Experimental and computational studies of enantioseparation of three profen enantiomers with a focus on quantification of the enantiomeric impurities present in the corresponding enantiopure S-profen drugs

Abstract

A rapid reversed-phase high performance liquid chromatographic (HPLC) methodology for chiral separation of three profen compounds has been developed and then applied to enantiomeric impurity testing of their corresponding enantiopure drugs. The assay is specific, allowing quantitation of the enantiomeric impurities at levels of 0.0078%, 0.0105%, and 0.0416% relative to S-ibuprofen, S-naproxen, and S-ketoprofen, respectively. In order to gain a better insight into the chiral recognition mechanisms of chiral profens on an FLM Chiral NQ(2)-RH column, molecular docking studies were carried out using AutoDock 4.0 software. It was found that hydrogen bonding, hydrophobic interactions, and π-π stacking were all involved in stereoselective interactions, and the calculated binding energy (BE) obtained reflected the binding strength of each enantiomer interacting with a chiral selector. The higher the BE value, the harder it was to elute the corresponding enantiomer, which also accorded with the enantiomer elution order observed in the actual enantiomeric separation. Additionally, thermodynamic analysis revealed that the enantioseparation process at 15–40°C was driven mainly by entropic contributions. The methodology was further validated according to the International Council for Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use (ICH) guideline Q2 (R1) and proved to be sensitive, linear, precise, and accurate for determining R-profen impurities in three commercially available single-enantiomer S-profen drugs. As expected, in the case of products acquired in actual pharmacies, the levels of all of the monitored impurities were found to be lower than the allowable impurity limits.