Band-broadening in packed chromatographic columns

In the past few years some problems have arisen as to the interpretation of experimental plate height data of packed chromatographic columns by means of the theory of VAN DEEMTER et al. These difficulties concern the “eddy diffusion” and “mass transfer” terms in particular. Light may be thrown on this matter by the recognition of unevenness of flow throughout the column cross-section as a major source of band-broadening. By considering the contribution to plate height resulting from the interaction between such flow profiles and various mechanisms of lateral transport of material it proved possible to solve the above-mentioned difficulties.     

Theoretical investigation of diffusion along columns packed with fully and superficially porous particles

Chromatographic columns packed with shell particles are now nearly twice more efficient than columns packed with conventional, fully porous particles. Shell particles are made of a solid core surrounded by a porous shell of constant thickness. Diffusion through the bed of packed columns is complex due to their heterogeneity. It involves diffusion through the external and the internal fluid, and surface diffusion. Six diffusion models are compared that combine these diffusion mechanisms. They involve the external porosity of the bed (?(e)), the ratio of the core to the particle diameters (ρ), and the ratio of the shell diffusivity to the bulk diffusion coefficient (Ω). Four different theoretical approaches were considered. They are based on (1) the additivity of the mass flux densities modulated by the obstruction factors caused by non-porous spherical inclusions; (2) the effective medium theory of Landauer; (3) the effective medium theory of Garnett for spherical inclusions; and (4) the probabilistic theory of Torquato (for binary composite materials only). The two Landauer models fail because they cannot account for the obstruction factor imposed by the presence of non-porous spherical inclusions. The ternary Garnett model (3) provides an excellent approximation of the actual diffusion mechanism but the most physically relevant model seems to be the one derived from a combination of the Garnett model for a binary core-shell particle and of the Torquato model for random dispersion of contacting spheres in a matrix. Accurate measurements of axial dispersion coefficients are needed to validate or reject the semi-empirical parallel diffusion models and to select the most appropriate one. The results of such measurements made with the peak parking method for various compounds are reported in the companion paper.     

High speed analysis in packed columns

Summary A general procedure is presented for predicting the minimum analysis time required to achieve a given chromatographic separation. By combining equations relating the residence time, resolution, and gas flow characteristics of a column, a direct computational procedure is evolved for determining the minimising values of: the inlet to outlet pressure ratio, the column length and the linear gas velocity. This technique is then utilised to discuss the relative sensitivity of the solution to the values of the terms , , C _g , C _L , and D and hence to determine their optimal settings with respect to the characteristic properties of the system. In particular, the feasibility of fast analyses (<6 sec) within the accessible limits of these parameters is explored, together with a quantitative assessment of the influence of the liquid loading, carrier-gas type and support particle size. The condition when permeability requirements demand a compromise between these factors is identified in terms of the expected flowrate restrictions.     

Study of electroosmotic flow in packed capillary columns

The aim of the work was to find the relationship between the structure of the stationary phase and the velocity of the electroosmotic flow (EOF) that it can generate. The attention was paid to the dependence of the electroosmotic mobility (microEOF) on such parameters as: (i) coverage density of a series of specially synthesized C18 stationary phases with/without end-capping, with monomeric/polymeric architecture; (ii) the length of the alkyl chain in the alkylamide (AP) bonded phase (the phases studied were AP with: C1, C5, C6, C7, C8, C12 and C18 alkyl chains) and the effect of the presence of amide and residual amine groups; (iii) the effect of the mobile phase composition on the EOF; (iv) the effect of pH on the EOF. The obtained results have shown that there is no direct relationship between silanol activity (Galushko test) and electroosmotic mobility for C18 phases. The deterioration of the EOF has been observed for AP phases at high pH values. This effect has been attributed to the presence of hydrolytic pillow, which is connected with the sorption of water from hydro-organic mobile phases.     

In-situ deactivation of packed columns for SFC

Residual adsorptive activity of reversed phase (RP) column packings used in supercritical fluid chromatography (SFC) can be significantly reduced by a dynamic in-situ silanization with diphenyltetramethyldisilazane (DPTMDS). RP-materials thus deactivated were characterized both chromatographically and by solid-phase ²⁹Si NMR.     

