Determination of Tipranavir in Human Plasma by Reverse Phase Liquid Chromatography With UV Detection Using Photodiode Array

Abstract

Tipranavir has recently received accelerated approval from the FDA. The initial clinical use of tipranavir will be for patients with prior virologic failure with the presence of key HIV-1 protease inhibitor mutations. In Phase III trials patients with greater virologic response also had higher trough tipranavir concentrations (BI product information 2005). In addition, hepatotoxicity was concentration-related with a higher incidence in those patients exceeding a trough plasma concentration of 48.2 microg/mL (80 microM). Therefore, tipranavir may be an HIV-1 protease inhibitor for which therapeutic drug monitoring (TDM) may be helpful in optimizing outcomes. To quantitate tipranavir concentrations in human plasma, a method using reversed phase high performance liquid chromatography (RP-HPLC) was validated. Detection was effected using a photodiode-array detector, scanning at a wavelength of 254 nm. This method allows for detection of tipranavir to a lower limit of quantitation of 0.390 microg/mL with an interday variation in control value ranging from 2.9 to 4.6%. The method is being used in a clinical therapeutic drug monitoring program that is ongoing in our laboratory.

High-performance liquid chromatography assay for the determination of the HIV-protease inhibitor tipranavir in human plasma in combination with nine other antiretroviral medications

“…Six assays [4,[12][13][14][15][16] have been previously published for measuring the plasma concentration of TPV. Crommentuyn et al [12] proposed an assay based on mass spectrometry detection using equipment that is not readily available in all laboratories.…”

“…Crommentuyn et al [12] proposed an assay based on mass spectrometry detection using equipment that is not readily available in all laboratories. Recently, methods using UV detection were developed by Giraud et al [4], Colombo et al [13], Dailly et al [14], Sparidans et al [15] and Keil et al [16]. However, the first two methods are time consuming and yield suboptimal peak shapes, and the other methods are limited to quantifying TPV only.…”

“…This method has lower LLOQ for TPV than the previous methods [4,[13][14][15][16]. Sensitivity can be an issue for pediatric samples, or other drug compartments of particular clinical interest, such as breast milk, cerebro spinal fluid (CSF) and genital tract secretions.…”

“…To quantify the HIV PIs and NNRTIs simultaneously, several high-performance liquid chromatography (HPLC) methods have been developed [7][8][9][10][11]. To date, several methods to quantify TPV have been published [4,[12][13][14][15][16]. However, some of these methods are somewhat cumbersome, and have limited selectivity.…”

The Design and Implementation of A5146, a Prospective Trial Assessing the Utility of Therapeutic Drug Monitoring Using an Inhibitory Quotient in Antiretroviral-Experienced HIV-Infected Patients

“…The SUNY at Buffalo Pharmacology Support Laboratory (UB PSL), one of six ACTG-funded pharmacology research laboratories, which was experienced in TDM of antiretroviral drugs and the pharmacokinetics of protease inhibitors [28][29][30][31][32][33][34][35] , was selected by virtue of meeting these specified criteria. Following the initial implementation of A5146, the UB PSL was able to generate realtime PI trough concentration results with an average turn-around time of 7 days from specimen receipt.…”