Shear-driven flow approaches to LC and macromolecular separations.
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Chromatography, Liquid↗
Biomedical subjects
Gert Desmet
Publications and source records attributed to Gert Desmet.
General rules for the optimal external porosity of LC supports.
We present a series of numerically calculated plate height and flow resistance data obtained for an idealized chromatographic support mimic with variable bed porosity (0.3 </= epsilon </= 0.9), yielding a unique insight into how the main chromatographic performance parameters can be expected to vary with the external bed porosity, unbiased by any differences in molecular diffusivity or retention factor. The influence of pore heterogeneity effects is considered as well. It is found that the product h(min)nu(opt) depends only very weakly on epsilon and on the degree of pore heterogeneity. It is also found that the minimal separation impedance E(min) decreases monotonically with epsilon. This shows that the minimal plate height increase that can be expected for large porosity systems is always more than compensated by their reduced flow resistance, in agreement with the current observations in real silica monolith columns. Using the computed plate height data in an optimization analysis, it is found that large porosity supports can always potentially yield shorter analysis times or larger plate numbers than small porosity supports but need submicrometer feature sizes to actually achieve this. Assuming a lower limit on the producible or useable structural feature sizes, it is found that small N separations can best be performed with a small porosity packing, whereas large N separations require a large porosity packing if the column length (L) is left free. A plot yielding epsilon(opt) as a function of the required plate number has been established, showing that roughly epsilon(opt) approximately log(N) in both the ordered and the disordered support cases. It is also shown that the maximal increase in peak capacity ever to be expected from the use of high porosity supports is a factor of 2 (if the mobile-phase viscosity can be kept constant), potentially to be increased by a factor of 1.5 by increasing the homogeneity of the packing.
Journal Article↗
Importance and reduction of the sidewall-induced band-broadening effect in pressure-driven microfabricated columns.
The influence of the detailed design of the sidewall region upon the over-all band-broadening in microfabricated packed-bed or collocated monolithic support structure (COMOSS) columns has been investigated using computational fluid dynamics (CFD) simulation techniques. It is shown that, under unretained solute conditions, very small structural variations of the order of only 5% of the particle diameter can give rise to a 4-fold increase of the band-broadening. A comprehensive study has been made to quantify this effect as a function of the fluid velocity, the particle diameter, the channel widths, and of course, the sidewall region design. Because the sidewall effect can be fully attributed to a mismatch between the flow rates in the column center and in the sidewall region, it is fortunately also quite straightforward to avoid it. A very simple design, yielding band-broadening values identical to that of a hypothetical sidewall-less column for all possible values of the flow velocity, the particle diameter, or the channel width is proposed.
Biotechnology↗
Theoretical comparison of the band broadening in nonretained electrically and pressure-driven flows through an ordered chromatographic pillar packing.
Using a well-validated computational fluid dynamics simulation method, based on a multi-ion transport model, a detailed analysis of the differences in band broadening between pressure-driven (PD) and electrically driven (ED) flows through perfectly ordered, identical chromatographic pillar packings has been made. It was found that, although the eddy-diffusion band-broadening contributions were nearly completely absent in the considered structure, the ED flow still yields much smaller plate heights than the PD flow. This difference could be fully attributed to the different ways in which the ED and PD velocity profiles reshape when passing through a tortuous pore structure with undulating cross section. Whereas in the PD case the parabolic tip of the band front is continually squeezed and extended each time it passes a pore constriction, the ED flow displays some kind of band front restoring mechanism, with which the fluid elements of the band front are (at least partly) laterally re-aligned after each pore constriction passage. This could be clearly visualized from a series of step-by-step images of the progression of a sharply "injected" species band moving through the packing under ED and PD conditions.
Journal Article↗
High-velocity transport of nanoparticles through 1-D nanochannels at very large particle to channel diameter ratios.
We explore the possibility of generating high-velocity flows of nanoparticles through flat-rectangular nanochannels, which are only 50% deeper than the diameter of the particles. Using the shear-driven flow principle, 200-nm particles can, for example, be transported through a 300-nm-deep channel at velocities up to 35 mm/s (upper limit of our current setup). Working under high-pH conditions, the velocity of the carboxylated nanoparticles still respects the small-molecule velocity law, despite the high degree of confinement to which the particles are subjected. The high degree of confinement is also found to lead to a reduced band broadening. When injecting sharply delimited particle plugs, the plate heights observed for the flow of 0.2-microm particles through a 0.3-microm channel (with plate heights of the order of 1-2 microm) are, for example, approximately 1 order of magnitude smaller than for the flow of 1.0-microm particles through a 1.4-microm channel. It is also found that the band broadening is, within its statistical variation, independent of the fluid velocity over a large range of particle velocities (5-35 mm/s). The flow method distinguishes itself from pressure-driven field-flow fractionation and hydrodynamic chromatography in that the mean particle velocity is independent of the particle size over the entire range of possible particle to channel diameter ratios.
Journal Article↗
Model column structure for the analysis of the flow and band-broadening characteristics of silica monoliths.
