A method for prediction ion-exchange isocratic capacity factors from two initial gradient runs is developed. This does not assume so-called linear solvent strength (LSS) conditions, which cause significant errors in k'(C) vs. C relationships in ion-exchange chromatography. The errors associated with this approach and the LSS model are examined. The present approach allows a more accurate prediction of isocratic capacity factors for ion-exchange chromatography. Experimental application of the method to a variety of compounds, including peptides, polynucleotides and polysaccharides, separated by ion-exchange chromatography is described.

Optimization of high-performance liquid chromatography for application to a cough medication (Tussagesic) and its decomposition and byproducts was performed. Special emphasis was placed on the optimization of all parameters that relate to the chemical selectivity of the separation process itself and on the final proof of the ruggedness of the optimized system. All analytes can be reliably determined and the response surface can be represented graphically. This provides a means for improved transfer of methods between laboratories and for efficient system documentation.

Computer simulation (DryLab software) as an aid for the development of gradient high-performance liquid chromatographic methods is reviewed. Several examples of its application are presented and the accuracy of such predictions is discussed.

Computer simulation (DryLab software) as a means of facilitating the development of isocratic high-performance liquid chromatographic methods is reviewed. The various features of computer simulation are discussed and several examples of its application are presented.

Computer simulations can be used to develop high-performance liquid chromatographic gradient elution methods. However, the usefulness of this approach depends on the accuracy of the resulting predictions. Possible sources of error in computer simulation for the prediction of separation based on gradient elution have been investigated. This has in turn led to recommendations for minimizing such errors. With suitable precautions it appears possible to make adequately reliable predictions of separation by gradient elution. Several examples with protein mixtures as samples are reported.

The effect of the gradient on high-performance liquid chromatographic separations has been examined from a theoretical standpoint, using computer simulations to visualize the effects of different variables. Samples to be separated by gradient elution can be classified according to their separation characteristics into three groups (referred to here as cases I, II and III). Each of these sample types responds differently to a change in gradient conditions.

Case III samples exhibit changes in band spacing when the gradient conditions are varied, and gradients composed of multiple linear segments are especially useful for controlling band spacing and resolution for such samples. The effect of different gradient conditions (starting %B, gradient steepness and gradient shape) on the separation of case III samples is examined in detail.