Particle packed columns and monolithic columns in high-performance liquid chromatography-comparison and critical appraisal

The review highlights the fundamentals and the most prominent achievements in the field of high-performance liquid chromatography (HPLC) column development over a period of nearly 50 years. After a short introduction on the structure and function of HPLC columns, the first part treats the major steps and processes in the manufacture of a particle packed column: synthesis and control of particle morphology, sizing and size analysis, packing procedures and performance characterization. The next section is devoted to three subjects, which reflect the recent development and the main future directions of packed columns: minimum particle size of packing, totally porous vs. core/shell particles and column miniaturization. In the last section an analysis is given on an alternative to packed columns-monolithic columns, which have gained considerable attraction. The challenges are: improved packing design based on modeling and simulation for targeted applications, and enhanced robustness and reproducibility of monolithic columns. In the field of miniaturization, particularly in chip-based nano-LC systems, monoliths offer a great potential for the separation of complex mixtures e.g. in life science.     

Evaluation of packed capillary liquid chromatography columns and comparison with conventional-size columns

Apart from extracolumn effects peak dispersion in liquid chromatographic columns is caused by the column inlet, the packed bed, and the column outlet. A strategy applicable for independent evaluation of the individual sources of column band broadening was developed on the basis of the linear extrapolation method (LEM). This method was applied to compare the performance of packed capillary LC columns from various commercial suppliers with conventional-size columns. The columns differed widely in their performance with respect to peak shapes and widths for standard substances. The capillary columns were found well packed, but in some cases overall performance would benefit from improving the design of the area between the packed bed and the connecting capillaries, containing frits as well as dead volumes.     

Experimental investigation of liquid chromatography columns by means of computed tomography

The efficiency of packed chromatographic columns was investigated experimentally by means of computed tomography (CT) techniques. The measurements were carried out by monitoring tracer fronts in situ inside the chromatographic columns. The experimental results were fitted using the equilibrium dispersive model (EDM) and varying the so called apparent axial dispersion coefficient. The additivity of the first and second central moments was exploited to estimate column efficiency in different regions of the column. The results showed that the columns under investigation offered a higher column efficiency in the centre compared to the wall region. Furthermore the void fraction in the vicinity of the walls was lower than that in the column core. For this reason the bands were conveyed faster in the central region of the column where the permeability was higher. This result is in good agreement with earlier findings.     

Photopolymerized sol-gel frits for packed columns in capillary electrochromatography

Porous sol-gel frits are fabricated in a capillary column by filling it with a solution of 3-(trimethoxysilyl)propyl methacrylate, hydrochloric acid, water, toluene (porogen), and a photoinitiator (Irgacure 1800) and exposing it to UV light at 365 nm for 5 min. The separation column (30 cm x 75 microm I.D.) contains between the inlet and outlet frits a 15-cm packed segment filled with 5-microm silica particles modified with the chiral compound (S)-N-3,5-dinitrobenzoyl-1-naphthylglycine. A detection window (1 mm long) is placed immediately after the outlet frit. To demonstrate the performance of this chiral separation column, mixtures of 16 different amino acids (three of which are not naturally occurring) derivatized with the fluorogenic reagent 4-fluoro-7-nitro-2,1,3-benzoxadiazole were separated by capillary chromatography. The enantiomeric separation of the column results in a resolution ranging from 1.21 to 8.29, and a plate height ranging from 8.7 to 39 microm.     

Viscous fingering in packed chromatographic columns: non-linear dynamics

Viscous fingering (VF) is a hydrodynamic instability that occurs in a chromatographic column when a less viscous fluid displaces another more viscous one. This instability is detrimental to separation techniques as it leads to distorted peaks and peak broadening. Nonlinear interactions between developing fingers lead to complex dynamics investigated in the present study by means of numerical simulations based on a simple model for miscible VF of finite samples. We review the properties of nonlinear VF and discuss the quantitative measures that can be applied both on such numerical as well as on experimental data to gain insight into the influence of the parameters of the problem on the nonlinear properties of the fingers and on the broadening of output peaks.     

On the stability of micro-LC columns (packed fused silica capillary columns)

The life expectancy and resistance to flow, pressure shocks, solvent gradients, and bending of packed fused silica capillary columns (Micro-LC) are discussed.     

Use of high-efficiency packed columns for gas-solid chromatography : V. Micropacked columns

Micropacked columns are constructed using graphitized carbon black as a support for gas-liquid-solid chromatography. Their effectiveness is evaluated in terms of Van Deemter plots, flow-rates, inlet pressure and column capacity. It is shown that such columns, by choosing an appropriate liquid phase, can be used for a wide range of temperatures and types of compound. Analytical applications connected with the analysis of complex organic mixtures in water are discussed. The problems of the analysis of air pollutants are also considered.     

