LC methods for acyclovir and related impurities determination.

Abstract

Acyclovir, guanine, and impurity A have been baseline separated with isocratic conditions at pH=3.0 and run time under 15 min by employing a SB-CN column from Agilent (150 mm x 4.6mm and 3.5 microm). Moreover, when run time was increased to 40 min six impurities (guanine, impurities A, F, G, Vir 3/4 and N(7)) plus acyclovir were separated in the same conditions. The mobile phase consisted of buffer A/acetonitrile 96:4 (v/v), being buffer A:25 mM H(3)PO(4) (Milli-Q H(2)O) brought to pH 3.0 with KOH. The same column provided separation for all the seven impurities described in pharmacopoeia, including impurity C, which coeluted with acyclovir in the previous conditions with a mobile phase prepared with 25 mM phosphoric acid (pH=1.8)/acetonitrile 96:4 (v/v). The method has been validated following ICH guidelines and it has demonstrated to be reliable for acyclovir and its impurities determination.