Liquid chromatographic assay for the antiviral nucleotide analogue tenofovir in plasma using derivatization with chloroacetaldehyde

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 μl plasma sample using protein precipitation with trichloroacetic acid. The fluorescent 1, N 6-etheno derivative is formed at 98 °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 usefullness of the assay was demonstrated for samples obtained from an HIV-infected patient treated with tenofovir.