We report on the use of commercial computational fluid dynamics software to study the band broadening in a perfectly ordered three-dimensional model structure, the so-called tetrahedral skeleton column (TSC), selected for its close geometrical resemblance to the specific pore network topology of silica monoliths. Van Deemter plots are presented for the case of a species flow through a non-porous skeleton and for the case of a retained component (k' = 1) in a porous skeleton (mesopore porosity epsilon = 0.6 in both cases). Using the flow domain as the characteristic scaling dimension, the TSC model yields reduced plate heights as small as h(min) = 0.8 and separation impedances as small as Emin = 120 for a retained component with k' = 1. The very small reduced plate heights for the TSC model can without any doubt largely be attributed to the perfect homogeneity of the considered model structure: the B and C terms are similar to those obtained in real silica monoliths with similar external porosity, whereas the A term is significantly (about a factor of 10) smaller. The present study hence suggests that further experimental work to obtain more homogeneous silica networks could yield large gains in reduced plate height and separation impedance. Comparing the three-dimensional TSC model with a 2D array of cylinders, it was found that the use of the domain size as the characteristic dimension in the reduced plate height expression is much more appropriate than the use of the skeleton size, hence validating earlier approaches adopted in the literature.
Pressure↗
A novel microstep device for the size separation of cells.
We report on a series of preliminary experiments investigating the applicability of a novel method for the size separation of nano- and microsized particles and cells. The working principle is based on the application of a shear-driven flow through stepwise tapered micro- or nanochannels. Size separations of mixtures of 0.5 and 1.0 microm carboxylated polystyrene beads as well as of binary mixtures of Staphylococcus aureus and Saccharomyces cerevisiae cells and of S. cerevisiae and Escherichia coli cells are demonstrated.
Escherichia coli↗
Exploiting the benefits of miniaturization for the enhancement of DNA microarrays.
The present study demonstrates that the best way to enhance DNA microarray assays, both in terms of analysis speed and in final spot intensity, is to dissolve the available molar amount of sample in the smallest possible buffer volume and to subsequently convect this solution continuously across the surface of the array. The presently proposed shear-driven flow system is pre-eminently suited for this task, as it allows to induce strongly enhanced lateral transport rates, independently of the degree of miniaturization of the hybridization chamber. This transport enhancement method, however, only increases the hybridization rate and not the final spot intensity, as neither can any of the other transport enhancement methods already proposed in literature. A series of experiments with synthetic single-stranded (ssDNA) samples and an accompanying mass balance analysis are presented to demonstrate these points.
DNA, Single-Stranded↗
A computational study of the porosity effects in silica monolithic columns.
We report on a theoretical study of the influence of the through-pore porosity on the main chromatographic performance parameters (reduced theoretical plate height, flow resistance, and separation impedance) of silica monoliths. To investigate this problem devoid of any structural uncertainties, computer-generated structural mimics of the pore geometry of silica monolithic columns have been studied. The band broadening in these synthetic monoliths was determined using a commercial Computational Fluid Dynamics (CFD) software package. Three widely differing external porosities (epsilon = 0.38, epsilon = 0.60, and epsilon = 0.86) are considered and are compared on the basis of an identical intra-skeleton diffusivity (Ds = 5 x 10(-10)m2/s), internal porosity (epsilon(int) = 0.5), and for the same phase retention factor (k' = 1.25). Since the data are obtained for perfectly ordered structures, the calculated plate heights and separation impedances constitute the ultimate performance ever to be expected from a monolithic column. It is found that, if silica monoliths could be made perfectly homogeneous, domain size-based reduced plate heights as small as h(min) approximately 0.8 (roughly independent of the porosity) and separation impedances as small as Emin approximately 130 (epsilon = 0.60) and Emin approximately 40 (epsilon = 0.86) should be achievable with pure water as the working fluid. The data also show that, although the domain size is a much better reduction basis than the skeleton size, the former is still not capable of bringing the van Deemter curves of different porosity columns into perfect agreement in the C term dominated velocity range. It is found that, in this range, large porosity monoliths can be expected to yield smaller domain size-based reduced plate heights than small porosity monoliths.
Chromatography↗
A correlation for the pressure drop in monolithic silica columns.
To gain insight into how the pressure drop in monolithic silica columns is determined by the microscopic details of the pore structure, a series of well-validated computational fluid dynamics simulations has been performed on a simplified model structure, the so-called tetrahedral skeleton column. From these simulations, a direct correlation between the pressure drop and two main structural properties (skeleton thickness and column porosity) of the monolithic skeleton could be established. The correlation shows good agreement with the experimental pressure-drop data available from the literature on silica monoliths, especially when a correction for the flow-through pore size heterogeneity is made. The established correlation also yields a much more accurate representation of the relation between the flow resistance and the bed porosity than does the Kozeny-Carman model, making it much better suited for porosity optimization calculations.
Journal Article↗
Experimental Van Deemter plots of shear-driven liquid chromatographic separations in disposable microchannels.
We present a new stationary phase coating method, yielding a monolayer of densely arrayed porous HPLC beads (d(p)=4 microm) for use in a disposable shear-driven flow LC system. The system is inherently suited for whole-column detection through the small voids between the individual particles of the layer. The chromatographic performance of the system has been characterized by performing a series of coumarin dye separation experiments (reversed-phase mode) and by measuring the theoretical plate height as a function of the mobile phase velocity. The resulting Van Deemter curve, yielding a value of about 90,000 plates/m near the u=u(opt) velocity, shows good agreement with the theoretical expectations, and hence constitutes the first full validation of the theory of shear-driven chromatography.
Chromatography, Liquid↗
Chromatographic explanation for the side-wall induced band broadening in pressure-driven and shear-driven flows through channels with a high aspect-ratio rectangular cross-section.
Based on a simple, chromatography-based analogy, a quantitatively exact explanation for the strong additional band broadening induced by the presence of the side-walls in flow channels with a large aspect-ratio rectangular cross-section is given and validated for two different flow types: pressure-driven and shear-driven flow.
Chromatography↗