Development and validation of enantioselective high performance liquid chromatographic method for Valacyclovir, an antiviral drug in drug substance

Abstract

A chiral high performance liquid chromatographic method was developed and validated for the enantiomeric resolution of Valacyclovir, l-valine 2-[(2-amino-1,6-dihydro-6-oxo-9h-purin-9-yl) methoxy] ethyl ester, an antiviral agent in bulk drug substance. The enantiomers of Valacyclovir were resolved on a Chiralpak AD (250 mm × 4.6 mm, 10 μm) column using a mobile phase system containing n-hexane: ethanol: diethylamine (30:70:0.1, v/v/v). The resolution between the enantiomers was found not less than four. The presence of diethylamine in the mobile phase has played an important role in enhancing chromatographic efficiency and resolution between the enantiomers. The developed method was extensively validated and proved to be robust. The limit of detection and limit of quantification of ( d)-enantiomer were found to be 300 and 900 ng/ml, respectively, for 20 μL injection volume. The calibration curve showed excellent linearity over the concentration range of 900 ng/ml (LOQ) to 6000 ng/ml for ( d)-enantiomer. The percentage recovery of ( d)-enantiomer was ranged from 97.50 to 102.18 in bulk drug samples of Valacyclovir. Valacyclovir sample solution and mobile phase were found to be stable for at least 48 h. The proposed method was found to be suitable and accurate for the quantitative determination of ( d)-enantiomer in bulk drugs substance. It can be also used to test the stability samples of Valacyclovir.