Determination of saquinavir in human plasma by high-performance liquid chromatography.
We developed and characterized a high-performance liquid chromatography (HPLC) assay for the determination of saquinavir, an HIV protease inhibitor, in human plasma samples. Extraction of plasma samples with diethyl ether resulted in quantitative recovery of both saquinavir and its stereoisomer Ro 31-8533 which was used as an internal standard. The assay was performed isocratically using 5 mM H2SO4 (pH 3.5) and acetonitrile (75.5:24.5, v/v) containing 10 mM tetrabutylammonium hydrogen sulfate (TBA) as a mobile phase, a Nucleosil 3C8 column kept at 45 degrees C and UV detection at 240 nm. Using this method, saquinavir and Ro 31-8533 can be separated from endogenous substances, and in the concentration range of 5-110 ng/ml the relative standard deviations for the determination of saquinavir were below 5%. The detection limit of saquinavir in human plasma was 1 ng/ml. The usefulness of the method was demonstrated by quantification of saquinavir in plasma of human subjects treated with 600 mg of saquinavir per os or 12 mg intravenously.
Ha
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Department of Internal Medicine, University Hospital, Zürich, Switzerland.
Follath
F
F
Bloemhard
Y
Y
Krähenbühl
S
S
D000284:Administration, Oral; D019380:Anti-HIV Agents / Q000008:administration & dosage / Q000097:blood*; D002851:Chromatography, High Pressure Liquid; D017320:HIV Protease Inhibitors / Q000008:administration & dosage / Q000097:blood*; D006801:Humans; D007275:Injections, Intravenous; D015203:Reproducibility of Results; D019258:Saquinavir / Q000008:administration & dosage / Q000097:blood*
D019380:Anti-HIV Agents; D017320:HIV Protease Inhibitors; D019258:Saquinavir