Determination of tenofovir in human plasma by high-performance liquid chromatography with spectrofluorimetric detection.
A sensitive high-performance liquid chromatography method was developed with spectrofluorimetric detection for the determination of tenofovir, a new HIV reverse transcriptase inhibitor, in human plasma. After precipitation of 200 microl of plasma samples by methanol and evaporation of the supernatant, fluorescent derivatized compounds were obtained by a 40-min incubation at 80 degrees C with chloroacetaldehyde 0.34% at pH 4.5. The assay was performed isocratically using 5 mM Na(2)HPO(4) (pH 6), containing tetrabutylammonium (TBA) chloride 5 mM, and acetonitrile (85:15, v/v) as mobile phase, and a Cluzeau C(8) plus satisfaction column maintained at 35 degrees C. Detection was performed at excitation and emission wavelengths set at 236 and 420 nm, respectively. In these conditions, tenofovir can be separated from adefovir, the internal standard, and endogenous substances. The method was found to be linear and has been validated over a concentration range of 5-1000 microg/l. The average coefficient of the limit of quantification (5 microg/l) was 5.38% and at this concentration, a signal-to-noise ratio of 500 was measured
Jullien
Vincent
V
Service de Pharmacologie Clinique, Université René Descartes, Groupe Hospitalier Cochin-Saint-Vincent-de-Paul, Assistance Publique-Hôpitaux de Paris, 82 avenue Denfert-Rochereau, 75674 Paris Cedex 14, France.
Tréluyer
Jean-Marc
JM
Pons
Gérard
G
Rey
Elisabeth
E
D000225:Adenine / Q000031:analogs & derivatives* / Q000097:blood*; D002138:Calibration; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D007202:Indicators and Reagents; D063065:Organophosphonates; D009943:Organophosphorus Compounds / Q000097:blood*; D012015:Reference Standards; D015203:Reproducibility of Results; D018894:Reverse Transcriptase Inhibitors / Q000097:blood*; D013050:Spectrometry, Fluorescence; D000068698:Tenofovir
D007202:Indicators and Reagents; D063065:Organophosphonates; D009943:Organophosphorus Compounds; D018894:Reverse Transcriptase Inhibitors; D000068698:Tenofovir; D000225:Adenine