Determination of 2'-beta-fluoro-2',3'-dideoxyadenosine, an experimental anti-AIDS drug, in human plasma by high-performance liquid chromatography.
2'-Beta-fluoro-2',3'-dideoxyadenosine (F-ddA, lodenosine) is an experimental anti-AIDS drug currently being evaluated in a Phase I clinical trial. A simple and specific HPLC method with UV detection, suitable for use in clinical studies, has been developed to determine both F-ddA and its deaminated catabolite, 2'-beta-fluoro-2',3'-dideoxyinosine (F-ddI) in human plasma. After inactivation of plasma HIV by 0.5% Triton X-100, the compounds of interest are isolated and concentrated using solid-phase extraction. Processed samples are separated by use of a pH 4.8 buffered methanol gradient on a reversed-phase phenyl column. The method has a linear range of 0.05-5 microg/ml (0.2-20 microM) and intra-assay precision is better than 8%. Analyte recovery is quantitative and plasma protein binding is minimal. In addition, drug and metabolite levels measured in Triton-treated human plasma remain stable for at least 5 months when samples are stored frozen without further treatment. Compound concentrations determined after samples are processed and then frozen for up to 1 month before analysis are also unchanged.
Laboratory of Medicinal Chemistry, Division of Basic Sciences, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892-4255, USA.
J S
JS
Roth
H
H
Ford
M
M
Tanaka
H
H
Mitsuya
J A
JA
Kelley
D000163:Acquired Immunodeficiency Syndrome / Q000097:blood / Q000821:virology; D019380:Anti-HIV Agents / Q000097:blood* / Q000378:metabolism; D001798:Blood Proteins / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D016048:Dideoxyadenosine / Q000031:analogs & derivatives* / Q000097:blood / Q000378:metabolism; D004355:Drug Stability; D006678:HIV / Q000187:drug effects / Q000254:growth & development; D006679:HIV Seropositivity / Q000097:blood / Q000821:virology; D006801:Humans; D011485:Protein Binding; D015203:Reproducibility of Results; D013056:Spectrophotometry, Ultraviolet
D019380:Anti-HIV Agents; D001798:Blood Proteins; C073072:lodenosine; D016048:Dideoxyadenosine