Validating a Numerical Simulation of the ConsiGma(R) Coater.

Continuous manufacturing is increasingly used in the pharmaceutical industry, as it promises to deliver better product quality while simultaneously increasing production flexibility. GEA developed a semi-continuous tablet coater which can be integrated into a continuous tableting line, accelerating the switch from traditional batch production to the continuous mode of operation. The latter offers certain advantages over batch production, e.g., operational flexibility, increased process/product quality, and decreased cost. However, process understanding is the key element for process control. In this regard, computational tools can improve the fundamental understanding and process performance, especially those related to new processes, such as continuous tablet coating where process mechanics remain unclear. The discrete element method (DEM) and computational fluid dynamics (CFD) are two methods that allow transition from empirical process design to a mechanistic understanding of the individual process units. The developed coupling model allows to track the heat, mass, and momentum exchange between the tablet and fluid phase. The goal of this work was to develop and validate a high-fidelity CFD-DEM simulation model of the tablet coating process in the GEA ConsiGma® coater. After the model development, simulation results for the tablet movement, coating quality, and heat and mass transfer during the coating process were validated and compared to the experimental outcomes. The experimental and simulation results agreed well on all accounts measured, indicating that the model can be used in further studies to investigate the operating space of the continuous tablet coating process.

Separations using a porous-shell pillar array column on a capillary LC instrument.

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.

Experimental optimization of flow distributors for pressure-driven separations and reactions in flat-rectangular microchannels.

We report on the results of an experimental study established to optimize the design of microfabricated flow distributors for use in pressure-driven separations and reactions in flat-rectangular channels. For this purpose, the performance of a wide variety of possible flow distributor designs etched in glass/silicon wafers was compared, using CCD camera detection to study the shape and variance of the bands eluting from them. The best performance was obtained with radially interconnected distributors with a diverging inlet section and filled with diamond-shaped pillars, oriented perpendicular to the main flow direction and with a high transversal over axial aspect ratio. It was found that the best distributor designs start with a diverging section containing some 10-12 subsequent rows of high aspect ratio pillars (with a transversal width making up 10-15% of the final channel width) and with a divergence angle selected such that the sloped side-walls run parallel with the sides of the diamond-shaped pillars. After this zone, one or more regions with pillars with a smaller aspect ratio should be provided to increase the number of exit points. To prevent the formation of dead zones in these subsequent zones, so-called distributor wedges can be used to prevent the formation of any dead zones in the wake of the large aspect ratio pillars of the preceding section.

Fabrication and chromatographic performance of porous-shell pillar-array columns.

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.

High-speed shear-driven flows through microstructured 1D-nanochannels.

A new flow type for the conduction of rapid chromatographic and macro-molecular separations in 1D nanochannels is reported. It combines the pressure-drop-less operation of shear-driven flows with the meandering flow paths that are present in ordered arrays of micro- and nanopillars. Similar to shear-driven flows in open channels, the achievable fluid velocity is quasi unlimited and is not affected by a pressure- or voltage-drop, while the axial dispersion in the microstructured pillar arrays remains surprisingly low. In the present paper, we report on a series of flow resistance and band broadening experiments that have been conducted to characterize the hydrodynamical properties of this new flow type. In addition, theoretical computational fluid dynamics (CFD) simulations have been performed to explain the observations. Good agreement between theory and experiment was obtained.

Estimation of surface desorption times in hydrophobically coated nanochannels and their effect on shear-driven and pressure-driven chromatography.

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.

Modeling the effect of species retention on the band broadening in perfectly ordered silica monolithic column mimics with variable external porosity and intra-skeleton diffusivity.

Computational fluid dynamics simulations of the band broadening in an idealized, and highly ordered 3-D model of silica monoliths are reported. As this high degree of order induces only a minimal eddy-dispersion, the band broadening is very sensitive to the intra-skeleton diffusivity and retention factor of the analytes. The simulations hence provide a maximal view on how the C(m)- and C(s)-contributions to the band broadening depend on the intra-skeleton diffusivity and retention factor. By comparing two model-structures with different external porosities, some interesting qualitative insights on the effect of the through-pore diameter are obtained as well. Because of the precisely known intra-skeleton diffusivity, the obtained plate height data also provide an ideal test case for the general plate height expression of chromatography. Writing out this model, identifying the geometrical parameters, and determining their value using a parameter fitting algorithm, a set of parameter values can be found which allows to accurately predict the retention factor dependency of the band broadening over a very broad range of mobile phase velocities. Remaining modeling problems are the apparent intra-skeleton diffusion dependency of the shape factor describing the intra-skeleton mass transfer, and the absence of mathematical expressions to predict the model shape factor values.

1 mm ID poly(styrene-co-divinylbenzene) monolithic columns for high-peak capacity one- and two-dimensional liquid chromatographic separations of intact proteins.

The LC performance of a 1x50 mm polymer monolithic column format was demonstrated with high-peak capacity one- (1D) and offline two dimensional (2D) LC separations of intact proteins. After optimizing the RP 1D-LC conditions, including column temperature, flow rate and gradient time, a peak capacity of 475 was achieved within a 2-h analysis. The suitability of the monolithic column was also demonstrated for fast 1 min protein separations yielding 1 s peak widths determined at half peak height. In addition, an offline 2D-LC method was developed using the micro-fraction collection capabilities of the autosampler allowing automatic fractionation of intact proteins after the weak-ion-exchange (WAX) separation, and re-injection of the fractions onto the second-dimension RP monolithic column. The best peak capacity-to-analysis time ratio was obtained when applying 10 min second-dimension RP gradients. At optimized conditions, the WAX/x/RPLC separation of intact Escherichia coli proteins was performed within 6 h yielding a maximum theoretical peak capacity of 4880.

