Evaluation and prediction of stereoisomerizations in comprehensive two-dimensional chromatography.

Abstract

Conformational and configurational changes such as isomerizations, epimerizations, diastereomerizations and, enantiomerizations are important for the investigation of a large variety of processes ranging from protein folding to the stereostability of drugs. Under optimized conditions, these processes lead to an elution profile characterized by a plateau formation between the two interconverting species in chromatographic separations in a certain temperature range. By temperature-dependent measurements and subsequent computer simulation of the experimental chromatograms, the forward and backward rate constants k1 and k(-1), the Gibb's energy DeltaG++, activation enthalpy DeltaH++, and entropy DeltaS++ can be obtained. Due to its high efficiency two-dimensional chromatography is able to resolve the time-dependent distribution of the two species in the second dimension, thereby yielding the precise ratio of stereoisomers. An algorithm for the simulation and evaluation of two-dimensional chromatographic experiments has been developed, based on the theoretical plate model, which allows the determination of rate constants and barriers of isomerization, epimerization, and enantiomerization processes from two-dimensional chromatographic experiments. In the present article a detailed description of the extended theoretical plate model required for the simulation, the methods available, and examples for the evaluation of complex experimental data and the prediction of the separation conditions to observe isomerization, epimerizations, and enantiomerizations in two-dimensional chromatography are given.