[Simultaneous quantitative determination of amprenavir and indinavir in human plasma by high-performance liquid chromatography].
A reversed-phase high-performance liquid chromatographic assay for the determination of the HIV protease inhibitors amprenavir (Agenerase) and indinavir (Crixivan) in human plasma is described, using a mobile phase consisting of 0.50 M phosphate buffer (adjusted to pH 5,5) - Milli-Q water - acetonitrile (120: 1,080: 800, v/v/v). A solid-phase extraction using C18 extraction columns (Discovery columns 100 mg, 1 ml Supelco) and a liquid-liquid extraction with 0.5 ml hydrogenocarbonate/carbonate buffer (adjusted to pH 10.6) and 6 ml methyl ter-butyl ether have been compared. The liquid-liquid extraction has been chosen to be easier and cheaper. The method has been validated over the range of 60 to 3,000 ng/ml for amprenavir and 20 to 3,000 ng/ml for indinavir using a 0.5 ml sample volume. The specificity, linearity, accuracy and precision have been studied. The limit of detection was respectively for amprenavir and indinavir 15 and 4 ng, and the limit of quantification was 60 and 20 ng/ml. Stability tests under various conditions were performed. This assay can readily be used in a hospital laboratory for the routine monitoring of plasma concentrations of amprenavir in HIV-infected patients. The trough plasma concentrations average has been determined in patients treated by amprenavir and indinavir for seven months.
Laboratoire de toxicologie et de pharmacocinétique, Hôpital Ambroise-Paré, 9, avenue Charles-de Gaulle, 92100 Boulogne-Billancourt.
S
S
Laussine
A
A
Roux
B
B
Delhotal-Landes
B
B
Flouvat
D019380:Anti-HIV Agents / Q000097:blood* / Q000737:chemistry; D002219:Carbamates; D002851:Chromatography, High Pressure Liquid / Q000191:economics / Q000295:instrumentation / Q000379:methods* / Q000592:standards; D016903:Drug Monitoring / Q000191:economics / Q000295:instrumentation / Q000379:methods / Q000592:standards; D005663:Furans; D015658:HIV Infections / Q000097:blood / Q000188:drug therapy; D017320:HIV Protease Inhibitors / Q000097:blood* / Q000737:chemistry; D006801:Humans; D019469:Indinavir / Q000097:blood* / Q000737:chemistry; D012680:Sensitivity and Specificity; D013449:Sulfonamides / Q000097:blood* / Q000737:chemistry; D013997:Time Factors
D019380:Anti-HIV Agents; D002219:Carbamates; D005663:Furans; D017320:HIV Protease Inhibitors; D013449:Sulfonamides; C095108:amprenavir; D019469:Indinavir