Modelling the relation between the species retention factor and the C-term band broadening in pressure-driven and electrically driven flows through perfectly ordered 2-D chromatographic media.

The band broadening that can be expected in perfectly ordered cylindrical pillar arrays has been calculated for a wide range of intra-particle diffusion coefficients (D(sz)) and zone retention factors (0

Performance evaluation of long monolithic silica capillary columns in gradient liquid chromatography using peptide mixtures.

A systematic study is reported on the performance of long monolithic capillary columns in gradient mode. Using a commercial nano-LC system, reversed-phase peptide separations obtained through UV-detection were conducted. The chromatographic performance, in terms of conditional peak capacity and peak productivity, was investigated for different gradient times (varying between 90 and 1320min) and different column lengths (0.25, 1, 2 and 4m) all originating from a single 4m long column. Peak capacities reaching values up to n=10(3) were measured in case of the 4m long column demonstrating the high potential of these long monoliths for the analysis of complex biological mixtures, amongst others. In addition, it was found that the different column fragments displayed similar flow resistance as well as consistent chromatographic performance in accordance with chromatographic theory indicating that the chromatographic bed of the original 4m long column possessed a structural homogeneity over its entire length.

A numerical study of the assumptions underlying the calculation of the stationary zone mass transfer coefficient in the general plate height model of chromatography in two-dimensional pillar arrays.

The present study investigates the validity of one of the key assumptions underlying the general plate height model of chromatography, i.e., the presumed independency of the individual band broadening contributions. More precisely, it is investigated under which conditions the mass transfer inside the stationary zone (e.g., porous pillars) is independent from the axial transport of species outside this zone, and how strongly any such dependency would affect the validity of the general plate height model of chromatography. For this purpose, detailed calculations of the species concentration distribution inside and outside the porous pillars of a computer-mimic of a porous pillar array column have been made. These simulations revealed a clear interplay between the mass transfer inside and outside the pillars, manifesting itself as an asymmetry of the species concentration distribution inside the pillars. The latter is in disagreement with the basic assumption used to calculate the value of the C(s)-term of the general plate height model. The asymmetry-effect is largest at low reduced velocities, high retention factors and high intra-pillar diffusion coefficients. Fortunately, these are conditions where the C(s)-term is relatively small, which might explain why the general plate height model of chromatography (and based on the symmetry assumption) can represent the band broadening in a porous pillar array within an accuracy on the order of some 1-2%.

High-efficiency liquid chromatography-mass spectrometry separations with 50 mm, 250 mm, and 1 m long polymer-based monolithic capillary columns for the characterization of complex proteolytic digests.

In this study, high-efficiency LC-MS/MS separations of complex proteolytic digests are demonstrated using 50 mm, 250 mm, and 1m long poly(styrene-co-divinylbenzene) monolithic capillary columns. The chromatographic performance of the 50 and 250 mm monoliths was compared at the same gradient steepness for gradient durations between 5 and 150 min. The maximum peak capacity of 400 obtained with a 50mm column, increased to 485 when using the 250 mm long column and scaling the gradient duration according column length. With a 5-fold increase in column length only a 20% increase in peak capacity was observed, which could be explained by the larger macropore size of the 250 mm long monolith. When taking into account the total analysis time, including the dwell time, gradient time and column equilibration time, the 50mm long monolith yielded better peptide separations than the 250 mm long monolithic column for gradient times below 80 min (n(c)=370). For more demanding separation the 250 mm long monolith provided the highest peak production rate and consequently higher sequence coverage. For the analysis of a proteolytic digest of Escherichia coli proteins a monolithic capillary column of 1m in length was used, yielding a peak capacity of 1038 when applying a 600 min gradient.

Parameters affecting the separation of intact proteins in gradient-elution reversed-phase chromatography using poly(styrene-co-divinylbenzene) monolithic capillary columns.

An experimental study was performed to investigate the effects of column parameters and gradient conditions on the separation of intact proteins using styrene-based monolithic columns. The effect of flow rate on peak width was investigated at constant gradient steepness by normalizing the gradient time for the column hold-up time. When operating the column at a temperature of 60 degrees C a small C-term effect was observed in a flow rate range of 1-4 microL/min. However, the C-term effect on peak width is not as strong as the decrease in peak width due to increasing flow rate. The peak capacity increased according to the square root of the column length. Decreasing the macropore size of the polymer monolith while maintaining the column length constant, resulted in an increase in peak capacity. A trade-off between peak capacity and total analysis time was made for 50, 100, and 250 mm long monolithic columns and a microparticulate column packed with 5 microm porous silica particles while operating at a flow rate of 2 microL/min. The peak capacity per unit time of the 50mm long monolithic column with small pore size was superior when the total analysis time is below 120 min, yielding a maximum peak capacity of 380. For more demanding separations the 250 mm long monolith provided the highest peak capacity in the shortest possible time frame.

Effect of the presence of an ordered micro-pillar array on the formation of silica monoliths.

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.