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The present work describes a rapid and sensitive advance liquid chromatographic
ultra high-pressure liquid chromatography (UHPLC) method
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 μm) 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 (
0.999), accurate (recovery, 97.0�100.2%) and precise (CV, ≤ 3.1%) in the selected
concentration range of 0.1�1.0 μg/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
, 3440.7�26.2 ng.hr/mL) and GCV-NDs
, 3380.5�29.3 ng.hr/mL) provided approximately 5 fold increase in the
relative ocular bioavailability compared with GCV solution (AUC
, 650.8�14.9 ng.h/
mL) and nearly 2.5 fold higher than the GCV-NPs (AUC
, 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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