Nevirapine quantification in human plasma by high-performance liquid chromatography coupled to electrospray tandem mass spectrometry. Application to bioequivalence study

Publisher Website
Google Scholar
Abstract
A rapid, sensitive and specific method to quantify nevirapine in human plasma using dibenzepine as the internal standard (IS) was developed and validated. The method employed a liquid–liquid extraction. The analyte and the IS were chromatographed on a C18 analytical column, (150 × 4.6 mm i.d. 4 µm) and analyzed by tandem mass spectrometry in the multiple reaction monitoring mode. The method had a chromatographic run time of 5.0 min and a linear calibration curve over the range 10–5000 ng ml−1 (r2 > 0.9970). The between‐run precision, based on the relative standard deviation for replicate quality controls was 1.3% (30 ng ml−1), 2.8% (300 ng ml−1) and 3.6% (3000 ng ml−1). The between‐run accuracy was 4.0, 7.0 and 6.2% for the above‐mentioned concentrations, respectively. This method was employed in a bioequivalence study of two nevirapine tablet formulations (Nevirapina from Far‐Manguinhos, Brazil, as a test formulation, and Viramune from Boehringer Ingelheim do Brasil Química e Farmacêutica, as a reference formulation) in 25 healthy volunteers of both sexes who received a single 200 mg dose of each formulation. The study was conducted using an open, randomized, two‐period crossover design with a 3 week washout interval. The 90% confidence interval (CI) of the individual ratio geometric mean for Nevirapina/Viramune was 96.4–104.5% for AUC(0–last), 91.4–105.1% for AUC(0–∞) and 95.3–111.6% for Cmax (AUC = area under the curve; Cmax = peak plasma concentration). Since both 90% CI for AUC(0–last) and AUC(0–∞) and Cmax were included in the 80–125% interval proposed by the US Food and Drug Administration, Nevirapina was considered bioequivalent to Viramune according to both the rate and extent of absorption. Copyright © 2002 John Wiley & Sons, Ltd.