Published:Journal of Chromatographic Science,
ISSN 0021-9665 Volume
47, Number 2, February 2009, pp. 140-148
Rapid and Specific Liquid Chromatographic Tandem
Mass Spectrometric Determination of Tenofovir in Human Plasma
and its Fragmentation Study
Manish Yadav1,2, Tulsidas Mishra1,
Puran Singhal1,
Sailendra Goswami1, and Pranav S. Shrivastav2, 1Bioanalytical Research Department, Veeda Clinical Research,
Ambawadi, Ahmedabad 3800015, India; 2Chemistry Department, School
of Sciences, Gujarat University, Ahmedabad 380009, India
A simple, specific, and high throughput liquid
chromatography tandem mass spectrometry method is developed for
the determination of tenofovir, a nucleotide reverse transcriptase
inhibitor, in human plasma using adefovir as internal standard.
Plasma samples are prepared by solid-phase extraction of the
analyte and internal standard using Waters Oasis MCX cartridges
(1 cc, 30 mg). The chromatographic separation is achieved on
a reversed-phase Chromolith, C18 analytical column (100 mm × 4.6
mm, 5 µm) under isocratic conditions. The mobile phase
consists of 0.5% formic acid in water and acetonitrile (90:10,
v/v) to give a run time of 1.8 min. The protonated precursor → product
ion transitions for tenofovir and IS are monitored on a triple
quadrupole mass spectrometer, operating in the multiple reaction
monitoring and positive ion mode. The fragmentation pathways
for tenofovir are studied by varying the collision energy (5–55
V) using nitrogen as CAD gas. A linear dynamic range of 3.1–1002.0
ng/mL is established using 0.2 mL plasma sample. The method is
fully validated for its sensitivity, selectivity, accuracy and
precision, matrix effect, recovery, stability, and dilution integrity.
It is applied to a bioequivalence study in 43 human subjects
after oral administration of 300 mg tablet formulation under
fasting conditions.
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