Detailed kinetic performance analysis of micromachined radially elongated pillar array columns for liquid chromatography.

The individual factors that determine the kinetic performance (B- and C-term band broadening and bed permeability Kv) of radially elongated pillar (REP) columns are studied. To this end, columns with REPs having 4 different aspect ratios (AR=9, 12, 15, 20) were characterized experimentally and by means of numerical simulations. A tortuosity and retention based plate height equation was established, enabling a good global fit for all studied conditions. The B-term plate height contribution appears to decrease with a factor equaling the square of the flow path tortuosity τ. Going from AR=12 to AR=20 (τ=5.7 and τ=9.0 respectively), this resulted in a shift in plate height expressed in axial coordinates from Hmin=0.42 µm to Hmin=0.25 for non-retained conditions and from H=0.77 µm to H=0.57 µm for a component with k=1.0. The obtained parameters were combined to predict optimal time-efficiency combinations for all possible channel lengths. This revealed an efficiency limit of N=10(7) plates for a non-retained component and N=7-8 × 10(6) for k=1 for a channel with an AR=20, corresponding to a channel length of 2.5m and a void time of 2.4h.

Mass-manufacturable polymer microfluidic device for dual fiber optical trapping.

We present a microfluidic chip in Polymethyl methacrylate (PMMA) for optical trapping of particles in an 80µm wide microchannel using two counterpropagating single-mode beams. The trapping fibers are separated from the sample fluid by 70µm thick polymer walls. We calculate the optical forces that act on particles flowing in the microchannel using wave optics in combination with non-sequential ray-tracing and further mathematical processing. Our results are compared with a theoretical model and the Mie theory. We use a novel fabrication process that consists of a premilling step and ultraprecision diamond tooling for the manufacturing of the molds and double-sided hot embossing for replication, resulting in a robust microfluidic chip for optical trapping. In a proof-of-concept demonstration, we show the trapping capabilities of the hot embossed chip by trapping spherical beads with a diameter of 6µm, 8µm and 10µm and use the power spectrum analysis of the trapped particle displacements to characterize the trap strength.

Merging Open-Tubular and Packed Bed Liquid Chromatography.

Experimental and theoretical proof is provided for the fact that a microfabricated packed bed column, which is uniformly filled with radially elongated pillars (REPs), can produce the same separation performance as nonpacked, open-tubular columns. These are generally recognized as the best possible chromatographic column format, offering the highest conceivable separation speed and efficiency. It is also demonstrated both experimentally and theoretically that, as long as pressure is not a limiting factor, the REP column format can even outperform the open-tubular column format, with significant gains in either speed or efficiency proportional to the tortuosity, τ, of the bed. Conducting chromatographic experiments on 4 cm long micromachined packed bed columns filled with radially elongated pillars, separation efficiencies corresponding to N = 160,000 theoretical plates (unretained analytes) and N = 70,000 theoretical plates were achieved, despite the relatively large interpillar distance (2.5 µm).

Assessment and numerical search for minimal Taylor-Aris dispersion in micro-machined channels of nearly rectangular cross-section.

A mathematical procedure using the Matlab® PDE toolbox to calculate the numerical constant appearing in the general Taylor-Aris expression for the dispersion in a laminar flow through open-tubular conduits with a variety of quasi-rectangular cross-sectional shapes is described. The procedure has been applied to assess the effect of some of the most frequently occurring etching imperfections (linear or curved tapering of the inter-pillar distance along the depth coordinate, occurrence of local notches) in etched pillar array columns. In addition, covering a broad range of possible geometries, a number of new shapes and optimal geometries to minimize the dispersion in open-tubular microchannels and pillar array columns have been proposed. Making a full shape-sensitivity study, it was also found that, whereas the proposed designs can theoretically reduce the dispersion up to a factor of 8, relatively small deviations from this ideal shape can however again dramatically increase the dispersion. Designers should therefore be very careful before implementing an optimized shape and should first aim at solving the etching imperfection problems.

Suppression of the sidewall effect in pillar array columns with radially elongated pillars.

