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Abstract

The present work describes a rapid and sensitive advance liquid chromatographic technique, ultra high-pressure liquid chromatography (UHPLC) method with UV detection to quantify antiviral drug ganciclovir (GCV) in rabbit aqueous humor. After deproteinisation with acetonitrile, gradient separation of GCV was achieved on a Waters Acquity BEH C18 (50 mm x 2.1 mm, 1.7 �¼m) column at 50�C. The mobile phase consisted of 0.1% trifluoroacetic acid in water (pH 3.5) and acetonitrile (95:5, v/v) at a flow rate of 0.45 mL/min. GCV analysis was performed at a wavelength of 254 nm with total run time of 3 min. Method was found to be selective, linear ( r 2 = 0.999), accurate (recovery, 97.0�100.2%) and precise (CV, â�¤ 3.1%) in the selected concentration range of 0.1�1.0 �¼g/mL. Detection and quantitation limit of GCV in aqueous humor were 3.0 and 10.0 ng/mL, respectively. The method was applied to compare aqueous humor levels of GCV after single topical instillation of GCV solution, GCV nanoparticles, GCV nanocomplexes and GCV niosomal dispersions. Topical instillation of GCV-NCs (AUC 0â��t , 3440.7�26.2 ng.hr/mL) and GCV-NDs (AUC 0â��t , 3380.5�29.3 ng.hr/mL) provided approximately 5 fold increase in the relative ocular bioavailability compared with GCV solution (AUC 0â��t , 650.8�14.9 ng.h/ mL) and nearly 2.5 fold higher than the GCV-NPs (AUC 0â��t , 1350.2�18.5 ng.h/mL). The results indicate that the nanocomplexes and niosomal dispersions increases ocular bioavailability of GCV and prolong its residence time in the eye.

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