Abstract

A validated reverse phase chiral liquid chromatographic method for the enantiomeric purity determination of nateglinide in bulk drug samples and pharmaceutical dosage forms

Author(s): A.Madhavi, A.Naidu

A simple isocratic reverse phase chiral HPLC method was developed for the enantiomeric purity determination of nateglinide in bulk drugs and dosage forms with a short run time of about 20min. Chromatographic separation of nateglinide and its enantiomer was achieved on a bonded macro cyclic glycopeptides (Chirobiotic-T column (2504.6)mmwith 5m particle size) stationary phase. Bonded macrocyclic glycopeptides stationary phase found to be enantioselective for L and D enantiomers of nateglinide with a resolution (RS) of greater than 3.4. The mobile phase used was a mixture of buffer and acetonitrile in the ratio of 70: 30 (v/v). Buffer consists of 5 mMof tetra-n-butyl ammoniumhydrogen sulfate, pH adjusted to 3.5 using diluted ammonia solutions (1 in 10). The test concentration is 1.0 mg mL-1 in diluent (6:4 (v/v) acetonitrile and water). This method is capable of detecting the L-nateglinide up to 0.06g wrt test concentration of 1000g mL-1 for a 10L injection volume. The drug was subjected to stress conditions of hydrolysis, oxidation, photolysis and thermal degradation. There is no interference of degradants with Dnateglinide and L-nateglinide. The developed RP-LC method was validated with respect to linearity, accuracy, precision and robustness. The percentage recovery of L-nateglinide of nateglinide in bulk drug samples and in dosage forms ranged from 93.0 to 102.0%. The test solution was found to be stable in the diluent for 48h after the preparation.