Development and Validation of RP-HPLC Method for the Determination of Ganciclovir in Bulk Drug and in Formulations

A simple, rapid, accurate, and precise gradient reversed-phase HPLC (RP-HPLC) method has been developed for the determination of ganciclovir (GNC) in pharmaceuticals. Chromatographic separation was carried out on inertsil ODS C18 (4.6？mm i . d × 2 5 0 ？mm, 5.0？μm) LC column using ammonium acetate buffer, sodium salt of hexane sulfonic acid as ion-pairing reagent in 1000？mL water, and acetonitrile (90？:？10) (v/v) as mobile phase at a flow rate of 1.0？mL？ m i n ？ 1 and with UV detection at 245？nm at column temperature (30°C). The runtime under these chromatographic conditions was 10？min. The method was linear over the range of 0.02–75？μg？ m L ？ 1 . The limits of detection (LOD) and quantification (LOQ) values were 4.1 and 20？ng？ m L ？ 1 , respectively. The method was successfully extended to study the effect on GNC upon treatment with 2？N NaOH, 2N HCl, and 5% H2O2 for 2？hrs at 80°C and upon exposure to UV (1200？K lux hrs) for 72？hrs and thermal (105°C) for 5？hrs. The proposed method was further applied to the determination of GNC in pharmaceuticals, with good percent recovery. The accuracy and the precision of the method were validated on intraday and interday basis in accordance with ICH guidelines. 1. Introduction Ganciclovir (GNC), chemically known as 2-amino-9- { [ ( 1 , 3 - d i h y d r o x y p r o p a n - 2 - l ) o x y ] m e t h y l } -6,9-dihydro-3H-purin-6-one (Figure 1), is a nucleoside analogue widely used in the treatment of cytomegalovirus infections. Figure 1: Structure of ganciclovir. It has proved to be effective against cytomegalovirus in immunocompromised patients, mainly in those with the acquired immunodeficiency syndrome (AIDS), congenital immunodeficiency, or in individuals following organ transplantation [1, 2]. Various techniques have been developed for the determination of GNC in pharmaceuticals. It is official in the United States Pharmacopoeia [3], which describes an HPLC method for its determination in injections and in oral suspension. The literature is enriched with several methods for the determination of GNC in pharmaceutical dosage forms including body fluids. The most extensively used technique for the quantitation of ganciclovir is HPLC, but most of the procedures using this technique are devoted to body fluids like plasma [4–14], plasma and tissues [15], serum [16], and blood samples [17]. There is only one report [18] dealing with the application of HPLC for the determination of pharmaceutical formulations, that is eye drops. GNC in bulk drug and in its formulations has been assayed by UV spectrometry by measuring the