Improved fluorometric high-performance liquid chromatographic assay for (-)-carbovir in rat blood and urine

Rory P. Remmel, Shu Hui Huang, David Hoff, Cheryl L. Zimmerman

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Carbovir is a carbocyclic guanosine analogue with potent in vitro activity against the human immuno-deficiency virus. All of the activity resides in the (-)-enantiomer. An ion-paired liquid chromatographic assay for (-)-carbovir was developed on a Spherisorb C8 column with fluorescence detection (275 nm excitation, 345 nm emission). Guanosine nucleosides are fluorescent at a pH < 2.5, and fluorescence detection resulted in a four-fold improvement in the limit of quantitation (0.039 μg/ml) compared to the previously developed assay with ultraviolet detection. Standard curves were processed with an internal standard at (-)-carbovir concentrations of 0.039-40 μg/ml in whole rat blood with a solid-phase extraction technique. Total variability was less than 16% at all concentrations and less than 10% at concentrations > 0.3 μg/ml. Within-day variability was less than 7.5% at concentrations > 0.3 μg/ml. Urine was analyzed directly after dilution and an diethyl ether wash to remove impurities. The total coefficients of variation were less than 10% from 0.5-20 μg/ml in urine. The concentrations of (-)-carbovir in rat blood were detectable for as long as 8 h after intravenous and oral doses of 20 and 60 mg/kg, respectively.

Original languageEnglish (US)
Pages (from-to)109-118
Number of pages10
JournalJournal of Chromatography B: Biomedical Sciences and Applications
Issue numberC
StatePublished - 1990

Bibliographical note

Funding Information:
This work was supported in part by Public Health Service Grant ROl AI28236 from the National Institute of Health. The authors wish to thank Dr.

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