Simulation of elution profiles in liquid chromatography—I: Gradient elution conditions, and with mismatched injection and mobile phase solvents

• An extension of the LC simulation method of Czok and Guiochon is described.
• Results for isocratic and gradient elution, overload and solvent mismatch are given.
• Excellent agreement is found between simulations, experimental results and theory.
• The simulations help to characterize effects of LC parameters on performance.
• The simulations are useful when departing significantly from ‘standard’ conditions.

High-performance liquid chromatography (HPLC) simulators are effective method development tools. The goal of the present work was to design and implement a simple algorithm for simulation of liquid chromatographic separations that allows for characterization of the effect of injection solvent mismatch and injection solvent volume overload. The simulations yield full analyte profiles during solute migration and at elution, which enable a thorough physical understanding of the effects of method variables on chromatographic performance. The Craig counter-current distribution model (the plate model) is used as the basis for simulation, where a local retention factor is assigned for each spatial and temporal element within the simulation. The algorithm, which is an adaptation of an approach originally described by Czok and Guiochon (Ref. [10]), is sufficiently flexible to allow the use of either linear (e.g., Linear Solvent Strength Theory) or non-linear models of solute retention (e.g., Neue-Kuss (Ref. [36])). In this study, both types of models were used, one for simulating separations of a homologous series of alkylbenzenes, and the other for separations of selected amphetamines. The simulation program was validated first by comparison of simulated retention times and peak widths for five amphetamines to predictions obtained using linear solvent strength (LSS) theory, and to results from experimental separations of these compounds. The simulated retention times for the amphetamines agreed within 0.02% and 2.5% compared to theory and experiment, respectively. Secondly, the program was evaluated for simulating the case where there is a compositional mismatch between the mobile phase at the column inlet and the injection solvent (i.e., the sample matrix). This work involved alkylbenzenes, and retention time and peak width predictions from simulations were within 1.5 and 6.0% of experimental values, respectively, even without correction for extra-column dispersion. The issues of sample/eluent solvent mismatch and solvent volume overload are especially important when considering the challenges of transferring eluent from the first to the second dimension in comprehensive two-dimensional liquid chromatography.