Liquid chromatographic assay for the antiviral nucleotide analogue tenofovir in plasma using derivatization with chloroacetaldehyde |
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Authors: | Sparidans Rolf W Crommentuyn Kristel M L Schellens Jan H M Beijnen Jos H |
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Institution: | Utrecht University, Faculty of Pharmaceutical Sciences, Department of Biomedical Analysis, Division of Drug Toxicology, Sorbonnelaan 16, 3584 CA Utrecht, Netherlands. r.w.sparidans@pharm.uu.nl |
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Abstract: | A sensitive and selective reversed-phase liquid chromatographic assay for tenofovir in human plasma has been developed and validated. Tenofovir was isolated from a 200 microl plasma sample using protein precipitation with trichloroacetic acid. The fluorescent 1,N(6)-etheno derivative is formed at 98 degrees C in the buffered extract with chloroacetaldehyde. This derivative was analysed using gradient ion-pair liquid chromatography and fluorescence detection at 254 nm for excitation and 425 nm for emission. In the evaluated concentration range (20-1000 ng/ml), the intra-day precision was 4% and the inter-day precision was 5-6%. An accuracy of between 97 and 110% was determined. The lower limit of quantification was 20 ng/ml with an inter-day precision of 11%, an intra-day precision of 12% and an accuracy of 103%. The assay is subject to interference from co-administered abacavir. The usefulness of the assay was demonstrated for samples obtained from an HIV-infected patient treated with tenofovir. |
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