ABSTRACT
A high-performance liquid chromatographic method for the quantitative determination of the HIV reverse transcriptase inhibitor lamivudine ((-)-2'-deoxy-3'-thiacytidine, 3TC, Epivir) in human plasma, saliva and cerebrospinal fluid is described. Lamivudine was extracted from samples using silica extraction columns prior to reversed-phase high-performance liquid chromatography with ultraviolet detection at 270 nm. The method has been validated over the range of 10 (lower limit of quantitation) to 5000 ng/ml using a 0.5-ml sample volume. Between-day and within-day precisions ranged from 3.5 to 9.0%. The assay has been used for the quantitative analysis of lamivudine in plasma and cerebrospinal fluid of HIV-1 infected patients.
Subject(s)
Anti-HIV Agents/analysis , Lamivudine/analysis , Reverse Transcriptase Inhibitors/analysis , Anti-HIV Agents/blood , Anti-HIV Agents/cerebrospinal fluid , Anti-HIV Agents/pharmacokinetics , Chromatography, High Pressure Liquid , HIV Infections/blood , HIV-1 , Humans , Lamivudine/blood , Lamivudine/cerebrospinal fluid , Lamivudine/pharmacokinetics , Reproducibility of Results , Reverse Transcriptase Inhibitors/blood , Reverse Transcriptase Inhibitors/cerebrospinal fluid , Reverse Transcriptase Inhibitors/pharmacokinetics , Saliva/chemistry , Saliva/virology , Sensitivity and Specificity , Spectrophotometry, Ultraviolet , Toxoplasmosis, Cerebral/cerebrospinal fluid , Toxoplasmosis, Cerebral/drug therapyABSTRACT
A sensitive high-performance liquid chromatographic assay has been developed to determine the levels of 3'-amino-3'-deoxythymidine (AMT), a cytotoxic metabolite of 3'-azido-3'-deoxy-thymidine (AZT, zidovudine), in human plasma. The sample pretreatment involved solid-phase extraction using cation-exchange extraction columns. Chromatography was carried out on a C8 column, using a mobile phase of methanol-0.01 M ammonium acetate (pH 5)-0.25 M sodium dioctylsulfosuccinate (60:40:4, v/v/v) and ultraviolet detection at 265 nm. The method has been validated, and stability tests under various conditions have been performed. The lower limit of quantitation is 5 ng/ml (using 500-microliters human plasma samples). The bioanalytical assay has been used for the determination of AMT in patients with AIDS who used AZT.