Enantiospecific Analysis of Mitiglinide Calcium by RP-HPLC with Precolumn Derivatization

OBJECTIVE To establish a chiral precolumn derivatization method for optical purity test of mitiglinide calcium.METHODS The enantiomers of mitiglinide calcium were derivatized with a precolumn chiral derivatization reagent S-(-)-1-(1-naphthyl) ethylamine (S-NEA). The diastereoisomers produced were separated on an Agilent Zorbax C18 (250 mm × 4.6 mm，5 μm) column with a mobile phase composed of acetonitrile-0.01 mol·L-1 phosphate buffer (pH 2.5) (50∶50). The flow rate was set at 1.5 mL·min-1 and the detection wavelength was set at 225 nm. RESTULTS The method was linear in the range of 0.30-3.0 μg·mL-1 for the R-mitiglinide calcium. The lower limit of detection for R-mitiglinide calcium was 0.1 μg·mL-1 and the lower limit of quantification was 0.3 μg·mL-1. The inter- and intra- day precision (RSD) was below 5.0% and the average recovery was 102.4%. CONCLUSION The derivatives are stable in the mobile phase. The method is sensitive and suitable for the test of optical purity of mitiglinide calcium.