Liquid chromatographic assay of carbovir, a carbocyclic nucleoside active against human immunodeficiency virus

Rory P. Remmel, Yoon Hee Yeom, Mei Hua, Robert Vince, Cheryl L. Zimmerman

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8 Scopus citations


Carbovir is a novel carbocyclic guanosine derivative that has potent in vitro activity against human immunodeficiency virus, the causative agent of acquired immunodeficiency syndrome (AIDS). Two methods of sample preparation were developed for the analysis of carbovir in rat blood. Solid-phase extraction on C18 extraction columns proved to be the most effective. Whole rat blood (200 μl) was diluted with 0.8 ml of distilled water containing the internal standard. After two freeze-thaw cycles to lyse the red blood cells and subsequent centrifugation at 13 000 g, the supernatant was loaded on the C18 extraction columns. Carbovir and the internal standard were eluted with methanol-water (60:40). The extract was evaporated and reconstituted in mobile phase and the samples were injected onto a high-capacity reversed-phase column. The compounds were detected at 252 nm. Other nucleosides that could be used in the treatment of AIDS such as zidovudine and acyclovir did not interfere. Standard curves were linear over the concentration range 0.156-28.0 μg/ml (r2>0.99). The within-day coefficient of variation was <7.6% at all concentrations (n=4). The between-day coefficient of variation ranged from 16.7 to 2.0% (n=14). The limit of sensitivity was 0.05 μg/ml with a 200-μl blood sample and the average extraction recovery was 74%. Carbovir was stable in rat blood for at least 4 h at 37°C. The assay was used to determine the blood levels of carbovir in a rat after a 20 mg/kg intravenous dose.

Original languageEnglish (US)
Pages (from-to)323-331
Number of pages9
JournalJournal of Chromatography B: Biomedical Sciences and Applications
Issue number2
StatePublished - 1989

Bibliographical note

Funding Information:
This work was supported in part by Public Health Service Grant ROl CA23263 from the National Cancer Institute. The authors gratefully acknowledge the assistance of Mr. Jay Brownell in the synthesis of large amounts of compound for the animal work and the technical assistance of Ms. Shu-Hui Huang.


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