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

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Analytical technologies in the biomedical and life sciences, 2006-01, Vol.830 (1), p.6-12
• Main Authors: Delahunty, Tom, Bushman, Lane, Fletcher, Courtney V.
• Format: Article
• Language: English
• Subjects: Adefovir (PMEA); Adenine - analogs & derivatives; Adenine - blood; Adenine - pharmacokinetics; Analysis; Analytical, structural and metabolic biochemistry; Biological and medical sciences; Calibration; Chromatography, High Pressure Liquid - methods; Fundamental and applied biological sciences. Psychology; General pharmacology; HIV Infections - blood; Humans; LC/MS/MS; Mass Spectrometry - methods; Medical sciences; Organophosphonates - blood; Organophosphonates - pharmacokinetics; Pharmacology. Drug treatments; Plasma; Reference Standards; Reverse Transcriptase Inhibitors - blood; Reverse Transcriptase Inhibitors - pharmacokinetics; Sensitivity and Specificity; Tenofovir; Tenofovir (TNF)
• ISSN: 1570-0232; 1873-376X
• DOI: 10.1016/j.jchromb.2005.10.015

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 × 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 μl 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.