Axial development and radial non-uniformity of flow in packed columns

Flow inhomogeneity and axial development in low-pressure chromatographic columns have been studied by magnetic resonance imaging velocimetry. The columns studied included (a) an 11.7-mm I.D. column packed with either 50 microm diameter porous polyacrylamide, or 99 or 780 microm diameter impermeable polystyrene beads, and (b) a 5-mm I.D. column commercially packed with 10 microm polymeric beads. The packing methods included gravity settling, slurry packing, ultrasonication, and dry packing with vibration. The magnetic resonance method used averaged apparent fluid velocity over both column cross-sections and fluid displacements greater than one particle diameter and hence permits assessment of macroscopic flow non-uniformities. The results confirm that now non-uniformities induced by the conical distributor of the 11.7-mm I.D. column or the presence of voids at the column entrance relax on a length scale of the column radius. All of the 11.7-mm I.D. columns examined exhibit near wall channeling within a few particle diameters of the wall. The origins of this behavior are demonstrated by imaging of the radial dependence of the local porosity for a column packed with 780 microm beads. Columns packed with the 99-microm beads exhibit reduced flow in a region extending from ten to three-to-five particle diameters from the wall. This velocity reduction is consistent with a reduced porosity of 0.35 in this region as compared to approximately 0.43 in the bulk of the column. Ultrasonicated and dry-packed columns exhibit enhanced flow in a region located between approximately eight and 20 particle diameters from the wall. This enhancement maybe caused by packing density inhomogeneity and/or particle size segregation caused by vibration during the packing process. No significant non-uniformities on length scales of 20 microm or greater were observed in the commercially packed column packed with 10 microm particles.     

Dense colloid transport in a bifurcating fracture

In this work, the transport of dense colloids through a water-saturated, bifurcating fracture is investigated using a constant spatial step particle tracking technique. The size of the constituents of a colloid plume is an important factor affecting the partitioning of dense colloids at the bifurcation. While neutrally buoyant colloids partition between daughter fractures in proportion to flow rates, dense colloids will preferentially exit fractures that are gravitationally downgradient, notwithstanding that the majority of the interstitial fluid may flow through the upper fracture. Comparison of the partitioning ratio between daughter fractures with the ratios of characteristic settling, diffusion, and advection time reveal that these parameters control how colloids behave at fracture bifurcations.     

Applications of heart cutting from packed to capillary columns

The power of capillary gas chromatography can be enhanced by selective fractionation of the sample. One way of doing this is heart cutting from a packed gas chromatographic column. This paper describes modification of a commercial instrument for such separations. Components of the system include an automated Deans switch and an electrically heated trap. The benefits of this arrangement are illustrated with chromatograms of naphtha, urinary aromatic acids, and wine volatiles.     

Viscous fingering in packed chromatographic columns: linear stability analysis

When a fluid is displaced by a less viscous one in a porous medium, a hydrodynamic instability appears leading to the formation of some kind of fingers of the upstream fluid invading the downstream one, hence the name "viscous fingering" (VF) given to this instability. In a LC column, such an instability is likely to appear at that of the two interfaces between the sample and the eluent which exhibits an unfavorable viscosity contrast. It leads to distorted peak shapes and contributes to peak broadening. This phenomenon has been observed for long in SEC and more recently in RPLC on elution peak shapes as well as with various methods of in-column visualization. A simplistic LC column model is described to explain the origin of the VF instability and its characteristics. The general principles for analyzing hydrodynamic instabilities are described and the results of the linear stability analysis performed by Tan and Homsy [C.T. Tan, G.M. Homsy, Phys. Fluids 29 (1986) 3549 [1]], at the onset of the VF phenomenon for a step interface between two fluids are here applied to typical operating conditions encountered in analytical LC. The most probable growth rate and wavelength (linked to the finger width) of the instability are given in terms of particle size and solute diffusion coefficient, with particular emphasis on the role of the carrier velocity. Previously published qualitative observations about VF in chromatography are examined and interpreted at the light of this theory. The role of the column geometry on the development of the instability, the possible sources of noise or fluctuations triggering the instability, and the various experimental situations in which a significant viscosity contrast is encountered in LC are discussed.