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A microfluidic platform or “microfluidic mapper” is demonstrated, which in a single experiment performs 36 parallel biochemical reactions with 36 different combinations of two reagents in stepwise concentration gradients. The volume used in each individual reaction was 36 nl. With the microfluidic mapper, we obtained a 3D enzyme reaction plot of horseradish peroxidase (HRP) with Amplex Red (AR) and hydrogen peroxide (H2O2), for concentration ranges of 11.7 μM to 100.0 μM and 11.1 μM to 66.7 μM for AR and H2O2, respectively. This system and methodology could be used as a fast analytical tool to evaluate various chemical and biochemical reactions especially where two or more reagents interact with each other. The generation of dual concentration gradients in the present format has many advantages such as parallelization of reactions in a nanoliter-scale volume and the real-time monitoring of processes leading to quick concentration gradients. The microfluidic mapper could be applied to various problems in analytical chemistry such as revealing of binding kinetics, and optimization of reaction kinetics. [ABSTRACT FROM AUTHOR]

We explored the possibility to perform high speed and high efficiency liquid chromatographic separations in channels with a sub-100 nm depth. The mobile phase flow through these nano-channels was generated using the shear-driven flow principle to generate high speed flows which were the equivalent of a 12 000 bar pressure-driven flow. It was found that the ultra-fast mass transfer kinetics prevailing in this range of small channel depths allow to drastically reduce the C-term contribution to band broadening, at least up to the upper speed limit of our current set-up (7 mm s−1 mobile phase velocity), leaving the inescapable molecular diffusion (i.e., B-term band broadening) as the sole detectable source of band broadening. Due to the greatly reduced mass transfer limitations, 50 000 to 100 000 theoretical plates could be generated in the span of 1 to 1.5 seconds. This is nearly two orders of magnitude faster than the best performing commercial pressure-driven UHPLC-systems. With the employed channel depths, we appear to have struck a practical lower limit for the channel miniaturization of shear-driven flows. Despite the use of channel substrates with the highest grades of optical flatness, the overall substrate waviness (on the order of some 5 to 10 nm) can no longer be neglected compared to the etched channel depth, which in turn significantly influenced the local retention factor and band broadening. [ABSTRACT FROM AUTHOR]

This article reveals the first liquid chromatography (LC) separations performed on a microfabricated pillar array column under pressure-driven conditions. The pillars were non-porous and produced using a Bosch-type deep reactive ion etch (DRIE) to pattern the surface of a silicon wafer and had a diameter of approximately 5 pm. Two different packing densities were compared: one similar to the packing density of a packed bed (external porosity of approximately 49%) and one similar to the packing density of monolithic columns (external porosity of approximately 70%). The most densely packed pillar array yielded plate heights between 0.9–4.9 μm (depending on the degree of retention of the employed coumarin test compounds). This is considerably smaller than for a packed bed column filled with particles with the same diameter, demonstrating the large potential advantages of ordered pillar array columns compared with the much more random structure of packed beds. The main obstacle to developing this new type of column technology at the moment is the lack of reproducible coating procedures. [ABSTRACT FROM AUTHOR]

Abstract: An experimental study of a micromachined non-porous pillar array column performance under non-retentive conditions is presented. The same pillar structure has been fabricated in cyclo-olefin polymer (COP) chips with three different depths via hot embossing and pressure-assisted thermal bonding. The influence of the depth on the band broadening along with the already known contribution arising from the top and bottom cover plates has been studied. The experimental results exhibit reduced plate heights as low as 0.2, which are in agreement with the previous experimental work. Moreover, the constant values of the reduced Van Deemter expression are also in accordance with the previous studies. A more exhaustive study of the C-term band broadening is also presented, showing that comparing the space between the pillars with different open tubular rectangular channels offers a good estimation of the C-term band broadening that is obtained experimentally. These experimental results, hence, confirm that micromachined pillar array columns fabricated in COP can achieve the same performance as the ones fabricated in silicon for the presently studied pillar channel design. [ABSTRACT FROM AUTHOR]

Abstract: We report on the possibility to perform HDC in micropillar array columns and the potential advantages of such a system. The HDC performance of a pillar array column with pillar diameter=5μm and an interpillar distance of 2.5μm has been characterized using both a low MW tracer (FITC) and differently sized polystyrene bead samples (100, 200 and 500nm). The reduced plate height curves that were obtained for the different investigated markers all overlapped very well, and attained a minimum value of about h min =0.3 (reduction based on the pillar diameter), corresponding to 1.6μm in absolute value and giving good prospects for high efficiency separations. The obtained reduced retention time values were in fair agreement with that predicted by the Di Marzio and Guttman model for a flow between flat plates, using the minimal interpillar distance as characteristic interplate distance. [Copyright &y& Elsevier]

Abstract: We report on the synthesis of siloxane-based monoliths in the presence of a two-dimensional, perfectly ordered array of micro-pillars. Both methyltrimethoxysilane- and tetramethoxysilane-based monoliths were considered. The obtained structures were analyzed using scanning-electron microscopy and can be explained from the general theory of surface-directed phase separation in confined spaces. The formed structures are to a large extent nearly exclusively determined by the ratio between the bulk domain size of the monolith on the one hand and the distance between the micro-pillars on the other hand. When this ratio is small, the presence of the pillars has nearly no effect on the morphology of the produced monoliths. However, when the ratio approaches unity and ascends above it, some new types of monolith morphologies are induced, two of which appear to have interesting properties for use as novel chromatographic supports. One of these structures (obtained when the domain size/inter-pillar distance ratio is around unity) is a 3D network of linear interconnections between the pillars, organized such that all skeleton branches are oriented perpendicular to the micro-pillar surface. A second interesting structure is obtained at even higher values of the domain size/inter-pillar distance ratio. In this case, each individual micro-pillar is uniformly coated with a mesoporous shell. [Copyright &y& Elsevier]

Building upon the micromachined column idea proposed by the group of Regnier in 1998, we report on the first high-resolution reversed-phase separations in micromachined pillar array columns under pressure-driven LC conditions. A three component mixture could be separated in 3 s using arrays of nonporous silicon pillars with a diameter of approximately 4.3 μm and an external porosity of 55%. Under slightly retained component conditions (retention factor k′ = 0.65-1.2), plate heights of about H = 4 μm were obtained at a mobile phase velocity around u = 0.5 mm/s. In reduced terms, such plate heights are as low as hmin = 1. Also, since the flow resistance of the column is much smaller than in a packed column (mainly because of the higher external porosity of the pillar array), the separation impedance of the array was as small as E = 150, i.e., of the same order as the best currently existing monolithic columns. At pH = 3, yielding very low retention factors (k′ = 0.13 and 0.23), plate heights as low as H = 2 pm were realized, yielding a separation of the three component mixture with an efficiency of N = 4000-5000 plates over a column length of 1 cm. At higher retention factors, significantly larger plate heights were obtained. More experimental work is needed to investigate this more in depth. The study is completed with a discussion of the performance limits of the pillar array column concept in the frame of the current state-of-the-art in microfabrication precision. [ABSTRACT FROM AUTHOR]