HPLC DETERMINATION OF ACYCLOVIR AND ZIDOVUDINE IN MATERNAL PLASMA, AMNIOTIC FLUID, FETAL, AND PLACENTAL TISSUES USING ULTRA-VIOLET DETECTION

Abstract

ABSTRACT A sensitive and reproducible HPLC assay has been developed and validated for the separation and analysis of acyclovir and zidovudine from plasma, amniotic fluid, placental homogenate, and fetal homogenate. Acyclovir (9-[(2-hydroxyethoxy)-methyl]-guanosine, ACV) is the oldest and most widely used compound to treat episodes of genital herpes (herpes simplex virus-2, HSV-2). Zidovudine (3-azido-3′deoxythymidine, AZT) is the premier reverse transcriptase inhibitor released for the treatment of human immunodeficiency virus (HIV). Both of these drugs have been used in pregnant women to prevent the vertical (mother-to-child) transmission of their respective viruses. This gradient HPLC assay aids in the quantitation of these drugs from the matrices associated with pregnancy (maternal plasma, amniotic fluid, fetal tissue, and placenta). The mobile phase consists of 30 mM acetate/ citrate buffer ( pH 3) and methanol. The plasma and amniotic fluid samples are prepared using a combination of protein precipitation and filtration, while the more complex tissues are prepared with the use of a solid-phase extraction (SPE). The method was validated in the calibration range of 0.1–100 μg/mL and showed precision (%Relative Standard Deviation; %RSD) and accuracy (%Error) of less than 15% for all matrices over three days. The assay was applied to a pharmacokinetic study involving the co-administration of ACV and AZT in the pregnant rat.