| Objective: To develop a rapid and simple HPLC method for the simultaneous determination of aciclovir,ganciclovir and penciclovir in plasma. Methods : The plasma samples were purified by solid –phase extraction cartridges.The components were separated on a C18 column with a mobile phase of 0.015mol·L-1 phosphate buffer (adjusted to pH3.14 with dilute phosphoric acid)–CH3OH(90:10) at a flow–rate of 1.0mL·min-1,temperature set at 40·C,and measured by fluorescence detection using an excitation wavelength of 270nm and an emission wavelength of 373nm.Guanine was chosen as the internal standard. Results: All components were eluted in 10 minutes and separated each other. The resolutions between guanine and ganciclovir, ganciclovir and aciclovir, aciclovir and penciclovir were 5.1, 4.0, 4.8, respectively. The linear equations were Y=0.024947+0.084400C ( r=0.9992 ) for ganciclovir , Y=0.016342+0.078968C(r=0.9999)for aciclovir and Y=0.038859+0.44034C(r=0.9998)for penciclovir,respectively.The linearity ranged from 0.10 to 50.0μg·mL-1 for both ganciclovir and aciclovir and from 0.04 to 20.0μg·mL-1 for penciclovir, respectively.At a signal-to-noise ratio of 3:1, the detection limit was 0.05μg·mL-1 for both ganciclovir and ciclovir and 0.01μg·mL-1 for penciclovir, respectively. The extraction recoveries of these drugs were all higher than 70% and method recoveries varied from 90% to 110%. The precision (RSD) of within-day and between-day were all less than 6%. Conclution: This methond is simple ,rapid ,accurate and sensitive .It can be applied to determine the concentrations of ganciclovir, aciclovir and penciclovir in plasma simultaneously and rapidly,and can be thus suitable for use in monitoring therapeutic drugs, studying pharmacokinetics and bioavailabilities in human.
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