Isomeric separation of Beraprost sodium using an α 1-acid glycoprotein column

Abstract

The separation of the four stereoisomers present in Beraprost sodium, a prostacyclin analogue, has been accomplished using an α 1-acid glycoprotein stationary phase (Chiral AGP column). The stereoisomers are baseline resolved with a runtime of less than ten minutes. This chiral separation is used to quantitate the four Beraprost sodium stereoisomers present in the bulk drug, tabletted formulations and in pharmacological and toxicological studies. The mobile phase variables that were found to have an effect on the stereoisomeric separation were studied and include: ionic strength, type and concentration of organic modifier, mobile phase pH and column temperature. Optimum stereoisomer separation was achieved on the Chiral AGP column. Calibration curves were linear for all four stereoisomers over the range of 0.024 to 4.04 μg/ml using fluorescence detection (correlation coefficients were greater than 0.999). A detection limit of 0.004 μg/ml was found for each stereoisomer. This assay has been used to determine the ratio of the four stereoisomers in the bulk drug as well as in the final formulated tablets.