Optimization of elution conditions and comparison of emerging biocompatible columns on the resolving power and detection sensitivity of oligonucleotides by ion-pairing reversed-phase liquid chromatography mass spectrometry.

A generic performance comparison strategy has been developed to evaluate the impact of mobile-phase additives (ion-pairing agent / counter ion systems), distinct stationary phases on resulting resolving power, and MS detectability of oligonucleotides and their critical impurities in gradient IP-RPLC. Stationary-phase considerations included particle type (core-shell vs. fully porous particles), particle diameter, and pore size.

Absolute configuration assignment of highly fluorinated carboxylic acids via VCD and MRR spectroscopy.

Chiral analysis has become a crucial step in studying the stereospecific synthesis of Active Pharmaceutical Ingredients (APIs). Both Vibrational Circular Dichroism (VCD) and Molecular Rotational Resonance (MRR) spectroscopy are capable of determining absolute configurations (ACs) via comparison of experimental and calculated data. In this regard, each technique has its own caveats.

Recent advances in analytical techniques for high throughput experimentation.

High throughput experimentation is a growing and evolving field that allows to execute dozens to several thousands of experiments per day with relatively limited resources. Through miniaturization, typically a high degree of automation and the use of digital data tools, many parallel reactions or experiments at a time can be run in such workflows. High throughput experimentation also requires fast analytical techniques capable of generating critically important analytical data in line with the increased rate of experimentation.

Practical constraints in the kinetic plot representation of chromatographic performance data: theory and application to experimental data.

It is demonstrated that the kinetic plot representation of experimental plate height data can also account for practical constraints on the column length, the peak width, the viscous heating, and the mobile-phase velocity without needing any iterative solution routine. This implies that the best possible kinetic performance to be expected from a given tested support under any possible set of practical optimization constraints can always be found using a directly responding calculation spreadsheet template. To show how the resulting constrained kinetic plots can be used as a powerful design and selection tool, the method has been applied to a series of plate height measurements performed on a number of different commercial columns for the same component (butyl-parabene) and mobile-phase composition.

Experimental validation of the tetrahedral skeleton model pressure drop correlation for silica monoliths and the influence of column heterogeneity.

This paper describes the use of computational fluid dynamics for the calculation of the flow resistance through computer-generated models resembling silica monoliths. This study was undertaken to determine the effect of skeleton heterogeneity on the flow resistance and, more precisely, to test the hypothesis that increased skeleton heterogeneity decreases the flow resistance. To evaluate the proposed model, 24 real silica monoliths have been prepared using the same method, covering a wide range of skeleton sizes (2.

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 View Article and Find Full Text PDF

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.

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.

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.

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).

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.

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.