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

Important note: Information in this article was accurate in 1991. The state of the art may have changed since the publication date.

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

J Chromatogr. 1990 Dec 14;534:109-18. Unique Identifier : AIDSLINE MED/91244921
Remmel RP; Huang SH; Hoff D; Zimmerman CL; Department of Medicinal Chemistry, College of Pharmacy,; University of Minnesota, Minneapolis 55455.

Abstract: Carbovir is a carbocyclic guanosine analogue with potent in vitro activity against the human immunodeficiency 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 less than 2.5, and fluorescence detection resulted in a four-fold improvement in the limit of quantitation (0.039 microgram/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 micrograms/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 greater than 0.3 microgram/ml. Within-day variability was less than 7.5% at concentrations greater than 0.3 microgram/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 micrograms/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.

Keywords: Analysis of Variance Animal Antiviral Agents/*ANALYSIS/BLOOD/URINE Chromatography, High Pressure Liquid/*METHODS Dideoxynucleosides/*ANALYSIS/BLOOD/URINE Fluorescence Rats Reproducibility of Results Sensitivity and Specificity Stereoisomers Support, U.S. Gov't, P.H.S. JOURNAL ARTICLE
910930
M9190691

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