Measurement and modeling of the equilibrium behavior of the Tröger's base enantiomers on an amylose-based chiral stationary phase.

The binary isotherms of the two enantiomers of Tröger's base were measured on a system made of pure 2-propanol as the mobile phase and of Chiralpak AD, a chiral stationary phase (CSP) based on amylose tri-(3,5-dimethylphenyl carbamate). The experimental data were acquired using both frontal analysis and the perturbation method. The results obtained are most unusual. The adsorption of the more-retained (-)-enantiomer is not competitive: the amount adsorbed onto the CSP at equilibrium with a constant concentration of the (-)-enantiomer is independent of the concentration of the (+) enantiomer. On the other hand, the adsorption of the less-retained enantiomer is cooperative: the amount of this (+)-enantiomer adsorbed by the CSP at equilibrium with a constant concentration of this enantiomer increases with increasing concentration of the (-)-enantiomer. Such a phenomenon has hardly ever been reported. A model equation is proposed that accounts well for all these isotherm data.

Experimental studies of pressure/temperature dependence of protein adsorption equilibrium in reversed-phase high-performance liquid chromatography.

The effect of the average pressure and temperature of the column on the adsorption equilibrium of insulin variants on a C8 bonded silica was studied in isocratic reversed-phase HPLC. Analytical injections of samples of four different insulins (bovine, porcine. Lys-Pro and human recombinant) were carried out at constant flow-rate but under increased average pressure. The temperature dependence of the retention parameters over the range 25-50 degrees C was studied under two different average column pressures (47 and 147 bar). Substantial increases of the retention time (up to 300%) were observed when the pressure and/or the temperature were increased. Similar adsorption-induced changes in the partial molar volume at constant temperature (deltaVm approximately 102 ml/mol) were found for all the variants studied. Furthermore, deltaVm was revealed to be practically independent of the temperature, which suggests that the temperature has no or very little influence on the mechanism of the pressure induced perturbations in the molecular structure of the solute. This conclusion was also derived from the observed temperature dependence of the logarithm of the retention factor (k) measured under different pressures. The relation between the temperature and In k was nonlinear with a parabolic shape. Moreover, the shapes of the plots corresponding to the low and high pressures were found to be exactly the same, except that the curves were vertically shifted, due to the difference between the two average column pressures. These results indicate that pressure and temperature affect the retention behavior of insulins in a different and separate way.

Comparative modeling of breakthrough curves of bovine serum albumin in anion-exchange chromatography.

The experimental results of a previous study of the mass transfer kinetics of bovine serum albumin (BSA) in ion-exchange chromatography, under nonlinear conditions, were reevaluated using the general rate model of chromatography. Solutions of this model were obtained numerically. The influences of axial dispersion, the resistance to mass transfer from the bulk mobile phase to the surface of the packing particles, and the intraparticle mass transfer resistances on the profiles of the breakthrough curves of BSA were investigated. The results obtained are compared to those of a previous investigation of the same data, using the simple transport-dispersive model and the lumped pore diffusion model. The results obtained show that the use of an oversimplified model for the analysis of chromatographic data can lead to erroneous interpretations of the experimental data and to misunderstandings of the fundamentals of the processes involved. Finally, a theoretical comparison between the properties and the range of application of the three models is provided.

Modeling of the sorption and diffusion processes of volatile organic air pollutants in grape fruits.

A mathematical model for the description of the sorption and diffusion processes of gaseous toluene and p-xylene in fruits of grape has been proposed. This model is based on the Fick's II low regarding a particle with a spherical shape (such as the berry of grapevine) and describes changes of air pollutant concentrations in different layers of the fruit, that is, the wax, peel, and pulp, during an exposure to contaminated air. The mass transfer coefficient and diffusion coefficients in the respective layers can be estimated using the experimental values. The theoretical data and the results from the exposure under steady-state laboratory conditions were compared and showed a good applicability of the proposed model for the prediction of volatile air pollutant partitioning in grape berries.

Modeling of the separation of the enantiomers of 1-phenyl-1-propanol on cellulose tribenzoate.

The competitive adsorption isotherms of rac-1-phenyl-1-propanol on cellulose tribenzoate were measured by competitive frontal analysis. The experimental data were fitted to four different isotherm models: Langmuir, Bilangmuir, Langmuir-Freundlich, and Tóth. The fittings of the experimental data to all four models were satisfactory. It was excellent in the case of the Langmuir-Freundlich and the Tóth models. Overloaded elution profiles calculated with the Tóth isotherm were in good agreement with the experimental profiles in all the different experimental conditions investigated. This work extends to the case of binary mixtures the equivalence between the general rate and the lumped pore diffusion models already demonstrated for pure compounds when the ratio between the Stanton and the Biot numbers exceeds 5. The adsorption energy distribution for the Tóth isotherm was also calculated.

Determination of mobile phase effect on single-component adsorption isotherm by use of numerical estimation.

Numerical estimation was used to determine adsorption isotherm parameters of a single-component in a normal-phase system. The distribution isotherm of methyl deoxycholate was described between the mobile phase containing hexane, ethyl acetate, methanol with varied concentration and a silica gel adsorbent. The effect of the mobile phase composition on the isotherm parameters and the band profiles was investigated. The results obtained were used to simulate the overload gradient elution. The validity of the method proposed was verified by comparison of the computer simulations with the experimental band profiles.

Adsorption/partition model of liquid chromatography for chemically bonded stationary phases of the aliphatic cyano, reversed-phase C8 and reversed-phase C18 types.

A novel approach was introduced to modeling solute retention in the liquid chromatography systems, employing silica-based aliphatic chemically bonded stationary phases of the cyano, reversed-phase C8 and reversed-phase C18 types, and the mixed binary eluents most frequently used in the reversed-phase and normal-phase chromatography modes (i.e. using the methanol-water and the 2-propanol-n-hexane liquid mixtures, respectively). This approach takes notice of the mixed (adsorption/partition) mechanism of solute retention, in which both, the adsorptive and the dispersive forces contribute to the overall energetics of this process. Performance of our new model was compared with that of the widely recognized and on a routine basis applied Schoenmakers approach, and it was found out that both models perform with a practically equal and outstanding accuracy.

Calculation of chromatographic band profiles with an implicit isotherm.

The numerical method for solving the mathematical model of chromatography process coupled with implicit isotherm has been proposed. The exemplary predictions of elution band profiles were performed for competitive adsorption data of 2-phenylethanol and 3-phenylpropanol on ODS-silica with methanol-water as the mobile phase. The simulations of chromatography process with various isotherm models taking into account lateral interactions in adsorbed phase and surface heterogeneity were discussed.