An important bottleneck of pillar array columns designed for liquid chromatography is that small deviations of the target 'magical distance' at the sidewall region leads to detrimental sidewall effects, due to local differences of linear velocities at the sidewall region versus other locations in the pillar bed. In the present study, we demonstrate that a lateral elongation of the pillar significantly increases the tolerance for offsets of the magical distance. By shifting the sidewall distance 600 nm for 2 pillar aspect ratio (AR) designs (AR=3 and 9), only minor sidewall effects on the measured plate heights could be observed for the AR=9 columns, while the plate height was roughly doubled when using the wrong versus the correct sidewall distance for the AR=3 columns. Technologically, this constitutes a huge advantage because small deviations (order of 100 nm) between the set and the finally achieved value for the inter-pillar distance are very common using mid-UV lithography based etching processes.

Integration of uniform porous shell layers in very long pillar array columns using electrochemical anodization for liquid chromatography.

Electrochemical anodization has been applied to grow porous shell layers of 300 nm (30 nm pores) in 5 µm diameter pillar array columns (PACs) with a spacing of 2.5 µm. Using turn structures preceded and followed by the flow distributor structures recently introduced by our group and filled with radially elongated pillars, columns with quasi unlimited channel lengths could be conceived. The uniformity of the porous PAC was assessed by determining local plate heights along the channel, which appeared to be constant. Minimal (absolute) plate heights (H) between 4 and 6 µm were obtained at optimal flow rates when imposing increasing retention factors. Upon measuring the surface area involved in chromatographic retention as an indicator of the available surface area, an increase in the surface area by a factor of about 30 compared to that of non-anodized pillars was found. On reconfiguring a commercial HPLC instrument to enable on-chip injections, 90% of the performance (expressed in theoretical plates) could be maintained for a 1 m column, while for a 25 cm column severe losses were still observed. As the corresponding pressure drop for optimal operation of retained components is on the order of 10 bar per m only, portable and cheaper HPLC devices with high efficiencies become realistically conceivable.

On the advantages of radially elongated structures in microchip-based liquid chromatography.

We report on the possibility to realize submicrometer plate heights using chromatographic pillar array columns filled with radially elongated diamond-shaped pillars, even when using a relatively large interpillar distance (2.5 µm) and axial pillar width (5 µm). It is demonstrated that the use of high aspect ratio radially elongated pillars which are 15 times wider in the radial than in the axial direction can lead to a fivefold reduction of the minimal plate height compared to beds filled with pillars with a similar interpillar distance but with an aspect ratio around unity (cylinders and diamonds).This increase in performance can be attributed to a decrease in longitudinal dispersion, reflected by a reduction of the B-term by a factor of about 25. Experiments were conducted at room temperature, as well as at elevated temperature (70 °C), where the B-term band broadening is known to be more critical. The main advantage of radially elongated pillar beds is that they enable a drastic reduction of the footprint of pillar array columns, allowing design of very long channels with a minimum of turns. Under retained conditions, a four-component laser dye mixture could be separated over a distance of only 1.5 mm.

A membrane microcontactor as a tool for integrated sample preparation.

A membrane microcontactor suitable to perform liquid-liquid extraction as well as evaporation in order to conduct enrichment steps in sample preparation of organ samples has been designed, fabricated, and characterized. Spacers of 100- or 200-µm high were constructed in a metal substrate with a channel width of 13 mm and the extraction kinetics in these channels was evaluated. The spacers were designed such that at the entrance and exit region a uniform flow distribution could take place and that a uniform flow profile could be guaranteed along the channel, hence allowing a large freedom in sample volume to be processed. The extraction and evaporation kinetic behavior of the device was first evaluated by extraction of a drug candidate (4-(2,5-dimethyl-pyrrol-1-y1)-2-hydroxybenzoic acid). To evaluate the device under more challenging working conditions, a homogenized mice kidney sample containing the drug candidate that was administered in life condition was cleaned and enriched with the extraction and evaporation modules and characterized by high-performance liquid chromatography, yielding an overall analysis time of 15-20 min per sample only. The system has the potential to be operated in a continuous fashion, making it appealing to be implemented in screening or high-throughput applications.