Sensitive assay for determining plasma tenofovir concentrations by LC/MS/MS.

TitleSensitive assay for determining plasma tenofovir concentrations by LC/MS/MS.
Publication TypeJournal Article
Year of Publication2006
AuthorsDelahunty T, Bushman L, Fletcher CV
JournalJ Chromatogr B Analyt Technol Biomed Life Sci
Date Published2006 Jan 2
KeywordsAdenine, Calibration, Chromatography, High Pressure Liquid, HIV Infections, Humans, Mass Spectrometry, Organophosphonates, Reference Standards, Reverse Transcriptase Inhibitors, Sensitivity and Specificity, Tenofovir

An LC/MS/MS assay for the determination of tenofovir (TNF) was developed and validated for use with the EDTA anticoagulated human plasma matrix. Heparin-treated plasma and serum matrices were also validated. After addition of adefovir as an internal standard, trifluoroacetic acid was used to produce a protein-free extract. Chromatographic separation was achieved with a Polar-RP Synergi, 2.0 mm x 150 mm, reversed-phase analytical column. The mobile phase was 3% acetonitrile/1% acetic acid, aq. Detection of TNF and the internal standard was achieved by ESI MS/MS in the positive ion mode using 288/176 and 274/162 transitions, respectively. The method was linear from 10 to 750 ng/ml with a minimum quantifiable limit of 10 ng/ml when 250 microl aliquots were analyzed. The usefulness of this LC/MS/MS method to routinely monitor plasma concentrations of TNF was demonstrated along with its ability to assist in the performance of pharmacokinetic studies.

Alternate JournalJ. Chromatogr. B Analyt. Technol. Biomed. Life Sci.
PubMed ID16269276
Grant ListR01 AI33835 / AI / NIAID NIH HHS / United States
U01 AI38858 / AI / NIAID NIH HHS / United States
U01 AI41089 / AI / NIAID NIH HHS / United States