Reverse Phase High Performance Liquid Chromatography Method for Quantification of Acyclovir in Rat Plasma

A highly sensitive and selective reverse phase high performance liquid chromatographic method has been developed and validated for the estimation of acyclovir in rat plasma. Centrifugation followed by protein precipitation was utilized for the separation of acyclovir from plasma. The chromatographic separation was performed using methanolwater (50:50, v/v) as an isocratic mobile phase whose pH was adjusted with orthophosphoric acid (pH 2.5), at a flow rate of 0.8 mL/min with an octa-decyl silane (C18) column (5 micron particle size, 250 x 4.60 mm id). Detection was carried out using a Waters 486 Tunable Absorbance Detector at 254 nm. Under these conditions, the retention time was 3.8 min in a run time of 6.00 min. The analytical method was linear over the concentration range of 0.4-2 µg/mL of acyclovir in rat plasma with a limit of quantification 52.70 while limit of detection was 17.39 ng/mL. The high recovery and low relative standard deviation confirm that the method was accurate, reproducible and can be successfully used to quantify acyclovir in plasma following administration of acyclovir in rats.