Quantification of tipranavir in human plasma by high-performance liquid chromatography with UV detection

A simple method for the quantification of tipranavir, a new non-peptidic protease-inhibitor, was developed. An internal standard, prazepam, was added to 100 mu l of plasma before a liquid-liquid extraction by 3 ml of tert-butyl methyl ether. The extracts were evaporated to dryness and reconstituted with 100 mu l of mobile phase before being injected in the chromatographic system. The separation was made on a C8 column using sodium acetate buffer (pH 5): methanol: acetonitrile (35:30:35, v/v/v) as mobile phase. The detection was performed at a wavelength of 260 nm. The method was linear and has been validated over a concentration range of 2-80 mgA. The mean precision and accuracy of the method were respectively, 10.5 and -9.1%. The mean recovery was 70.8%. (c) 2005 Elsevier B.V. All rights reserved.