We report on the possibility to achieve ultra high efficiencies (order of 1 million theoretical plates) in liquid chromatography in a relatively short time of 20 min (elution time of unretained marker). This was achieved using a micropillar array column with optimized pillar diameter (5 μm) and interpillar distance (2.5 μm) to operate close to the Knox and Saleem limit of micropillar array columns in the region of the 1 million theoretical plate mark under the prevailing pressure restriction (350 bar in the present study). The obtained efficiency was slightly affected (some 15 to 20% around the optimal flow rate) by the turns that were inevitably needed to arrange a 3 m long column on a 4 in. silicon wafer.

We report on a new approach to obtain highly homogeneous silica-monolithic columns, applying a sol-gel fabrication process inside a rectangular pillar-array column (1 mm in width, 29 microm in height and 33.75 mm in length) having a cross-sectional area comparable to that of a 200 microm diameter circular capillary. Starting from a silicon-based pillar array and working under high phase-separation-tendency conditions (low poly(ethylene glycol) (PEG)-concentration), highly regular silica-based chromatographic systems with an external porosity in the order of 66-68% were obtained. The pillars, 2.4 microm in diameter, were typically clad with a 0.5 microm shell layer of silica, thus creating a 3.4 microm total outer pillar diameter and leaving a minimal through-pore size of 2.2 microm. After mesopore creation by hydrothermal treatment and column derivatization with octyldimethylchlorosilane, the monolithic column was used for chip-based liquid-chromatographic separations of coumarin dyes. Minimal plate heights ranging between 3.9 microm (nonretaining conditions) and 6 mum (for a retention factor of 6.5) were obtained, corresponding to domain-size-reduced plate heights ranging between 0.7 and 1.2. The column permeability was in the order of 1.3 x 10(13) m(2), lower than theoretically expected, but this is probably due to obstructions induced by the sol-gel process in the supply channels.

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We investigated the achievable separation performance of a 9-cm-long and 1-mm-wide pillar array channel (volume = 0.6 μL) containing 5 μm diameter Si pillars (spacing 2.5 μm) cladded with a mesoporous silica layer with a thickness of 300 nm, when this channel is directly interfaced to a capillary LC instrument. The chip has a small footprint of only 4 cm × 4 mm and the channel consists of three lanes that are each 3 cm long and that are interconnected using low dispersion turns consisting of a narrow U-turn (10 μm), proceded and preceded by a diverging flow distributor. Measuring the band broadening within a single lane and comparing it to the total channel band broadening, the additional band broadening of the turns can be estimated to be of the order of 0.5 μm around the minimum of the van Deemter curve, and around some 1 μm (nonretained species) and 2 μm (retained species) in the C-term dominated regime. The overall performance (chip + instrument) was evaluated by conducting gradient elution separations of digests of cytochrome c and bovine serum albumin. Peak capacities up to 150 could be demonstrated, nearly completely independent of the flow rate.

The present paper provides a detailed analysis of the analyte-wall adsorption effects in nanochannels, including a random walk study of the analyte-wall collision frequency, and uses these insights to estimate wall desorption times from chromatographic experiments in nanochannels. Using coumarin dye analytes and using a methanol/water mixture buffered at pH 3 in 120-nm deep channels, the surface desorption times on naked fused-silica glass were found to be maximally of the order of 60 to 150 mus, while they were found to be on the order of 100 to 500 mus on a hydrophobically coated wall. These nonzero adsorption and desorption times lead to an additional band broadening when conducting chromatographic separations. Shear-driven flows, requiring a noncoated moving wall and a stationary coated wall, intrinsically turn out to be more prone to this effect than pressure-driven or electro-driven flows for example. The present study also shows that, interestingly, the number of analyte-wall collisions increases with the inverse of the channel depth and not with its second power, as would be expected from the Einstein-Smoluchowski relationship for molecular diffusion.

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.