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Analysis of various nucleosides in plasma using solid phase extraction and high-performance liquid chromatography
D L Walters1, D L Jacobs, J E Tomaszewski
1Battelle, Columbus, OH 43201, USA.
Journal of Pharmaceutical and Biomedical Analysis
|March 4, 2000
Summary
A validated reversed-phase high-performance liquid chromatography (HPLC) method quantifies HIV-1 antiviral nucleosides and their metabolites in animal plasma. This analytical method supports drug development by enabling accurate measurement in biological matrices.
Area of Science:
- Analytical Chemistry
- Pharmacology
- Biochemistry
Background:
- The National Cancer Institute (NCI) screens nucleoside analogs for human immunodeficiency virus type 1 (HIV-1) antiviral activity.
- Antiviral drug candidates undergo preclinical testing in animal models to assess toxicity and pharmacokinetics.
- Accurate quantification of nucleosides in biological samples is crucial for supporting drug development studies.
Purpose of the Study:
- To modify and validate a sensitive reversed-phase high-performance liquid chromatography (HPLC) method for quantifying specific nucleosides and their metabolites.
- To support preclinical antiviral drug development by providing reliable analytical methodology for plasma samples.
- To adapt the validated HPLC method for similar nucleoside compounds.
Main Methods:
- Validation of a reversed-phase HPLC method for quantifying 6-chloro-2',3'-dideoxyguanosine (6ClddG), 6-chloro-2',3'-dideoxyinosine (6ClddI), 2',3'-dideoxyguanosine (ddG), 2',3'-dideoxyinosine (ddI), 2'-fluoro-2',3'-dideoxyara-adenosine (betaFlddA), and 2'-beta-fluorodideoxyinosine (betaFddI).
- Analysis performed in rat and dog plasma using appropriate anticoagulants (EDTA or heparin).
- Inclusion of pentostatin (4 microg ml(-1)) in plasma samples to inhibit in-vitro deamination, with calibration standards prepared across defined concentration ranges.
Main Results:
- Validated HPLC methods were established for the target nucleosides and their metabolites in rat and dog plasma.
- Calibration standards were prepared within specific ranges (e.g., 0.1-10 microg ml(-1) for betaFlddA and betaFddI).
- The validated methods were successfully applied to analyze plasma samples from various animal studies.
Conclusions:
- The modified and validated reversed-phase HPLC method provides a sensitive and adaptable analytical tool for quantifying antiviral nucleosides and their metabolites in plasma.
- This methodology is essential for supporting pharmacokinetic and efficacy studies in preclinical drug development.
- The method's successful application in animal studies demonstrates its utility and reliability.