packed with 1.5-microm porous particles

The evolution of chromatography has led to the reduction in the size of the packing materials used to fabricate HPLC columns. The increase in the backpressure required has led to this technique being referred to as ultrahigh-pressure liquid chromatography (UHPLC) when the column backpressure exceeds 10000 psi (approximately 700 bar). Until recently, columns packed with sub-2-microm materials have generally fitted into two classes; either short (less than 5 cm) columns designed for use on traditional HPLC systems at pressures less than 5000 psi (350 bar), or capillary columns (inner diameters less than 100 microm). By using packing materials with diameters <2 microm to fabricate UHPLC columns, there is an increase in efficiency and a decrease in the analysis time that are directly proportional to the size of the packing material. In order to realize and exploit the increase in efficiency, however, the columns must maintain lengths typically associated with analytical columns (15-25 cm). We have packed 1 mm diameter, 150 mm in length columns with 1.5 microm packing material, and evaluated their performance in UHPLC. The pressure required to achieve optimum linear velocities in plots of plate height versus linear velocity was in the vicinity of 1104 bar (16000 psi). The 1.5 microm particle-packed column was compared with the more traditional 150 mm long analytical columns packed with 3 microm materials. This column showed an efficiency that was approximately twice that observed with the 3 microm packed column and a concomitant reduction in the analysis time, theoretically predicted.     

Short communication performance of octadecylsilylated monolithic silica capillary columns of 530 microm inner diameter in HPLC

Monolithic silica capillary columns were successfully prepared in a fused silica capillary of 530 microm inner diameter and evaluated in HPLC after octadecylsilylation (ODS). Their efficiency and permeability were compared with those of columns pakked with 5-microm and 3-microm ODS-silica particles. The monolithic silica columns having different domain sizes (combined size of through-pore and skeleton) showed 2.5-4.0-times higher permeability (K= 5.2-8.4 x 10(-14) m2) than capillary columns packed with 3-mm particles, while giving similar column efficiency. The monolithic silica capillary columns gave a plate height of about 11-13 microm, or 11 200-13 400 theoretical plates/150 mm column length, in 80% methanol at a linear mobile phase velocity of 1.0 mm/s. The monolithic column having a smaller domain size showed higher column efficiency and higher pressure drop, although the monolithic column with a larger domain size showed better overall column performance, or smaller separation impedance (E value). The larger-diameter (530 microm id) monolithic silica capillary column afforded a good peak shape in gradient elution of proteins at a flow rate of up to 100 microL/min and an injection volume of up to 10 microL.     

一种多通道匀浆装填毛细管色谱柱的新装置及应用

针对目前毛细管色谱柱装柱效率低、不同批次装填的毛细管色谱柱之间性能差异大的问题,我们发展了一种多通道匀浆装填毛细管色谱柱的新装置。该装置以液相色谱泵提供压力、采用磁力搅拌保持匀浆液均匀分散,一次可装填多达6根毛细管色谱柱。以牛血清白蛋白(BSA)的胰蛋白酶酶切肽段混合物为样本,选择峰容量、蛋白覆盖率、3个特定离子的保留时间以及毛细管色谱柱柱压为指标,在毛细管液相色谱-质谱联用系统上对装填的反相毛细管色谱柱的性能进行了评价。分别考察了一次装填的6根毛细管色谱柱、两次装填的12根毛细管色谱柱以及一次装填1根与一次装填6根毛细管色谱柱的性能及稳定性。实验结果表明:同一批次装填的6根毛细管色谱柱的性能相近;不同批次装填的12根毛细管色谱柱的峰容量和覆盖率没有明显的区别,但保留时间和毛细管色谱柱柱压的稳定性较差;一次装填1根和一次装填6根毛细管色谱柱柱性能的稳定性与两次分别装填6根毛细管色谱柱的稳定性相近,即采用本装置可显著提高毛细管色谱柱的装填效率且每次装填毛细管色谱柱的数量不会对柱性能产生影响。     

A study of the efficiency of monolithic silica gel capillary columns for gas chromatography

The efficiency and dynamic characteristics of seven silica-gel-based monolithic capillary columns were analyzed by separating on them a mixture of five light hydrocarbons. For helium carrier gas flowing at an optimum velocity, the height equivalent to a theoretical plate was found to be 0.15–0.20 mm, values comparable to those typical of packed capillary columns. An analysis of the Van Deemter curves for the columns under study demonstrated that the main contribution to the smearing of the chromatographic zone comes from the diffusional processes in the mobile phase while the mass transfer between the mobile and stationary phases plays only a minor role. At the same time, the parameter A in the Van Deemter equation, which characterizes the degree of column packing uniformity, was found to be negative. This result contradicts the classical theory of chromatography and calls for further studies of monolithic capillary columns.     

Use of electrokinetic measurements for characterization of columns used in capillary electrochromatography

The separation mechanism in capillary electrochromatography (CEC) is a hybrid differential migration process, which entails the features of both high-performance liquid chromatography (HPLC) and capillary zone electrophoresis (CZE), i.e., chromatographic retention and electrophoretic migration. The focus of this paper is on the use of electrokinetic data, such as current, electroosmotic flow (EOF) and column efficiency measurements, that are readily available, for an improved understanding of CEC separations. A framework is presented here for the use of this data for evaluation of a variety of performance parameters including, conductivity ratio, interstitial EOF mobility, porosity, and zeta potential. This framework is applied for characterization of two monolithic columns with different chemistry that were manufactured in-house. The above-mentioned performance parameters were calculated for the two columns and it is found that the poly(VBC-EGDMA-SWNT) monolithic column with the GPTMS-PEI coating offers a significantly improved flow distribution in comparison to the poly(VBC-EGDMA) monolithic column. This observation is confirmed by performing separation of peptides on the two columns and height equivalent of a theoretical plate (HETP) measurements on the resulting peaks. It is shown that following our approach leads to an improved understanding of the separations achieved with the columns and to better column design.     

Capillary electrochromatography using columns packed with a supercritical fluid carrier

Summary Columns for capillary electrochromatography may be prepared by packing reversed phase silica-based material using a supercritical fluid carbon dioxide carrier. Procedures for the in-situ manufacture of frits and UV detection windows, and the wetting of columns are described. The columns were employed in two commercial instruments (and a home-built system), and their properties investigated during the separation of standard mixtures of test compounds. The columns are highly efficient and durable, with reduced plate heights of 1.0–1.4. The repeatability of retention time, peak area and peak height was measured. The influence of applied voltage and column temperature and of electrokinetic injection parameters was explored, along with other practical considerations.