HPLC enantiomer separation of a chiral 1,4-dihydropyridine monocarboxylic acid

Abstract

An enantioselective anion exchanger based on tert-butylcarbamoylquinine as chiral selector and thiol-modified silica as chromatographic support was applied for the enantiomer separation of the shortacting calcium antagonist clevidipine after its hydrolysis to methyl 4-(2′,3′-dichlorophenyl)-2,6-dimethyl-1,4-dihydropyridine-3,5-dicarboxylate. This hydrolyzed derivative of clevidipine is the primary metabolite and precursor of the parent drug. Method development included the steps of optimization of the composition of the mobile phase (pH, type and content of organic modifier, polar organic mode), screening of various structural analogs of above mentioned CSP, and evaluation of the effect of the flow rate. These detailed studies gave insight into the operational mode of the CSP, which is basically an enantioselective anion exchange mechanism. The polar organic mode turned out to be advantageous with regard to enantioselectivity and resolution. The optimized method makes use of an eluent composed of 0.125% acetic acid in acetonitrile with flow rate of 1 ml/min at a constant temperature of 25 °C, and allows the separation of the both enantiomers with enantioselectivity α of 1.25 and a resolution R S of 3.0 within 10 min. On the above mentioned quinine carbamate phase the ( R)-enantiomer is stronger retained, while a change to the corresponding quinidine carbamate CSP allows the reversal of the elution order.