Enantiomeric separation of pidotimod by HPLC using chiral stationary phase

An HPLC method for the determination of the pidotimod enantiomers using amylose tris（3,5-dimethylphenylcarbamate) as chiral stationary phase was developed to control the quality of bulk drug.The separation of pidotimod and its enantiomers were performed on a CHIRALPAK^?IA(250 mm×4.6 mm,5 μm)column at the temperature of 25 °C.The mobile phase consisted of methyl tert-butyl ether-acetonitrile-trifluoroacetic acid(35∶65∶0.2);the flow rate of the mobile phase was 1.0 mL/min and the detection wavelength was 210 nm.Under the established conditions,pidotimod and its enantiomers could be separated completely.This method could be used for the determination of the pidotimod enantiomers.