Determination of the enantiomerization energy barrier of some 3-hydroxy-1,4-benzodiazepine drugs by supercritical fluid chromatography

The first-order kinetic equation for irreversible reactions was used to determine the enantiomerization barrier of some of 3-hydroxy-1,4-benzodiazepine enantiomers by supercritical fluid chromatography (SFC). The racemates of lorazepam, oxazepam and temazepam were separated by SFC on chiral (R,R)-Whelk-O1 column with supercritical carbon dioxide containing 12.5% methanol and 0.5% diethylamine as a mobile phase. Peak areas of enantiomers prior to (AA0, AB0) and after the separation (AA, AB), used for calculation of the enantiomerization barrier, were determined by computer-assisted peak deconvolution of peak clusters from the chromatograms. It was demonstrated for the first time that using a model for a four-peak cluster produces height precise results, and most closely approximates the published results. The kinetic equation for irreversible reactions was used to determine apparent enantiomerization rate constants. The dependence of the apparent enatiomerization barrier (ΔGappA→B, ΔGappB→A) on temperature was used to determine apparent activation enthalpy (ΔHappR→S, ΔHappS→R) and entropy (ΔSappR→S, ΔSappS→R) for all studied benzodiazepines.